Discrimination and classification of extra virgin olive oil using a chemometric approach based on TMS-4,4'-desmetylsterols GC(FID) fingerprints of edible vegetable oils.

A single out-line HPLC-GC (FID) analytical method is applied to acquire the chromatographic fingerprint characteristic of the TMS-4,4'-desmetylsterol derivative fraction of several marketed edible vegetable oils in order to identify and discriminate the most valuable extra-virgin olive oils from the other vegetal oils (canola, corn, grape seed, linseed, olive pomace, peanut, rapeseed, soybean, sesame, seeds (non-specified composition but usually a blend of corn and sunflower) and sunflower). The natural structure of the preprocessed data undergoes a preliminary exploration using principal component analysis and heat map-based cluster analysis. A partial least squares-discriminant model is first trained from 53 oil samples (only 3 latent variables) and externally validated from 18 test oil samples. No classification errors are found and all the test samples are correctly classified. Additional classification models are also built in order to discriminate among vegetables-oil families and excellent results have been also achieved.

Gas chromatographic-mass spectrometric characterization of thebaol, an opium constituent, and its structural analogs.

A GC-MS method is described for the characterization of thebaol, a component of opium poppy. The method includes preliminary sample derivatization to TMS, TBDMS, TFA, PFP and HFB substituted products. Fragmentation of resulting derivatives is unique under electron ionization, and proceeds via consecutive loss of two radicals that violate the "even-electron rule". Peaks of [M-2CH3]+. and [M-C4H9-CH3]+. ions show maximum intensities in the spectra of trimethyl- and tert-butyldimethylsilyl-thebaols. Elimination of perfluoroalkyl and methyl radicals from M+. is characteristic for TFA, PFP and HFB thebaols. The same fragmentation peculiarity is characteristic for derivatives prepared from related natural compounds containing vicinal 2-methoxyphenol moieties. The unique fragmentation of trialkylsilyl and perfluoroacyl derivatives of thebaol can be successfully used for thebaol determination within complex mixtures. This is part 4 from the series "Analytical derivatives in mass spectrometry", parts 1, 2 and 3 see [1-3].

Stability comparison of two dermal emulsions containing Hippophae Rhamnoides L. oil.

Two formulations of multiple emulsion (ME-1 & ME-2) containing Hipophae rhamnoides L. oil were prepared. Along with emulsifiers (Abil EM 90, Synperonic F127), Magnesium and Zinc were used as stabilizer in ME-1 and ME-2 respectively. Both formulations were prepared using Two-Step Method and after preparation ME-1 and ME-2 were stored at different storage conditions i.e., 4°C, 25°C, 40°C, 40°C+ 75% RH for four weeks for stability evaluation. At 4°C ME-1 and ME-2 showed phase inversion and were excluded from further stability evaluation. While ME-1 and ME-2 both were stable at 25°C for a period of four weeks as no color change, phase separation and liquefaction occurred. ME-1 and ME-2 at 40°C, 40°C+ 75% RH exhibited change in color, liquefaction and phase separation. The decrease in viscosity and globule size whereas increase in electrical conductivity and pH were observed at 40°C, 40°C+ 75% RH for a period of four weeks. Multiple emulsion from Hippophae rhamnoide L. oil are unstable at refrigeration and high temperature conditions. These formulations must be placed at room temperature to increase their shelf life.

Product Selectivity of Esterification of L-Aspartic Acid and L-Glutamic Acid Using Chlorotrimethylsilane.

TMSCI works as an acid catalyst precursor for selective esterification of L-aspartic and L-glutamic acids in the presence of primary, secondary and tertiary alcohols. Although excess TMSCI was required for the completion of esterification, the resulting alkyl TMS ether could be azeotropically removed by simple evaporation with alcohol.

Development of cetyl dimethicone based water-in-oil emulsion containing botanicals: Physical characteristics and stability.

The aim of current research was to develop a water-in-oil emulsion containing grape seed extract for application in cosmeceuticals. Finally grinded dried grape seeds powder was extracted with hydro alcoholic mixture. Emulsions consisting of different concentrations of cetyl dimethicone (Abile EM90), the nonionic emulsifier, liquid paraffin as oily phase and water as aqueous phase were developed. Color, odor, pH, viscosity, liquefaction, phase separation, centrifugation and thermal stability of the formulated emulsions were observed at various storage temperatures i.e. 8±0.5°C, 25±0.5°C, 40±0.5°C and 40°C±0.5°C with 70% RH. The stable formulation consist of 16% mineral oil, 4% of ABIL EM 90(®), 4% grape seeds extract, 1% rose oil and 75% distilled water. All the results derived from this study showed good stability over the three months study period which indicates w/o emulsion can be used as carrier of 4% grape seeds extract to enhance desired effects when applied topically.

