report 2002

Transcrição

report 2002

Laboratório Associado
para a Química Verde
Tecnologias e Processos Limpos
REPORT
2002
ii
Executive Summary
REQUIMTE (Rede de Química e Tecnologia) results from a long-standing
collaboration between two university based research Centers – “CQFB: Centro de Química
Fina e Biotecnologia da Universidade Nova de Lisboa” and “CEQUP: Centro de Química da
Universidade do Porto”.
The association of the two centers was recognized as the Laboratório Associado para
a Química Verde by the Portuguese Ministério de Ciência e do Ensino Superior in November
of 2001, and was formerly chartered as a nonprofit scientific organization in January of 2003.
The scientific expertise and complementary knowledge available, put together by
CQFB and CEQUP, has been focused on the topic GREEN CHEMISTRY – CLEAN
TECHNOLOGIES AND PROCESSES with a wide range of tools and from different
perspectives.
In general terms, the existing competencies will be drawn from the areas identified as
expertise fields within REQUIMTE: chemistry, (micro)biology, biochemistry and molecular
biology, molecular modeling, bio(catalysis) and reaction mechanisms, (bio)conversion and
bioremediation, transport phenomena, separation processes, sensor development,
monitoring and control.
The available expertise will be used to implement the use of cleaner products and
cleaner technologies, preventing pollution at its source. By working with manufacturers,
governmental agencies, consumers and general public new ideas and new attitudes can be
implemented. REQUIMTE can act as a promoter and is available to answer questions and
problems placed from outside. REQUIMTE will look forward to assist manufacturers in order
to develop cooperative efforts to design and redesign products and processes that will reduce
life cycles environmental impacts. Activities wil be implemented to provide reliable information
on the environmental benefits, performances and economic feasibility of green chemistry and
clean processes.
The year 2002 was the first in which REQUIMTE was fully operational - a difficult but
profitable time: frequent meetings of the board of directors (monthly), creation of formal
incentives to strengthen the cooperation between researchers of the two Centers (seed
money for joint projects) and the recruitment of the first seven researchers (Investigadores
Auxiliares) to carry out activities under the contract of Laboratório Associado.
Laboratório Associado para a
Química Verde
iii
Mission Statement
a.
REQUIMTE, which is recognized as the Laboratório Associado para a Química Verde by the
Portuguese Ministério de Ciência e do Ensino Superior, is a voluntary association of two research
Centers which have freely opted to collaborate in research and postgraduate activities, whilst still
being fully committed to their respective Universities.
b.
REQUIMTE considers itself to have made a very important contribution to the way in which
Chemistry, Biology and Chemical Engineering being carried out in Portugal is viewed internationally
— as judged by the quality and quantity of its scientific publications and also by the number and
quality of ongoing research projects.
c.
REQUIMTE will focus the activities of its reaserchers to implement the principles of Green
Chemistry.
d.
REQUIMTE also aims to optimize internal collaboration and to identify the best international
strategic partners.
e.
REQUIMTE, whilst wishing to preserve its well-founded and successful roots in traditional chemical,
biological and engineering sciences, intends fully to strengthen its international research in
innovative and novel topics and to become a pool of attraction for young and well established
national and international scientists.
f.
REQUIMTE views its involvement in the postgraduate training of young people as a very important
function. REQUIMTE policy is to select projects in such a way that students working within the
organization will gain, in the course of their researches, the skills required by the employment
market.
g.
REQUIMTE is presently the largest chemical network in Portugal, with approximately 350
researchers, of which more than 130 hold a Ph.D. degree.
h.
REQUIMTE provides an optimal environment to explore the synergies of the their expertise in
answering the needs of the productive sector and providing specialized services and consulting.
i.
REQUIMTE is currently pursuing an environmentally driven reasearch in association with the
chemical industry to develop closed loop industrial processes.
j.
REQUIMTE is working to foster public awareness of key chemical and biochemical concepts, to
help understand the costs and benefits of technology in the modern world and to develop a
balanced global appreciation of environmental issues.
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REQUIMTE in 2002
2002 was the first year in which REQUIMTE operated as a “Laboratório Associado”,
although the cooperation existed since 1996. The strategic plan was to focus expertises in
Analytical, Biological, Inorganic, Organic, Physical and Theoretical Chemistry and in
Chemical Engeeniring in a contemporary unifying concept – that of Green Chemistry.
2002 was the first year when the funding contract with FCT-MCES was operational and
REQUIMTE hired seven new researchers and seven technicians in order to carry out
activities included in the contract with the Ministry. Fourteen calls for application were open
and more than one hundred candidates have applied. The profiles of the seven new
researchers are provided in Anex.
The scientific production was also considered a key aspect of the activity of
REQUIMTE, and the total number of articles and chapters in books has grown to 181, much
to the dedication of its graduate students, 17 of which have completed their doctoral thesis
and 9 their Master Thesis in 2002.
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People at REQUIMTE
Board of Directors
Baltazar de Castro (UP)
Isabel Moura (UNL)
Manuel Nunes da Ponte (UNL)
Co-ordinating Comittee of the Scientific Council
Ana Lobo (UNL)
Baltazar de Castro (UP)
Isabel Moura (UNL)
José Costa Lima (UNL)
José Moura (UNL)
Manuel Nunes da Ponte (UNL)
Maria de Lourdes Basto (UNL)
Maria Rangel (UNL)
Advisory Comittee
Prof. Hartmut Frank
Environmental Chemistry & Ecotoxicology, Universität Bayreuth, D - 95440 Bayreuth, Deutschland
Ecotoxycology; Analytical organic chemistry; Protein and peptide chemistry; Chiral synthesis
Prof. William Wakeham
Imperial College of Science, Technology and Medicine, Sherfield Building, London SW7 2AZ, UK
Chemical engineering
Prof. William B. Motherwell
University College London, Christopher Ingold Laboratories, 20 Gordon St., London WC 1 H OAJ, UK
Organic synthesis; Reaction mechanisms in organic chemistry
Prof. Luigi Marzilli
Emory University, Chemistry Department, 1515 Pierce Drive, Atlanta, GA 30322, USA
Spectroscopy; Inorganic synthesis; B12 chemistry; Bioinorganic chemistry; Metal based drugs
Prof. Jean L. Rivail
Université Henri Poincaré, Laboratoire de Chimie Théorique, B.P. 239, 54506 Vandoeuvre-lesNancy, France
Theoretical chemistry
Prof. Marek Trojanowicz
University of Warsaw, Faculty of Chemistry, Pasteura 1, PL-02-093,WARSAW, Poland
Analytical Chemistry
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RESEARCH REPORT
2002
AREA I
NATURAL PRODUCTS: SCREENING AND SYNTHESIS
3
TOPIC 1
ORGANIC SYNTHESIS AND CHEMISTRY OF NATURAL PRODUCTS
The following main four major topics of interest have evolved in the team:
1 The area of natural products which continues to attract the interest of the research workers of this
group includes: a) the study of Portuguese flora - the Ulex family - to identify flavonoids with biological
activity, and - Euphorbia species - which led to the isolation of new macrocyclic jatrophane diterpene
polyesters with cytotoxic activity against human cancer cell lines MCF-7 (breast), NCI-H460 (lung) and
SF-268 (CNS), as well as significant multidrug resistance activity; b) the study of Tunisian medicinal
plants (through a collaboration with the University of Monastir); c) the study of medicainal plants from
Guiné-Bissau to identify active principles, such as stritosamide,which was evaluated
pharmacologically.
2 The aim of the efforts in organic synthesis is to devise simple and novel ways of producing
biomolecules or derivatives thereof of established pharmacological interest. Synthesis of biologically
active alkaloids: among the indolo[2,3-a]carbazoles, the alkaloids of the staurosporine type are among
the most highly researched ones in view of their dramatic effect as PKC inhibitors. The aglycone of
staurosporinone, itself a natural product, was synthesised in the shortest synthesis yet published. A
reinvestigation showed that in the reaction of bis-indole and sulfur several dimeric molecules were
obtained, and that the monomeric unstable dithiete was formed in an equilibrium with the dimers. This
dithiete was used in a previous synthesis of the alkaloid arcyriaflavin A. A full account of a new
approach to the physostigmine related desoxyeseroline, involving a 3,3-sigmatropic rearrangement of
a hydroxamic acid derivative, was published.
3 In the mechanistic area, sigmatropic rearrangements of enehydroxylamines were found to be
essentially intramolecular processes by using deuterium labelled precursors and finding no evidence
for cross-coupled products. The reaction was extended to give a variety of heterocycles and promises
to become an important one in the chemistry armamentarium. The fact that in these rearrangements
no atoms are added or lost means that they may represent an important method from a ‘green
chemistry’ point of view. A new method for aziridination of electrodeficient olefins was devised using
Pd catalysis and sodium N-bromo-p-toluenesulfonamide. Investigation of the analogs N-chloro
compounds were however discouraging. An enantiomeric version of the reaction is still being studied.
Radical holding chiral substituents were generated by photolysis of thiohydroxamic acids derivatives
and added to electrodeficient olefins to produce gamma-amino acids.
4 Computational methods were developed in response to practical chemical problems. Two chirality
codes were developed recently that represent: a) the chirality generated by chiral carbon atoms; or b)
the chirality of 3D structure considered as a rigid set of protons (atoms) with properties (atomic
properties). It was demonstrated that such a code could be successfully applied to the prediction of
the enantiomeric selectivity in chemical reactions and in chiral chromatography using artificial neural
networks. In the last year the the chirality codes were used for the prediction of the absolute
configuration of enantiomeric alcohols, on the basis of the 1H NMR chemical shift differences of their
Mosher’s esters derivatives. The predictions were done by counterpropagation neural networks.
Chirality codes were also calculated for all the products in a database of 15212 enantioselective
chemical reactions. These codes will be used in database searches of similar chirality codes, for the
planning of enatioselective synthesis. A new system was built for the fast estimation of NMR chemical
shifts of CHn protons, which is based on automatic knowledge acquisition from a data set of
examples. The relationship between protons in defined molecular structures and the corresponding
1H NMR chemical shift was established by counterpropagation neural networks (CPG NNs), which
used topological, physicochemical and geometric descriptors for hydrogen atoms as input, and the
chemical shift of the corresponding proton as output. In the last year the method was improved by
using backpropagation neural networks instead of CPG NNs. A new method for representing chirality
(previously developed in the group) was further improved and successfully applied to the prediction of
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enantioselectivety in chemical reactions and chiral chromatography. New descriptors for protons were
also developed and used as input to neural networks in order to predict 1H NMR chemical shifts. A
web gateway to the simulator was implemented for on-line 1H NMR chemical shifts’ prediction from
molecular structure. It was also proposed to represent the individual positions in DNA sequences by
virtual potentials generated by other bases of the same sequence. This is a compact representation of
the neighbourhood of a base. The distribution of the virtual potentials over the whole sequence can be
used as a representation of the entire sequence (SEQREP code). To evaluate its biological
significance, the SEQREP code was used for training Kohonen self-organizing maps (SOMs) in two
applications: a) detection of Alu sequences, and b) classification of sequences encoding for HIV-1
envelope glycoprotein (env) into subtypes A-G. It was demonstrated that SOMs clustered sequences
belonging to different classes into distinct regions. Possible areas of application of SEQREP codes
include functional genomics, phylogenetic analysis, detection of repetitions, database retrieval, and
automatic alignment. [Software: SEQREP code, and for training Kohonen SOMs available from http://
www.dq.fct.unl.pt/qoa/jas/seqrep].
TOPIC 2
SELECTIVE SYNTHESIS AND STRUCTURAL CHEMISTRY
1 Ecologically friendly asymmetric chemistry using sugars and non-polluting solvents Under this
topic directed toward the valorisation of sucrose by incorporating it in polymers, its conversion to
natural compounds and its possible application as a chiral auxilliary, we have developed a selective
derivatisation of sucrose ,which allowed the selective synthesis of galloylated compounds as potential
antioxidants, as well as unsaturated monomers and their corresponding polymers . During this period
research was carried out in the area of asymmetric synthesis and catalysis New methods for the
synthesis of potential novel catalysts containing N and O atoms were synthesized and their ability to
catalyse a model reaction, the enantioselective alkylation of benzaldehyde by diethylzinc, was
investigated. Also palladium catalysed transformations are of great use to organic chemists and we
have found that the addition of diethylamine can improve the Stille coupling reaction using unsaturated
organotin compounds and also reduce considerably the amount of reduced products even using
saturated tin components. Thus we have been able to synthesise important intermediates in a
proposed synthesis of coriandrina and also complete syntheses of some polyoxygenated natural
products which have both saturated and unsaturated side chains.
2 Development of new synthetic methodologies in asymmetric chemistry mainly using non-classic
media and dynamic kinetic resolution, with a strong support from computational molecular modeling
and NMR studies. We studied the stereoselective behavior of nucleophilic substitution reactions of
aab initio level, with HF and DFT models, we were able to propose a new model for the reaction
mechanism, and to propose new chiral auxiliaries with expected better performance. In order to find
experimental evidence for the proposed model we have explored and developed new techniques in
NMR with special focus on PFG-diffusion measurements. We also studied the preparation and use of
ionic liquids in selective transport and as an organic reaction in asymmetric catalysis and biocatalysis.
3 Organometallic chemistry. The effect of the trimethylsilyl (TMS) as solubiliser group of ligands and
their corresponding metal-complexes in supercritical carbon dioxide (scCO2) has been analysed using
the easily available TMS-substituted complexes (hð5-Me3SiC5H4)MoO2Cl , (hð5-Me3SiC5H4)2ZrCl2 ,
and (hð 5 -Me 3 SiC 5 H 4 )Co(CO)I 2 . For comparison purposes, the solubility measurements were
performed in parallel with the analogous non-substituted derivatives. In all cases an increase in the
solubility of the TMS-substituted complexes was observed, being the influence of the TMS group in
the chemical behaviour of the complexes small. Complex (hð5-Me3SiC5H4)MoO2Cl has been used as
catalyst for two homogeneous catalysed processes in scCO2: the oxidation of PPh3 using molecular
oxygen as oxidant and the epoxidation of cyclohexene using t-butylhydroperoxide (TBHP). The
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molecular structure of (hð5-Me3SiC5H4)Co(CO)I2 has been determined by X-ray crystallography (see
figure).
C9
Si
C7
C8
Co
O
C
I3'
2.74
I3
I2
3.41
I1
TOPIC 3
PHYSICAL ORGANIC CHEMISTRY / RADICAL CHEMISTRY
1 Study of radical induced oxidation of cynnamic and caffeic acids. Photolytic generation of hydroxyl and
sulphate radicals. In situ reaction of radicals with cynnamic acid, caffeic acid, 3-phenylacetic acid and
derivatives. Identification of products by HPLC and mass spectrometry. Characterization of transient
radicals by ESR spectroscopy. Preparative scale photolytical production of unknown products for
further identification.
2 Reaction calorimetry in the scale-up of a chemical process. Multi-step synthesis of pirlindole
chlorohydrate. Optimization of each synthetical step. Characterization of intermediates. Calorimetric
study of the reactions. Development of analytical methods and quality control assays.
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AREA 2
FOOD SAFETY
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TOPIC 4
ANALYTICAL AND BIOORGANIC CHEMISTRY
1 In the area of natural products, the activity of this group have been developed in the scope of the on
going projects, and was mainly directed to the separation, identification and characterization of sugar
fractions and resin glycosides from natural extracts.
2 In the area of food safe and quality our main activity focus the characterization of volatile
compounds in food and different pine varieties.
3 In the area of environmental control and bioremediation our main goal was the study of some
steroid compounds in order to evaluate their role as potential markers of fecal pollution.
TOPIC 5
AUTHENTICITY OF FOOD PRODUCTS AND ANALYSIS OF UNDESIRABLE COMPONENTS
Our studies were directed to quality control, safety and authenticity of foodstuffs in order to answer the
public awareness and consumer’s demands.
1 Chemical, and biochemical methodologies have been developed, implemented and applied in the
monitoring of some food products. These studies included the evaluation of nitrogen compounds
(biogenic amines, 4-(5)-imidazole), proteins (caseins, α-lactoalbumin, ß-lactoglobulin), free and total
amino acid enantiomers, organic acids (lactic, acetic, tartaric, citric, malic, succinic, propionic), fatty
acids (cis/trans isomers), polycyclic aromatic hydrocarbons, sterolic composition and aromatic
compounds.
2 These parameters were used as an approach to the chemical characterization of “Protected Origin
Denomination” products and others. Also, being monitored along the processing steps, provided
useful information in order to control/prevent adulterations in the food industry.
3 Several food matrices were studied namely dairy products, oils and fats, wine, carbonated beverages,
coffee and nuts (walnut and hazelnut).
4 The applied methodologies were HPLC (UV, diode-array, RI, Fluorescence and ELSD detection) and
HRGC (MS, FID, ECD and NPD)..
TOPIC 6
PHYTOCHEMISTRY
1 Methodologies for the separation and identification of phenolic compounds were developed and
applied, in order to define the qualitative and quantitative chemical profiles of olive fruits from several
cultivars, quince fruit (seeds, pulp and peel), Centaurium erythraea (small centaury) flowering tops
and sage shoots. This work has provided relevant data:
a Olive fruit cultivars exhibited a similar phenolic profile, with hydroxytyrosol and oleuropein as the
major compounds; and with rutin and luteolin 7-O-glucoside as the two main flavonoids. The
anthocyanic profile was affected by the maturation index, with lower index corresponding to lower
anthocyannins concentration or absence of these compounds.
b Quince fruit pulps contained mainly caffeoylquinic acids and one quercetin glycoside, while the
peels presented the same caffeoylquinic acids, and several flavonol derivatives. The highest
content of phenolics was found in peels. Quince fruit seeds presented a different phenolic
composition, in which several C-glycosyl flavones were found.
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c The technique developed for small centaury may be considered complimentary to the one
proposed by the European Pharmacopoeia for the characterization of this species, since nine
xanthones were unequivocally identified. The quantification of these compounds could be explored
as a tool for its botanical classification at a subspecies or varietal level.
d In sage shoots fifteen phenolic compounds were identified, five of them had been previously
detected in field grown plants; so, in vitro systems may be a good alternative for phenolic
compound production.
2 The organic acids composition of quince fruit and jam was also studied. The results suggested that
their levels and ratios might be useful for the determination of percent fruit content of quince jams.
Additionally, the citric acid value could be used in the differentiation between homemade and
industrially manufactured quince jams.
3 The antioxidant capacity of infusions of Lippia citriodora, Hypericum androsaemum, Cynara
cardunculus and C. erythraea was evaluated. All infusions proved to be scavengers of superoxide
radical and hydroxyl radical. H. androsaemum, C. cardunculus and C. erythraea effect on superoxide
radical is also due to their capacity for the inhibition of xanthine oxidase. In a general way L. citriodora
and H. androsaemum infusions presented higher antioxidant activity, although with a pro-oxidant
effect observed for hydroxyl radical when in high concentrations. With the exception of C.
cardunculus all the other infusions exhibited scavenging activity towards hypochlorous acid.
4 The hepatoprotective activity of H. androsaemum infusions was also assessed in vitro and in vivo.
The results suggested that the protective effect was related to its free radicals scavenging and metal
chelating capacities, although a toxic effect was also noticed. The biological effects observed might be
related with the phenolic compounds present in each infusion and with their interactions with the other
constituents.
5 The study of some constituents of seeds and leaves of walnuts (Juglans regia) and hazelnuts (Corylus
avelana) from several Portuguese varieties was also undertaken. During this year, the fatty acid and
phytosterol composition of walnuts collected during the 2001 harvest was already analyzed. While the
fatty acid profile revealed a very similar profile for the several analyzed varieties, the phytosterol
composition showed some differences among the varieties.
TOPIC 7
TOXICITY MECHANISMS
Studies on the mechanisms of toxicity were continued:
1 In vitro studies for evaluation of the antioxidant properties of plant extracts traditionally used in human
diet;
2 In vivo and in vitro studies of biotransformation and toxicological evaluation of the anphetaminic
derivatives;
3 In vitro studies, using isolated rat cardiomyocytes, for evaluation of cardiotoxicity of compounds;
4 Implementation of accurate and sensitive atomic absorption spectrometry for quantification of heavy
metals present in food packaging materials.
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TOPIC 8
PHYSICOCHEMICAL CHARACTERIZATION OF FOOD PRODUCTS
1 Development and chemical, structural and rheological characterisation of proteinpolysaccharide mixed aqueous systems. The enzymatic hydrolysis of a whey protein concentrate
(WPC) was done using three different enzymes (pepsin, trypsin and alcalase). For each enzyme,
different temperatures, times of hydrolysis and pHs were tested. The hydrolysates were characterised
by RP-HPLC: peptides formed during hydrolysis were separated in function of their polarity. It was
observed that, depending on the enzyme used, the peptidic profiles as well as the degree of
hydrolysis of a-lactalbumin and b-lactoglobulin, the main proteins in whey, differed in the hydrolysates.
The thermal gelation of 10% w/w aqueous solutions of the hydrolysates obtained after 15 minutes of
hydrolysis, at pH 7, was monitored by dynamic rheological measurements at a fixed frequency and
their mechanical spectra were determined at 80ºC and after quenching at 20ºC. The hydrolysates
obtained with pepsin and trypsin gave weaker gels than those obtained with WPC while the
hydrolysate of alcalase did not form a gel, in the conditions of the experiments. (Financed by
Fundação para a Ciência e Tecnologia, through contract POCTI / QUI / 36452/2000)
2 Rheological behaviour and morphology of protein/polysaccharide mixed systems. The effect of
Cassia gum galactomannan on the thermal gelation of a whey protein isolate (WPI) at 80°C has been
monitored by rheological dynamic measurements at fixed frequency and the mechanical spectra of
the gels have been recorded at 80°C and after quenching to 20°C. The mechanical spectra were
analysed quantitatively by fitting a Cole-Cole model to the storage and loss compliance versus
frequency data. The concentration of WPI was 110 g/L and that of Cassia gum varied from 0 to 11 g/L.
At low concentration, the presence of the galactomannan enhanced the aggregation rate and the
strength of the protein gel but did not modify intrinsically the gelation process and the structure of the
system. At higher galactomannan concentrations, the gels displayed substantially lower moduli values
and the shape of the mechanical spectra was modified. This was tentatively interpreted as the
consequence of a depletion-induced liquid-liquid phase separation occurring during the course of the
aggregation process.
3 Use of galactomannan/starch mixtures in low-oil food emulsions stabilisation. The purification
of tara gum (a galactomannan) by precipitation with isopropanol (from an aqueous solution) and its
chemical characterisation were done. Moreover, the rheological behaviour of aqueous solutions of this
gum was studied using a controlled stress rheometer: flow curves and frequency sweeps
(“mechanical spectra”) were obtained at different concentrations of the gum. The behaviour was
typical of a macromolecular solution.
