Electrospray ionization tandem mass spectrometry of labdane-type acid diterpenes.

Copaifera (Leguminoseae) species produce a commercially interesting oleoresin that displays several biological activities, including antimicrobial and anti-inflammatory properties. Labdane-type diterpenes are the main chemical constituents of these oleoresins, and copalic acid is the only compound that has been detected in all Copaifera oleoresins. In this study, we investigate some aspects of the gas-phase fragmentation reactions involved in the formation of the product ions from the deprotonated compounds (-)-ent-copalic acid (1), (-)-ent-3ß-hydroxy-copalic acid (2), (-)-ent-3ß-acetoxy-copalic acid (3), and (-)-ent-agathic acid (4) by electrospray ionization tandem mass spectrometry (ESI-MS/MS) and multiple stage mass spectrometry (MSn ). Our results reveal that the product ion with m/z 99 is common to all the analyzed compounds, whereas the product ion with m/z 217 is diagnostic for compounds 2 and 3. Moreover, only compound 4 undergoes CO2 (44 u) and acetic acid (60 u) elimination from the precursor ion. Thermochemical data obtained by computational chemistry at the B3LYP/6-31G(d) level of theory support the proposed ion structures. These data helped us to identify these compounds in a crude commercial Copaifera langsdorffii oleoresin by selective multiple reaction monitoring (MRM). Finally, a precursor ion scan (PIS) strategy aided screening of labdane-type acid diterpenes other than 1 to 4 in the same Copaifera oleoresin sample and led us to propose the structures of 8,17-dihydro-ent-agathic acid (5) and 3-keto-ent-copalic acid (6), which have not been previously reported in Copaifera oleoresins.

New insights in the chemical composition of benzoin balsams.

Benzoin balsam is an anthropic exudate obtained from the bark of several species of Styrax trees that is mainly used as a perfume fixative as well as a flavouring agent. Benzoe tonkinensis Laos (also commercialized under the denomination Siam benzoin balsam) displaying characteristic vanilla notes and already being largely used to flavour all kinds of edible goods, was intended to be proposed by Agroforex Company to the Codex Committee on Food Additives for evaluation as a food additive. For this purpose, the present paper reports the phytochemical characterisation of both the volatile and non-volatile fractions of benzoin balsams and the quantitation of some of the major components by gas and liquid chromatography techniques. Four coniferyl and two morinol derivatives were characterised for the first time in Benzoe tonkinensis Laos. Finally, two liquid chromatographic methods used to easily discriminate Siam from Sumatra balsam (also known as Benzoe sumatranus Indonesia) were developed.

Balsam of Peru: past and future.

Balsam of Peru (BOP) is a well-known contact allergen that is one of the most prevalent in the United States. For some patients allergic to BOP, external avoidance of fragrance is not enough to eliminate their dermatitis. A BOP avoidance diet has been shown to help many of these patients. This article reviews BOP allergens found in food. A special patch test series of allergens found in BOP along with data regarding which foods contain specific BOP constituents makes it possible to design potential targeted, refined, and simplified diets for BOP systemic contact dermatitis.

Natural resins and balsams from an eighteenth-century pharmaceutical collection analysed by gas chromatography/mass spectrometry.

Historical nomenclature has not always been unequivocally associated with the botanical origin of natural resins. The availability of natural resins has changed throughout the centuries and so have their trade names. Furthermore, adulterations and lack of knowledge have led to variations in the composition of the products traded under the same name. This investigation aims at a new understanding of the interrelation between the historical and modern terms for natural resins. Different Pinaceae and Pistacia resins, mastic, sandarac, copaiba balm and burgundy pitch from Vigani's Cabinet, a 300-year-old pharmaceutical collection owned by Queens' College, Cambridge (UK) were investigated. Related reference materials from modern collections were analysed together with natural resins derived from reliable botanical sources. The analytical method was gas chromatography/mass spectrometry (GC-MS) with and without derivatisation with trimethylsulfonium hydroxide. This technique provided detailed molecular compositions of the studied materials, which in turn led to particular data profiles of the materials. Marker molecules of Copaifera, Pinaceae, Cupressaceae and Pistacia resins were identified. By comparing the GC-MS data profiles to the reference samples, a clearer picture of the connection between nomenclature and botanical origin was obtained. With the aid of the marker molecules and data profiles, it was then possible to clarify the nomenclature of the aged resins sampled from Vigani's Cabinet.

[Pattern of contact sensitization to paraphenylenediamine and its detection in hair dyes]. / Patrón de sensibilización por contacto a parafenilendiamina y su detección en tintes capilares.

