HRGC/FID/NPD and HRGGC/MSD study of Colombian ylang-ylang (Cananga odorata) oils obtained by different extraction techniques

Steam distillation (SD), simultaneous distillation-solvent extraction (SDE), and supercritical (CO₂) extraction (SFE) were used to isolate volatile secondary metabolites from fresh, totally mature flowers of Colombian ylang-ylang (Cananga odorata). The various extracts were analyzed by capillary chromatography (DB-1, DBWAX, 60 m columns) using FID, NPD or MSD (EI, 70 eV). Kováts indexes, mass spectra, or standard substances were employed for compound identification. 51, 70, and 73 compounds at concentrations above 100 ppb were detected in the SD, SDE, and SFE extracts, respectively. The main constituents of these extracts were linalool (20.7, 28.0, and 16.5%), germacrene-D (10.1, 3.1, and 20.3%) benzyl benzoate (14.1, 2.9, and 3.9%), benzyl acetate (9.6, 17.0, and 6.2%), caryophyllene (3.1, 2.9, and 3.9%), and p-methylanisole (6.8, 6.1, and 2.7%). 85% of the composition of SDE extracts was represented by oxygenated compounds. Heavy hydrocarbons (C_n >20) and fatty acids were found only in the SFE extracts, which also had a higher content of nitrogenated compounds (phenylacetonitrile, 4-methylbenzaldoxime, indole, 2-phenyl-nitroethane, and methyl anthranilate) and sesquiterpenes (43% vs 19.5% in SD and 8.1% in SDE) and 1.5 – 2 times lower concentration of monoterpenes and light oxygenated compounds than the SD (49.7%) and SDE (64.5%) extracts.     

HRGC and GC–MS analysis of essential oil from colombian ylang-ylang (Cananga odorata Hook fil. et Thomson,forma genuina)

Samples of essential oil from Colombian ylang-ylang trees were analyzed by means of HRGC, HRGC–MS, IR and ¹H- and ¹³C-NMR. 57 components were detected, 51 of which were positively identified. Camphene and anethol were identified in ylang-ylang essential oil for the first time. Among the composition-determining variables studied (extraction time, part of the flower, and flower freshness), the extraction time and the flower condition (fresh versus dry) were found to have the largest incidence in the quality of the essential oil.     

Chemical Profiles,In Vitro Antioxidant and Antifungal Activity of Four Different Lavandula angustifolia L. EOs

Lavandula angustifolia L., known as lavender, is an economically important Lamiaceae due to the production of essential oils (EOs) for the food, cosmetic, pharmaceutical and medical industries. The purpose of this study was to determine the chemical composition of EOs isolated from four inflorescences of L. angustifolia L. collected in different geographical areas: central-southern Italy (LaCC, LaPE, LaPS) and southern France (LaPRV). The essential oils, obtained by steam distillation from plants at the full flowering stage, were analyzed using gas chromatography coupled with mass spectrometry (GC-MS). More than 70 components identified in each sample showed significant variability among the main constituents. The four EOs analyzed contained the following as main component: linalool (from 30.02% to 39.73%), borneol (13.65% in LaPE and 16.83% in La PS), linalyl acetate (24.34% in LaCC and 31.07% in LaPRV). The EOs were also evaluated for their in vitro antifungal activity against two white rot fungi (Phanerochaete chrysosporium and Trametes cingulata) as potential natural biodeteriogens in the artworks field, and against Sclerotium rolfsii, Botrytis cinerea and Fusarium verticilloides responsible for significant crop yield losses in tropical and subtropical areas. The results confirm a concentration-dependent toxicity pattern, where the fungal species show different sensitivity to the four EOs. The in vitro antioxidant activity by DPPH assay showed better scavenging activity on LaCC (IC₅₀ 26.26 mg/mL) and LaPRV (IC₅₀ 33.53 mg/mL), followed by LaPE (IC₅₀ 48.00 mg/mL) and LaPS (IC₅₀ 49.63 mg/mL). The potential application of EOs as a green method to control biodeterioration phenomena on a work of art on wood timber dated 1876 was evaluated.     