4 During this period on-going studies on the characterisation and modelling of osmotic dehydrated
pumpkin. Experimental determinations on dehydration kinetics and shrinkage, change of colour and
mechanical properties. New methodologies for on-line observation of under-the-microscope
dehydration of fruits were tested, involving on-line image acquisition, to be further submitted to an
image analysis of the process. The research activity was developed in the framework of the following
projects:
a Predictive methods for physical properties of foods (PPFA). New experimental developments
to measure the porosity of high moisture content food and similar materials. (CYTED project
XI.12).
b High moisture content tropical fruits: processing, modified atmosphere packaging and
quality assessment. We have pursued on-going studies on experimental measurement of
mechanical properties and texture of osmotic dehydrated fruits (CAPES/GRICES, with University
of Campinas, Brazil).
c Technology for food grade biodegradable films in Ibero-America.
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AREA 3
ENVIRONMENTAL CONTROL AND (BIO)REMEDIATION
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TOPIC 9
ENVIRONMENTAL CONTROL
1 Pesticides Analysis. The knowledge of the concentration and distribution of pesticides or of their
degradation products in the environment is receiving a great deal of attention because of their toxic
character. Prevention of the negative effects of these substances requires a systematic monitoring of
their remains in agricultural products, waters and soils. Gas chromatographic methods have been
developed to study the adsorption of pyrethroids (a major group of insecticides widely used in the
world) in cork and to control pesticides residues in wine. Procedures for determining several groups of
pesticides, in environmental waters, based on their reduction/oxidation or/and adsorption are being
optimized using conventional carbon electrodes, gold and mercury film microelectrodes, as well as,
potentiometric detectors. In order to increase the sample throughput and reduce the human effort,
flow injection analysis (FIA) systems with amperometric detectors are being also implemented.
2 Laboratory Waste Management. Chemistry teaching and research work requires the performance of
a series of laboratory experiments, in which several hazardous and/or toxic compounds are usually
involved. Specially for being performed in a routine fashion, the waste generated by these experiences
may represent a serious risk in environmental terms. This situation demands urgent reactions from
school and institutions that should present proper solutions to the pollution problem inherent to its own
functioning. In this sense, students, researchers and school staff of the Instituto Superior de
Engenharia do Porto (ISEP), have been playing a major role in implementinga laboratory waste
management program. This project is based on the reduction, reusing and recycling of each disposal,
before detoxification or destruction, and finally secure storage or safe disposal of wastes.
3 Soils characterization and remediation. Major remediation techniques currently available for soils
that are contaminated with heavy organic compounds, include processes like incineration, thermal
description, soil washing/solvent extraction, disposal at hazardous land fills or vapour extraction.
While each of these techniques has some advantages, there are concerns regarding their usage
either from pollutant characteristics and environmental or economic points of view. Vapour extraction
and the use of mixed solvents are the selective cleanup techniques considered. Several factors
influence the effectiveness of soil remediation so they must be evaluated. Cyclo hexane and toluene
were the pollutants studied when vapour extraction was applied. The influence of soil water and
natural organic matter content on the results obtained for the limits of remediation through soil vapour
extraction was estimated.
For soil solvent washing, the solubility of the organic pollutant in the solvent and miscibility of the
solvent with water are important factors. Since water was present in soil as moisture, water was
considered as a component of the extracting solvent in ternary mixtures of water: acetone and
ethylacetate. Ex situ soil solvent washing may have a low effective cost when soil solvent washing is
followed by the recovery of solvent. After separating the pollutant from the wash solution, the mixed
solvent was recycled by feeding the wash solution to distillation column.
4 Speciation of heavy metals in sediments: application to the Ave river basin. The evaluation of
the heavy metal (total and speciation) pollution of the Ave river basin was performed. The comparison
of results obtained in this study with other works previously performed in the basin suggests a
decreasing tendency of the heavy metal pollution in the studied area of the basin. This fact is probably
due to the implementation of the municipal wastewater treatment plants. Chromium was the main
responsible for the heavy metal contamination; the speciation of chromium in the sediments has
shown that this metal was mainly in the oxidable and residual fractions. Multivariate analysis
(correlation matrix and factor analysis) suggests that a significant amount of chromium is associated
with the organic matter and is due to an anthropogenic input.
5 Interaction between metal ions and buffers for the environmental and biological pH ranges.
The behaviour of solutions containing, independently 3-(N-tris[hydroxymethyl]methylamine)-2hydroxypro-propanesulfonic acid (TAPSO) or 3-([1,1-dimethyl-2-hydroxyethyl)amine]-2-
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hydroxypropanesulfonic acid (AMPSO), and copper(II) were studied by two analytical techniques,
direct current polarography (DCP) and glass electrode potentiometry (GEP), at fixed total ligand to
total copper(II) concentration ratios and various pH values, at 25ºC and ionic strength 0.1 M KNO3.
Combined interpretation of data from DCP and GEP indicated the formation of six main species. Five
of these complexes are reported for the first time.
TOPIC 10
ANALYTICAL METHODOLOGIES
1 Electroanalysis. Analysis of active substances in pharmaceutical preparations and biological fluids is
mainly performed by chromatographic methods. However, electrochemical techniques can provide
advantages over these methods by being faster, cheaper and more sensitive. Several voltammetric
and FIA methods, using either glassy carbon or hanging mercury drop electrodes, were successfully
developed for the determination of various active substances in pharmaceutical preparations. In
addition, after an appropriate liquid-liquid extraction and based on the reduction of antidepressants
and their metabolites at the hanging mercury drop electrode, it was possible to apply a polarographic
technique to antidepressant drug monitoring in biological fluids. Furthermore, the electrochemical
activity of drugs of abuse and their metabolites at a glassy carbon electrode allowed the proposal of
several oxidation mechanisms. These electrochemical studies allow assessing the physiological
behaviour and influence on normal body functions of the respective substances and obtaining
information concerning their absorption, distribution, biotransformation and elimination in order to
determine its bioavalability and pharmacokinetic profile in humans.
TOPIC 11
DEVELOPMENT OF TRANSDUCERS
New trends were investigated based on exploitation of different transducing schemes such us optical and
electrochemical. Studies on immobilization techniques were used for the monitoring low levels of drugs,
food and pesticides. Studies were developed concerning the construction of flow-through voltametric
electrodes dedicated to the implementation of manifolds with multi-side detection. In more detail the
following results can be referred:
1 A glucose biosensor with the immobilization by sol-gel technology has been constructed and
evaluated. The glucose biosensor was based on the encapsulation of GOX within a sol-gel glass
prepared with 3-aminopropyltriethoxy silane, 2-(3,4-epoxycyclohexyl)-ethyltrimetoxy silane and HCl.
2 Study of the electrochemical behaviour of the herbicide Asulam by cyclic and square wave voltametry.
Based on the obtained results an analytical procedure for the determination of the pesticide in waters
with a limit of the detection of 10-8 M was developed.
3 Construction and evaluation of a tubular voltametric detector dedicated to a new strategy for the
automation of the standard addition method in flow systems with amperometric detection. The
usefulness of the proposed detection system was evaluated in the determination of ascorbic acid in
fruit juices.
4 A tubular periodate selective electrode without inner reference solutions was been developed for
glycerol determination in distilled spirits, based on the oxidation of this polyol by periodate.
5 Application of the determination of diacetyl by adsorptive stripping voltametry after derivatization with
o-phenylenediamine to the beer control with an automatic system.
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TOPIC 12
AUTOMATION AND INSTRUMENTATION
Several continuous flow systems and dedicated equipment were developed and applied in automatic
laboratorial determinations or in on-line control of industrial processes. Special attention has been
dedicated to procedures based on the multicommutated flow methodologies and to the new concept of
multi-pumping flow analysis. In this theme the following activities can be stressed:
1 The concept of multicommutated developed by our research group in collaboration with CENA has
been reviewed in order to permit a proper evaluation of the characteristics and potentialities of the
related flow systems, to assist methodological implementation and to present the similarities with
other existing strategies.
2 A novel strategy for the implementation of flow based analytical procedures using micro-pumps has
been proposed. The pumps are switched individually or in combination, in order to create a pulsed
flow stream through the analytical path and are the only active devices acting simultaneously as liquid
propelling units, sample insertion ports and commutating elements.
3 A flow procedure based on the multicommutation concept has been developed for the flow titration of
ascorbic acid in pharmaceutical preparations using potentiometry with an iodide tubular electrode as
detector.
4 Two independent methods for the determination of diclofenac were simultaneously implemented in a
automatic analytical system, based on the concept of sequential injection analysis, providing real time
assessment of the results quality.
5 For the first time a multisyringe flow injection analysis system with potentiometric detection has been
reported. The methodology developed was applied to the determination of chloride in mineral, tap and
wastewaters and the results were compared with those obtained by sequential injection analysis
reported in literature.
6 An automated multicommutated flow system for the spectrophotometric determination of
clomipramine in pharmaceutical preparations was developed. The method was based on the oxidation
by ammonium monovanadate in acidic medium yielding a coloured product. The reaction
development was enhanced when the binary sampling approach was exploited to insert the sample
into the carrier stream.
7 A multi-task flow system based on the concept of sequential injection analysis of potassium
clavulanate in pharmaceutical formulations was developed. In addition to in-line preparation of the
solutions submitted to the analysis, the system simultaneously detected the clavulanate anion and the
potassium cation.
8 A sequential injection analysis system for the determination of Pd in human urine by electrotermal
atomic absorption spectrometry has been developed. The system runs fully independent of the auto
sampler of the equipment. The automatic procedure was based on the retention of the complex
formed between Pd and diethyldithiocarbamate on a column of silica C18 and elution with ethanol
directly into the graphite furnace.
9 An flow injection system with multisided amperometric detection was developed for sequential
determination of two analytes with a single sample injection and a single detector. The flow manifold
was applied in the determination of salicylic and acetylsalycilic acids in pharmaceutical preparations.
10 The usefulness of sonochemical formation of species has been evaluated on the perspective of
reducing or elimination of the reagent over the concept of green analytical chemistry and applied to
the determination of Cr(III).
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11 Development of a versatile potentiometer that works with an electrode array in flow systems. The
potentialities of the instrument were demonstrated by monitoring an array of seven flow-through ionselective electrodes for Ag+ and three electrodes for Cl-, Ca2+ and K+.
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AREA 4
CLEAN PROCESSES AND TECHNOLOGIES
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TOPIC 13
THERMODYNAMICS AND SEPARATION PROCESSES
1 Fundamental thermodynamic research:
a Phase equilibrium study of the binary systems [C4mim][N(Tf)2]+ isobutanol; [C4mim][PF6] + ethanol
and [C4mim][PF6]+H2O, and the ternary system [C4mim][PF6] + ethanol + H2O.
b Studies of supercritical CO2 – induced phase changes in (ionic liquid + water + ethanol) solutions.
Application to Biphasic Catalysis.
c Measurement of the densities of [C4mim][PF6], [C4mim][N(Tf)2] and of the densities and excess
volumes of [C4mim][N(Tf)2]+ [C4mim][BF4], [C2mim][N(Tf)2]+ [C10mim][N(Tf)2] and [C6mim][N(Tf)2]+
[C10mim][N(Tf)2].
2. Phase behaviour study of multicomponent systems:
a Acoustic technique was applied to investigate the critical behaviour of the binary systems CO2 +
perfluorinated surfactant (Krytox).
b Theoretical modelling of critical data using the Peng-Robinson Equation of State through the
calculation of critical and pseudo-critical lines.
c Vapour-liquid equilibrium of the binary systems:
• CO2 and citrus oil; VLE measurements were performed at temperatures between 323.15 K and
343.15 K and at pressures up to 12 MPa.
• C2H6 and citrus oil; VLE measurements were performed at temperatures between 323.15 K and
343.15 K and at pressures up to 12 MPa.
• CO2 and newly synthesised surfactants; solubility data and cloud points was obtained.
• CpCo(CO 2) + CO 2; p,T isopleths were measured by a synthetic method. Nine different
compositions ranging from 17.56 mol% to 94.23 mol% of CO2 were measured up to 15 MPa.
d Theoretical modelling of the experimental phase equilibrium compositions of CpCo(CO2) + CO2
with the PREOS and fitting two temperature-dependent interaction parameters.
e Theoretical modelling of the vapour liquid equilibria of the ternary system methyl oleate / squalene
/carbon dioxide, measured at pressures up to 25 MPa and in the temperature range of 313 – 343K,
by using, (i) the Soave-Redlich-Kwong equation of state with the Mathias-Klotz-Prausnitz mixing
rule; (ii) polynomial correlations developed in terms of the mass parameters solute solubility and
selectivity of the solvent.
f
Design and construction of a new high pressure spectroscopic cell for phosphorescence and
fluorescence. High-pressure safety tests were already performed.
g Design and assembling of a new high pressure apparatus for the preparation of systems to be
studied using high pressure NMR spectroscopy.
h The high pressure NMR technique was used to provide structural information in systems including
polymers, peptides in CO 2. High pressure NMR spectra were obtained to provide structural
information on mixtures of different surfactants and CO2.
3 Development of clean reaction / separation processes involving alternative solvents:
a Diethylene glycol dimethacrylate (DEGDMA) was monitored by DRS and DSC upon polymerisation
with and without initiator (AIBN) at different temperatures. The resulting polymer was essayed in
CO2 supercritical impregnation experiments together with a liquid crystal (E7 from Merck) to
evaluate the feasibility of a PDLC (polymer dispersed liquid crystal).
b The relaxation associated with the glass transition of the monomers with increased length of the
ethylene chain TriEGDMA and TetraEGDGMA were studied by DRS.
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c The DRS experiments were also applied in the characterisation of the relaxation processes of 3
different biomaterials: chitosan (in collaboration of U. Minho), hydroxypropylcellulose and fructose
(in collaboration with IST).
d The dispersion polymerisation of diethylene glycol dimethacrylate (DEGDMA) was undertaken in
supercritical carbon dioxide using different surfactants, as a stabilizer, and AIBN as initiator. The
effect of the initial concentration of surfactant, initiator, monomer and reaction pressure on the
morphology of the resulting polymer was studied.
e The efficiency of alternative extractors such as the static mixers in supercritical fluid separation
processes was evaluated by experimentally measuring the hydrodynamics and mass transfer
kinetics of a Kenics static mixer when applied for a case study: the fractionation of methyl oleate
mixtures from squalene by means of supercritical carbon dioxide. CFD techniques were applied to
develop a geometric and fluid dynamics model of the Kenics static mixer employed in this study.
This work was performed in collaboration with Prof. José Paulo Mota from CQFB.
f
A dynamic simulation model of a countercurrent supercritical fluid packed extraction column was
developed that takes into account momentum, heat and mass balances along the packing height
of the column as a function of the macro variables, extraction time and packing coordinates. This
model was validated against experimental data collected in the SCF countercurrent column plant.
The model system methyl oleate / squalene / carbon dioxide was used in this study.
TOPIC 14
BIOCATALYSIS IN NONAQUEOUS SOLVENTS
We proceeded with ongoing studies of serine hydrolases in nonconventional media, namely supercritical
fluids and organic solvents. We also looked at the extraction of other proteins using supercritical fluids.
1 Strategies for improving enzyme activity, stability and selectivity in nonaqueous media.
Enzyme performance is often impaired in supercritical CO2. We were able to enhance enzyme activity
in this medium via changes in acid-base conditions, by using ion-exchange materials (solid H+/Na+
buffer pairs and a zeolite), which. These were selected based on the response ofof an organo-soluble
acid-base indicator. The amount of concentration of ion-exchange materials had an important effect
on the catalytic activity of subtilisin and the amount could be correlated with the indicator response in
a CO2/ethane mixturethis was related to the protonation and hydration states of the enzyme. We also
studied the combined effects of water activity and enzyme ionization state on the activity and
enantioselectivity of cutinase immobilized on different supports in organic media. The choice of
substrate was based on modelling studies. The organo-soluble indicator was used to monitor the
parameter pH-pNa that was again controlled with solid H+/Na+ buffer pairs. We looked at how the
approaches used affected the yields of conversion, the competition between hydrolysis and
transesterification and cutinase enantioselectivity. We further demonstrated that salt hydrate pairs,
commonly used as in-situ water activity buffers for enzymatic reactions in nonaqueous media, can
have important acid-base effects on enzymes in these media. Finally, we showed that providing acidbase control is extremely important for the evaluation of true water activity effects on enzyme activity
in nonaqueous solvents.
2 Micelle formation. We proceeded with studies aimed at determining the effect of several
experimental parameters on the behavior of subtilisin encapsulated in reverse micelles of aerosol OT
(AOT) in supercritical ethane, by developing a factorial design. We also determined the size of the
micelles formed at some of the experimental conditions tested for reaction, by using dynamic light
scattering. We kept looking for surfactants appropriate for forming micelles in supercritical CO2 that
might allow enzymatic reactions. We started a study of enzyme catalysis in such systems, using
pepsin.
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3 Macroemulsions in CO2. We applied the concept of macroemulsions to protein extraction with
supercritical CO2. This concept was tested in an apparatus whose main piece of equipment is a static
mixer. We solved analytical problems related to the quantification of protein and the measurement of
activity of the target protein. We had the opportunity to participate in a number of experiments
performed with this apparatus that demonstrated the usefulness of the methodology for protein
extraction.
4 Aroma synthesis. We started operating the continuous set-up for the synthesis of geranyl acetate.
We looked at the separation of the reaction mixture components and at the effect of operation
conditions on reaction yield.
TOPIC 15
PHOTOCHEMISTRY AND SUPRAMOLECULAR CHEMISTRY
The research developed by our group during the year 2002 concerns the development of chemical
sensors based on supramolecular (photo)chemistry. Taking into account that Supramolecular chemistry
is based on the “bottom up” approach, i. e., from simple to complex systems, our approximation to the
field of chemical sensors always begins with the synthesis of several molecules containing receptor units
(at the moment, polyamine chains or macrocycles) and signalling units (fluorescent moieties, such as
naphthalene, anthracene, pyrene, etc).
In general, fluorescent sensors are very promising systems due their high sensitivity. Following the
“bottom-up” approach, a simple chemical sensor can be elaborated towards different functions, at the
expenses of an increasing level of complexity. For example, a fluorescent chemosensor can transduce
the coordination of a given substrate in a luminescent signal but it could also store this information in an
optical memory, so it could be read later and only then erased. In certain systems, elementary
movements were explored, originating components for the building of simple molecular machines.
During the year 2002, the Photochemistry and Supramolecular Chemistry group focused most of his
effort in the European network “Molecular-Level Devices and Machines”. The research activity of our
group during 2002 results from the involvment of all group members in local based research as well as
national and international collaborations. In national collaborations, the ITQB (J. C. Lima) was our most
active partner. From international collaborations, several articles were published with the participation of
the University of Bolonha (Italy), Florence (Italy), Amsterdam (Holland), Bonn (Germany), Valencia
(Spain), Santiago de Compostela (Spain) (C. Lodeiro) and São Paulo (Brasil) (J. C. Lima).
TOPIC 16
CHEMICAL REACTION ENGINEERING AND CATALYSIS
Several studies were developed, as follows:
1 Activated carbon supported catalysts were prepared by the incipient wetness method in order to study
the adsorption of SO2. Various transition and alkali metal salts were tested. In situ X ray diffraction
essays were also carried out in order to identify the phases present during reactions. The catalyst
mixture effect was also studied in the adsorption of SO2. Synergistic effects were observed for the
binary mixtures. The most promising system was V+Cu.
2 Cr (III) and Cr (VI) adsorption studies were performed in acidic and basic activated carbons at
different pH in batch systems. Obtained results showed that chromium adsorbs better in acidic
carbons. Those experimental results were afterwards modulated (Langmuir, BET, Toth, RadkePrausnitz isotherms). Those activated carbons were also characterised by XPS, TPD and N2
adsorption (77K).
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3 Polymeric catalytic membranes consisting of molibdophosphoric acid immobilized in dense polymeric
matrixes such as polyvinyl alcohol and polydimethylsiloxane have been tested in the hydration
reaction of alpha-pinene. The strong dependence between catalytic activity and the polymer crosslinking shows clearly that the pinene conversion is diffusion controlled.
4 Homopolymers of N-isopropylacrylamide (PNIPAAM) with different chain lengths and coplymers
PNIPAAM/Vinylsaccharide (copoli(N-isopropylacrylamide/1-deoxy-1-methacrylamido-D-glucitol) with
different compositions were synthesised and characterised. Liquid-liquid equilibria studies of aqueous
solutions of all these polyacrylamides were performed in a wide range of (T,p) experimental conditions
(300 < T/K < 460, 0 < p/bar < 700). An He/Ne laser light scattering technique was used for the
determination of cloud-point (T, p, x) conditions. The experimental results were used to assist in the
determination of computed values at temperatures beyond experimental accessibility, which are
obtained by the application of a modified Flory-Huggins model. The model also estimates the excess
properties of these solutions. Due to the intrinsic self-associating nature of these systems, all studied
solutions show a Lower Critical Solution Temperature (LCST). Both modeling results and H/D isotope
substitution effects suggest also the existence of Upper Critical Solution Temperatures (UCST),
therefore closed-loop type phase diagrams. However, these upper-temperature branches are
experimentally inaccessible. Pressure effects were particularly interesting.
5 A novel method to prepare supported catalysts using a plasma torch was carried out using physical
mixtures of a metal (Pd, particles or molecular precursor) and a traditional support material (alumina).
Catalytic studies of the selective isomerisation of 1-butene indicate that catalysts created with this
novel method are more selective than catalysts prepared on the same support by incipient wetness.
Characterisation studies suggest that the plasma generated catalysts consist of nano-scale metal
particles on highly modified support material.
6 Under projects in cooperation with other two groups of the same CQFB scientific area, was performed
characterisation of polymers, namely polyhydroxyalkanoates obtained by biosynthesis and
polyacrilates obtained in supercritical CO2.
TOPIC 17
MEMBRANE ENGINEERING
1 Food Quality and Processing
a Concentration of fruit juices by osmotic evaporation. Osmotic evaporation is a membrane
technique with fascinating advantages over conventional concentration processes, since it works at
low temperature and pressure, thus avoiding thermal and mechanical damage of the solutes. The
effect of temperature, concentration and osmotic agent’s nature and of the hydrodynamic
conditions on the water transport through the membrane was evaluated. Resistance to water vapor
transfer through the membrane can be described by molecular and Knudsen resistances. The
effect of the nature and of the characteristics of the hydrophobic membrane–porosity, thickness
and pore diameter – on these two mechanisms was evaluated.
b Membrane Processing and Monitoring. Two main projects were developed in this field: 1 – study
of the recovery of aroma compounds from diluted aqueous streams by pervaporation; 2 –
development of integrated pre-enrichment techniques for sample discrimination by electronic
sensorial systems (electronic nose). The project on the recovery of aromas by pervaporation was
focused on the understanding of the molecular interactions between target solutes and the
membrane material. Real-time monitoring of the permeating stream, by on-line mass spectrometry,
allowed us to describe the process of solute permeation in steady-state and in transient operating
conditions. Pervaporation was also integrated with electronic sensorial systems, in order to
enhance discrimination of wine samples, in an automated mode. This approach allowed us to
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successfully use the electronic nose for sample discrimination, even when they present high
ethanol content.