BACKGROUND: One of the greatest challenges in occupational dermatology is the identification of chemical substances used by patients in their work in order to determine their allergenic potential. Numerous techniques have been described for the identification of allergenic compounds. These tests must be sensitive, specific, and safe. We describe a study to detect the presence of paraphenylenediamine (PPD) in hair dyes that are commercially available in Spain. MATERIAL AND METHODS: We undertook an experimental study involving qualitative and semiquantitative detection of PPD in hair dyes sold in Spain. The qualitative technique we used was a previously described colorimetric method involving dilution of the dye with isopropyl alcohol followed by addition of a reagent solution (1g of vanilla in 15 ml of isopropyl alcohol and 7.5 ml of hydrochloric acid). A quantitative study was then done in which the dye was extracted in 96% ethanol and subjected to 1-dimensional thin-layer chromatography. RESULTS: A total of 15 brown and 12 blonde dyes were analyzed. PPD was identified in all of the brown dyes analyzed, irrespective of whether it was indicated (n = 12) or not (n = 3) in the composition. PPD was found in 6 of the 9 blonde dyes that indicated it in the composition and 2 of the 3 in which it was not indicated. Semiquantitative analysis by thin-layer chromatography revealed that the concentration of PPD in brown hair dyes (mean, 3%) was higher than in blonde dyes (mean, 0.1-0.3%). CONCLUSIONS: The presence of PPD in hair dyes is related to the color of the dye. It is consistently present in darker dyes and at low levels in blonde dyes. This study highlights the clinical and epidemiological importance of identifying allergens in dermatology, particularly in occupational dermatology.

Analytical investigation of styrax and benzoin balsams by HPLC- PAD-fluorimetry and GC-MS.

An HPLC and GC study has been conducted on the aromatic oleoresins styrax and benzoin produced by several American, Mediterranean and East-Asian trees, and widely used in ancient civilisations for their therapeutic and odoriferous properties. Initial experiments were performed by HPLC-PAD-fluorimetry for the analysis of several aromatic components, and then completed by GC-MS for the characterisation of both aromatic and triterpenic derivatives. In this work, it was crucial to isolate from fresh natural exudates, and to characterise by two-dimensional NMR, some of the major constituents in order to extend the standard molecular pool prior to chromatographic identifications. This study reveals coniferyl benzoate as an excellent distinctive fluorescent biomarker of Siam benzoin substrate. It also confirms that fluorimetric-coupled detection is a powerful analytical tool for the identification of compounds in Hamamelidaceae extracts that are almost undetectable by UV. GC-MS was successfully applied to the determination of the botanical origin of Sumatra benzoin, and to the identification of lupeol [3beta-lup-20(29)-en-3-ol] for the first time in such balsam-type materials.

Morfoanatomia de folhas de bálsamo: Sedum dendroideum Moc. et Sessé ex DC, Crassulaceae / Leaf morpho-anatomy of Sedum dendroideum Moc. et Sessé ex DC, Crassulaceae

Sedum dendroideum Moc. et Sessé ex DC, Crassulaceae é uma espécie suculenta, conhecida popularmente como bálsamo. Na medicina tradicional, suas folhas são empregadas para tratar inflamações de pele, contusões e problemas gástricos, em razão das ações emoliente e cicatrizante. Com o objetivo de fornecer subsídios morfológicos e taxonômicos, realizou-se estudo morfoanatômico das folhes. Observou-se que as folhas são simples, sésseis e alternas e apresentam formato oboval, levemente assimétrico, com ápice obtuso, base decurrente e margem lisa. A epiderme é uniestratificada e possui células com paredes anticlinais delgadas e onduladas. Os estômatos são anisocíticos, estando presentes em ambas as faces. Mesofilo homogêneo e numerosos idioblastos com conteúdo fenólico foram evidenciados.

A study of amber and copal samples using FT-Raman spectroscopy.

FT-Raman spectra were collected from fossil resins originating from a variety of geographical locations. The spectral profiles of most of the fossil resins could be related to modern resins containing diterpenoid components with predominantly labdane skeletons. The spectra collected from the fossil resin sample from Borneo differed from other fossil resins and was found to contain triterpenoid components. The differences in the spectral profile of fossil resins containing diterpenoid components are shown to relate to differences in level of maturation rather than geographical origin. FT-Raman spectra of fossil resins cannot be used to distinguish source although the degree of maturation can be used as an indicator to narrow the range of possible geographical origins.