Sediment Matrix Induced Response Enhancement in the Gas Chromatographic–Mass Spectrometric Quantification of Insecticides in Four Different Solvent Extracts from Ultrasonic and Soxhlet Extraction

The performance of ultrasonic and Soxhlet extraction using hexane, dichloromethane, ethylacetate/methyl-tert-butylether (1/3, v/v) and hexane/acetone (1/1, v/v) for the analysis of seventeen insecticides in sediments was evaluated. The contents of the extracts differed severely. The extracts obtained with ethylacetate/methyl-tert-butylether (1/3, v/v) and hexane/acetone (1/1, v/v) were dark yellow to green, whereas the extracts obtained with dichloromethane and hexane were light yellow and clear respectively. This is due to higher solubility of matrix compounds in ethylacetate/methyl-tert-butylether (1/3, v/v) and hexane/acetone (1/1, v/v). High loads of coextracted matrix compounds lead to matrix effects in the evaporation step of GC–MS measurements. This is known as matrix induced response enhancement effect. Matrix effects and recoveries were checked by analysis of spiked sediments. The suitable choice of extraction method in connection with an appropriate solvent separates the analytes from matrix compounds. Matrix effects are reduced and recoveries of spiked samples are improved.Revised: 6 January and 2 May 2005     

Chemical Constituents of the Essential Oil from Ecuadorian Endemic Species Croton ferrugineus and Its Antimicrobial,Antioxidant and α-Glucosidase Inhibitory Activity

Croton ferrugineus Kunth is an endemic species of Ecuador used in traditional medicine both for wound healing and as an antiseptic. In this study, fresh Croton ferrugineus leaves were collected and subjected to hydrodistillation for extraction of the essential oil. The chemical composition of the essential oil was determined by gas chromatography equipped with a flame ionization detector and gas chromatography coupled to a mass spectrometer using a non-polar and a polar chromatographic column. The antibacterial activity was assayed against three Gram-positive bacteria, one Gram-negative bacterium and one dermatophyte fungus. The radical scavenging properties of the essential oil was evaluated by means of DPPH and ABTS assays. The chemical analysis allowed us to identify thirty-five compounds representing more than 99.95% of the total composition. Aliphatic sesquiterpene hydrocarbon trans-caryophyllene was the main constituent with 20.47 ± 1.25%. Other main compounds were myrcene (11.47 ± 1.56%), β-phellandrene (10.55 ± 0.02%), germacrene D (7.60 ± 0.60%), and α-humulene (5.49 ± 0.38%). The essential oil from Croton ferrugineus presented moderate activity against Candida albicans (ATCC 10231) with an MIC of 1000 μg/mL, a scavenging capacity SC₅₀ of 901 ± 20 µg/mL with the ABTS method, and very strong antiglucosidase activity with an IC₅₀ of 146 ± 20 µg/mL.     

Phytochemical Characterization of Rhus coriaria L. Extracts by Headspace Solid-Phase Micro Extraction Gas Chromatography,Comprehensive Two-Dimensional Liquid Chromatography,and Antioxidant Activity Evaluation

Rhus coriaria L. (Anacardiaceae), commonly known as sumac, has been used since ancient times for many different applications, and nowadays is used mostly as a spice obtained from its in the Mediterranean and the Middle ground fruits and employed for flavoring and garnishing food, predominantly Eastern regions. Traditionally, sumac has been also used in popular medicine for the treatment of many ailments including hemorrhoids, wound healing, diarrhea, ulcers, and eye inflammation. Sumac drupes are indeed rich in various classes of phytochemicals including organic acids, flavonoids, tannins, and others, which are responsible of their powerful antioxidant capacity, from which treatment of many common diseases such as cardiovascular disease, diabetes, and cancer could benefit. In this work we evaluated the influence of fruit ripeness, conservation, and processing. To this aim, a phytochemical characterization of six different samples of Rhus coriaria L. was carried out. Specifically, headspace solid-phase micro extraction gas chromatography coupled to mass spectrometry and comprehensive two-dimensional liquid chromatography coupled to photodiode array and mass spectrometry detection, were employed. A total of 263 volatile compounds, including terpene hydrocarbons, acids, and aldehydes, as well as 83 polyphenolic compounds, mainly gallic acid derivatives, were positively identified. All samples showed a significant antioxidant activity by means of oxygen radical absorbance capacity, in line with their polyphenolic content and composition. Such findings set a solid ground to support the utilization of this plant as an attractive target for novel nutraceutical approaches and for drug discovery.     