2 Clean Technologies
a Membrane Processing. The ultimate goal of this project is the design of clean membrane
processes for recovery, separation and purification of high added-value products. The main
scientific focus in this area is the understanding and mathematical modeling of the transport
process of the species involved across porous and non-porous membranes, with a special
emphasis on water transport in liquid membranes, transport of solutes through ionic liquids’
supported liquid membranes, transport of hydrophobic molecules through non-porous membranes
and transport of electrically charged species in ion-exchange membranes. This project has been
focused on the study of membrane processes for clean and selective recovery of biological
products (chiral compounds, aromas and proteins) from dilute streams. Different membrane
processes have been studied, namely liquid membrane extraction using selective carriers (chiral
selectors), pervaporation for integrated reaction and recovery of solutes from dilute aqueous
streams and from ionic liquids, ultrafiltration for fractionation of proteins with pharmaceutical
interest, and nanofiltration for recovery and fractionation of antioxidant compounds from
agroindustrial waste streams.
b Production of polyhydroxyalkanoates. PHAs are thermoplastics synthesized by bacteria, having
similar material properties of polypropylene. Contrary to the petroleum based plastics, PHAs are
biodegradable and are produced from renewable resources (sugars and fatty acids) and then their
utilization have no impact in the environment. Conditions that lead to the optimum production of
PHAs by mixed microbial cultures were studied in the last year. Sequencing batch reactors (SBR)
were used to improve the yield of PHA production. Under the optimum conditions used, the amount
of PHA produced per cell dry weight reached 80%, which is similar to the value reported for pure
cultures. Characterization of these biopolymers confirmed identical properties of those of
polypropylene. It was possible to define the composition of the biopolymer, and consequently its
properties, by using different types of substrates. The microbial culture presented in the reactor
was identified by using molecular probes (Fluorescence in situ Hybridization-FISH) and their
metabolism is currently being studied.
c Adsorbed Natural Gas. Adsorbed Natural Gas (ANG) is the storage technology that achieves the
best compromise between compression costs and on-board storage capacity. We have looked at
some of the key issues for ANG technology, which are the efficient management of heat effects
and the development of filtering systems to protect the main gas storage vessel by trapping
undesirable components during the filling phase and releasing them in a controlled manner on the
emptying part of the cycle.
d Hybrid gas separation process. A new hybrid gas separation process combining membrane
permeation and pressure swing (PSA) adsorption has been developed. The coupled process
increases the efficiency of the pressurization and high-pressure adsorption steps, thereby
improving separation performance as compared to a standalone PSA.
e Viscous fluid mixing by chaotic advection. Viscous fluid mixing by chaotic advection in both
time-periodic and spatially-periodic 3-D prototype flows have been studied experimentally and
theoretically. For the case of periodically forced flows we have shown that there is an optimum
modulation frequency that leads to better mixing. The use of analytical tools, such as stretching
calculations and tracer tracking methods, confirm that the optimised protocol does result in very
effective mixing.
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f
Adaptive software sensors. This work addressed the application of software sensors for reaction
kinetics in stirred tank bioreactors. The application of adaptive observer-based estimators was
extensively studied. Several theoretical aspects concerning stability, tuning and dynamics of
convergence were subject of detailed analysis. The application of the theory was illustrated with a
baker´s yeast fermentation process. The results clearly confirmed the theoretical developments.
g Adaptive control of dissolved oxygen concentration in bioreactors. In many industrial
fermentation processes oxygen availability is the main limiting factor for product production. The
main purpose of this study was to derive an effective closed-loop control solution for the addition of
the carbon source when the oxygen transfer rate is close to the maximum. The main results
showed that DO concentration may be precisely controlled with a model reference adaptive
controller and that the productivity of a penicillin production process could be significantly
improved.
h Modelling cells reaction kinetics with modular artificial neural networks. Modular neural
networks are able to perform task decomposition and are commonly applied for speech recognition
and image analysis. The objective of this work was to demonstrate the applicability of these types
of networks in complex biological systems. The main results showed that modular networks are
able to discriminate between metabolic pathways during the training phase and are able to develop
expertise in describing the kinetics of each of the pathways
3 Bioremediation
a Membrane Bioreactors. The bioreactors studied included cell recycle using ultra/microfiltration
membranes (e.g., for drinking water denitrification), membrane supported biofilm reactors using
non-porous hydrophobic membranes (e.g., for detoxification of industrial waste waters
contaminated with organochlorinated pollutants), supported liquid membrane bioreactors using
ionic liquids (e.g., for treatment of pharmaceutical effluents), and development of ion-exchange
membrane bioreactors for removal and degradation of ionic micropollutants from drinking water
supplies (e.g., for water denitrification and removal of perchlorate). This project involved also two
major concerns: the development of new, on-line monitoring techniques, namely 2D-fluorometry
and confocal microscopy for monitoring of biofilms; and the development of mathematical models
of solute transport across the permselective membrane and biofilm.
b Wastewater Treatment using biological processes. Nutrient removal was successfully achieved
in a SBR operated with short intermittent aeration cycles. Different operating conditions were used
and the efficiency of ammonia, phosphorus and carbon removal evaluated. This process allows for
a 40% reduction on the aeration time and consequently on the treatment plant operating costs.
Biodegradation of pesticides was studied using mixed cultures. The microbial consortium used
proved able to completely mineralise molinate from water streams.
TOPIC 18
BIOINORGANIC AND PROTEIN ENGINEERING
1 IRON-SULFUR PROTEINS AND PROTEIN ENGINEERING
a NMR solution structures of two mutants of desulforedoxin. The differences in geometry at the
metal centres in the two known [Fe-4S] proteins rubredoxin (Rd) and desulforedoxin (Dx) are
postulated to be a result of the different spacing of the C-terminal cysteine pair in the two proteins.
In order to address this question, two mutants of Desulfovibrio gigas Dx with modified cysteinyl
spacing were prepared and their solution structures have been determined by NMR.The solution
structure of the muttants were solved by NMR.
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b Formation of a Stable Cyano-Bridged Dinuclear Iron Cluster in Superoxide Reductases.
Spectroscopic evidence for the formation of a dinuclear cyano-bridged adduct after K(3)Fe(CN)(6)
oxidation of the superoxide reductases neelaredoxin from Treponema pallidum and
desulfoferrodoxin from Desulfovibrio vulgaris was reported.
c Biochemical and Spectroscopic Characterization of a New Iron-Sulfur Protein Isolated from
Desulfovibrio desulfuricans ATCC 27774. A novel iron-sulfur protein was purified from the
extract of Desulfovibrio desulfuricans (ATCC 27774).The purified protein is a 13.4kDa dimmer with
a polypeptide chain containing 60 amino acids residues, with 8 cysteines that coordinate the two
[4Fe-4S] clusters. 1D and 2D 1H NMR studies are reported on the oxidized 2x[4Fe-4S] clusters of
the Desulfovibrio desulfuricans ferredoxin ATCC 27774 (Dd27Fd).
2 MOLYBDENUM AND TUNGSTEN CONTAINING ENZYMES
a Gene sequence and the 1.8 A crystal structure of the tungsten-containing formate
dehydrogenase. Desulfovibrio gigas formate dehydrogenase is the first representative of a
tungsten-containing enzyme from a mesophile that has been structurally characterized. The protein
was purified and spectroscopically characterized.
b Biochemical and spectroscopic characterization of an aldehyde oxidoreductase isolated
from Desulfovibrio aminophilus. We have purified and characterized an aldehyde
oxidoreductase from Desulfovibrio aminophilus (Dam), a sulfate reducing bacterium. The enzyme
is a homodimer (monomer ca 100 kDa), and contains a molybdenum centre and two [2Fe-2S]
clusters. Its UV-visible and EPR spectra in native and reduced states shows large similarities with
other AORs from Desulfovibrio gigas (Dg), Desulfovibrio desulfuricans and Desulfovibrio
alasakensis (2-4).
c Nitrate Reductase from Pseudomonas nautica. The heat solubilized and detergent solubilized
nitrate reductases were purified from the cytoplasmic membranes of Pseudomonas nautica. The
entire enzyme complex is composed of three subunits having Mr of 130,000 Da, 65,000 Da and
20,000 Da.
3 HYDROGENASES
a
O ENDOR detection of a solvent-derived Ni-(OH(x))-Fe bridge in the hydrogenase from D.
gigas. The nature and chemical characteristics of the Ni-X-Fe bridging ligand in Desulfovibrio
gigas (Dg) and Desulfovibrio vulgaris Miyazaki (DvM) were studied by 35 GHz CW (17)O ENDOR
measurements on the Ni-A form of the enzyme, exchanged into H(2)(17)O, on the active Ni-C (S =
(1)/(2)) form prepared by H(2)-reduction of Ni-A in H(2)(17)O, and also on Ni-A formed by
reoxidation of Ni-C in H(2)(17)O.
17
4 EPR AND SPIN LABEL STUDIES
a Bilirubin directly disrupts membrane lipid polarity and fluidity, protein order, and redox
status in rat mitochondria. EPR spectroscopy analysis of isolated rat mitochondria was used to
test the hypothesis that UCB physically interacts with mitochondria to induce structural membrane
perturbation, leading to increased permeability, and subsequent release of apoptotic factors. It was
concluded that apoptosis induced by UCB may be mediated, at least in part, by physical
perturbation of the mitochondrial membrane.
5 OTHER STUDIES
a Membrane structural changes support the involvement of mitochondria in the bile saltinduced apoptosis of rat hepatocytes and Perturbation of membrane dynamics in nerve
cells is an early event during bilirubin-induced apoptosis
b The presequence of murine 5-Aminolevulinate Synthase
c The murine 5-Aminolevulinate Synthase, the first enzyme of the heme biosynthetic pathway
d High-Pressure NMR spectroscopy of polymers and biopolymers in CO2 emulsions
e Vif (virion infectivity factor) structure and localization during HIV-2 infection.
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TOPIC 19
BIOCHEMISTRY AND BIOPHYSICAL OF PROTEINS
1 Denitrification Enzymes and Reduction of Nitrite to Ammonia. During this year severall studies
were performed on the N2O reductase from P.nautica 617. The amino acid sequence of the gene
cluster necessary to the expression of the protein is in progress. Spectroscopic methods combined
with density functional calculations are used to develop a detailed bonding description of the m(4)sulfide bridged tetranuclear Cu-Z cluster in N2O reductase. The ground state of Cu-Z has the 1Cu(II)/
3Cu(I) configuration. The single electron hole dominantly resides on one Cu atom (Cu-I) and partially
delocalizes onto a second Cu atom (Cu-II) via a Cu-I-S-Cu-II sigma/sigma superexchange pathway
which is manifested by a Cu-II —> Cu-I intervalence transfer transition in absorption. The dominantly
localized electronic structure description of the Cu-Z site results from interactions of Cull with the two
additional Cu’s of the cluster (Cu-III/Cu-IV), where the Cu-Cu electrostatic interactions lead to hole
localization with no metal-metal bonding. The substrate binding edge of Cu-Z has a dominantly
oxidized Cu-I and a dominantly reduced Cu-IV. The electronic structure description of Cu-Z provides a
strategy to overcome the reaction barrier of N2O reduction at this Cu-I/Cu-IV edge by simultaneous
two-electron transfer to N2O in a bridged binding mode. One electron can be donated directly from
Cu-IV and the other from Cull through the Cu-II-S-Cu-I sigma/sigma superexchange pathway. A
frontier orbital scheme provides molecular insight into the catalytic mechanism of N2O reduction by
the Cu-Z cluster.
The nitrite reductase from D.desulfuricas 27774 is an enzyme with two type of subunities (NrfA and
NrfH) was studied by electrochemistry and the development of a biosensor is on the way.The number
of hemes per subunity in NrfH was determined (4 hemes of the type c). NrfA contains 5 hemes c and
its structure was determined by X-ray crystallography. These considerations enabled the revision of a
vast amount of existing spectroscopic data on the Nrfha complex that was not originally well
interpreted due to the lack of knowledge on the heme content and the oloigomeric enzyme status.
Based on EPR and Mossbauer parameters and their correlation to structural information recently
obtained from X-ray crystallography on the NrfA structure we proposed the full assignment of midpoint
reduction potentials values to the individual hemes. NrfA contains the high-spin catalytic site (-80 mV).
In NrfA is also located a quite unusual high redox potential (+150mV9/low-spin bis-His coordinated
heme, considered to be the site where electrons enter.
2 Peroxidases. The production of cytochrome c peroxidase (CCP) from Pseudomonas (Ps.) stutzeri
(ATCC 11607) was optimized by adjusting the composition of the growth medium and aeration of the
culture. The protein was isolated and characterized biochemically and spectroscopically in the
oxidized and mixed valence forms. The activity of Ps. stutzeri CCP was studied using two different
ferrocytochromes as electron donors: Ps. stutzeri cytochrome C-551 (the physiological electron donor)
and horse heart cytochrome c. These electron donors interact differently with Ps. stutzeri CCP,
exhibiting different ionic strength dependence. The CCP from Paracoccus (Pa.) denitrificans was
proposed to have two different Ca2+ binding sites: one usually occupied (site I) and the other either
empty or partially occupied in the oxidized enzyme (site II). The Ps. stutzeri enzyme was purified in a
form with tightly bound Ca2+. The affinity for Ca2+ in the mixed valence enzyme is so high that Ca2+
returns to it from the EGTA which was added to empty the site in the oxidized enzyme. Molecular
mass determination by ultracentrifugation and behavior on gel filtration chromatography have revealed
that this CCP is isolated as an active dimer, in contrast to the Pa. denitrificans CCP which requires
added Ca2+ for formation of the dimer and also for activation of the enzyme. This is consistent with
the proposal that Ca2+ in the bacterial peroxidases influences the monomer/dimer equilibrium and the
transition to the active form of the enzyme. Additional Ca2+ does affect both the kinetics of oxidation
of horse heart cytochrome c (but not cytochrome C-551) and higher aggregation states of the enzyme.
This suggests the presence of a superficial Ca2+ binding site of low affinity.
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3 Ferrochelatase. Resonance Raman (RR) spectroscopy is used to examine porphyrin substrate,
product, and inhibitor interactions with the active site of murine ferrochelatase (EC 4.99.1.1), the
terminal enzyme in the biosynthesis of heme. The enzyme catalyzes in vivo Fe2+ chelation into
protoporphyrin IX to give heme. The RR spectra of native ferrochelatase show that the protein, as
isolated, contains varying amounts of endogenously bound high- or low-spin ferric heme, always at
much less than 1 equiv. RR data on the binding of free-base protoporphyrin IX and its metalated
complexes (Fe(III), Fe(II), and Ni(II)) to active wild-type protein were obtained at varying ratios of
porphyrin to protein. The binding of ferric heme, a known inhibitor of the enzyme, leads to the
formation of a low-spin six-coordinate adduct. Ferrous heme, the enzyme’s natural product, binds in
the ferrous high-spin five-coordinate state. Ni(II) protoporphyrin, a metalloporphyrin that has a low
tendency toward axial ligation, becomes distorted when bound to ferrochelatase. Similarly for freebase protoporphyrin, the natural substrate of ferrochelatase, the RR spectra of porphyrin-protein
complexes reveal a saddling distortion of the porphyrin. These results corroborate and extend our
previous findings that porphyrin distortion, a crucial step of the catalytic mechanism, occurs even in
the absence of bound metal substrate. Moreover, RR data reveal the presence of an amino acid
residue in the active site of ferrochelatase which is capable of specific axial ligation to metals
4 Iron-sulfur proteins. Desulfovibrio gigas desulforedoxin (Dx) consists of two identical peptides, each
containing one [Fe-4S] center per monomer. Variants with different iron and zinc metal compositions
arise when desulforedoxin is produced recombinantly from Escherichia coli. The three forms of the
protein, the two homodimers [Fe(III)/Fe(III)]Dx and [Zn(II)/Zn(II)]Dx, and the heterodimer [Fe(III)/
Zn(II)]Dx, can be separated by ion exchange chromatography on the basis of their charge differences.
Once separated, the desulforedoxins containing iron can be reduced with added dithionite. For NMR
studies, different protein samples were prepared labeled with N-15 or N-15 + C-13. Spectral
assignments were determined for [Fe(II)/Fe(II)]Dx and [Fe(II)/Zn(II)]Dx from 3D N-15 TOCSY-HSQC
and NOESY-HSQC data, and compared with those reported previously for [Zn(II)/Zn(II)]Dx.
Assignments for the C-13(alpha) shifts were obtained from an HNCA experiment. Comparison of H-1N-15 HSQC spectra of [Zn(II)/Zn(II)]Dx, [Fe(II)/Fe(II)]Dx and [Fe(II)/Zn(II)]Dx revealed that the
pseudocontact shifts in [Fe(II)/Zn(II)]Dx can be decomposed into inter- and intramonomer
components, which, when summed, accurately predict the observed pseudocontact shifts observed
for [Fe(II)/Fe(II)]Dx. The degree of linearity observed in the pseudocontact shifts for residues greater
than or equal to8.5 Angstrom from the metal center indicates that the replacement of Fe(II) by Zn(II)
produces little or no change in the structure of Dx. The results suggest a general strategy for the
analysis of NMR spectra of homo-oligomeric proteins in which a paramagnetic center introduced into
a single subunit is used to break the magnetic symmetry and make it possible to obtain distance
constraints (both pseudocontact and NOE) between subunits.
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AREA 5
CATALYSTS, SOLVENTS AND NON-TOXIC COMPOUNDS
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TOPIC 20
NOVEL HETEROGENEOUS CATALYSTS
1 Metalossurfactants. As part of the ongoing project on “Design of novel metalossurfactants with
catalytic properties: physical-chemical, morphologic and reactivity properties” we have characterized
the morphology of the aggregates formed in aqueous solution by [Fe(RR’bpy)2(CN)2] and RR’Bpy,
where R and R’ are alkyl chains with 1, 5, 7, 9, 13 and 17 carbon atoms. The morphological
characterization was performed by several complementary microscopic techniques (HR-TEM, FESEM and cryo-SEM) and by dynamic light-scattering. Some of the studies have been performed in the
University of Nijmegen, Netherlands, in collaboration with Prof. R. Nolte and Dr. M. Feiters. The
results obtained show a remarkable dependence of the morphology of the metallo-aggregates with the
length of the alkyl chain.
2 Metal Nanoassemblies. This work is being developed in collaboration with Prof. J. Shelnutt, Sandia
National Laboratories, USA. The main purpose of the work is to develop chemical methods to
produce metal nanoparticles with controlled shape and size. We have been able to prepare
nanoassemblies of Pt nanoparticles at the surface of micelles and vesicles, containing a variable
number of Pt nanoparticles. These nano-assemblies are extraordinarily stable in solution and, unlikely
Pt-sols, do not precipitate over long periods of time (several monthes). These nano-assemblies also
show a high reactivity as catalysts for hydrogen production, with no need for use of electron-relay
intermediates, and a strong resistance towards poisoning.
3 Platinum complexes with cytotoxic effects. This work is performed in collaboration with Dr. Paula
Marques, University of Coimbra, and its main purpose is to prepare Pt complexes with biogenic
polyamines, to test their cytoctoxic effects and correlate them with the structure/spectroscopic
properties of the complexes. Several complexes have been prepared, and some of them show very
promising cytotoxic activity.
4 Novel catalysts by heterogenisation of metal complexes. Manganese and Nickel complexes with
Schiff base ligands (salen type) have been synthesised by methodologies developed in our lab and
characterised by several techniques: elemental analyses, mass spectra, cyclic voltammetry , EPR,
FTIR and UV-Vis spectroscopies. Several X-ray diffraction studies have been made where crystals
were obtained. The catalytic activity of these complexes in the epoxidation of styrene was evaluated in
CH2Cl2 and CH3CN, using NaOCl and PhIO as oxygen sources. The experiments were done at room
temperature and reaction time, the styrene conversion, chemical selectivities of products (epoxide and
benzaldehyde) and respective yields were determined by GC-FID. All the complexes showed catalytic
activity in the experimental conditions used, although the data shown that these complexes are more
efficient when PhIO was used as the oxygen source in CH3CN. The complexes were anchored onto
activated carbons and were encapsulated in fauzasite zeolites (NaX and NaY) and pillared clays
(PILCs) using several strategies already developed in our lab. The complexes were anchored onto
microporous activated carbons (obtained from NORIT) using linking agents such as cyannuric
chloride or by direct reaction between the groups on the ligands and the surface carbon groups. The
complexes were encapsulated in zeolites and PILCs by the flexible ligand method where the ligand is
made to diffuse within the pores of the material, that was previously modified with manganese or
nickel acetate. All the materials were characterised by elemental analyses, XPS, SEM (EDS), XRD,
nitrogen adsorption, temperature programmed desorption (TPD), termogravimetry, EPR (Mn
complexes), FTIR and Uv-vis spectroscopies. The catalytic properties of the new catalysts were
determined in the heterogeneous epoxidation of styrene in CH2Cl2 and CH3CN, using NaOCl and
PhIO as oxygen sources, using equivalent experimental conditions of those used in homogeneous
phase. The same catalytic properties were evaluated (reaction time, styrene conversion, chemical
selectivities of products, epoxide and benzaldehyde and respective yields). For these catalysts we
have evaluated the leaching of the active phase and their reusabibility. All the catalysts showed
catalytic activity: the reaction times were larger than those observed in homogeneous phase, the
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styrene conversion decreased, but chemical selectivities were quite similar to those of homogeneous
media. Nickel catalysts showed partial leaching of the complexes when encapsulated in zeolites, but
Mn complexes immobilised in activated carbons and PILCs dis not show leaching of Mn centres and
could be reused until 3 times without loose significant catalytic efficiency.
5 Conducting films for electrocatalysis. Metal salen complexes (M=Cu, Ni, Pd and Pt) were prepared
by methods developed in our lab and were characterised by standard techniques. These monomers
were electropolymerised in CH3CN at Pt electrodes by potentiodynamic methods, using different
supporting electrolytes. For some of the monomers, the polymerisation process was studied using
several techniques: coulometry, UV-vis spectroscopy, electrochemical microbalance (EQCM) and
elipsometry. The data suggested nucleation as a first step followed by consolidation/reticulation
processes. The films were characterised by several techniques: cyclic voltammetry, rotating disk
electrode voltammetry, chronoamperometry, SEM, AFM, in-situ elipsometry, EQCM coupled to probe
beam deflection (PBD), in-situ FTIR and UV-Vis, electroacoustic and electrochemical impedance,
EPR. The films showed, in general, high electrochemical stability (which depends strongly on the
substituents in the aldehyde moiety), behaves as electrochromic materials, and show a semiconductor
behaviour. The films can act as electrochemical mediators towards electrochemical probes
(ferrocene) and depending on their thickness can behave as rigid films or viscoelastic materials.