Determination of terpenoid indole alkaloids in hairy roots of Rhazya stricta (Apocynaceae) by GC-MS.

INTRODUCTION: Rhazya stricta Decne. (Apocynaceae) is a medicinal plant rich in terpenoid indole alkaloids (TIAs), some of which possess important pharmacological properties. The study material including transgenic hairy root cultures have been developed and their potential for alkaloid production are being investigated. OBJECTIVE: In this study, a comprehensive GC-MS method for qualitative and quantitative analysis of alkaloids from Rhazya hairy roots was developed. METHODS: The composition of alkaloids was determined by using GC-MS. In quantification, the ratio between alkaloid and internal standard was based on extracted ion from total ion current (TIC) analyses. RESULTS: The developed method was validated. An acceptable precision with RSD ≤ 8% over a linear range of 1 to 100 µg/mL was achieved. The accuracy of the method was within 94-107%. Analysis of hairy root extracts indicated the occurrence of a total of 20 TIAs. Six of them, pleiocarpamine, fluorocarpamine, vincamine, ajmalicine and two yohimbine isomers are reported here for the first time in Rhazya. Trimethylsilyl (TMS) derivatisation of the extracts resulted in the separation of two isomers for yohimbine and also for vallesiachotamine. Clearly improved chromatographic profiles of TMS-derivatives were observed for vincanine and for minor compounds vincamine and rhazine. CONCLUSION: The results show that the present GC-MS method is reliable and well applicable for studying the variation of indole alkaloids in Rhazya samples.

Gas chromatography-mass spectrometry microanalysis of alpha- and gamma-tocopherol in plasma and whole blood.

BACKGROUND: Assessing vitamin E status in humans is critical for nutritional evaluation and verification of clinical and biological compliance of supplemented subjects. An accurate analytical method for measuring the two main vitamin E isoforms, i.e. α- and γ-tocopherol (α- and γ-TOH) in small volumes of plasma can facilitate the application of this analysis to clinical trials and in situations where a limited amount of sample is available. METHODS: We have developed a micro method, which uses only 5 µL plasma, based on isotope dilution, trimethylsilation and GC-MS. The method was validated according to the guidelines of the International Conference on Harmonization of analytical procedures. The method was also applied to 5 µL of whole blood for the potential use in conditions were the availability of specimens is limited. RESULTS: Accurate quantitation of α-TOH and γ-TOH was achieved at levels ≥ 0.417 µM and ≥ 0.007 µM, respectively. Within-day coefficient of variation was 1.31% and 4.70% for α-TOH and γ-TOH, respectively. Between-day coefficient of variation was 1.32% and 2.88% for α-TOH and γ-TOH, respectively. Recovery, assessed at three concentration levels, ranged 98-103% and 100-102% for α-TOH and γ-TOH, respectively. The method allowed the detection of α-TOH and γ-TOH in 5 µL whole blood and in membranes of red blood cells washed from 5 µL of blood as well. The analytical performance was assessed in plasma from a cohort of Italian healthy subjects (n = 205). The mean plasma concentrations were 28.01 ± 6.31 and 0.68 ± 0.48 µM (mean ± SD) for α-TOH and γ-TOH, respectively. Alpha-TOH correlated with total cholesterol (r = 0.617, p < 0.0001) and triglycerides (r = 0.420, p < 0.0001) while γ-TOH correlated modestly with total cholesterol (r = 0.213, p < 0.0001) but not with triglycerides. γ-TOH, but not α-TOH, was significantly lower in smokers than in non-smokers (0.72 ± 0.50 vs. 0.56 ± 0.37, µM, mean ± SD, p = 0.017). Given the high sensitivity, the method allowed to be applied to 5 µM whole blood without specific modification. CONCLUSIONS: This micro-method represents an analytical advancement in α- and γ-TOH assay that is available to accurately verify the nutritional status and compliance after supplementation in large-scale settings, and to measure the two vitamers in conditions where sample availability is limited.

