New porphyrins tailored as biodiesel fluorescent markers

This work reports the synthesis, characterization, and evaluation of new porphyrins tailored to become biodiesel fluorescent markers. The compounds were obtained by the synthetic modification of the commercially available porphyrin 5,10,15,20-meso-tetrakis(pentafluorophenyl)porphyrin (TPPF(20)) using ethanol and hexadecan-1-ol (cetylic alcohol) as nucleophilic reagents. The stability of the marked biodiesel fuel solutions was investigated every 15 days for a total period of 3 months, and under different storage temperature and light exposure condition, simulating the conventional stock conditions. The influence of the different substituents of the porphyrins on the fluorescence properties of the biodiesel fuel markers was also assessed. The resulting porphyrins were highly soluble in biodiesel fuel and displayed strong fluorescence characterized by two strong emission bands. The fluorescent markers did not affect the biodiesel physical properties and were stable in storage conditions for at least 3 months at a concentration of 4 ppm. The best storage condition was found to be absence of light and 6 degrees C; the limit of detection by photoluminescence technique had magnitude of 10(-13) mol L(-1). The synthesized porphyrins were characterized by nuclear magnetic resonance ((1)H-NMR and (19)F-NMR), mass spectrometry (HRMS), ultraviolet visible absorption spectroscopy, and photoluminescence spectroscopy. (C) 2011 Elsevier Ltd. All rights reserved.

Purification and structural characterization of compounds isolated from Cystoseira abies-marina

6th Spanish-Portuguese-Japanese Organic Chemistry Symposium, Lisboa, de 18 a 20 de Julho de 2012 (Poster Communication).

Abordagem integrada na análise química de amostras ambientais

Dissertação para obtenção do Grau de Doutor em Química

Identificação e quantificação de adutos covalentes formados entre a valina Nterminal da hemoglobina e o fármaco nevirapina em doentes de HIV: Desenvolvimento e validação de um método analítico por espectrometria de massa de alta resolução

A nevirapina é o fármaco antirretroviral mais utilizado nos países em desenvolvimento para combater o vírus da imunodeficiência humana (HIV). Apesar dos seus efeitos benéficos, a nevirapina tem sido associada a casos de hipersensibilidade cutânea e hepatoxicidade graves ou mesmo fatais. Estudos recentes sugerem o envolvimento da bioactivação do seu metabolito de Fase I, 12-OHNevirapina, a espécies electrofílicas capazes de formar adutos covalentes com proteínas. A identificação dos adutos formados, por esta via de bioactivação, com a valina N-terminal da hemoglobina (Hb) isolada de doentes sob terapêutica com a nevirapina foi já efectuada. A técnica analítica utilizada envolveu a utilização do método de derivatização N-alquilo Edman que, por reacção com fenilisotiocianato (PITC), permite destacar o aduto formado com o resíduo N-terminal da Hb, sob a forma de uma hidantoina, que foi posteriormente analisado e quantificado por cromatografia líquida acoplado à espectrometria de massa de tandem por ionização de electrospray. O presente trabalho teve como objectivo inicial optimizar o procedimento experimental envolvido no procedimento de N-alquilo Edman, tendo-se inicialmente comparado a sensibilidades obtidas mediante a utilização de dois agentes derivatizante: o PITC e o isotiocianato de fluoresceína (FTIC). Este processo inicial permitiu estabelecer que a utilização do PTIC como agente de derivatização conduzia a melhores resultados, tendo-se prosseguido com o desenvolvimento e a validação do método analítico para a quantificação deste aduto por cromatografia líquida acoplado a espectrometria de massa de alta resolução (LC-HRMS). O método analítico desenvolvido apresentou recuperações entre 36 e 57 %, supressão de sinal de 19 % e um limite de quantificação (LOQ) de 1,4 ppb (média da exactidão = 98 %; CV = 5 %). A metodologia validada foi depois aplicada na quantificação de adutos de nevirapina em duas amostras de Hb isolada de doentes de HIV e os níveis de adutos obtidos foram de 95 e 92 pmol/g de Hb.

Test methods: anabolics.

In the International Olympic Committee (IOC) accredited laboratories, specific methods have been developed to detect anabolic steroids in athletes' urine. The technique of choice to achieve this is gas-chromatography coupled with mass spectrometry (GC-MS). In order to improve the efficiency of anti-doping programmes, the laboratories have defined new analytical strategies. The final sensitivity of the analytical procedure can be improved by choosing new technologies for use in detection, such as tandem mass spectrometry (MS-MS) or high resolution mass spectrometry (HRMS). A better sample preparation using immuno-affinity chromatography (IAC) is also a good tool for improving sensitivity. These techniques are suitable for the detection of synthetic anabolic steroids whose structure is not found naturally in the human body. The more and more evident use, on a large scale, of substances chemically similar to the endogenous steroids obliges both the laboratory and the sports authorities to use the steroid profile of the athlete in comparison with reference ranges from a population or with intraindividual reference values.