6 Chemosensors based on transition metal complexes. Nickel and copper complexes with Schiff
base ligands functionalised with groups that can act as coodination sites for representative and
lanthanide cations were prepared by methodologies developed in our laboratory. The functionalities
(crown ether and pseudo crown ethers groups, flexible pendent arms bearing oxygen and nitrogen
atoms) were introduced in the aldehyde moiety and imine bridge of the Schiff base. These complexes
were characterised by several techniques: elemental analyses, mass spectra, EPR, FTIR and UV-Vis
spectroscopies and cyclic voltammetry. Cation recognition studies were performed in several solvents,
using cyclic voltammetry and UV-vis spectroscopy and for the complexes that polymerise in CH3CN,
recognition studies were also done in the respective films using cyclic voltammetry, in-situ FTIR and
UV-Vis and electroacoustic impedance. In solution, it was possible to determine equilibrium constants
for the complexation of lanthanides and alkaline-earth cations for the majority of the metal complexes
by Uv-vis spectroscopy. Cyclic voltammetry allowed the semi-quantiative evalutation of the interaction
of all the cations tested (+1, +2 and +3). The polymeric films adsorded all the metal cations tested (+1,
+2 and +3), although they exhibited different behaviour. This technique also showed that depending
on the experimental conditions used, metal ion can be de-complexed and the film can be reused for
recognition of other metal ions. IR and UV-Vis spectroscopies indicated the film cation coordination
sites and the influence of their presence in the electronic structure of the films. Equilibrium constants
for the adsorption of Barium were determined by electroacoustic impedance.
TOPIC 21
THEORETICAL ELECTROCHEMISTRY AND METAL CATALYSIS
Electrochemistry interfacial processes and metal catalysis phenomena play a major role in many areas of
(bio)chemistry and physics, with important pharmaceutical and industrial applications. Our group is
particularly interested in simulating these types of phenomena at a microscopic level. We have been
using both numerical simulation techniques, such as Molecular Dynamics (MD) simulations, and
quantum mechanical methods (e.g., Density Functional Theory (DFT)). A detailed description concerning
our main present interests, which have derived from years 2000/2001, follows.
1 Interfacial phenomena. Several liquid interfaces (e.g., water|nitrobenzene and water|2-heptanone),
which are commonly used on bench experiments and on large-scale industrial processes were
studied thoroughly. MD simulations were carried out to analyse in detail these interfaces at a
molecular level. A careful choice of the inter and intramolecular potentials used on those simulations,
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and, in some cases, the development of new ones (by resorting to quantum calculations) was needed.
The interfaces were found to be molecularly sharp, highly corrugated and broadened by capillary
waves. A novel description of the dynamics of liquid interfaces was achieved, which help in clarifying a
subject controversial for more than one decade. In addition, possible changes in the interface’s
properties that may be induced by the addition of an electrolyte to both solvents (as usual in
experiments) or the change of temperature have been evaluated.
The process of ion transfer across a liquid|liquid interface has also been studied. The potential of
mean force for the transfer of small ions was calculated by non-classical MD simulations. The results
show that the transfer process is an activationless process, and its mechanism is now well
understood.
Considering other more complex ions and new liquid|liquid interfaces with different miscibilities will
extend this work further and some of this work is now being finalized. The inclusion of a prototype
biological membrane between the two phases is presently being considered and the first publication
has been prepared.
2 Metal Catalysis. Since 1995 we have been exploring the applicability of DFT methods, together with
the cluster model approach, to the study of catalytic processes by metallic surfaces. The first systems
considered were methanol and ethylene (as well as of its reaction products or intermediates)
adsorbed on copper, silver and gold surfaces. These three surfaces were chosen due to the
anomalous interaction of the methanol and ethylene species on silver when compared with the
generality of other transition metal surfaces. Satisfactory results concerning conformations, energetics
and vibrational frequencies were attained. The adsorption of acetylene, which is involved in several
typical elementary catalytic reactions, was also studied. The obtained results are in excellent
agreement with recent Scanning Tunnelling Microscopy results. In addition the mechanism for the
oxidation of methanol in copper, gold and platinum was investigated, and some of its steps have been
elucidated. At the same time, preliminary work is going on for the reactions of methane and carbon
dioxide on supported ruthenium-based catalysts (namely the dry reforming of methane and the
production of methanol).
Theoretical studies are also being pursued that aim to predict how strongly chiral metal surfaces may
differentiate between enantiomers of small adsorbed organic molecules. Specifically, we want to
assess the enatiospecific adsorption properties of D,Lglucose on kinked chiral gold(321)R,S surfaces.
Gold surfaces exhibit a lower poisoning and the study of the electro-oxidation of glucose is quite
important in the development of glucose sensors for in vivo and in vitro measurements of glucose
concentration in human blood (diabetes).
The results support the usefulness of the cluster model approach in studies of heterogeneous
catalysis involving transition metal catalysts, so that this work, if continues to be successful as it
appears to be now, may be extended in future to new reactions and different metals.
THEME 22
PROTEIN CRYSTALLOGRAPHY
1 Enzymes containing the molybdopterin cofactor. We have pursued our studies on different
molybdopterin-containing enzymes and the most important result has been the fully refined crystal
structure of the W-containing formate dehydrogenase.
a W-Formate Dehydrogenase from Desulfovibrio gigas. The Desulfovibrio (D.) gigas formate
dehydrogenase (DgW-FDH) is the first representative of a tungsten-containing enzyme isolated
from a mesophile to be structurally characterized. It is a heterodimer of 110 and 24 kDa subunits.
While the large subunit is homologous to E. coli FDH-H and to D. desulfuricans ATCC 27774
Nitrate Reductase, and harbors the W active site and one [4Fe-4S] center, no structure of a small
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subunit ortholog is available. The small subunit contains three [4Fe-4S] clusters. The structural
homology of with E coli FDH-H shows that the essential residues at the active site are conserved
and include the Se-Cys158, His159 and Arg407. The buried active site is accessible via a positively
charged tunnel, while product release may be facilitated, for H+ by a chain of water molecules and
protonable amino acid side chains and for CO2 through a hydrophobic channel conserved in FDHs.
b Aldehyde oxidoreductase from Desulfovibrio gigas (MOP). New crystallographic studies were
carried out in preparations of the enzyme treated with enzyme inhibitors. From all forms analysed,
crystals treated with high concentrations of arsenite show that the molecule binds directly to the Mo
atom, displacing the hydroxyl ligand. Refinement is under progress.
c Isoquinoline Oxidoreductase (IsoOr) from Brevundimonas diminuta 7 (an enzyme from the
Xanthine Oxidase family). With a new purification protocol more homogeneous samples of IsoOr
were obtained. Recently, single crystals were obtained which will be the starting point for the
crystallographic analysis of this enzyme, in its native and substrate/inhibitor-bound forms.
2 Heme-containing enzymes
a Di-haem Cytochrome c Peroxidases (CCP): Structural basis for the mechanism of Ca2+
activation of the from Pseudomonas nautica 617. The crystal structure of this CCP was
obtained at two different pH values, presenting structural differences, which allow a glimpse at the
calcium activation mechanism. Form IN, obtained at pH 4.0 (P3121) and refined at 2.2 Å resolution
presents a closed conformation where the peroxidatic haem adopts a six ligand coordination,
hindering the peroxidatic reaction to take place. Form OUT, obtained at pH 5.3 (P6422), refined at
2.4 Å resolution, shows an open, active conformation, due to the unbinding of the distal histidine
which provides peroxide access to the active site. This “activated” form has a bound Ca2+ ion,
which is essential for enzymatic activation.
b Membrane bound cytochrome c nitrite reductase from Desulfovibrio desulfuricans at 2.3 Å
(NIR)- The relevance of the two calcium sites in the structure of the catalytic subunit (NrfA).
The gene encoding cytochrome c nitrite reductase (NrfA) from Desulfovibrio desulfuricans ATCC
27774 was sequenced and the crystal structure of the enzyme was determined to 2.3 Å resolution.
In comparison to homologous structures, it presents structural differences mainly located at the
regions surrounding the putative substrate inlet and product outlet, and includes a well defined
second calcium site with octahedral geometry, coordinated to propionates of hemes 3 and 4, and
caged by a loop non-existent in the previous structures. The highly negative electrostatic potential
in the environment around hemes 3 and 4 suggests that the main role of this calcium ion may not
be electrostatic but structural, namely in the stabilization of the conformation of the additional loop
that cages it and influences the solvent accessibility of heme 4. The NrfA active site is similar to
that of peroxidases with a nearby calcium site at the heme distal side nearly in the same location
as occurs in the class II and class III peroxidases.
3- Prostate kallikrein isolated from stallion seminal plasma: a homologue of human PSA– A
project of protein identification and sequencing by crystallography. The protein was purified as a
contaminant of HSP3, a horse seminal protein of the cysteine-rich secretory protein (CRISP) family,
from Equus caballus sperm. At the stage of crystallization and structure solving, the nature of the
protein was unknown. Prostate-specific kallikrein, a member of the gene family of serine proteases,
was initially discovered in semen and is the most useful serum marker for prostate cancer diagnosis
and prognosis. The crystal structure at 1.42 Å resolution of horse prostate kallikrein (HPK) is the first
structure of a serine protease purified from seminal plasma. It shares extensive sequence homology
with human prostate-specific antigen (PSA), including a predicted chymotrypsin-like specificity as
suggested by the presence of a serine residue at position S1 of the specificity pocket. In contrast to
other kallikreins, HPK shows a structurally distinct specificity pocket. Its entrance is blocked by the
kallikrein loop, suggesting a possible protective or substrate selective role for this loop. The HPK
structure seems to be in an inactivated state and further processing might be required to allow the
binding of substrate molecules. Crystal soaking experiments revealed a binding site for Zn2+ and Hg2+,
two known PSA inhibitors.
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THEME 23
TRANSPORT IN BIOLOGICAL SYSTEMS
1 Cystic Fibrosis. The correlation between Water Transport and Cystic Fibrosis Transmembrane
Conductance Regulator (CFTR) channel has been reported in oocytes expressing CFTR and in
airway epithelia when activated by cAMP. In order to further establish these relations, we designed the
following experimental strategy: apply patch clamp techniques to study ionic channels in isolated
membrane patches, and stopped flow techniques to study water transport in membrane vesicles
prepared from the same cell lines.
a Electrophysiological studies in in inside out patches. The activation mechanism of CFTR by
phosphorylation of the R domain is known to depend on PKA, the main kinase responsible for its
activation. Yet, it is not completely clear if in vivo the PKA enzymes that activate CFTR are soluble
or anchored to the cell membrane. In excised inside out patch clamp experiments, with multiple
channels in each patch of CACO2 cell membrane, it was possible to observe a chloride
conductance activated by cAMP in the presence of ATP. Since the membrane patch was nominally
cytoplasm free, the results obtained may suggest that PKA anchored to the cell membrane is
responsible for the activation of CFTR.
b Water Transport in membrane vesicles containing CFTR channel. The results obtained above
suggested that the activation of the CFTR could be obtained in vesicle preparations from the same
cell lines. We performed water permeability studies in membrane vesicles prepared from: Caco2, a
cell line that express normally the CFTR channel, used as positive control; CHO that normally does
not express the CFTR channel, used as negative control; CHO transfected with the normal CFTR
gene. The activation energy (Ea) measured for these different vesicle populations under different
experimental conditions was found to be very high for all the preparations tested. Vesicles from the
Caco2 cell line had an Ea of the order of 15 Kcal.mol-1 that didn’t change when the vesicles were
pre-incubated with CFTR activators (cAMP, ATP, IBMX). For the CHO vesicles, Ea measured in
control conditions (without activators) was similar on both transfected and non-transfected cell
lines and around 12.5 Kcal.mol-1. These results do not show a correlation between the activity of
the CFTR channel and water transport in membrane vesicle preparations.
2 A Minimal Mathematical Model of Calcium Homeostasis (in collaboration with J. F. Raposo, L. G.
Sobrinho from IPO, Lisboa). A mathematical model of calcium homeostasis was built. In the model the
controlling factors were the plasma concentrations of calcium, PTH and calcitriol and the effector
organs were the parathyroids, the bone, the kidney and the intestine. Other factors can be added as
the need arises. The model was aimed at simulating what happens in a single individual but its
parameters and variables can be adjusted to the corresponding published average values.
Simulations of published observations in humans undergoing the infusion of calcium or its chelators
were done. With a single exception, these simulations provided a good fit to the data. The response of
the system to extrinsic perturbations was characterized by simulating chronic infusions of calcium,
PTH and calcitriol. Finally the steady state response to perturbations in some of its parameters (the
secretory mass of the parathyroids and the affinity and/or sensitivity of the calcium, PTH and calcitriol
receptors) and to renal failure was also investigated in an attempt to analyze the pathogenesis of
clinical hypo or hypercalcemias. In its present form the model cannot be used to base clinical
decisions in individual cases. However it requires modest computational resources and clinicians with
a modest mathematical background can manipulate it. It is a useful tool for the analysis of general
mechanisms of the diseases of calcium metabolism and for the design of clinical experiments aimed
at characterizing these diseases. The model can also be the core of future auto - adaptive extensions
to be used in individual patients.
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TOPIC 24
BIOMIMETIC SYSTEMS AND SIMULATION
MOLECULAR MODELLING. We are generally interested in the molecular modelling of biological and
chemical systems. Accordingly, we have been working on systems such as proteins, cyclodextrins,
lanthanide (III) chelates and small peptides. Many of these studies are carried out in close collaboration
with experimentalists. We have been using both molecular simulations and quantum mechanics
techniques, as well as quantum mechanics/molecular mechanics (QM/MM) hybrid methods, to perform
these studies. A more detailed description concerning our main present interests, which have derived
from years 2001/2002 follows:
1 Fight against cancer
a Molecular simulations on lanthanide (III) chelates and host-guest complexes with γ-cyclodextrins:
the aim is to design new contrast agents for Magnetic Resonance Imaging, an important tool for
clinical diagnosis (e.g., tumour therapy). This project is being pursued in close collaboration with
NMR and calorimetry experts (Prof. Carlos Geraldes and Margarida Bastos from the Universities
of Coimbra and Porto, respectively, both in Portugal) and is part of the COST Programme, having
been financed by the Fundação para a Ciência e Tecnologia, Portugal, project PRAXIS/PCEX/C/
QUI/67/96.
b Study of the catalytic mechanism of enzymatic reactions, such as P450 1A2 (an enzyme
involved in the metabolic pathway of carcinogenesis) and RNR (ribonuclease reductase, an
enzyme thought to be involved in cancer), resorting to both quantum mechanics and QM/MM
methods. These studies are being carried out in collaboration with two other computational
chemists (Profs. Cristina Menziani and Leif Eriksson from the Universities of Modena, Italy, and
Uppsala, Sweden, respectively) and being financed by the National Foundation for Cancer
Research, U.S.A. (P450 1A2) and by the Fundação para a Ciência e Tecnologia, Portugal (RNR)
via project POCTI/35376/QUI/2000.
c Screening of 3,5 billion small molecules as potential inhibitors of 16 proteins, directly involved
in cancer, which are current targets of the pharmaceutical industry. This project is part of the work
carried out by the NFCR Centre for Computational Drug Design, based at Oxford, and directed by
Prof. Graham Richard of which Maria João Ramos is an Associate Director, being financed by the
National Foundation for Cancer Research, U.S.A.
2 Drug Design
a Modelling of mimetic modifications of bioactive peptides by inclusion of novel synthetic amino
acids. This project is being worked on in close collaboration with an organic chemist (Prof. Hernâni
Maia from the University of Minho, Portugal) and being financed by the Fundação para a Ciência e
Tecnologia, Portugal, via project POCTI/35380/QUI/2000.
b Design of protein novel inhibitors for P450 1A2 and RNR, using the knowledge acquired on the
transition states obtained during the study of the catalytic pathways of the respective enzymatic
reactions mentioned in part I. The collaborations mentioned in I(ii) apply also here.
c Design of new potential therapeutic agents for the treatment of acquired immunodeficiency
syndrome (AIDS) by investigating the binding modes of different anti-AIDS chelators like ATA, HPH
and other analogs to achieve a better design of anti-HIV metal chelators. This project is being
carried out in close collaboration with Dr. Rakesh Sharma, an experimental organic chemist from
Delhi University, in India.
d Design of new antimalarial compounds based on electrostatic profiles of established drugs. This
study is being performed in collaboration with Prof. Madalena Pinto from the Faculty of Pharmacy
at the University of Porto, Portugal.
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ARTIFICIAL CHEMISTRY. A different theme dedicated to the simulation of biological systems has been
initiated recently using concepts of artificial chemistry. Artificial Chemistry is defined as the result of
multiple and autonomous agent interaction, typically in large variety and quantity. The motion in phase
space of these agents is often conditioned by a complex environment. The set agents/
rules_of_interaction/environment is a formal system that may be isomorphic to a material system, in a
certain degree. Generally, the Artificial Chemistry formal systems use the most general principles that
govern real systems and aplly it to abstract agents in an attempt to obtain similar results and an insight of
which principles are in fact relevant to the functioning of the real system. In Artificial Chemistry the goal is
not so much to model the agents (to model accurately molecules, for example) but to generate a formal
system that is isomorphic to a generally complex, and sometimes unpredictable, system like a biological
system, from a cell to an ecosystem. The work being developed is inspired in the general principles of
Lewis acid-base reactions. These reactions are the base of nucleophilic additions and substitutions for
the synthesis of the most important biological molecules using ATP. In developing a formal system the
agents have therefore different virtual donor and acceptor properties and interact using the well stirred
reactor abstract dynamics. Bonding occurs, with some probability, when a donor finds an acceptor in the
well stirred reactor forming polyagents. It is being build a software tool to implement this kind of formal
system. Presently, the tool enables to define a mixture of a large number of agents of ten different kinds
with up to four bonding sites each. Each site may have an arbitrary donor or acceptor strength. The basic
implementation done up to now enables to create systems with interesting properties like (1) the
establishment of equilibrium states for polyagent quantities, (2) the mimetization of Brønsted-like acidbase reactions and of (3) competitive SN1-type reaction mechanisms. Some important properties like
catalysis, self-regulation and polyagent recognition need the definition of a 3D placement of bonding sites
in each kind of agent, giving the polyagents conformational property. Another crucial capability that the
software tool must have is to be able to implement an open formal system, allowing for agent flux through
the well stirred reactor. These two features are to be developed during 2003.
TOPIC 25
BIOPHYSICS OF ORGANIZED MEDIA
The determination of partition coefficients, for fluoroquinolones, tuberculostatic and benzodiazepines in
DMPG and DMPC liposomes, have been performed, with and without phase separation, by
spectrophotometry, derivative spectroscopy, and fluorimetry. The results have shown that drug partition is
controlled by size, charge and fluidity of the organized media and by the hydrophilicity and acid/base properties
of the drugs.
Location studies have been performed in the same liposomes systems by fluorimetry and NMR. Quenching
fluorescence studies were performed to assess drug location using several stearic acids as fluorescence
probes, for the drugs that are non fluorescent, and using KI as a quenching agent for fluorescent drugs.
Anisotropy fluorescence studies were also performed to determine changes in membrane fluidity using the
fluorescent fluidity probes, 1,6-diphenyl-1,3,5-hexatriene (DPH) and trimethylammonium-diphenylhexatriene
(TMA-DPH). These results show that the degree of incorporation depends on the electrostatic/hydrophobic
characteristics of the liposomes and on the hydrophobic characteristics and acid/base properties of the
drugs. The NMR results corroborate these assumptions. For fluoroquinolones it was possible to conclude
that, the mechanism by which these drugs permeates through the phospholipid bilayer must include an
electrostatic adsorption at the interface region and this association must be the first step that governs their
mechanism of interaction with bacterial natural membranes.
We have also begun to use these liposomes systems as membrane models to understand the role of lipid
composition on the mechanism of action of some membrane proteins transporters. The proteins that we are
using are the P-glycoprotein multidrug transporter and some pore proteins, porins, responsible for the
uptake and transport of fluoroquinolones and tuberculostatics in bacteria.
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TOPIC 26
METAL BASED DRUGS
Activities within this area have been focused in two themes:
1 Design of orally active insulin-mimetic metal complexes. The design of such compounds involves
the synthesis of specific ligands, synthesis of the corresponding metal complexes and evaluation of its
chemical properties, toxicity and insulin-like performance. A set of chelators of the 3-hydroxy-4pyridinone type and exhibiting variable hydrophilic/lipophilic balance have been synthesized and
characterized. The corresponding metal complexes with Zn(II), Cu(II), Ni(II) and Co(II) have also been
synthesized and characterized in the solid state and in solution as has been done previously for the
parent vanadyl complexes. Protonation and stability constants are being determined by potentiometric
and spectrophotometric methods and speciation diagrams established. The behaviour of this
complexes in aqueous solution as a function of pH and metal:ligand ratio is compared with that of the
vanadyl complexes. Evaluation of the insulin-like action of vanadium complexes was made in
collaboration with Prof. Hiromu Sakurai (Kyoto University) and the Toxicology group at REQUIMTE.
2 Design of functionalized siderophores to target infection processes. The design of new chelators
is crucial for treatment of diseases associated with iron overload and also to prevent the growth of
undesirable bacteria. Among the latter, pathogenic bacteria are associated to a great variety of
infectious processes that lead to well known diseases such as TUBERCULOSIS and AIDS. To
understand the action of the siderophores it is necessary to be sure that the molecules are able to
penetrate de vacuolar cavities of macrophages and also to have a way of mapping its presence in cell
media. Synthetic pathways to functionalize known siderophores like desferrioxamine or enterobactin
as well as new hexadentate chelators are being established in order to obtain molecules able to target
infection processes and remove iron. The work has been initiated in the last trimester of the year and
at the moment a desferrioxamine derivative has been modified in order to be anchored to amino
dextran type molecules.