Generic Role of Polymer Supports in the Fine Adjustment of Interfacial Interactions between Solid Substrates and Model Cell Membranes.

To understand the generic role of soft, hydrated biopolymers in adjusting interfacial interactions at biological interfaces, we designed a defined model of the cell-extracellular matrix contacts based on planar lipid membranes deposited on polymer supports (polymer-supported membranes). Highly uniform polymer supports made out of regenerated cellulose allow for the control of film thickness without changing the surface roughness and without osmotic dehydration. The complementary combination of specular neutron reflectivity and high-energy specular X-ray reflectivity yields the equilibrium membrane-substrate distances, which can quantitatively be modeled by computing the interplay of van der Waals interaction, hydration repulsion, and repulsion caused by the thermal undulation of membranes. The obtained results help to understand the role of a biopolymer in the interfacial interactions of cell membranes from a physical point of view and also open a large potential to generally bridge soft, biological matter and hard inorganic materials.

Oxidative trifluoromethylation and trifluoromethylthiolation reactions using (trifluoromethyl)trimethylsilane as a nucleophilic CF3 source.

The trifluoromethyl group is widely prevalent in many pharmaceuticals and agrochemicals because its incorporation into drug candidates could enhance chemical and metabolic stability, improve lipophilicity and bioavailability, and increase the protein bind affinity. Consequently, extensive attention has been devoted toward the development of efficient and versatile methods for introducing the CF3 group into various organic molecules. Direct trifluoromethylation reaction has become one of the most efficient and important approaches for constructing carbon-CF3 bonds. Traditionally, the nucleophilic trifluoromethylation reaction involves an electrophile and the CF3 anion, while the electrophilic trifluoromethylation reaction involves a nucleophile and the CF3 cation. In 2010, we proposed the concept of oxidative trifluoromethylation: the reaction of nucleophilic substrates and nucleophilic trifluoromethylation reagents in the presence of oxidants. In this Account, we describe our recent studies of oxidative trifluoromethylation reactions of various nucleophiles with CF3SiMe3 in the presence of oxidants. We have focused most of our efforts on constructing carbon-CF3 bonds via direct trifluoromethylation of various C-H bonds. We have demonstrated copper-mediated or -catalyzed or metal-free oxidative C-H trifluoromethylation of terminal alkynes, tertiary amines, arenes and heteroarenes, and terminal alkenes. Besides various C-H bonds, aryl boronic acids proved to be viable nucleophilic coupling partners for copper-mediated or -catalyzed cross-coupling reactions with CF3SiMe3. To further expand the reaction scope, we also applied H-phosphonates to the oxidative trifluoromethylation system to construct P-CF3 bonds. Most recently, we developed silver-catalyzed hydrotrifluoromethylation of unactivated olefins. These studies explore boronic acids, C-H bonds, and P-H bonds as novel nucleophiles in transition-metal-mediated or -catalyzed cross-coupling reactions with CF3SiMe3, opening new viewpoints for future trifluoromethylation reactions. Furthermore, we also achieved the oxidative trifluoromethylthiolation reactions of aryl boronic acids and terminal alkynes to construct carbon-SCF3 bonds by using CF3SiMe3 and elemental sulfur as the nucleophilic trifluoromethylthiolating reagent. These oxidative trifluoromethylation and trifluoromethylthiolation reactions tolerate a wide range of functional groups, affording a diverse array of CF3- and CF3S-containing compounds with high efficiencies, and provide elegant and complementary alternatives to classical trifluoromethylation and trifluoromethylthiolation reactions. Because of the importance of the CF3 and SCF3 moieties in pharmaceuticals and agrochemicals, these reactions would have potential applications in the life science fields.

The use of trimethylsilyl cyanide derivatization for robust and broad-spectrum high-throughput gas chromatography-mass spectrometry based metabolomics.