Analysis and quantification of vitamin D metabolites in serum by ultra-performance liquid chromatography coupled to tandem mass spectrometry and high-resolution mass spectrometry: a method comparison and validation.

RATIONALE: The aim of the work was to develop and validate a method for the quantification of vitamin D metabolites in serum using ultra-high-pressure liquid chromatography coupled to mass spectrometry (LC/MS), and to validate a high-resolution mass spectrometry (LC/HRMS) approach against a tandem mass spectrometry (LC/MS/MS) approach using a large clinical sample set. METHODS: A fast, accurate and reliable method for the quantification of the vitamin D metabolites, 25-hydroxyvitamin D2 (25OH-D2) and 25-hydroxyvitamin D3 (25OH-D3), in human serum was developed and validated. The C3 epimer of 25OH-D3 (3-epi-25OH-D3) was also separated from 25OH-D3. The samples were rapidly prepared via a protein precipitation step followed by solid-phase extraction (SPE) using an HLB μelution plate. Quantification was performed using both LC/MS/MS and LC/HRMS systems. RESULTS: Recovery, matrix effect, inter- and intra-day reproducibility were assessed. Lower limits of quantification (LLOQs) were determined for both 25OH-D2 and 25OH-D3 for the LC/MS/MS approach (6.2 and 3.4 µg/L, respectively) and the LC/HRMS approach (2.1 and 1.7 µg/L, respectively). A Passing & Bablok fit was determined between both approaches for 25OH-D3 on 662 clinical samples (1.11 + 1.06x). It was also shown that results can be affected by the inclusion of the isomer 3-epi-25OH-D3. CONCLUSIONS: Quantification of the relevant vitamin D metabolites was successfully developed and validated here. It was shown that LC/HRMS is an accurate, powerful and easy to use approach for quantification within clinical laboratories. Finally, the results here suggest that it is important to separate 3-epi-25OH-D3 from 25OH-D3. Copyright © 2012 John Wiley & Sons, Ltd.

High-resolution mass spectrometry applied to the study of metabolome modifications in various chicken tissues after amoxicillin administration

The performance of high resolution accurate mass spectrometry (HRMS) operating in full scan MS mode was investigated for the quantitative determination of amoxicillin (AMX) as well as qualitative analysis of metabolomic profiles in tissues of medicated chickens. The metabolomic approach was exploited to compile analytical information on changes in the metabolome of muscle, kidney and liver from chickens subjected to a pharmacological program with AMX. Data consisting of m/z features taken throughout the entire chromatogram were extracted and filtered to be treated by Principal Component Analysis. As a result, it was found that medicated and non-treated animals were clearly clustered in distinct groups. Besides, the multivariate analysis revealed some relevant mass features contributing to this separation. In this context, recognizing those potential markers of each chicken class was a priority research for both metabolite identification and, obviously, evaluation of food quality and health effects associated to food consumption.

Quantitative performance of a quadrupole-orbitrap-MS in targeted LC-MS determinations of small molecules.

High-resolution mass spectrometry (HRMS) has been associated with qualitative and research analysis and QQQ-MS with quantitative and routine analysis. This view is now challenged and for this reason, we have evaluated the quantitative LC-MS performance of a new high-resolution mass spectrometer (HRMS), a Q-orbitrap-MS, and compared the results obtained with a recent triple-quadrupole MS (QQQ-MS). High-resolution full-scan (HR-FS) and MS/MS acquisitions have been tested with real plasma extracts or pure standards. Limits of detection, dynamic range, mass accuracy and false positive or false negative detections have been determined or investigated with protease inhibitors, tyrosine kinase inhibitors, steroids and metanephrines. Our quantitative results show that today's available HRMS are reliable and sensitive quantitative instruments and comparable to QQQ-MS quantitative performance. Taking into account their versatility, user-friendliness and robustness, we believe that HRMS should be seen more and more as key instruments in quantitative LC-MS analyses. In this scenario, most targeted LC-HRMS analyses should be performed by HR-FS recording virtually "all" ions. In addition to absolute quantifications, HR-FS will allow the relative quantifications of hundreds of metabolites in plasma revealing individual's metabolome and exposome. This phenotyping of known metabolites should promote HRMS in clinical environment. A few other LC-HRMS analyses should be performed in single-ion-monitoring or MS/MS mode when increased sensitivity and/or detection selectivity will be necessary.

Controle de autenticidade de óleos de copaíba comerciais por cromatografia gasosa de alta resolução

Copaiba oil is a resin extracted from the trunk of trees of Copaifera species which grow in Brazil where it is widely used in popular medicine as an anti-inflammatory, antiseptic anti-bactericidal, diuretic, dermatological, expectorant, and anti-infective. The comparative study of the composition of commercial copaiba oils was carried out by high resolution gas chromatography (HRGC) and high resolution gas chromatography-mass spectrometry (HRGC-MS) analysis. The commercial oils were compared with authentic oils, collected from the southeast and northern regions of Brazil. Sixteen commercial oils were analysed and two of them revealed adulterations with fatty acids. Using the chromatographic profile of authentic copaiba species oils, it is possible to analyse commercial oils and determine the authenticity of these materials.