Química Verde
ANNEXES
A - 2
ANNEX I
RESEARCH TEAM
Química Verde
A - 3
A
MEMBERS OF STAFF
Química Verde
A - 4
Ana M. Lobo
Baltazar Manuel Romão Castro
Conceição Gomez Gonzalez Beça
Hugo Gil Ferreira
Isabel Moura
Jose Alberto Nunes Ferreira Gomes
Jose Luis Fontes Costa Lima
José Moura
Luis Sousa Lobo
Manuel Nunes da Ponte
Margarida Alice Ferreira
Maria Lurdes Souteiro Bastos
Pedro Brito Correia
Sundaresan Prabhakar
Professora Catedrática
Professor Catedrático
UNL
FCUP
FEUP
UNL
UNL
FCUP
FFUP
UNL
UNL
UNL
FFUP
FFUP
UNL
UNL
Abel José de Sousa Costa Vieira
Alberto Manuel Carneiro Sereno
Alberto Nova Araujo
Ana Cristina Moreira Freire
Duarte José da Costa Pereira
Fernando Pina
João Paulo Goulão Crespo
Jose Fernando Magalhâes Cardoso
Karin Tonnies Gil Ferreira
Maria Conceição B. S. M Montenegro
Maria Conceição S. S. Rangel Gonçalves
Maria João Ribeiro Nunes Ramos
Maria João Romão
Maria Pilar Figueroa Gonçalves
Maria Teresa Barros
Rosa Maria Moreira Seabra Pinto
Susana Filipe Barreiros
Teresa Maria Fonseca de Moura
Professor Associado
Professor Associado
Professor Associado
Professora Associada
Professor Associado
Professor Associado
Professor Associado
Professor Associado
UNL
FEUP
FFUP
FCUP
FCUP
UNL
UNL
ICBAS-UP
UNL
FFUP
ICBAS-UP
FCUP
UNL
FEUP
UNL
FFUP
UNL
UNL
Cristina Maria Delarue Alvim Matos
Maria Leonor Oliveira Madureira Pinto
Maria Carmo Veiga Fernandes Vaz
Professora Coordenadora
ISEP
ISEP
ISEP
Agostinho Almiro Almeida
Alberta Paula Gameiro Santos
Alexandre Lopes Magalhães
Alice Pereira
Ana Aguiar Ricardo
Ana M. Lourenço
Ana Maria Martelo Ramos
Andre Alberto Sousa Melo
Ângela M. S. Relva
Anjos Macedo
António Gil de Oliveira Santos
António Jorge Parola
Carlos Alberto Mateus Afonso
Cristina Costa
Eduarda Graças Rodrigues Fernandes
Elvira Maria M. Gaspar
Eulália Fernanda Carvalho Pereira
Felix Dias Carvalho
Fernando Manuel Gomes Remião
Helena Maria Vieira Monteiro Soares
Henrique Guedes
Isabel Coelhoso
Isabel Maria Ligeiro da Fonseca
Isabel Maria Viegas Oliveira Ferreira
J. Carlos Lima
João Aires de Sousa
João Luis Machado Santos
João Paulo Noronha
Professor Auxiliar
Professora Auxiliar
Professor Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professor Auxiliar
Professora Auxiliar
Professor Auxiliar
Professor Auxiliar
Professor Auxiliar
Professor Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professor Auxiliar
Professor Auxiliar
Professora Auxiliar
Professor Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professor Auxiliar
Professor Auxiliar
Professor Auxiliar
Professor Auxiliar
FFUP
FCUP
FCUP
UNL
UNL
UNL
UNL
FCUP
UNL
UNL
UNL
UNL
UNL
UNL
FFUP
UNL
FCUP
FFUP
FFUP
FEUP
UNL
UNL
UNL
FFUP
UNL
UNL
FFUP
UNL
Química Verde
A - 5
João Sotomayor
Joquim Silvério Marques Vital
Jorge Lampreia
Jose Augusto Caldeira Pereira
Jose Oliveira Fernandes
Luisa M. Ferreira
M. João Melo
Madalena Dionísio
Manuela Pereira
Marco R.G. da Silva
Maria Ascenção Reis
Maria Beatriz Prior Pinto Oliveira
Maria Beatriz Guerra Junqueiro
Maria Gabriela Teles Cepeda Ribeiro
Maria Lúcia Sousa Saraiva
Maria Natália Dias Soeiro Cordeiro
Maria Salete Reis Dias Rodrigues
Maria Teresa Avilés Perea
Paula Cristina Branquinho Andrade
Paula S. Branco
Paulina Mata
Paulo Mota
Pedro M. Abreu
Pedro Manuel Alexandrino Fernandes
Pedro Simões
Pedro Tavares
Rui Alexandre Santos Lapa
Rui Oliveira
Professor Auxiliar
Professor Auxiliar
Professor Auxiliar
Professor Auxiliar
Professor Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professor Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professor Auxiliar
Professor Auxiliar
Professor Auxiliar
Professor Auxiliar
Professor Auxiliar
Professor Auxiliar
Professor Auxiliar
UNL
UNL
UNL
ICBAS-UP
FFUP
UNL
UNL
UNL
UNL
UNL
UNL
FFUP
FFUP
FCUP
FFUP
FCUP
FFUP
UNL
FFUP
UNL
UNL
UNL
UNL
FCUP
UNL
UNL
FFUP
UNL
Ermelinda Manuela Jesus Garrido
Professora Adjunta
ISEP
Carla Manuela Soares Matos
Catarina Isabel Guerra Rodrigues
Cristina Maria C. Morais Couto
Cristina Maria Cavadas Morais Couto
Ivone Valente Oliveira
João Luís Tavares de Matos Gomes
Jorge Caldeira
Lígia Maria da Silva Rebelo Gomes
Luísa Lima Gonçalves
Maria Alexandra Bernardo
Maria da Graça Soveral Rodrigues
Maria Helena Reis Prado de Castro
Rita Isabel Lemos Catarino
Sara Isabel Xavier Candeias
Stephane Besson
Professor Auxiliar
Professor Auxiliar
Professor Auxiliar
Professor Auxiliar
Professora Auxiliar
Professor Auxiliar
Professor Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professora Auxiliar
Professor Auxiliar
UFP
ISCSN
ISCSN
ISCSN
ISCSN
UFP
ISCSS
ISCSN
ISCSS
ISCSS
FFUL
ISCSN
UFP
U. Losófona
U. Losófona
Ana Maria Philips
Elisa Pais Rocha Gonçalves
Eurico Cabrita
Isabel Mafra
Marcela Alves Segundo
Maria Manuel Marques
Owen Catchpole
Serguey Bursakov
Uwe Pischel
Investigador Principal
Investigadora Principal
Investigador Auxiliar
Investigadora Auxiliar
UNL
UP
REQUIMTE
REQUIMTE
REQUIMTE
REQUIMTE
REQUIMTE
REQUIMTE
REQUIMTE
Manuel Rui Azevedo Alves
Jorge Manuel Pinto Jesus Garrido
Maria Isabel Branco Alves Martins
Maria Teresa de Oliva Teles Moreira
Patrícia Valentão
Olivia Maria de Castro Pinho
Susana Isabel Pereira Casal
Abel José Assunção Duarte
Professor Coordenador
Professor Adjunto
Professora Adjunta
Professora Adjunta
Assistente
Assistente
Assistente
Assistente
IPVC
ISEP
ISEP
ISEP
FFUP
FCNAUP
FFUP
ISEP
Química Verde
A - 6
Hendrikus Petrus Antonius Nouws
José Tomás Soares de Albergaria
Maria de Fátima de Sá Barroso
Maria Goreti Ferreira Sales
Maria Isabel Brito Limpo de Serra
Maria João Dantas Ramalhosa Ferreira
Maria Manuela Barbosa Correia
Olga Manuela Matos de Freitas
Rita Isabel Simões Ferreira
Simone Barreira Morais
Salomé de Sousa Teixeira
Valentina Maria Fernandes Rodrigues
Maria Elisa Amorim Matos Fernandes Soares
Eulalia Maria Bernardino Mendes
Maria Isabel Almeida Cardoso
Carla Rodrigues
Cecília Bonifácio
João Fraga
Nelson Brito
Rui Viegas
Maria Aurora Soares da Silva
Sérgio Cruz Monteiro de Morais
Assistente
Assistente
Assistente
Assistente
Assistente
Assistente
Assistente
Assistente
Assistente
Assistente
Assistente
Assistente
Assessora Principal
Assessora
Assessora
Téc. Sup. 2.ª Classe
Téc. Informática
Téc. Informática
Encarregada de trabalhos
Encarregado de trabalhos
ISEP
ISEP
ISEP
ISEP
ISEP
ISEP
ISEP
ISEP
IPVC
ISEP
ISEP
ISEP
FFUP
FFUP
FFUP
REQUIMTE
REQUIMTE
REQUIMTE
REQUIMTE
REQUIMTE
ISEP
ISEP
KEY
UNL
FCUP
FFUP
FCNAUP
ISEP
ISCSN
UFP
Universidade Nova de Lisboa
Universidade do Porto - Faculdade de Ciências
Universidade do Porto - Faculdade de Farmácia
Universidade do Porto - Fac.Ciências da Nutrição
Instituto Superior de Engenharia do Porto
Instituto Superior de Ciências de Saúse (Norte)
Universidade Ferbando Pessoa
FFUL
FEUP
ICBAS-UP
Universidade de Lisboa - Faculdade de Farmácia
Universidade do Porto - Faculdade de Engenharia
Universidade do Porto - Inst. Ciências Biomédicas
IPVC
Inst. Politécnico de Viana do Castelo
ISCSS
Instituto Superior de Ciências de Saúse (Sul)
U. Lusófona Universidade Lusófona
Química Verde
A - 7
B
POST-DOCTORAL FELLOWS
Química Verde
A - 8
Ana Luisa Carvalho
Anders Thapper
Carla Sousa
Carlos Brondino
Carlos Lodeiro
Elsa Henriques
Françoise Auchère
Iwona Biernacka
João Miguel Dias
José Trincão
Krasimira Petrova
Lalit Sharma
PauloLemos
Pavel Izak
Rakesh Kumar
Roeland Boer
Rui Duarte
Shuwen Yao
Smilja Todorovic
Svetlana Lyubchik
Svetlozar Velizarov
Thierry Michaud
Thomas Schäfer
Vanya Bogdanova Kurteva
Yuri Binev
Química Verde
A - 9
C
Ph. D. STUDENTS
Química Verde
A - 10
Adelaide de Sousa
Adriana Martins Pimenta
Akapong Suwattanamala
Alexandra Antunes
Ana Cláudia Barreira Nunes
Ana Pamplona
Ana Rosa Silva
Andrea Carneiro
Axel Müller
Branca Maria Cardoso Monteiro da Silva
Bruno Nunes
Carina Madalena Martins Machado
Carla Portugal
Carlos Cunha
Carmen Pinheiro
Célia Peres
Christophe Siquet
Cristina Cordas
Cristina Correia
Cristina Manuela Pinto Vieira Lopes
Cristina Matos
Daniel Antunes dos Santos
David de Andrade Sousa Costa
Emilia Maria Gonçalves Santos
Eugénia Nogueiro
Fernando Siñeriz de Paz
Filomena Freitas
Francisco SIlva
Gabriela Almeida
Gundula Wolf
Helena Suzana da Costa Machado Ferreira
Hugo Santos
Inês Cabrito
Isabel Cristina Timóteo
Isabel Esteves
Joana Andrêa Soares Amaral
João Adalberto Amaral Lourenço
João Alexandre Velho Prior
João Miguel Reis de Aguiar Navarro y Rosa
Jorge Alexandre S. Pereira
Jorge Dias
Jorge Rebelo
José Eduardo dos Santos Félix Castanheiro
José Esperança
José luis dos Santos
Karine Lopes Marques
Laila Hassano Ribeiro
Ludwig Krippahl
Luís Alexandre Fernandes Cobra Branco
Luis Mayor Lopez
Luisa Serafim
Magda Basto
Manuela Frasco
Márcia Cláudia Dias Carvalho
Marco Preto
Margarida Moncada
Maria de Fátima Assunção Lucas
Maria Luisa Soares Silva
Maria M. Santos
Maria Teresa Viciosa Plaza
Mariana Duarte
Mário Eusébio
Mário Gomes
Marlene Susana Dionísio Lucio
Marta Andrade
Marta Corvo
Marta Filipa Teixeira Ribeiro
Marta Morais Saraiva de Andrade
Nuno Cerqueira
Nuno Milhazes
Patrícia Neves
Patrícia Raleiras
Paula Cristina de Almeida Jerónimo
Paula Cristina de Azevedo Gomes Pinto
Paula Garcia
Paulo Glória
Pedro Manuel da Cunha Catalão Pires dos
Santos
Pedro Miguel Pimenta Gois
Pedro Rodrigues Marques Maia
Pedro Vidinha
Raquel Fortunato
Rita Noronha
Rui Ruivo
Rute Fonseca
Sara Cristina Silva Cunha
Sérgio Alves
Sílvia Garcia
Sofia Antunes
Sofia Nunes
Sofia Rocha Pauleta
Susana Gaudêncio
Susana Pereira
Tânia Costa
Teresa Alves
Teresa Casimiro
Teresa Maria de Jesus Teixeira
Teresa Santos Silva
Vasco Bonifácio
Victor Alves
Vineet Pande
Wancheng Sittikijyothin
Química Verde
A - 11
D
M. Sc. STUDENTS
and
other research students
Química Verde
A - 12
Ana Coelho
Ana Isabel Farinha
Ana Vinha
Angélica Alves da Silva
Carla Beatriz Rodrigues Veiros
Cristina Maria Rodrigues Ferreira Alves
Delfina de Vasconcelos
Joana Maria Fernandes Meireles
M. Sc. Students
Luís Carlos Matos
Luis Miguel do Carmo
Luis Neto
Maria Manuela Mendes Pinto
Maria Virgínia Gomes Mota
Mário Cardoso
Susana Maria Ferreira de Melo Abreu
Paula Alexandra Carvalho Rodrigues
Other Research Students
Alexandre Carvalho
Ana Cristina Cruz
Ana Rita Duarte
Ana Sofia Cruz
Ana Sofia Pinto
Angelo Figueiredo
Carla Rosa
Carlota Macedo
Celina Santos
Duarte Paulo Martins Torres
Elsa Maria Pereira
Elsa Soares
Hugo Santos
Iva Costa
Luís Pinto
Márcia Guilherme
Maria Adelaide Miranda
Maria João Amorim
Marta Lopes
Nádia Ribeiro
Pedro Nunes
Ricardo Branco
Sandro Soares
Silvia Alexandra Lopes Fernandes
Soraia dos Santos
Susana Nascimento
Valdemar Figueira
Vânia Maria Alves Gomes Chibante
Química Verde
A - 13
E
Profiles of the new researchers
contracted under the Associate
Laboratory Program
Química Verde
A - 14
Dr. Owen John Catchpole was born on the 6th of July 1963. He has a Bachelor of Chemical Engineering,
Hons (1st), by the University of Canterbury, NZ, (1986), and a Ph. D. in Chemical Engineering, by the University
of Birmingham, UK, (1991). In 2002 he received the New Zealand Royal Society Silver Medal by his notable
research work. The present research/professional activities are concerned with Supercritical fluid (SCF)
extraction, scale-up and design of pilot plants; Catalytic reaction engineering; and Herbal processing (steam
distillation, solvent extraction).
As major research and development achievements it should be cited the design, construction,
commissioning and operation of New Zealand’s first commercial scale SCF extraction facility; the development
of patented technology for removing cholesterol from butter, and for producing low cholesterol meat products by
SCF extraction; and the invention of a novel process using gas and supercritical anti-solvent technology to
separate mixtures of natural products. He his author/co-author of chapters in three books on supercritical extraction,
5 patents/patent applications, 19 journal papers, 40 refereed publications.
Dr. Sergey Bursakov graduated in Soil Science and Agrochemistry with specialization in Microbiology
and has got his Master of Science in 1983 in Faculty of Soil Science, Department of Soil Biology Department
of Soil Biology of M.V.Lomonosov Moscow State University, Russia. He defended PhD Thesis in Biochemistry
in 1987 with Highest Honors in A.N.Bach Institute of Biochemistry Russian Academy of Science. He worked
from 1996 in Department of Chemistry New University of Lisbon as visiting professor in the lab. Professor
J.J.G. Moura (1995 – 1997), PhD scientist – in the lab. Professor I.Moura (1997 – 2002). At the end of 2002
he joined the Requimte – Laboratório Associado – CQFB as an Auxiliary Researcher in area of protein
biochemistry.
Dr.S.Bursakov have teaching and scientific experience and coordination MS and PhD Thesis. He is
involved in several projects and has published 28 articles, and 42 communications. Scientific interests:
enzymology, spectroscopy of metalloenzymes, biochemistry and physiology plants and microorganisms.
Special interest focused on the metabolism of sulfate reducing bacteria and several molybdenum (nitrate
reductase, aldehide oxido reductase, formate dehydrogenase), tungsten (formate dehydrogenase), cobalt
and zinc (ATP sulfurylase, ADP kinase) containing enzymes.
Dr. Eurico J. Cabrita graduated in Applied Chemistry (Organic Chemistry) in 1994 in Faculdade de
Ciências e Tecnologia (Universidade Nova de Lisboa), where he obtained his Ph.D. degree in Organic
Chemistry in 1999. In 2000 he joined the Institut für Analytische Chemie in Leipzig, Germany, as a posdoctoral research fellow. There he worked in the development and application of NMR diffusion based
techniques in organic chemistry. In 2002 and while still in Leipzig he started collaboration with researchers
from the Centro de Química Fina e Biotecnologia of FCT-UNL in a multidisciplinary project, involving, molecular
modeling, NMR spectroscopy, and organic synthesis, for the rationalization of the induction of asymmetry in
organic chemistry. At the end of 2002 he joined the Requimte - Laboratório Associado - Centro de Química
Fina e Biotecnologia where he is currently an Auxiliary Researcher in NMR. His research interests include
the application and development of new techniques in NMR, with special focus to the study and understanding
of reaction mechanisms, molecular recognition and intermolecular interactions in catalytic and/or asymmetric
processes, the study of reactive intermediates and the conjugation of NMR, molecular modeling and organic
synthesis as a tool to the study of reaction mechanisms.
Dr. Uwe Pischel is a German citizen, who graduated in Chemistry at the Humboldt-University Berlin
(1998) and obtained his PhD in Chemistry from the University of Basel (2001) working in mechanistic
organic photochemistry and the development of fluorescent probes. In June 2002 he went for a postdoctoral
stay to the CSIC Institute of Chemical Technology at the Polytechnical University of Valencia, which was
supported by a prestigious Research Fellowship of the Deutsche Forschungsgemeinschaft. There Dr. Pischel
was involved in the exploration of chiral recognition in excited state processes. His curriculum vitae includes
Química Verde
A - 15
23 publications, often in international leading journals (J. Am. Chem. Soc., Angew. Chem. Int. Ed.). In 2002
Dr. Pischel was awarded the Prize of the Faculty of Philosophy and Natural Sciences of the University of
Basel and in 2003 the Albert-Weller-Prize of the Photochemistry Group of the German Chemical Society. In
the beginning of 2003 he joined the group of Inorganic Chemistry/Materials Chemistry at REQUIMTE/
University of Porto. His current research interests include the development of new luminescent probes and
materials based on transition metal complexes.
Dr. Isabel Mafra graduated in Food Engineering in 1992 in Escola Superior de Biotecnologia
Universidade Católica Portuguesa. After some experience in the industry, she joined the Faculdade de
Engenharia da Universidade do Porto in 1994 where she worked in research projects related to the
development of analytical methods (HPLC-fluorescence, GLC) for food analysis. She obtained a Master in
Biological Engineering in 1997 in Universidade do Minho. In 1997 she joined the Departamento de Química
Universidade de Aveiro where she carried out research work in a FAIR project (OLITEXT FAIR CT 97-3053)
in the study of olive pulp cell wall polysaccharides related to changes with ripening and processing of some
european varieties suitable to table olives. In 2002 she obtained her PhD in Chemistry (Biochemistry) in
Universidade de Aveiro. In the same year and same institution she started to work as a pos-doctoral researcher
in the application of NMR to the structural characterisation of polysaccharides. At the beggining of 2003 she
joined the REQUIMTE Laboratório Associado – CEQUP as an Auxiliar Researcher. The reasearch work
has been carried out in Faculdade de Farmácia Universidade do Porto and relates to the development and
application of Molecular Biology methods, namely PCR techniques, for the evaluation of PDO (Protected
Designation of Origin) food, food autenticity and detection of GMO (Genetically Modified Organisms) in
food. Her research interests include the development and applicaton of PCR tecniques in GMO, cheeses,
oil-bearing crops and derived food and feed stuffs.
The scientific and technical profile of Doctor Maria Manuel Barata Marques is clearly centered in the
synthesis of natural products. In fact both her Ph.D. Work (concluded in 2001) as well as her postdoctoral
work with J. Mulzer, in Vienna, focused in the development of new synthetic methodologies for heterocycles
and natural products of biological importance. From her postgraduate CV one may draw attention to the
areas of ‘green chemistry’ (Venice Summer School), mass spectrometry (Jennings, FCT-UNL) and NMR
(Zaragoca). One expects that Dr. MMB Marques will become an important addition to the scientific human
resources of REQUIMTE.
Química Verde
A - 16
ANNEX II
PUBLICATIONS
Química Verde
A - 17
A
Scientific Articles
and
Book Chapters
Química Verde
A - 18
High-pressure phase-behaviour monitoring for reaction processes improvement
A. Aguiar-Ricardo; T. Casimiro, C.M.M. Duarte, H.M. Woods; R.M.T. Griffiths; S.M. Howdle; N.Ribeiro
Chem. Eng. Transactions, 2002, Vol. 2, 591-596
Determination of chloride by multisyringe flow injection analysis and sequential injection analysis
with potentiometric detection
A. Andrade-Eiroa, J. A. Erustes, R. Forteza, V. Cerdá, J.L.F.C. Lima
Anal. Chim. Acta, 467, 25-33 (2002)
Generation dependent photoactivity and pH sensitiviy of dendrimers: methyl orange-substituted
poly(prophyleneamines)
A. Dirksen, E. Zuidema, C. Kauffmann, A. Roque, R. M. Williams, F. Pina, F. Vötgle, L. De
Macromolecules, 2002, Vol. 35, 2743-2747
Proton transfer in anthocyanins and related flavylium salts. Determination of ground-state rate
constants with nanosecond laser flash photolysis
A. L. Macanita; P. F. Moreira; J. C. Lima; F. H. Quina; C. Yihwa; C. Vautier-Giongo
J. Phys. Chem., 2002, Vol. 106, 1248-1255
Simultaneous potentiometric and flurimetric determination of diclofenac in a sequencial injection
analysis system
A. M. Pimenta, A. N. Araújo, MJ.C.B.S.M. Montenegro
Anal. Chim. Acta, 470, 185-194 (2002)
Hygrogenation of pinene in carbon dioxide: comparasion between two and three phase
A. Milewska; D. Gourgouillon; D. Chouchi; I. Fonseca; M. Nunes da Ponte
Superc. Fluids Chem. Reactivit, 2002, 129-132
Immobilisation of amine functionalised nickel(II) Schiff base complexes onto activated carbon
treated with thionyl chloride
A. R. Silva, M. Martins, M. M. A. Freitas, A. Valente, C. Freire, B. de Castro, J. L. Figueiredo
Microporous and Mesoporous Materials, 2002, 55, 275-284.