Reproducible and quantitative gas chromatography-mass spectrometry (GC-MS)-based metabolomics analysis of complex biological mixtures requires robust and broad-spectrum derivatization. We have evaluated derivatization of complex metabolite mixtures using trimethylsilyl cyanide (TMSCN) and the most commonly used silylation reagent N-methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA). For the comparative analysis, two metabolite mixtures, a standard complex mixture of 35 metabolites covering a range of amino acids, carbohydrates, small organic acids, phenolic acids, flavonoids and triterpenoids, and a phenolic extract of blueberry fruits were used. Four different derivatization methods, (1) direct silylation using TMSCN, (2) methoximation followed by TMSCN (M-TMSCN), (3) direct silylation using MSTFA, and (4) methoximation followed by MSTFA (M-MSTFA) were compared in terms of method sensitivity, repeatability, and derivatization reaction time. The derivatization methods were observed at 13 different derivatization times, 5 min to 60 h, for both metabolite mixtures. Fully automated sample derivatization and injection enabled excellent repeatability and precise method comparisons. At the optimal silylation times, peak intensities of 34 out of 35 metabolites of the standard mixture were up to five times higher using M-TMSCN compared with M-MSTFA. For direct silylation of the complex standard mixture, the TMSCN method was up to 54 times more sensitive than MSTFA. Similarly, all the metabolites detected from the blueberry extract showed up to 8.8 times higher intensities when derivatized using TMSCN than with MSTFA. Moreover, TMSCN-based silylation showed fewer artifact peaks, robust profiles, and higher reaction speed as compared with MSTFA. A method repeatability test revealed the following robustness of the four methods: TMSCN > M-TMSCN > M-MSTFA > MSTFA.

[Water-soluble non-alkaloid chemical constituents contained in Corydalis yanhusuo by trimethylsilyl derivatization GC-MS].

OBJECTIVE: To study the water-soluble non-alkaloid chemical constituents of Corydalis yanhusuo. METHOD: The 80% alcohol extracts of C. yanhusuo passed through DA201 macroporous resin. Eluted fractions were collected and passed though 732 # cation exchange resin. Water eluate was collected, dried and derived with trimethylsilane. Gas Chromatography/Mass Spectrometry and NIST 2005 library were adopted for MS/MS mass spectrogram to infer the compound structure. RESULT: Sixteen compounds were tentatively identified from about fifty peaks detected by GC-MS and identified as hydroxyl and carboxyl polar compounds. CONCLUSION: These sixteen compounds were found for the first time in C. yanhusuo. The results provide scientific basis for in-depth development of C. yanhusuo.

Characteristic fragmentation patterns of trimethylsilyl and trimethylsilyl-oxime derivatives of plant disaccharides as obtained by gas chromatography coupled to ion-trap mass spectrometry.

The characteristic fragmentation pattern of six reducing and two non reducing type disaccharides-(neohesperidose, acuminose, sambubiose, rutinose, vicianose, primverose, and two arabinosyl-inositols) has been described. These saccharides have not been previously identified by on-line chromatographic techniques. Unambiguous specific characteristics of the TMS (oxime)s such as mass distribution, syn/anti oximes ratios and elution order proved to be associated with their reducing or non reducing character, with their aldosyl property and with the position of their O-glycosidic linkages. The practical utility of the mass fragmentation study of these rare disaccharides was demonstrated, at the first time, by the simultaneous, on-line identification and quantification of the acuminose, vicianose, primverose and arabinosyl-inositol contents of tea leaves, from green and black tea blends of Indian and Chinese origin.

Gas chromatographic retention indices of trimethylsilyl derivatives of terpene alcohols.

This paper presents the experimentally determined retention indices (RI(TMS)) for a set of 75 silylated terpenols (33 monoterpenols and 42 sesquiterpenols). The attempt was made to assess the dependence of RI(TMS) on RI (for non-silylated terpenols) and on RI(Ac) (for acetylated terpenols). Satisfactory linear regression parameters (RI(TMS)=b(0)+b(1)RI) were observed for tertiary substituted monoterpenols and primary or secondary substituted sesquiterpenols. The mass spectra of silylated terpenols that were not found in the available literature are in Supplementary information.

Phytosterols, unsaturated fatty acid composition and accumulation in the almond kernel during harvesting period: importance for development regulation.

The evolution of the composition of sterols and squalene during the maturation of the fruit of three cultivars (Achaak, Perlees and Mazetto) almond (Prunus amygdalus Batsh) was investigated. At the same time the evolution of oleic, linoleic and linolenic fatty acids was also studied. The qualitative and quantitative analyses were made by GC-MS and GC-FID. The present study is based on three axes: The first one is the structural and molecular identification of compounds sterolic and squalene, which are based on the principal of cleavage and the fragmentation characteristic of each peak provided by mass spectrometry. The second axis is interested in the physiological phenomenon of phytosterols accumulation: biosynthesis, evolution, and their relation with squalene as well as their interconversion. The third axis is an emergence of a relationship, which seems to exist, between the biosynthetic compounds of the glyceridic fraction of almond oil (mainly fatty acids) and those of the unsaponifiable fraction (particularly sterols). This relation may be established by 24-methylene cholesterol.