Ácido ent-16alfa,17-diidroxicauran-19-óico isolado de Ouratea semiserrata e os desafios estereoquímicos dos carbonos quirais C-4 e C-16

Phytochemical investigation of the leaves and branches of a specimen of Ouratea semiserrata led to the isolation and characterization of ent-16alpha,17-dihydroxykauran-19-oic acid, along with other natural products. This diterpenoid and its derivatives were used to unambiguous ¹H and 13C chemical shifts assignments and to indicate some mistake data described in the literature as consequence mainly of the stereochemicals of the chiral carbons C-4 and C-16. The HRMS spectra were also analysed.

Influência do material do alambique na composição química das aguardentes de cana-de-açúcar

This paper describes variations in the profile of the main volatile organic compounds present in Brazilian sugar cane spirits distilled in copper and stainless steel distillers. The main organic compounds: aldehydes, ketones, carboxylic acids, alcohols and esters, were determined through High Performance Liquid Chromatography (HPLC) and High Resolution Gas Cromatography (HRGC). The spirits produced in copper distillers exhibit higher contents of aldehydes with respect to the ones produced in stainless steel. The inverse is true with respect to the higher alcohol and ester contents. No significant variation has been observed for the carboxylic acids.

Principais substâncias responsáveis pelo aroma de mangas comerciais brasileiras identificadas por cromatografia gasosa de alta resolução/olfatometria/espectrometria de massas

Six Brazilian commercial mangoes were analysed by HRGC-O-AEDA-MS, viz., Carlota, Haden, Espada, Coração de boi, Rubi and Tommy Atkins. All them showed ethyl butanoate as the main aroma character impact compound by AEDA evaluation. The ethyl esters of 2 and 3-methylbutanoic acids are also important, the main contribution in Carlota variety being 2(S) enantiomer. In Rubi variety, both 2(R) enantiomer and 3-methyl isomer contributes to the caprylic fruity note observed. In four varieties, viz., Haden, Espada, Rubi and Tommy Atkins, d-3-carene showed to be the second impact aroma compound presented. In Tommy Atkins variety, a-pinene also has a significant contribution, mainly due to its (1R,5R)(+)-enantiomer.

Estado da arte da cromatografia gasosa de alta resolução e alta temperatura

The developments in stationary phase synthesis and capillary column technology, have opened new perspectives in analysis of high molecular mass compounds (³600 daltons) and thermolabile organic compounds by High Temperature High Resolution Gas Chromatography (HT-HRGC). HT-HRGC is a new analytical borderline and its application to the analysis of high molecular mass compounds is still in its infancy. The apolar and medium polar gum phases can now be operated at temperatures up to 400-480ºC, being used for the analysis of n-alcanes up to C-100, lipids, oligosaccharides, industrial resins, polyglycerols, cyclodextrins, porphyrins, etc. This technique should play a leading role as a powerful tool, for many different analysis types, in multidisciplinary fields of Science.

Análise dos álcoois, ésteres e compostos carbonílicos em amostras de óleo fúsel

Analysis of alcohols, esters and carbonyl compounds were performed using HRGC and HPLC techniques in samples of fusel oils from three different Brazilian alcohol distilleries. High content of isoamyl alcohol (390 g.L-1), isobutyl alcohol (158 g.L-1), ethyl alcohol (28,4 g.L-1), methyl alcohol (16,6 g.L-1) and n-propyl alcohol (11,9 g.L-1) were found. These compounds represent 77 ± 8 % of the approximated weight of a liter of fusel oils. The obtained results show the feasibility of using fusel oils as low-cost raw material for the synthesis of chemicals.

Diterpenóides com esqueleto cleistantano de Vellozia aff. carunculares martius ex seubert (Velloziaceae)

From the hexane extract of leaves, stem and roots, of Vellozia aff. carunculares, fourteen cleistanthane diterpenoids were detected by HRGC-MS, through the analysis of their fragmentation patterns. Comparison of their MS spectra with those of authentic standards previously isolated from other species of the family Velloziaceae allowed the identification, with a high degree of confidence, of eigth known cleistanthane diterpenoids. Of these, four were isolated and identified from their MS, IR and ¹H and 13C NMR spectra, as the known cleistantha-8,11,13-trien-7-one, cleistantha-8,11,13-trien-3,7-dione, cleistantha-1,8,11,13-tetraen-3,7-dione and the first reported 6-alpha-hydroxy-cleistantha-8,11,13-trien-7-one. The triterpenoid beta-amyrin was also isolated.