Photochromic propierties of 3-Methyl substituted flavylium salts
A. Roque; C. Lodeiro; F. Pina; M. Maestri; R. Ballardini and V. Balzani
Eur. J. Org. Chem., 2002, Vol. 16, 2699-2709
Crystal structure of a prostate kallikrein isolated from stallion seminal plasma: a putative PSA
A.L. Carvalho; L. Sanz; D. Barettino; A. Romero; J.J. Calvete; M.J. Romão
J. Mol. Biol., 2002, Vol. 322, 325-337
A sequencial injection analysis system for potassium clavulanate determination using two
potentiometric detectors
Adriana M. Pimenta, Alberto N. Araújo, Maria Conceição B.S.M. Montenegro
J. Pharm. Biom. Anal., 30, 931-937 (2002)
As limitações da linguagem no ensino da mecânica quântica
Alexandre Magalhães
Boletim da SPQ, 2002, 86, 70-72
Silicon and Iron levels in tissues of animals treated with a “ferrimagnetic ceramic” with
oncotherapeutic potential (anti-tumor) value
Almeida, T., Soares, M. E., Cavalheiro, J., Bastos, M. L.
J. Trace Elem. Med. Biol., 2002, 16, 255-259
Separation and quantification of the major casein fractions by reverse-phase high-performance
liquid chromatography and urea-polyacrylamide gel electrophoresis detection
Ana C.A. Veloso, Natércia Teixeira, Isabel M.P.L.V.O. Ferreira
J. Chromatogr. A, 2002, 967, 209-218
Detecção de adulterações em produtos alimentares contendo leite e/ou proteínas lácteas
Ana Cristina A. Veloso, Natércia Teixeira, Isabel M.P.L.V.O. Ferreira e Margarida A.
Quim. Nova, 2002, 25, 4, 609-615
Química Verde
A - 19
Potentiometric flow injection determination of glycerol in distilled spirits
Ana Paula G. Gervasio, Eduardo P. Borges, Elias A.G. Zagatto, Boaventura F. Reis, Rui A.S. Lapa, José
L.F.C. Lima
J. Agric. Food Chem, 50, 74 - 77 (2002)
Automatic potentiometric flow titration procedure for ascorbic acid determination in
pharmaceutical formulations
Ana Paula S. Paim, Cristina M.N.V. Almeida, Boaventura F. Reis, Rui A.S. Lapa, Elias A.G. Zagatto, José
L.F.C. Lima,
J. Pharm. Biom. Anal, 28, 1221 - 1225 (2002)
Heterogenization of a Functionalized Copper(II) Schiff Base Complex by Direct Immobilization
onto an Oxidized Activated Carbon
Ana R. Silva, M.ª Madalena A. Freitas, Cristina Freire, Baltazar de Castro, José Luís Figueiredo
Langmuir, 2002, 18, 8017-8024.
Hydrogen evolution and consumption in AOT-isooctane reverse micelles by Desulfovibrio gigas
hydrogenase
Andrade SLA, Moura JJG
Enzyme and Microbial Technolog, 2002, Vol. 31, 398-402
Photochemistry of nickel salen based complexes and relevance to catalysis
B. de Castro, R. Ferreira, C. Freire, H. Garcia, E. J. Palomares, M. J. Sabater
New J. Chem., 2002, 26, 405-410.
Zinc-Substituted Desulfovibrio gigas desulforedoxins:resolving subunit degenaracy with nonsymmetric pseudocontact shifts
B.Goodfellow; S.Nunes; F.Rusnak;I.Moura;C.Ascenso;J.J.G.Moura;B.F.Volkman
Protein Sci., 2002, Vol. 11, 2464-2470
Reductive activation of aerobically purified Desulfovibrio vulgaris hydrogenase: Mössbauer
characterization of the catalytic H cluster
B.H. Huynh, P. Tavares, A.S. Pereira, I. Moura, J.J.G. Moura
Power of Anaerobes, 2002,
The adsorption of ethylene on the (110) surfaces of copper, silver and platinum: a DFT study
Bernardo C. G. P. M., Gomes J. A. N. F.
J. Mol. Struct. (Theochem), 2002, 159-169
Study of the azidoacetic acid adsorption in Ag (111)
C. A. Dias; R. Carrapa, M. T. Barros; T. A. Gasche; O. M. N. D. Teodoro; M.L.Costa, M. H. Cabral; A. M.
C. Moutinho
Vacuum, 2002, Vol. 64, 445
Synthesis of 4-Aryl-butylamine Fluorescent Probes
C. A. M. Afonso, J. P. S. Farinha
J. Chem. Research, 2002, 584-586
Biogeociclos: Uma visão molecular das enzimas e dos mecanismos envolvidos nos ciclos dos
elementos - Parte I: Biogeociclo do Azoto
C. Carneiro; J.J.G. Moura
Bol. Soc. Port. Quím., 2002, Vol. 86, 67-82
Manipulation of the Reactivity of a Synthetic Anthocyanin Analogue in Aqueous Micellar
C. V.-Giongo, C. Yihwa, P. F. Moreira, Jr., J. C. Lima, A. A. Freitas, M. Alves, F. H. Quina, A. L. Maçanita
Langmuir, 2002, Vol. 18, Nr. 26, 10109-10115
Phase Equilibrium for Capsaicin + Water + Ethanol + Supercritical Carbon Dioxide
C.M.M. Duarte; M. Crew; T. Casimiro; A. Aguiar-Ricardo, M. Nunes da Ponte
J. Supercritical Fluids, 2002, Vol. 22, 87-92
Química Verde
A - 20
Second and Third Virial Coefficients of Three Binary Mixtures Containing Xenon, at 273 K.
Comparison between, Xe+C2H4, Xe+C2H6 and Xe+CO2
C.M.M.Duarte; C. Menduiña; A. Aguiar-Ricardo; M. Nunes da Ponte
Phys. Chem. Chem. Phys., 2002, Vol. 4, 4709-4715
Bilirubin directly disrupts membrane lipid polarity and fluidity, protein order and redox status in
rat mitochondria
C.M.P.Rodrigues; S.Sola; M.A.Brito; D.Brites; J.J.G.Moura
J. Hepat., 2002, Vol. 36, 335-341
Perturbation of membrane dynamics in nerve cells as an early event during bilirubin-induced
apoptosis
C.M.P.Rodrigues; S.Sola; R.E.Castro; P.A.Laires; D.Brites; J.J.G.Moura
Journal of Lipid Research, 2002, Vol. 43, 885-894
Identification of 4-methylthioamphetamine and some of its metabolites in mice urine by GC-MS,
after acute administration
Carmo, H., Boer, D., Remião, F., Carvalho, F., Reys, L. A., Bastos, M. L.
J. Anal. Toxicol., 2002, 26, 228-232
Effect of 3,4-methylenedioxymethamphetamine (“ecstasy”) on body temperature and liver
antioxidant status in mice. Influence of ambient temperature
Carvalho, M., Carvalho, F., Pereira, M.L., Pires-das-Neves, R., Bastos, M.L.
Arch. Toxicol., 2002, 76, 166-172
Role of metabolites in MDMA (ecstasy)-induced nephrotoxicity: an in vitro study using Rat and
Human renal proximal tubular cells
Carvalho, M., Hawksworth, G., Milhazes, N., Borges, F., Monks, T., Fernandes, E., Carvalho, F., Bastos, M.L.
Arch Toxicol., 2002, 76, 581–588
Interaction of grepafloxacin with large unilamellar liposomes: partition and fluorescence studies
reveal the importance of charge interactions
Catarina Rodrigues, Paula Gameiro, Salette Reis, J.L.F.C. Lima, Baltazar de Castro
Langmuir, 2002, 18, 10231-10236.
Citrate selective electrodes for the flow injection analysis of softdrinks, beers and
pharmaceuticals products
Cristina M. F. Ribeiro, Cristina Delarue-Matos, M. Goreti F. Sales, M. Carmo V. F. Vaz
Anal. Chim. Acta, 2002, 471, 41-49
Application of amperometric sol-gel biosensor to flow injection determination of glucose
Cristina Couto, Alberto Araújo, Conceição Montenegro, Jarbas Rohwedder, Ivo Raimundo, Célio Pasquini
Talanta, ,56, 997 - 1003 (2002)
Quantification of organic acids in grape musts and port wines
Cunha,S.C., Fernandes, J.O., Faria, M.A., Ferreira, I.M.P.L.V.O., Ferreira, M.A.
Cienc. Tecnol. Aliment., 2002, 3, 4, 212-216
In vitro study of the insulin-mimetic behaviour of vanadium(IV,V) coordination compounds
D. Rehder, J. Pessoa, C. Geraldes, M. Castro, T. Kabanos, T. Kiss, B. Meier, G. Micera, L. Pettersson, M.
Rangel, A. Salifoglou, I. Turel, D. Wang
J. Biol. Inorg. Chem., 2002, 7, 384-396
Molecular Dynamics Study of the Calcium Ion Transfer Across the Water/Nitrobenzene
dos Santos D. J. V. A., Gomes J. A. N. F.
ChemPhysChem, 2002, 3, 946-951
Fatty acid composition of Portuguese spreadable fats with emphasis on trans isomers
Duarte Torres, Susana Casal, M.B.P.P. Oliveira
Eur. Food Res. Technol., 2002, 214, 108-111
HR DOSY as a new tool for the study of chemical exchange phenomena
E. J. Cabrita; S. Berger
J. Magn. Reson., 2002, Vol. 40, S122-S127
Química Verde
A - 21
High resolution DOSY NMR with spins in different chemical surrounding: Influence of particle
exchange
E. J. Cabrita; S. Berger, P. Bräurer, J. Kärger
J. Magn. Reson., 2002, Vol. 157, 124-131
Viability and release of complexing compounds during accumulation of heavy metals by a
brewer’s strain
E.V. Soares, A.P.R.S. Duarte, R.A. Boaventura, H.M.V.M. Soares
Appl. Microbiol. Biot., 2002, 58, 836-841
Use of Saccharomyces cerevisiae for Cu2+ removal from solution: the advantage of using a
flocculent strain
E.V. Soares, G. De Coninck, F. Duarte, H.M.V.M. Soares
Biotechnol. Lett., 2002, 24, 663-666
Addition of nutrients induce a fast loss of flocculation in starved cells of Saccharomyces
Eduardo V. Soares, Abel A. Duarte
Biotechnol. Lett., 2002, 24, 1957-1960
Electrochemical determination of methyltin compounds
H.M.V.M. Soares, C.M.M. Machado, M.G.R.T. Barros
Mikrochim. Acta, 2002, 138, 43-48
Reductive electrochemical study of nickel(II) complexes with N2O2 Schiff base complexes and
spectroscopic characterisation of reduced species. Reactivity towards CO
F. Azevedo, C. Freire, B. de Castro.
Polyhedron, 2002, 21, 1695-1705.
Internal Dynamics of Poly(Methylphenylsiloxane) Chains as Revealed by Picosecond Time
Resolved Fluorescence, as a function of chain dimension
F. B. Dias; J. C. Lima; I. F. Piérola; A. Horta; A. L. Maçanita
Long Range Electron Transfer Quenching in Polyamine Chains Bearing a Terminal Naphthalene
Unit
F. Pina; J. C. Lima; C. Lodeiro; J. Seixas de Melo; P. Diaz; M. T. Albelda; E. Garcia-España
J. Phys. Chem., 2002, Vol. 106, 8207-8212
Superoxide reductase activities of neelaredoxin and desulfoferrodoxin metalloproteins
F. Rusnak, C. Ascenso, I. Moura and J.J.G. Moura
Methods in Enzymology, 2002, Vol. 349, 243-258
Multicommutation in flow analysis: concepts, applications and trends
Fábio R. P. Rocha, Boaventura F. Reis, Elias Zagatto, José L.F.C. Lima, Rui Lapa, João L.M. Santos
Anal. Chim. Acta, 468, 119-131 (2002)
2-Styrylchromones as novel inhibitors of xanthine oxidase. A structure-activity study
Fernandes, E., Carvalho, Silva, A.M.S., Santos, C.M.M., Pinto, D.C.G.A., Cavaleiro, J.A.S., Bastos, M.L.
J. Enz. Inhib. Med. Chem., 2002, 17(1),45-48
The reduction of ribonucleotides catalyzed by the enzyme ribonucleotide reductase
Fernandes, P. A., Eriksson, L. A., Ramos, M. J.
Theor. Chem. Acc., 2002, 108, 352-364
Zeolite molecular sieves have dramatic acid-base effects on enzymes in nonaqueous media
Fontes, N.; Partridge, J.; Halling, P.J.; Barreiros, S.
Biotechnol. Bioeng., 2002, Vol. 77, 296-305
Stoichiometric Production of Hydrogen Peroxide and Parallel Formation of Ferric Multimers
through Decay of the Diferric-Peroxo Complex, the First Detectable Intermediate in Ferritin
Mineralization
G. N. L. Jameson, W. Jin, C. Krebs, A. S. Pereira, P. Tavares, X. Liu, E. C. Theil; B. H.
Biochemistry, 2002, 41(45); 13435-13443
Química Verde
A - 22
Unraveling the substrate-metal binding site of ferrochelatase: An x-ray absorption spectroscopy
study
G.C. Ferreira; R. Franco; A. Mangravita; and G.N. George
Biochemistry, 2002, Vol. 41, Nr. 15, 4809-4818
Phenolic antioxidant compounds produced by in vitro shoots of sage (Salvia officinalis L.)
Gomes, P.C.; Seabra, R.M.; Andrade, P.B.; Ferreira, M.F.
Plant Sci., 2002, 162, 981-987
Gene Sequence and the 1.8 Å Crystal Structure of the Tungsten-containing Formate
Dehydrogenase from Desulfovibrio gigas
H. Raaijmakers; S. Macieira; J. M. Dias; S. Teixeira; S. Bursakov; R. Huber; J. J. G. Moura; I. Moura; M.
J. Romão
Structure Fold Des., 2002, Vol. 10, 1261-1272
Plasma generation of supported metal catalysts
H. Shim, J. Phillips, I. M. Fonseca, S. Carabineiro
Appl. Catal. A, 2002, Vol. 237, Nr. 1-2, 41-51
Enhancement of enzyme activity in supercritical carbon dioxide via changes in acid-base
Harper, N.; Barreiros, S.
Biotechnol. Prog., 2002, Vol. 18, 1451-1454
Eletroanalytical study of the pesticide asulam
Henri P.A. Nouws, Cristina Delerue-Matos, José Lima, E. Manuela Garrido, Pieter Vincke, Noel A. Maes
Int. J. Environ. An. Chem., 82, 69-76 (2002)
Thermodynamics of liquid (nitrogen + ethane)
Henrique J. R. Guedes; John A. Zollweg; Eduardo J. M. Filipe; Luís F. G. Martins; Jorge C. G. Calado
J. Chemical Thermodynamics, 2002, Vol. 34, Nr. 5, 669-678
Simulation of a new hybrid membrane/pressure swing adsorption process for gas separation
I. A. A. C. Esteves; J. P. B. Mota
Desalination, 2002, Vol. 148, 275-280
Discrimination of Varietal Olive Oils of the Portuguese Cultivars Cobrançosa, Madural and Verdeal
Based on their Fatty Acids Composition
J. A. Pereira, M.B.P.P. Oliveira, S. Casal, M. R. Alves
Acta Hort. 2002, 586. ISHS, 591- 594
Quimio-informática. Conteúdos que urge ensinar
J. Aires de Sousa
JATOON: Java Tools for Neural Networks
J. Aires-de-Sousa
Chemom. Intellig. Lab. Syst., 2002, Vol. 61, Nr. 1-2, 167-173
Prediction of enantiomeric selectivity in chromatography. Application of conformation-dependent
and conformation-independent descriptors of molecular chirality
J. Aires-de-Sousa; J. Gasteiger
J. Molec. Graphics and Model., 2002, Vol. 20, Nr. 5, 373-388
Prediction of 1H NMR Chemical Shifts Using Neural Networks
J. Aires-de-Sousa; M. Hemmer; J. Gasteiger
Anal. Chem., 2002, Vol. 74, Nr. 1, 80-90
Crystallization and characterization of Smaug: a novel RNA-binding motif
J. B. Green; T.A. Edwards; J. Trincao; C. R. Escalante; R. P. Wharton; A. K. Aggarwal
Biochem. Biophys. Res. Comm., 2002, Vol. 297, Nr. 5, 1085-1088
Color Stabilization of Anthocyanins: Effect of SDS Micelles on the Acid-Base and Hydration
Kinetics of Malvidin 3-Glucoside (Oenin)
J. C. Lima; C. Vautier-Giongo; A. Lopes; E. Melo; F. H. Quina; A. L. Maçanita
J. Phys. Chem., 2002, Vol. 106, 5851-5859
Química Verde
A - 23
A minimal Mathematical Model of calcium homeostasis.
J. F. Raposo; L. G. Sobrinho; H. Gil Ferreira.
J. Clin. Endoc. Metab., 2002, Vol. 87, 4330-4340
Oxidative behaviour of apomorphine and its metabolites
J. M. P. J. Garrido, C. Delerue Matos, M. F. M. Borgues, T. R. A. Macedo, A. M. OliveiraBioelectrochemistry, 2002, 55, 113-114
Electroanalytical determination of codeine in pharmaceutical preparations
J. M. P. J. Garrido, C. Delerue-Matos, M. F. M. Borges, T. R. A. Macedo, Ana M. OliveiraAnal. Lett., 2002, 35, 2487-2498
Ground and excited state properties of polyamine chains bearing two terminal naphthalene units.
Evidence for light induced movements
J. S. de Melo; M. T. Albelda; P. Díaz; E. García-España; C. Lodeiro; S. Alves; J. C. Lima; F. Pina; C. Soriano
J. Chem. Soc. Perkin Trans. 2, 2002, Vol. 5, 991-998
Intramolecular addition of acyldiazenecarboxylates onto double bonds in the synthesis of
heterocycles
J. V. Prata, D. T. S. Clemente, S. Prabhakar, A. M. Lobo, I. Mourato, P. S. Branco
J. Chem. Soc. Perkin Trans I, 2002, 513-528
Influence of olive storage period on oil quality of three Portuguese Cultivars of Olea europea,
Cobrançosa, Madural, and Verdeal Transmontana
J.A. Pereira, S. Casal, A. Bento, M.B.P.P. Oliveira
J. Agric. Food Chem., 2002, 50, 6335-6340
Sulfate Activation
J.J.G. Moura, S. Bursakov, I. Moura, O. Gavel and I. Moura
Encyclop. of Catalysis, 2002,
Crystallization And Preliminary X-Ray Analysis Of Two PH-Dependent Forms Of Cytocrome C
Peroxidase From Pseudomonas Nautica
J.M. Dias; C. Bonifácio; T. Alves; J.J.G. Moura; I. Moura; M J. Romão
Acta Crystallogr. D, 2002, Vol. 58, 697-699
Determination of polyphenols in wine by reaction with 4-aminoantipyrine and photometric flowinjection analysis
Jan W. Schoonen, M. Goreti F. Sales
Anal. Bioanal. Chem., 2002, 372, 822-828
Synthetic, Spectroscopic and X-ray Crystallographic Studies of [1,2,7,8]tetrathiacyclododecino[4,3-b:5,6-b’:10,9-b’’:11,12-b’’’]tetraindoles
Janosik, T.; Bergman, J.; Romero, I.; Stensland, B.; Stalhandske, C.; Marques, M. M. M.; Santos, M. M.
M.; Lobo, A. M.; Prabhakar, S.; Duarte, M. F.; Florêncio, M. H.;
Eur. J. Org. Chem., 2002, 1392-1396
Design and development of a multichannel potentiometer for monitoring an electrode array and
its application in flow analysis
Jarbas J.R. Rohwedder, Celio Pasquini, Ivo M. Raimundo Jr., M.Conceição B.S.Montenegro, Alberto N.
Araújo, Cristina M.C.M.Couto
J.Automated Methods & Management in Chem., 24 (11) 105-110 (2002)
New insights into the oxidation pathways of apomorphine
Jorge Garrido, Cristina Delerue-Matos, Fernanda Borges, Artur Silva, Tice Macedo, Ana M. Oliveira-Brett
J. Chem. Soc. Perkin Trans, 2, 2002, 1713-1717
Automated spectrophotometric determination of clomipramine on a multicommutated flow
José L.F.C. Lima, João A.V. Prior, Boaventura F. Reis, João L.M. Santos, Elias A.G. Zagatto
Anal. Chim. Acta, 467, 75-81 (2002)
Química Verde
A - 24
Spectrophotometric determination of phytic acid in plant extracts using a multi – pumping flow
system
Josiane M. T. Carneiro, Elias Zagatto, João L. M. Santos, José L. F. C. Lima
Anal. Chim. Acta, 474, 161-166 (2002)
As aminas aromáticas no contexto da carcinogénese química
L. L. Gonçalves
Double Critical Phenomena in (Water + Polyacrylamides) Solutions
L. P. N. Rebelo; Z. P. Visak; H. C. de Sousa; J. Szydlowski; R. Gomes de Azevedo; A. M. Ramos; V.
Najdanovic-Visak; M. Nunes da Ponte; J. Klein
Analysis of the Microbial Community Structure and Function of a Laboratory Scale Enhanced
Biological Phosphate Removal Reactor
Levantesi C., L.S. Serafim, G. R. Crocetti, P. C. Lemos, S. Rosseti, L. L. Blackall, M.A.M. Reis, V. Tandoi
Environ. Microbiol., 2002, Vol. 4, Nr. 10, 559-569
Recent Developments in the Synthesis of Biologically Active Indole Alkaloids
Lobo, A. M.; Prabhakar, S.;
J. Het. Chem., 2002, 429-436
Catalytic Asymmetric Dihydroxylation of Olefins Using a Recoverable and Reusable OsO42- in
Ionic Liquid [bmim][PF6]
Luís C. Branco, Carlos A. M. Afonso
Chem. Commun., 2002, 3036-3037
High Selective Transport of Organic Compounds by Using Supported Liquid Membranes Based
on Ionic Liquids
Luís C. Branco, João G. Crespo, Carlos A. M. Afonso
Angew. Chem. Int. Ed., 2002, Vol. 41, 2771-2773
Studies on the Selective Transport of Organic Compounds by Using Ionic Liquids as a Novel
Supported Liquid Membranes
Luís C. Branco, João G. Crespo, Carlos A. M. Afonso
Chem. Eur. J., 2002, Vol. 8, 3865-3871
Preparation and Characterization of New Room Temperature Ionic Liquids
Luís C. Branco, João N. Rosa, Joaquim J. Moura Ramos, Carlos A. M. Afonso
Chem. Eur. J., 2002, Vol. 8, 3671-3677
A radical approach towards indolizidine 167B
M. C. Corvo; M. M. A. Pereira
Tetrahedron Lett., 2002, Vol. 43, 455-458
Hydroformylation of alkenes in supercritical carbon dioxide catalysed by rhodium
trialkylphosphine complexes
M. F. Sellin; I. bach; J. M. Webster; F. Montilla; V. Rosa; T. Avilés; M. Poliakoff; D. J. Cole-Hamilton
J. Chem. Soc., Dalton Trans., 2002, 4569-4576
Micelle effects on multistate/multifunctional systems based on photochromic flavylium
compounds. The case of Luteolinidin.