Influência da pasta Sultan sobre o processo de reparo em alvéolo dental infectado: análise histológica em ratos / The influence of the Sultan's paste over delayed healing of wound extraction: a histological study in rats

O processo de reparo do alvéolo infectado de ratos (Rattus norvegicus, albinus, Wistar) foi avaliado após a limpeza cirúrgica e irrigaçäo com soro fisiológico seguida do preenchimento da loja com a pasta Sultan. Para tanto, todos os animais utilizados (8 ratos doadores e outros 60 animais experimentais) tiveram o incisivo central superior direito extraído e, alveolite provocada experimentalmente, nos animais doadores e naqueles dos Grupos I, II, III e IV. Os 60 ratos experimentais constituíram os seguintes Grupos: I. Alveolite sem tratamento; II. Limpeza cirúrgica e irrigaçäo; III. Pasta Sultan, apenas; IV. Limpeza cirúrgica e irrigaçäo mais pasta Sultan. Os animais, em número de cinco em cada grupo, foram sacrificados aos 6, 15 e 28 dias pós-operatórios e as peças obtidas incluídas em parafina, cortadas e coradas com hematoxilina e eosina para análise em microscopia óptica. Os resultados foram submetidos a análise qualitativa. Com base nos resultados foi possível concluir que: (1) O grupo da alveolite teve o reparo alveolar comprometido quanto a sua cronologia; (2) O grupo da limpeza cirúrgica seguida da irrigaçäo alveolar foi o que apresentou resultados superiores; (3) a pasta Sultan näo deve ser utilizada como curativo de preenchimento alveolar para o tratamento da alveolite

Estudo preliminar da compatibilidade biológica dos cimentos de óxido de zinco e copaíba e óxido de zinco e eugenol: avaliaçäo histológica de implantes subcutâneos em ratos / Preliminary study of the biological compatibility of a zinc oxide-copaiba and zinc oxide-eugenol cements: histological evaluation of subcutaneous implants in rats

No tecido conjuntivo subcutâneo de 20 ratos foram implantados tubos de polietileno preenchidos com o cimento do óxido de zinco e copaíba (Grupo I) e óxido de zinco e eugneol (Grupo Controle II). Decorridos os períodos de 7, 15, 30 e 60 dias, biópsias foram realizadas e as peças cirúrgicas foram cortadas seriadamente e coradas com hematoxilina e eosina. Aos 7 dias para o Grupo I, houve discreta presença de células inflamatórias, necrose de contato e degradaçäo de fibras colágenas, näo sendo observada a presença de neutrófilos ou de células gigantes. O cone capsular formado junto à abertura tubular apresentou discreta amplitude. Todos estes eventos histopatológicos foram ligeiramente superiores para o Grupo II nos primeiros períodos. Com o decorrer dos períodos, para os dois grupos, todos os eventos analisados reduziram em quantidade, e no último período o tecido conjuntivo da área principal de análise apresentava características histológicas de normalidade, permanecendo o tubo de polietileno totalmente envolto por um tecido fibroso denso. Estes achados sugeriram que o material do Grupo I (OZ copaíba) foi menos irritante que o material do Grupo II (Controle), quando implantados no tecido conjuntivo subcutâneo do rato

Dermatitis from propolis.

22 patients with dermatitis from propolis were studied. In all the 21 tests with propolis were positive, and in 19 they were also positive to balsam of Peru. The chromatogram of the balsam and propolis show marked similarity and 3 identical peaks could be recognised in both substances. Among the patients sensitive to balsam of Peru and propolis, 12 were tested with some common components; 3 were positive to cinnamyl cinnamate, 2 to vanillin and 1 to benzyl cinnamate. Chromatograms of the 3 propolis samples from the Warsaw region were very similar, but not identical. Some of the patients were tested with 35 essential oils and eugenol. Sensitivity to clove oil was common.

Infrared spectroscopy for tracing of topically applied ointment vehicles and active substances on healthy skin.

Infrared spectroscopy was used to trace active substances and ointment vehicles applied on the skin. Vaseline and lanoline could be traced after 8 hrs but not olive oil. From the active substances, ethyl-4-amino-benzoate (5 per cent), clioquinol (5 per cent), parabenes (15 per cent), 5,7-dichlor-8-hydroxy-2-methyl-chinolin (5 per cent), balsam of Peru (25 per cent) and pyroleum pini (12 per cent) could be traced 1 hr after application but had disappeared after 8 hrs. Ethylenediamine (1 per cent), chlorcresol (1 per cent), pyroleum lithantracis (5 per cent), were not traceable after 1 hr, and curiously neither neomycine sulphate in spite of its high concentration (20 per cent). The reaction of the skin surface lipids, after application of different substances, was deduced from the spectra. Clioquinol and pyroleum lithantracis seem to give rise to hydrolysis of the triglycerides, the free fatty acids being clearly identifiable. It is felt that infrared spectroscopy can be used as an effective method to trace different substances such as potent allergens on healthy or diseased skin.