Gas chromatographic-mass spectrometric fragmentation study of phytoestrogens as their trimethylsilyl derivatives: identification in soy milk and wastewater samples.

An analytical method for the identification of eight plant phytoestrogens (biochanin A, coumestrol, daidzein, equol, formononetin, glycitein, genistein and prunetin) in soy products and wastewater samples was developed using gas chromatography coupled with ion trap mass spectrometry (GC/MS-MS). The phytoestrogens were derivatized as their trimethylsilyl ethers with trimethylchlorosilane (TMCS) and N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA). The phytoestrogens were isolated from all samples with liquid-liquid extraction using ethyl acetate. Daidzein-d(4) and genistein-d(4) labeled standards were used as internal standards before extraction and derivatization. The fragmentation patterns of the phytoestrogens were investigated by isolating and fragmenting the precursor ions in the ion-trap and a typical fragmentation involved the loss of a methyl and a carbonyl group. Two characteristic fragment ions for each analyte were chosen for identification and confirmation. The developed methodology was applied to the identification and confirmation of phytoestrogens in soy milk, in wastewater effluent from a soy-milk processing plant, and in wastewater (influent and effluent) from a treatment plant. Detected concentrations of genistein ranged from 50,000 microg/L and 2000 microg/L in soy milk and in wastewater from a soy-plant, respectively, to 20 microg/L and <1 microg/L for influent and effluent from a wastewater treatment plant, respectively.

Effect of the cross-linking silane concentration in a novel silane system on bonding resin-composite cement.

OBJECTIVE: Four experimental blends of an organo-functional silane monomer with a non-functional cross-linking silane monomer (a novel silane system) were evaluated as adhesion promoters in an experiment in which a resin-composite cement was bonded to silica-coated titanium. MATERIAL AND METHODS: 3-Acryloyloxypropyltrimethoxysilane (as constant 1.0 vol%) was blended with 1,2-bis-(triethoxysilyl)ethane, where its concentration was 0.1, 0.2, 0.3, or 0.5 vol%. Titanium slides (n=20) were grit-blasted, silica-coated, and silanized with four experimental silane solutions, with a pre-activated silane Cimaratrade mark (VOCO, Germany) as control. After silanization, resin-composite cement stubs (Bifixtrade mark QM; VOCO, Germany) were photo-polymerized. The shear bond strength was measured after dry storage (24 h) or after thermo-cycling (6000 cycles between 5 degrees C and 55 degrees C). The resin stub failure mode was determined. RESULTS: Statistical analysis (ANOVA) showed that type of storage (p <0.05) and concentration of cross-linker silane (p<0.005) both significantly affected the shear bond strength. The highest shear bond strength was obtained with a blend of 1.0 vol% 3-acryloyloxypropyltrimethoxysilane+0.3 vol% 1,2-bis-(triethoxysilyl)ethane, 15.9 MPa (standard deviation SD 3.4 MPa) for both the thermo-cycled group and after dry storage (24 h), 14.3 MPa (SD 4.1 MPa) (n=8/group). The lowest values were obtained with Cimaratrade mark silane 7.3 MPa (SD 2.2 MPa) in dry storage and 7.9 MPa (SD 2.0 MPa) obtained with 1.0 vol% 3-acryloyloxypropyltrimethoxysilane+0.1 vol% 1,2-bis-(triethoxysilyl)ethane. The failure type was mainly cohesive. CONCLUSION: A novel silane system with an optimal concentration of the cross-linking silane may produce significantly higher shear bond strength between silica-coated titanium and resin-composite cement compared to a pre-activated silane product.

Sensitive and rapid method to quantify icaritin and desmethylicaritin in human serum using gas chromatography-mass spectrometry.