M. J. Melo, S. Moura, M. Maestri, F. Pina
J. Mol. Structure, 2002, Vol. 612, 245-253
Cytotoxic effects of metal complexes of biogenic polyamines. I. Platinum(II) spermidine
compounds: prediction of their antitumour activity
M. P. Marques, T. Girão, Maria C. Pedroso de Lima, A. Gameiro, E. Pereira, P. Garcia
Biochim. Biophys. Acta, 2002, 1589, 63-70.
Química Verde
A - 25
Metal complexes with an achiral symmetrical helicand. Crystal structure of an
ennatiomerically pure (M)-Zn(II) monohelicates.
M. R. Bermejo; M. Vázquez; J. Sanmartín; A: M. García-Deibe; M. Fondo; C. Lodeiro.
New. J. Chem., 2002, Vol. 10, Nr. 26, 1365-1370
Synthesis of Optically Active Monomers and Copolymers Derived from Protected 6-O-Acryloyl
Sucrose
M. T. Barros and F. Sineriz
Synthesis, 2002, 1407
An Expedient Stereoselective Synthesis of Polysubstituted Piperidin-2-ones
M. T. Barros; M. A. Januário-Charmier; C. D. Maycock; T. Michaud
Tetrahedron, 2002, Vol. 58, 1519
Determination of free formaldehyde in foundry resins as 2,4-dinitrophenylhydrazone by high
performance liquid chromatography
M. T. Oliva-Teles, P. Paiga, C. M. Delerue-Matos, M. C. M. Alvim Ferraz
Anal. Chim. Acta, 2002, 467, 97-103
Dielectric studies of the nematic mixture E7 on a hydroxypropylcellulose substrate
M. T. Viciosa; A. M. Nunes; A. Fernandes; P. L. Almeida; M. H. Godinho; M. Dionísio
Liq. Crys., 2002, Vol. 29, Nr. 3, 429-441
Metal complexes with a chiral N4 symmetrical Schiff base. Crystal structures of the ligand and its
Cu(II) and Ni(II) “mono—helicates”
M. Vazquez, M. R. Bermejo, J. Sanmartin, A. M. García-Deibe, C. Lodeiro, J. Mahía
J. Chem. Soc., Dalton Trans., 2002, Vol. 6, 870-877
Oxidation of ferrocene derivatives at a poly[Ni(saltMe)] modified electrodes
M. Vilas Boas, E. M. Pereira, C. Freire e A. R. Hillman
J. Electroanal. Chem., 2002, 558-539, 47-58.
Étude biologique et chimique de la plante Prasium major poussant en Tunisie. Structure d’un
heteroside steroidique et d’un steroide. Identification de constituents de la fraction
M.A. Mahjoub; H. Ben Jannet; P.J.M. Abreu;Z. Mighri
J. Soc. Chim. Tun., 2002, Vol. 4, Nr. 11, 1419
O ENDOR detection of a solvent-derived Ni-(OH(x))-Fe bridge that is lost upon
activation of the hydrogenase from Desulfovibrio gigas
M.Carepo; D.L.Tierney; C.D.Brondino; T.C.Yang; A.Pamplona; J.Telser; I.Moura; J.J.G.Moura;
B.M.Hoffman
J. Am. Chem. Soc., 2002, Vol. 124, 281-286
17
Construção de um bio-eléctrodo específico para determinação de nitritos
M.G. Almeida, P. Tavares e J.J.G. Moura
Molybdenum Enzymes in Reactions Involving Aldehydes and Acids
M.J. Romão, C.A. Cunha, C.D. Brondino and J.J.G. Moura
Marcel Dekker, 2002, Vol. 39, 539-570
Switching from Intramolecular Energy Transfer to Intramolecular Electron Transfer by the Action
of pH and Zn2+ Co-ordination
M.T. Albelda; P. Díaz; E. Garcia-España; J. C. Lima; C. Lodeiro; J. Seixas de Melo; A. J. Parola; F. Pina;
C. Soriano
Chem. Phys. Letters, 2002, Vol. 353, 63-68
Synthesis and Some Reactions of 2-Acyl-2-Alkyl-1,3-Dithiolane 1,1-Dioxides
M.T. Barros; Antonio S. Henriques; Alcino J. Leitao; C.D. Maycock
Helv. Chim. Acta, 2002, Vol. 58, 4059
Stereoselective Radical Reactions of Some Tartaric and Glyceric Acid Derivatives
M.T. Barros; C.D. Maycock; M.R. Ventura
Org. Letters, 2002, Vol. 4, 2035
Química Verde
A - 26
A Synthesis of Aziridines from alfa-Iodocycloenones
M.T. Barros; C.D. Maycock; M.R. Ventura
Tetrahedron Lett., 2002, Vol. 43, 4329
Dupuytrens Disease: Particularities of the Portuguese Population
Marques, M.; Silva, MA;. Cordeiro, MNDS;. Pinho, C; Martins, A; Amarante., J
Cir. Plást. Iberolatinoamer, 2002, 125, 28
Theoretical study of the amino acid interactions in the binding region of the trypsin-pancreatica
trypsin inhibitor complex
Melo, A., Ramos, M. J.
J. Mol. Struct. (Theochem), 2002, 580, 251-262
Characterization of Reactive Intermediates by Diffusion-Ordered NMR Spectroscopy: A Snapshot
of the Reaction of 13CO2 with [Cp2Zr(Cl)H
N. E. Schloerer; E. J. Cabrita; S. Berger
Angew. Chem. Int. Ed., 2002, Vol. 41, 107-109
A study of the thermal decomposition of 2-azidoethanol and 2-azidoethylacetate by ultraviolet
photoelectron spectroscopy and matrix isolation infrared spectroscopy
N. Hooper; L. J. Beeching; J. M. Dyke; A. Morris; J. S. Ogden; A. A. Dias; M. L. Costa; M. T. Barros; M. H.
Cabral; A. M. C. Moutinho
J. Phys. Chem., 2002, Vol. 106, 9968
Solid-Phase Microextraction in Combination with GC/MS for Quantification of the Major Volatile
Free Fatty Acids in Ewe cheese
O. Pinho, I.M.P.L.V.O. Ferreira, and M. A. Ferreira
Anal.Chem. 2002, 74, 5199-5204
Carbohydrates from Detarium microcarpum bark extract
P. Abreu; A. Relva
Carbohydr. Res., 2002, Vol. 337, 1663-1666
Clomipramine induces the opening of Na+ channels in the frog skin epithelium.
P. Alves; I. Graça; T. Moura
Pharmacology and Toxicology, 2002, Vol. 90, 161-168
Energy transfer between polyamine chains bearing naphthalene terminal units and K3[Co(CN)6 ]:
an example of a molecular photoreactor
P. Arranz; M. T. Albelda; E. Garcia-España; J. C. Lima; C. Lodeiro; J. Seixas de Melo; A. J. Parola; F.
Pina
Flavonoids from ‘Ulex airensis’ and ‘Ulex europaeus’ ssp. ‘europaeus’
P. Máximo; A. Lourenço; S. S. Feio; J. C. Roseiro
Journal of Natural Products, 2002, Vol. 65, Nr. 2, 175-178
A new prenylisoflavone from ‘Ulex jussiaei’
P. Máximo; A. Lourenço; S. S. Feio; J. C. Roseiro
Z. Naturforsch., 2002, Vol. 57c, 609-613
Cancro, uma patologia tão antiga quanto o homem.
P. S. Branco
Bol. Soc. Port. Quím., 2002, 49-53
Fractionation of Lipid Mixtures by Subcritical R134a in a Packed Column
P.C. Simões, O.J.Catchpole
Ind. Eng. Chem. Res., 2002, Vol. 41, Nr. 2, 267-276
µ4-sulfide bridged tetranuclear Cu-Z in N2O reductase: spectroscopy, electronic structure and
mechanism
P.Cheng; I.Cabrito; J.J.G.Moura; I.Moura; E.I. Solomon
Abst. Pap. Am. Chem. Soc., 2002, Vol. 224, 258-258
Química Verde
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Spectroscopic and electronic structure studies of the u4 sulfide bridged tetranuclear CuZcluster
in N2O reductase:molecular insight into catalytic mechanism
P.Cheng; I.Cabrito; J.J.G.Moura; I.Moura; E.I.Solomon
J. Am. Chem. Soc., 2002, Vol. 124, 10497-10507
Electronic structure description of the u4-sulfide bridged tetranuclear Cu(Z) center in
N2Oreductase
P.Cheng; S.G.DeBeer; I.Cabrito; W.E.Antholine; J.J.G.Moura; I.Moura; B.Hedman; K.O.Hodgson;
E.I.Solomon
J. Am. Chem. Soc., 2002, Vol. 124, 744-745
Automatic flow system with voltammetric. Detection for diacetyl monitoring during brewing
process
Pedro G. Rodrigues, José A. Rodrigues, Aquiles A. Barros, Rui A.S. Lapa, José L.F.C. Lima, J.M.
Machado Cruz, A.A. Ferreira
J. Agric. Food Chem, 50,(13), 3647 - 3653 (2002).
Antioxidant Properties of trans-e-Viniferin as Compared to Stilbene Derivatives in Aqueous and
Nonaqueous Media
Privat, C.; Telo, J.P.; Bernardes-Genisson, V.; Vieira, A.J.S.C.; Souchard, J.-P.; Nepveu, F.
J. Agr. Food Chem., 2002, Vol. 50, Nr. 5, 1213
Stability, dynamics of convergence and tuning of observer-based kinetics estimator
R. Oliveira, E. C. Ferreira, S. Feyo de Azevedo
J. Process Control, 2002, Vol. 12, 311-323
Fractionation of Edible Oil Model Mixtures by Supercritical Carbon Dioxide in a Packed Column –
Part I: Experimental Results
R. Ruivo, M.J. Cebola, P.C. Simões, M. Nunes da Ponte
Fractionation of Edible Oil Model Mixtures by Supercritical Carbon Dioxide in a Packed Column –
Part II: A Mass Transfer Study
R. Ruivo, M.J. Cebola, P.C. Simões, M. Nunes da Ponte
Cu 2+-Induced isoproterenol oxidation into isoprenochrome in adult rat calcium-tolerant
cardiomyocytes
Remião, F., Carvalho, M., Carmo, H., Carvalho, F., Bastos, M. L.
Chem. Res. Toxicol., 2002, 15, 861-869
Encapsulation of Copper(II) Complexes with Pentadentate N2O2 Schiff BaseLigands in a Pillared
Layered Clay
Rita Ferreira, Cristina Freire, Baltazar de Castro, Ana Paula Carvalho, João Pires da Silva, Manuela
Brotas de Carvalho
Eur. J. Inorg. Chem., 2002, 3032-3038.
Relocation of a tubular voltammetric detector for standard addition in FIA
Rita I.L. Catarino, M. Beatriz Q. Garcia, José L.F.C. Lima, E. Barrado, M.Vega
Electroanal., 14, 11 (2002)
Sequential determination of salicylic and acetylsalicylic acids by amperometric multisite
detection flow injection analysis
Rita I.L. Catarino, M. Beatriz Q. Garcia, Rui A.S. Lapa, José L.F.C. Lima, Enrique Barrado
J. AOAC Int., 85 (6) 1253-1259 (2002)
Development of a FIA system with amperometric detection for bentazone determination in
estuarine waters
Rodrigo P. A. G. Cerejeira, Cristina Delerue-Matos, M. Carmo V. F. Vaz
Anal. Bioanal. Chem., 2002, 373, 295-298
Química Verde
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Multi – pumping in flow analysis: concepts, instrumentation, potentialities
Rui A.S. Lapa, José L.F.C. Lima, Boaventura F. Reis, João L.M. Santos, Elias A.G. Zagatto
Anal. Chim. Acta, 466, 125-132 (2002)
Sequencial Preconcentration for the ETAAS. Determination of Pb in human Urine
Rui C. C. Costa, Agostinho A. Almeida, Alberto N. Araújo, Valdinete L. Silva
At. Spectrosc., 23 (1), 17 - 23 (2002)
Extraction of Edible Oils at High Pressure Conditions – Dynamic and Steady State Studies
Ruivo, R.; Paiva, A.; Cebola, M.J.; Simões, P.
Gas chromatographic-mass spectrometric quantification of 4-(5-)-methylimidazole in roasted
coffee after ion-pair extraction
S. Casal, J.O. Fernandes, M.B.P.P. Oliveira, M.A. Ferreira
J. Chromatogr. A, 2002, 976, 285-291
Determination of biogenic amines in coffee by an optimised liquid chromatographic method
S. Casal, M.B.P.P. Oliveira, M.A. Ferreira
J. of Liq. Chrom. & Rel. Techn., 2002, 25, 16, 2535-2549
Structural changes in the calcium-dependent activation of the di-heme cytochrome c peroxidase
of Paracoccus pantotrophus
S.R.Pauleta; Y.Lu; C.F.Goodhew; Y.Qiu; I.Moura; G.W.Pettigrew; J.A.Shelnutt
Biophys. J., 2002, Vol. 82, 63-63
Membrane structural changes support the involvement of mitochondria in the bile salt-induced
apoptosis of rat hepatocytes
S.Sola; M.A.Brito; D.Brites; J.J.G.Moura; C.M.P.Rodrigues
Clinical Science, 2002, Vol. 103, 475-485
HPLC/UV determination of organic acids in fruit juices and nectars
Sara C. Cunha, José O. Fernandes, Isabel M.P.L.V.O. Ferreira
Eur. Food Res. Technol., 2002, 214, 67-71
Detection and Visualization of Storage Polymers in Phosphate Removal Microorganisms
Serafim L. S., P. C. Lemos, C. Levantesi, V. Tandoi, H. Santos, M. A. M.Reis
J. Microbiol. Methods, 2002, Vol. 51, Nr. 1, 1-18
Effect of pH Control on EBPR Stability and Efficiency, Water Science and Technology
Serafim L. S., P. C. Lemos, M. A. M. Reis
Water Science and Technol., 2002, Vol. 46, Nr. 4-5, 179-184
Change in Metabolism of PHA Accumulation by Activated Sludge Modifying Operating Conditions
Serafim L. S., P. C. Lemos, M. A. M. Reis.
Water Science and Technol., 2002, Vol. 46, Nr. 1-2, 353-356
Phenolic profile of quince fruit (Cydonia oblonga Miller) pulp and peel
Silva, B.M.; Andrade, P.B.; Ferreres, F.; Domingues, A.L.,Seabra, R.M. e Ferreira, M.A.
J. Agric. Food Chem., 2002, 50, 4615-4618
Study of the organic acids composition of quince (Cydonia oblonga Miller) fruit and jam
Silva, B.M.; Andrade, P.B.; Mendes, G.C.; Seabra, R.M., Ferreira, M.A.
J. Agric. Food Chem., 2002, 50, 2313-2317
Chromium (III) Determination with 1,5-Diphenylcarbazide based on the oxidative effect of chlorine
radicals generated from CCl4 sonolysis in aqueous solution
Sivanildo da Silva Borges, Mauro Korn, J. L. F. Costa Lima
Anal. Sci., 18, 1361 - 1366 (2002)
Implementation of AAS methods for the Quantification of Hg and hexavalent chromium in
recycled paper used for food packaging
Soares M.E., Carneiro C., Bastos, M.L.
At. Spectrosc., 2002, 23 (5), 148-153
Química Verde
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Synthesis, X-ray structures, electrochemistry, magnetic properties, and theoretical studies of the
novel monomeric [CoI2(m-dppfO2)] and polymeric chain [CoI2(m-dppfO2)n].
T. Avilés; A. Dinis; J.O. Gonçalves; V. Félix; M.J. Calhorda; A. Prazeres; M.G.B. Drew; H. Alves; R.T.
Henriques; V. da Gama; P. Zanello; and M. Fontani.
Solubility measurements of polymerisation stabilisers in scCO2
T. Casimiro; T. Costa; R. Griffiths; S. Howdle; A. Aguiar-Ricardo
Vanadate oligomers interaction with phosphorylated myosin
T.Tiago; M.Aureliano; R.O.Duarte; J.J.G.Moura
Inorg. Chim. Acta, 2002, Vol. 339, 317-321
Speed of sound in liquid cyclic alkanes at temperatures between (283 and 343) K and pressures
up to 20 MPa
Toshiharu Takagi; Taro Sakura; Henrique J. R. Guedes
J. Chemical Thermodynamics, 2002, Vol. 34, Nr. 5, 1943-1957
Mass Transfer in Osmotic Evaporation: Effect of Pocess Parameters
V. D. Alves;I.M. Coelhoso
J. Membrane Sci., 2002, Vol. 208, Nr. 1-2, 171-179
Methoxylated xanthones in the quality control of small centaury (Centaurium erythraea) flowering
tops
Valentão, P.; Andrade, P.B.; Silva, E.; Vicente, A.; Santos, H. R.; Bastos, M.L.; Seabra, R.M.
J. Agric. Food Chem., 2002, 50, 460-463
Antioxidant activity of Hypericum androsaemum infusion: scavenging activity against superoxide
radical, hydroxyl radical and hypochlorous acid
Valentão, P.; Fernandes, E.; Carvalho, F.; Andrade, P.B.; Seabra, R.M.; Bastos, M. L.
Biol. Pharm. Bull., 2002, 25(10), 1320-1323
Studies on the antioxidant activity of Lippia citriodora infusion: scavenging effect on superoxide
radical, hydroxyl radical and hypochlorous acid
Biol. Pharm. Bull., 2002, 25(10), 1324-1327
Antioxidative properties of cardoon (Cynara cardunculus L.) infusion against superoxide radical,
hydroxyl radical and hypochlorous acid
J. Agric. Food Chem., 2002, 50, 4989-4993
Integrated Transport and Reaction in an Ion Exchange Membrane Bioreactor
Velizarov S., M.A.M. Reis, J.G. Crespo
Desalination, 2002, Vol. 149, 205-210
Ion Exchange Membrane Bioreactor for Selective Removal of Nitrate from Drinking Water: Control
of Ion Fluxes and Process Performance
Velizarov S.; J.G. Crespo; M.A.M. Reis
Biotechnol. Prog., 2002, Vol. 18, 296-302
Removal of Nitrate from Water in an Ion Exchange Membrane Bioreactor
Velizarov S.; J.G. Crespo; M.A.M. Reis
Water Science and Technol., 2002, Vol. 2, Nr. 2, 161-167
Phase behaviour of room temperature ionic liquids solutions: an unusually large co-solvent effect
in (water + ethanol)
V. Najdanovic-Visak; José Esperança; Luís Rebelo; Manuel Nunes da Ponte; Henrique J. R. Guedes;
Kenneth R. Seddon; Jerzy Szydlowsky
Phys. Chem. Chem. Phys., 2002, Vol. 4, 1701-1703
Química Verde
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Development and evaluation of an HPLC/DAD method for the analysis of phenolic compounds
from olive fruits
Vinha, A.F.; Silva, B.M.; Andrade, P.B.; Seabra, R.M.; Pereira, J.A.; Oliveira, M.B.
J. Liq. Chrom. Rel. Technol., 2002, 25 (1), 151-160
Optical and Spectroscopic Methods for Biofilm Monitoring
Wolf G., J.G. Crespo, M.A.M. Reis.