The prenyl-flavones, icaritin and desmethylicaritin, are bioactive compounds from the traditional Chinese medicinal herb, Epimedium, extracts of which can enhance bone health in animal models. In order to examine their bioavailability in humans, we have developed and validated a sensitive method to quantify icaritin and desmethylicaritin in human sera, using gas chromatography-mass spectrometry. The serum samples were extracted with ethyl acetate and then derivatized with BSTFA in pyridine (4:1). With genistein as internal standard, calibration curves with good linearity (R(2)>0.99) within the concentration range of 0.15-10nM in the selective ion monitoring mode were obtained. The limits of detection and quantization were 11 and 33 pM for icaritin, and 23 and 70 pM for desmethylicaritin, respectively; inter- and intra-assay variabilities were <15%, and accuracies were between 89 and 110%. Icaritin, but not desmethylicaritin, was detected from 1h, increasing to a peak at 8h (1.51+/-1.6 nM) in sera of human volunteers after ingestion of an aqueous decoction of Epimedium. This sensitive method can be used to quantify serum levels of icaritin and desmethylicaritin for pharmacokinetic studies.

Microwave-accelerated derivatization for the simultaneous gas chromatographic-mass spectrometric analysis of natural and synthetic estrogenic steroids.

A rapid microwave-accelerated derivatization process for the GC-MS analysis of steroid estrogens, estrone (E1), 17beta-estradiol (E2), estriol (E3), 17alpha-ethynylestradiol (EE2) and mestranol (MeEE2), was developed. Under microwave irradiation, the five estrogenic hormones studied were simultaneously derivatized with N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA)+trimethylchlorosilane (TMCS) in pyridine solution. Effects of irradiation time (15-120 s) and power level (240-800 W) on the yield of the derivatization were investigated. The derivatization under the irradiation of 800 W microwave for 60s produced comparable results when compared with the conventional heating process in a sand bath for 30 min at 80 degrees C in terms of derivatization yield, linearity and precision for all steroid hormones tested. The calibration curves are linear between 3.00 and 3.00 x 10(2) microg mL(-1). The square of the regression coefficients (R(2)) range from 0.979 to 1.000. The applicability of the method was evaluated on spiked river and distilled water samples at two concentrations, 25.0 and 2.00 x 10(2) ng mL(-1). The recoveries obtained by using microwave heating (60s, 800 W) were similar to those by conventional heating. When combined solid-phase extraction (SPE) with the application of the microwave-accelerated derivatization proposed here, the detection limits of 0.02-0.1 ng L(-1) for the steroid hormones have been achieved. The results demonstrated that microwave-accelerated derivatization is an efficient and suitable sample preparation method for the GC-MS analysis of estrogenic steroids.

High amounts of 17-methylated anabolic-androgenic steroids in effervescent tablets on the dietary supplement market.

In numerous studies it has been demonstrated that several nutritional supplements contain prohormones not declared on the label. In the current study two products (effervescent tablets) containing high amounts of the 17-methylated anabolic androgenic steroids metandienone (product 1: 16.8 mg/tablet) and stanozolol (product 2: 14.5 mg/tablet) were identified. Additionally in both products norandrostenedione was detected, in product 2 with minor amounts of several other steroids. The substances identified can cause enormous health risks. In addition, the use of the analyzed tablets can lead to positive doping results for metabolites of the respective steroids in sports. This study again shows the insufficient surveillance of the production and trade of dietary supplements. Consumers should be aware of the enormous health and doping risks connected with the use of such products. For GC-MS identification of the analytes the trimethylsilyl derivatives of the steroids and the mixed N-t-butyldimethylsilyl,O-trimethylsilyl derivatives were used. The quantitation of metandienone, norandrostenedione, and stanozolol was performed using HPLC-DAD.

Characterisation of wax works of art by gas chromatographic procedures.

To identify the various natural and synthetic substances used by sculptors at the end of the 19th century, several contemporary reference samples were investigated by high temperature gas chromatography (HT GC) and HT GC-MS. Using specific chromatographic conditions and minimising sample preparation, we could separate, detect and identify a wide range of biomolecular markers covering a great variety of molecular weights and volatilities, with a minimum amount of sample, in a single run. Beeswax, spermaceti, carnauba, candellila and Japan waxes as well as pine resin derivatives, animal fats, paraffin, ozokerite and stearin, used as additives in wax works of art, were chemically investigated. In the case of low volatile compounds, transbutylation was performed. The structure of long-chain esters of spermaceti was elucidated for the first time by HT GC-MS analysis. Such a method was then carried out on 10 samples collected on a statuette of Junon by Antoine-Louis Barye (Louvre Museum, Paris, France) and on a sculpture by Aimé-Jules Dalou (Musée de la Révolution Française, Vizille, France). The analytical results obtained provide new data on the complex recipes elaborated by sculptors at the end of the 19th century.