Re/view Environ.Sci. Bio/tech., 2002, Vol. 1, Nr. 3, 227-251
Interactions of protoporphyrin and its metal derivatives with the active site of mouse
Y.Lu; A.Sousa; R.Franco; A.Mangravita; G.C.Ferreira; I.Moura; J.A.Shelnutt
Biophys. J., 2002, Vol. 82, 2186-2186
Binding of protoporphyrin IX derivatives to the active site of wild-type mouse ferrochelatase at
low porphyrin-to-protein ratios
Y.Lu;A.Sousa;R.Franco;A.Magravita;G.C.Ferreira;I.Moura;J.Shelnutt
Biochemistry, 2002, Vol. 41, 8253-8262
A. Bencini, M. A. Bernardo, A. Bianchi, E. García-España, C. Giorgi, S. Luís, F. Pina, B. Valtancoli
Sensing Cations and Anions by Luminescent Polyamine Receptors in Solution
in Advances in Supramolecular Chemistry, Cap. 3, ed. G. Gokel, Cerberus Press, Vol. 8, 79-130 (2002)
A. J. S. Rodrigo; J. P. B. Mota; E. Saatdjian
Chaotic mixing and heat transfer in time-periodic 3-D flows
in Proceedings 12th International Heat Transfer Conference, ed. J. Taine, Elsevier Science, 489-494 (2002)
A. Lefèvre; J. P. B. Mota; A. J. S. Rodrigo; E. Saatdjian
Chaotic advection and heat transfer enhancement in Stokes flows (Keynote lecture)
in Proceedings 12th International Heat Transfer Conference, ed. J. Taine, Elsevier Science, 157-170 (2002)
A.M. Lourenco; P. Máximo; L. M. Ferreira; M. M. A. Pereira
Indolizidine and quinolizidine alkaloids structure and bioactivity
in Studies in Natural Products Chemistry, Bioactive Natural Products, Part H, ed. Atta-ur- Rahman, Elsevier,
Vol. 27, 233-298 (2002)
C. Rodrigues, P. Gameiro, S. Reis, J. L. F. C. Lima. B. Castro
Partitioning and Thermodynamics of Chlordiazepoxide in n-octanol/Buffer and Liposome Systems
in: Biophysical Chemistry - Membranes and Proteins, eds. Richard Templer and Robin Leatherbarrow,
Royal Society of Chemistry, 243-247 (2002)
E. M. Gaspar; J. P. Noronha
HPLC-ESI-MS: An useful alternative to FAB-MS in the analysis of intact resin glycosides from
Convolvulaceae plants
in Natural Products in the New Millennium: Prospects and Industrial Application, ed. Rauter, A.P., Palma,
F.B., Ara, Kluwer Academic Publishers, NL, PSE, 119-124 (2002)
I. A. A. C. Esteves; J. P. B. Mota
Modeling of an integrated membrane/PSA process for gas adsorption
in Fundamentals of Adsorption, ed. K. Kaneko, IK International, Vol. 7, 918-925 (2002)
J. P. B. Mota; A. J. S. Rodrigo
Thermal management of onboard natural gas adsorption storage systems
in Fundamentals of Adsorption, ed. K. Kaneko, IK International, Vol. 7, 125-132 (2002)
M.J.Romao; C.A. Cunha; C.D. Brondino; J.J. Moura
Molybdenum enzymes in reactions involving aldehydes and acids.
in Metal Ions in Biological Systems, Cap. 39, Vol. —, Nr. —, 539-570 (2002)
Química Verde
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P. Georgieva; R. Oliveira; S. Feyo de Azevedo
Instrumentation and Process Control - Process Control
in Encyclopedia of Dairy Sciences, ed. H. Roginski; J.W. Fuquay; P.F., Academic Press, 1392-1401 (2002)
R. Oliveira R; P. Georgieva; S. Feyo de Azevedo
Instrumentation and process control – Instrumentation
in Encyclopedia of Dairy Sciences, ed. J.W.H. Roginski; P.F. Fuquay, Academic Press UK, 1401-1410 (2002)
T. Sousa, E. Fernandes, M. Morato, F. Carvalho, A. A. Teixeira
Antioxidant enzyme activity in mesenteric artery from DPSPX-hypertensive rats
in: The Microcirculation and Vascular Biology. Proceedings of the 22nd Meeting of the European Society for
Microcirculation, 267-270. (Eds. Tooke J, Shore A, Whatmore J), Mondozzi Editore (2002)
Química Verde
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B
Dissertations
Química Verde
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Ph. D. Dissertations
Caracterização bioquímica e espectroscópica de enzimas contendo molibdénio em organismos
redutores de sulfato
Susana Andrade (Supervisor: J.J.G. Moura)
Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa, 2002
Caracterizacao estrutural e bioquímica de uma protease de serina (calicreina)
A. L. Carvalho (Supervisors: M. J. Romão; A. Romero)
Caracterização estrutural e funcional de proteínas contendo centros mononucleares de ferro não
hémico
Carla Ascenso (Supervisors: J.J.G. Moura; I. Moura)
Desnitrificação Biológica de Águas de Captação com Reactores de Membranas
Ana Maria Barreiros (Supervisors: M.A.M.Reis; J.G.Crespo)
Estrutura-Função de Hidrogenases
Marta Carepo (Supervisor: J.J.G. Moura)
Funcionalização de Enecarbamatos Endocíclicos
Marta Norton de Matos (Supervisor: Carlos A. M. Afonso)
Isolation of the genes of aldehyde oxidoreductase and formate dehydrogenase
Sofia Maciera (Supervisors: J.J.G. Moura; R. Huber)
Metaloenzimas envolvidas na redução do nitrato e do nitrito
Carla Carneiro (Supervisors: J.J.G. Moura; I. Moura)
Proteínas contendo molibdénio e tungsténio
Rui Duarte (Supervisor: J.J.G. Moura)
Síntese de Moléculas Polifuncionais Regiosselectivamente Substituídas
F. Siñeriz (Supervisor: M. T. Barros)
Structure and Function of Nitrate Reductase and of di-haem Cytochrome c Peroxidase
Joao M. Dias (Supervisors: Maria J. Romão; R. Huber)
Química Verde
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Desenvolvimento de Sistemas Automáticos Baseados em Técnicas de Fluxo Contínuo para
Monitorização de Parâmetros Ambientais e Industriais
Ivone Valente de Oliveira (Supervisor: Rui Alexandre Santos Lapa)
Faculdade de Farmácia da Universidade do Porto, 2002
Desenvolvimento de Sistemas Automatizados de Fluxo com Detecção Voltamétrica Multilocalizada
Rita Isabel Lemos Catarino (Supervisor: Maria Beatriz V. N. Q. Guerra Junqueiro)
Cardiotoxicidade do Isoproterenol e dos Produtos da sua Oxidação
Fernando Manuel Gomes Remião (Supervisor: Maria de Lourdes Bastos)
Isolamento e Caracterização de Proteínas Envolvidas na Via Desnitrificante em Pseudomonas
Chlororaphis
Dora Raquel da Silva Pinho (Supervisor: Baltazar de Castro)
Faculdade de Ciências da Universidade do Porto, 2002
Estudos de Partição e Localização de Fármacos em Lipossomas por Técnicas Espectroscópicas
Catarina Isabel Guerra Rodrigues (Supervisors: Baltazar de Castro, Paula Gameiro)
Faculdade de Ciências da Universidade do Porto 2002
Complexos de Metais Imobilizados em Alumino-silicatos Porosos - caracterização e aplicações
em Catálise
Rita Isabel Simões Pinto Ferreira (Supervisors: Baltazar de Castro, Cristina Freire)
Química Verde
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M. Sc. Dissertations
ICP-MS na Análise de águas: Análise directa de águas engarrafadas e avaliação de uma estratégia
de colheita de àguas naturais em mini-colunas
Suzana Maria de Castro Almeida Tavares (Supervisor: Agostinho Almiro Almeida)
Desenvolvimento de Metodologias Analíticas Baseadas em Sistemas Multicomutados para o Controlo
de Metais Pesados em Águas Residuais
Artur Manuel Bordalo Machado Figueirinha (Supervisor: Rui Alexandre Santos Lapa)
Evolução da proteólise do queijo de cabra transmontano por RP-HPLC/UV
Lara Alexandra Martins Marques Magalhães (Supervisors: Isabel Ferreira; Beatriz Oliveira)
Hidrocarbonetos aromáticos policíclicos em óleos vegetais
Vitor Hugo da Costa Gomes Moreira Teixeira (Supervisor: Beatriz Oliveira; Susana Casal)
Estudo Teórico do Mecanismo Catalítico da Enzima Piruvato Formato Liase
Maria de Fátima Assunção Lucas (Supervisor: M. J. Ramos)
Aquecimento global e degradação da camada de ozono
Paula Maria Coelho Pereira França (Supervisors: Gabriela Ribeiro; M.ª João Sinde)
Ensino/aprendizagem do equilíbrio químico na disciplina de Química do 12º ano - Uma abordagem
experimental
José Carlos Pinto de Oliveira Lopes (Supervisors: Gabriela Ribeiro; Aquiles de Barros)
Entalpias de reacção: uma proposta de abordagem laboratorial para o Ensino Secundário
Joaquim Fernando Neto Brandão Coelho (Supervisors: Gabriela Ribeiro; M.ª Dores Ribeiro da Silva)
Concepções dos alunos do 11º ano sobre soluções e processos de dissolução
Anabela Ferreira da Silva Veiga (Supervisors: Gabriela Ribeiro; Paula Gameiro)
Química Verde
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ANNEX III
RESEARCH PROJECTS
Química Verde
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Molecular Level Devices and Machines
European network RTN1-1999-00138
Coordinator : Fernando Pina
Adsorbed Natural Gas System with Guard-Bed Device
ANGUARD: EU ENK6-CT2000-00053
Coordinator: José Paulo Mota
The Mechanism, Specificity and Inhibition of Enzymes belonging to the Xantine Oxidase family:
Medical and Industrial Applications
European Research Network TMR: HPRN-CT-1999-00084 (2002/2004)
Coordinator: Maria João Romão
Structures and Reaction Mechanisms of Pteridine Related Enzymes
European Research Network TMR: FMRX-CT98-0204 (1998/2002)
New approach to waste recovery into selective adsorbents of heavy metals
Nato Science for Peace Program nº 977984/2002
NATO Coordinator: José Paulo Mota, Portuguese Coordinator: Isabel Fonseca
Intercalation as a new approach for obtaining highly efficient pi adsorbents and supports
catalysts
INTAS_00-00750/2000
Coordinator: Isabel Fonseca
Preparação de aziridinas quirais de utilidade sintética
POCTI / QUI / 37373 / 2001; € 30 335
Coordinator: Paula S. Branco
Métodos Sintéticos para N-heterociclos baseados em rearranjos sigmatrópicos
POCTI / QUI / 36456 / 2000; € 59 855
Coordinator: S. Prabhakar
Nova metodologia sintética de gama-amino ácidos quirais ùteis na concepçâo de novos
fàrmacos
POCTI/QUI/37423/2001; € 38 900
Coordinator: Maria Manuela Araújo Pereira
Molecular-Level Devices and Machines
POCTI/QUI/32442/99; € 119 711.5
Coordinator : Fernando Pina
Building blocks for photoactive molecular cages
POCTI/QUI/38637/2001; € 38 600
Coordinator: António Jorge Parola
Química Verde
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Photophysics and photochemistry of anthocyanins
POCTI/QUI/33679/99; € 45 889.41
Coordinator: João Carlos Lima
Estudos Estruturais e Mecanísticos de Enzimas Chaves do Ciclo do Azoto
PRAXIS/QUI/10087/98 (20 000 contos)
Coordinator: Isabel Moura
Removal of Cu, Cr, As and creosote from impregnated wood waste aiming its recycling
POCTI/32927/AGR/2000
Coordinator: Alexandra Ribeiro, DCEA, UNL
Collaborators (CQFB): Margarida Gonçalves, Mercedes Tojeiro
Desenvolvimento de um sistema de informaçâo e gestâo ambiental para o estuàrio do Sado
POCTI/33137/BSE/99-00
Coordinator: Maria Helena Costa, DCEA/IMAR, UNL
Collaborators (CQFB): Margarida Gonçalves, Marco Gomes da Silva, Luz Fernandes
Compostos esterólicos como potenciais marcadores da poluição fecal e sua correlação com
indicadores biológicos
PNAT/1999/BIO/15024
Coordinator: José Filipe Santos Oliveira, GDEH/UBIA, UNL
Collaborators (CQFB): Margarida Gonçalves, Marco Gomes da Silva, Nuno Lobato
Authenticity of portuguese natural products development of analytical methods to elucidate and
monitor chirality of volatile compounds
POCTI/AGR/11116/1998.
Coordinator: Ana Maria Ferreira da Silva Costa Freitas Drumond Ludovice
Collaborators: Ângela M. S. Relva and Marco Gomes da Silva
New carbohydrate-based compounds from ethnopharmacological plants: Bioactivity against
pathogenic yeasts
POCTI/1999/FCB/35863
Coordinator: Elvira Maria Mendes Sardão Monteiro Gaspar
Collaborators: João Paulo Noronha
Enzimas chave da activação do sulfato e do metabolismo energético. Novas proteínas de Co não
corrinóides e Zn-ATP sulfurilases, cinases do adenilato
POCTI/QUI/35384/2000; € 79 807
Bacterial cytochrome c peroxidases – activation, enzymatic mechanism and structure
POCTI/QUI/42309/2001; € 48 200
Enzimas chave da reduçâo do nitrato. Redutase do óxido nítrico e redutase do óxido nitroso – as
duas enzimas terminais
POCTI/BME/42265/2001; € 102 734
Química Verde
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Production of copolymers PHB/PHV by mixed cultures
POCTI/35675/BIO/2000 (ITQB, IBET, FCT/UNL)
Senior researcher from FCT/UNL: Ana Maria Ramos
A crystallographic contribution to the understanding of the structure and function of Mo-enzymes
POCTI/BME/35078/99; € 120 833.79
Phase-behaviour and microscopic characterisation of micro/macroemulsions In CO2. New
strategies for polymerisation and enzymatic catalysis
POCTI/35429/2000; € 199.519;
Coordinator: Ana Isabel Nobre Martins Aguiar de Oliveira Ricardo.
High Pressure NMR spectroscopy of polymers and biopolymers in CO2 emulsions
POCTI/QUI/42313/2001; € 58 450
Coordinator: Maria dos Anjos Macedo
Computational fluid mechanics and process dynamics of supercritical fluid extraction columns
with structured packings and static mixers
POCTI/2000/EQU/34957; 01/10/2000 a 01/10/2003; €64.844;
Coordinator: Pedro Miguel Calado Simões.
Structure-function relationships of serine hydrolases in non-conventional media
PRAXIS XXI/P/BIO/14314/1998 (10/2000 – 03/2003)
Coordinator (CQFB): Susana Barreiros
Engenharia de Metaloproteínas: Uma Aproximação Pós-Genómica
POCTI / BME / 36152 / 99; € 124 699.47
Coordinator: José J.G. Moura
Estudos Electroquímicos Dinâmicos em Proteínas de Transferência Electrónica
POCTI / QUI / 42277 / 2001; € 53 100
Coordinator:José J.G.Moura
A Structural Study of the Presequence of 5-Aminolevulinate Synthase (ALAS), the First Enzyme of
the Heme Biosynthetic Pathway
POCTI/BIA/10086/1998. (2001 – 2002)
Coordinator: Maria dos Anjos Macedo
NMR structural studies of the active center of 5-aminolevulinate synthase, the first enzyme of the
mammalian heme biosynthetic pathway
POCTI/BME/39184/2001; € 23 175.00
Coordinator. Maria dos Anjos Macedo
Orientation of proteins by liquid crystals
POCTI/QUI/42279/2001; € 40 800
Coordinator: Francisco Jorge Caldeira
Química Verde
A - 40
Vif (virion infectivity factor) structure and localization during HIV-2 infection
POCTI/ESP/36103/99; € 104 747.56
Collaborator: Francisco Jorge Caldeira
Development of New and Improved Processes for Palladium Catalysed Cross-Coupling Reactions
for Natural Products Synthesis.
POCTI/QUI/36536/99, € 39 903.83
Coordinator: Maria Teresa Barros
Rationalising Stereoselectivity in Organic Synthesis: From Theory to Practice
POCTI/QUI/42983/2001, € 80 040
Cordinator: António Gil de Oliveira Santos
Viscous fluid mixing and chaotic heat transfer in spatially- and time-periodic flows
POCTI/EME/40057/2001, € 40 000
Coordinator: José Paulo Mota
Structural and mechanistic studies of fatty acid desaturases. Looking for reactivity of diiron
clusters
POCTI/QUI/37413/2001; € 69 090
Coordinator: Pedro Tavares
Bioreactor de Membrana de Permuta Iónica para Desnitrificação de Água
PRAXIS/P/BIO/14065/1998; € 59 606
Coordenador: Maria Ascensão Reis
Reacção e Transporte num Reactor de Membranas de Permuta Iónica para Tratamento de Água
Potável
POCTI/EQU/39482/2001; € 61 875
Coordenador: Maria Ascensão Reis
Changes in the molecular dynamics followed by dielectric spectroscopy during the formation of a
polymer dispersed liquid crystal
POCTI/CTM/37435/2001; € 40 941
Coordinator: Madalena Dionísio
Resolução de misturas racémicas de substâncias activas em contactores de membranas
POCTI/FCB/43942/2001; € 75000
Coordinator: Prof. M. Helena Marques (FFUL)
Collaborators: Isabel Maria Coelhoso, João G. Crespo; C. M. Afonso
Engenharia de Proteínas aplicada a enzimas degradadoras de nitrilos
POCTI/CEG/2508/95, € 43 395
Coordinator: Jorge Lampreia
Protease de Plasmodium chabaudi como alvo na terapia da malária
POCTI/43637/BME/2000, € 4 987
Química Verde
A - 41
As proteases de parasitas da malária como alvo na quimioterapia
POCTI/ESP/42223/2001, € 7 570
Desenvolvimento e caracterização química, estrutural e reológica de sistemas aquosos mistos
proteína-polissacarídeo
POCTI / 1999 / QUI / 36452 (CEQUP and UM); € 124 820 (to CEQUP)
Coordinator:(CEQUP) Maria Pilar Gonçalves
Valorization of Salvia officinalis, Melissa officinalis, Mentha piperita and Lavandula angustifolia for
obtention of added value products with antimicrobial and antioxidant activities.
POCTI/AGR/43482/2001(CEQUP and UM); € 22 446 (to CEQUP)
Coordinator: Manuel Fernandes Ferreira (Universidade do Minho)
Coordinator: Rosa Seabra (CEQUP)
Preparação em Linha de Amostras de Pesticidas e sua Determinação em Águas
POCTI POCTI/AGR/35287/00; € 70 699
Coordinator: (CEQUP) Maria do Carmo Vaz
Gestão de Resíduos de Laboratório
POCTI/MGS/38801/2001; € 55.000.00
Coordinator: (CEQUP) Goreti Sales
Conducting/redox polymer networks based on transition metal complexes
POCTI/QUI/ 32821/99; (CEQUP, CIQ(UP) and U.M.) € 82 715 (to CEQUP)
Coordinator: (CEQUP) Ana Cristina Freire
Pillared Clays functionalised with transition metal complexes as new asymmetric catalysts
POCTI/QUI/ 42931/2001 (CEQUP and UM) € 17 100 (to CEQUP)
Coordinator (CEQUP): Manuela Botas de Carvalho
Síntese, estrutura, propriedades, especiação em solução e estudos biológicos de compostos de
vanádio com potencial para o tratamento de Diabetes
POCTI/QUI/35368/99 (IST, CNC, CEQUP); € 38 700 (to CEQUP)
Coordinator: (CEQUP) Maria Conceição Rangel
Design of novel metallo-surfactants with catalytical relevance
POCTI/QUI/38605/01 (CEQUP and UNL), € 25.300 (to CEQUP)
Coordinator: (CEQUP) Baltazar de Castro
Avaliação do efeito modulador do ambiente lipídico da membrana na interacção de fármacos com o
transportador de fármacos P-glicoproteina
POCTI/QUI/34308/99; € 68 834
Coodinator: (CEQUP) Costa Lima
Estudo da interacção de tampões aplicáveis em gamas de pH observadas em amostras ambientais
e biológicas com iões metálicos
POCTI/QUI/39950/2001; € 42.800
Coordinator: (CEQUP) Helena Soares
Química Verde
A - 42
Estudos Experimentais e Teóricos em Catalizadores Modelo e Suportados: Catalíticos Com
Relevância Industrial e Ambiental
POCTI/ QUI/33765/99 (CEQUP); € 9.300.00
Coordinator: (CEQUP) Natália Correia
Modelling inhibitor mechanisms in redical enzymes: QM/CM approach
POCTI /QUI/35376/2000 (CEQUP) € 134.675.43
Coordinator: (CEQUP) João Ramos
Amphetamines and physical exercise. An hazardous combination?
POCTI/ FCB/43562/2001 (CEQUP and UA) € 25.312 (to CEQUP)
Coordinator: (UA) Francisco Amado
Coordinator: (CEQUP) Félix Carvalho
Influence of Procyanidin Strutures on their Ability to Complex with Anthocyanins-A Molecular
Interpretation of Colour
POCT/ QUI/40124/2001 (ADFCUP and CEQUP); € 70.270 (to CEQUP)
Coordinator : (ADFCUP) Victor Freitas
Coordinator : (CEQUP) André Melo
Alteração Endócrinas e Enzimática na Solha (Platichthys Flesus) e na taínha ( Mugil Cephalus)
Expostos a Cotaminantes Orgãnicos e Metais Pesados no Estuário do Douro, e seu Uso como
Indicadores de Poluição
PDCTM/P/MAR/15280/1999 (CIMAR and CEQUP); €41.100 (to CEQUP)
Coordinator : (CIMAR) Maria Armanda Henriques
Electrocatalytic Reactivity of Gold Chiral Surfaces
POCTI/ QUI/ 41704/2001 (FCUP and CEQUP) ; € 20.250 (to CEQUP)
Coordinator: Ana Maria Martins
Coordinator: (CEQUP) Natália Correia
Hepatoxicity and cardiotoxicity studies of methylenedioxymentamphetamine and metabolites in
Wistar Rats
POCTI/ 36099/FCB/2000 (FFUP, UA and CEQUP), € 65.061 (to CEQUP)
Estudo da autenticidade de queijo de ovelha “Terrincho” e de queijo de cabra Transmontano
AGRO Med. 8/Acção 8.1 Contrato 26;Proj. 2001090072347; € 13.467 (to CEQUP)
(Esc. Sup. Agrária Bragança, Assoc. Produtores de Queijo Transmontano ANCRAS e CEQUP)
Coordinator: ( Esc. Sup. Agrária de Bragança) António Manuel Peres
Coordinator: (CEQUP) Isabel Ferreira
Desenvolvimento e Aplicação de Metodologias Expeditas de Controlo e Segurança no Sector AgroAlimentar
AGRO Med. 8/Acção 8.1 Contrato. 273; Proj. 2001090072925, € 25.348 (to CEQUP)
(Universidade Católica Portuguesa, CEQUP e DRAEDM)
Coordinator: (U. Católica Portuguesa) António Rangel
Coordinator: (CEQUP) José Costa Lima
Química Verde
A - 43
Aumento do teor de enxofre em betumes industriais
Projecto I&DT financiado pela Galp Energia; € 60 000
Coordinator: Jose Paulo Mota
Estudo da capacidade adsorvente da rolha de cortiça relativamente a fungicidas provenientes do
vinho
Projecto I&DT financiado pela Amorim & Irmãos; € 33 265
Coordinato: Cristina Delerue Alvim de Matos
Cientistas de Palmo e Meio
Coordinator: Prof. Paulina Mata
Comportamento reológico e morfologia de sistemas mistos proteína/polissacarídeos
ICCTI/CAPES
Coordinator: M.ª Pilar Gonçalves
Étude des interactions macromoleculaires entre proteines de transfert d’electrons
ICCTI – Embaixada da França (Contrato nº 316 C2)
Coordinator: José J.G.Moura
Analyse des interactions protéines-protéines par arrimage moléculaire sous contraintes RMN
Centre National de la Recherche Scientifique / ICCTI (PICS Nº 1392)
Coordinator: José J.G.Moura
Fruit juice concentration by osmotic evaporation in membrane contactors
Bilateral Cooperation Project ICCTI/OMFB P9/01 Portugal- Hungary
Isabel Maria Coelhoso/K. Belafi-Bako
Development of membrane systems for chiral separations based on molecular recognition
mechanisms
Integrated Action E-39/02 Portugal- Spain
Isabel Maria Coelhoso/Cristina Palet Ballus
Determinação electroquímica de espécies de interesse clínico e farmacêutico. Sistema
potenciométrico automático para determinações de macro e micro-nutrientes em nutrição parental.
ICCTI / CAPES (423 / CAPES)
Coordinator: M.ª Conceição Montenegro
Contributo da monitorização baseada em sensores ópticos e potenciométricos na eficiência do
processo produtivo e minimização do impacto ambiental da galvanoplastia industrial
ICCTI
Coordinator: Alberto Nova Araújo
Automação de análises selectoras e discriminatórias
GRICES (GRICES/CNPq)
Coordinator: Rui Lapa
Química Verde
A - 44
Desenvolvimento de métodos automáticos de análise em fluxo aplicados à determinação de amostras
de interesse farmacêutico e ambiental
CRUP (E - 23/01)
Coordinator: José L.F.C. Lima
Desenvolvimento de mini-sondas e de sistemas automáticos visando análises de baixo impacto
ambiental.
ICCTI/CAPES
Coordinator: José L.F.C. Lima
Química Verde

report 2002

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