Optimization of the Extraction and Analysis of Natural Androgen Steroids and Their Metabolites in Urine by GC/MS and GC/FID

A multiresidue method was developed for screening, quantification, and confirmation of nine natural androgen steroids and their metabolites in urine. Steroids were first extracted from urine by solid phase extraction, enzymatically deglucuronated, re-extracted using a liquid/liquid extraction for purification, and finally acetylated for GC/MS and GC/FID analysis. Each step of sample preparation, as well as analysis, was optimized: solid phase extraction, liquid/ liquid extraction, and derivatization reaction … Therefore, a rugged sample preparation procedure was developed leading to extracts of sufficient purity (recoveries >66% and few matrix compounds). The whole methodology allowed reliable detection and quantification of the nine steroids at low concentration levels. Linearity and repeatability were established and were found to be satisfactory (R² > 0.996, RSD < 11%). Finally, the method was applied to quantify compounds of interest in real samples collected from healthy volunteers and patients treated with 4-androstenedione or dehydroepiandrosterone.     

Chemical Profile,In Vitro Biological Activity and Comparison of Essential Oils from Fresh and Dried Flowers of Lavandula angustifolia L.

The chemical composition of essential oils (EOs) from dried and fresh flowers of Lavandula angustifolia L. (lavender), named LA 2019 and LA 2020, respectively, grown in central Italy was analyzed and compared by GC and GC-MS. For both samples, 61 compounds were identified, corresponding to 97.9% and 98.1% of the total essential oils. Explorative data analysis, performed to compare the statistical composition of the samples, resulted in a high level of global similarity (around 93%). The compositions of both samples were characterized by 10 major compounds, with a predominance of Linalool (35.3–36.0%), Borneol (15.6–19.4%) and 1,8-Cineole (11.0–9.0%). The in vitro antibacterial activity assay by disk diffusion tests against Bacillus subtilis PY79 and Escherichia coli DH5α showed inhibition of growth in both indicator strains. In addition, plate counts revealed a bactericidal effect on E. coli, which was particularly noticeable when using oil from the fresh lavender flowers at the highest concentrations. An in vitro antifungal assay showed that the EOs inhibited the growth of Sclerotium rolfsii, a phytopathogenic fungus that causes post-harvest diseases in many fruits and vegetables. The antioxidant activity was also assessed using the ABTS free radical scavenging assay, which showed a different antioxidant activity in both EOs. In addition, the potential application of EOs as a green method to control biodeterioration phenomena on an artistic wood painting (XIX century) was evaluated.     

Development and Comprehensive SPE-UHPLC-MS/MS Analysis Optimization,Comparison, and Evaluation of 2,4-Epibrassinolide in Different Plant Tissues

A determination method for trace 24-epibrassinolide (EBL) in plant tissues was developed using ultra-high-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS). The plant tissue samples were extracted using a methanol–formic acid solution, and the corresponding supernatant was purified with ODS C18 solid-phase extraction column. The extracts were separated using a Zorbax Eclipse Plus C18 (2.1 mm × 50 mm, 1.8 μm) column with methanol and 0.1% formic acid as the mobile phase. The ion source for the mass spectrometry was an electrospray ionization source with positive ion mode detection. The linear range of the target compound was 0.7~104 μg/kg, the limit of detection (LOD) was 0.11~0.37 μg/kg, the limit of quantification (LOQ) was 0.36~1.22 μg/kg, the recovery rate was 84.0~116.3%, and the relative standard deviation (RSD%) was 0.8~10.5. The samples of maize plumule, brassica rapeseed flower, and marigold leaf were detected using the external standard method. The optimization of the extraction method and detection method of EBL improved the detection sensitivity, laid a foundation for the artificial synthesis of EBL, improved the extraction rate of EBL, and provided a theoretical basis for the study of EBL in many plants.     

Evaluation of the effects of crude glycerol on the production and properties of novel polyhydroxyalkanoate copolymers containing high 11‐hydroxyoctadecanoate by Cupriavidus necator IPT 029 and Bacillus megaterium IPT 429

Crude glycerol (CG), a by‐product from biodiesel production, is a carbon source with potential as feedstock for polyhydroxyalkanoate (PHA) production. PHAs are biological macromolecules synthesized by microorganisms as intracellular carbon and energy storage granules. PHA production and its properties were investigated using Cupriavidus necator IPT 029 and Bacillus megaterium IPT 429 cultivated with CGs from different origins. The highest PHA extraction percentage (71.56% [w/v]) occurred when C. necator IPT 029 metabolized CG 3 (from the processing of biodiesel from castor bean oil). The gas chromatography–mass spectrometry analyses revealed novel PHA constituents as building blocks of medium (3‐hydroxytetradecanoate) and long (11‐hydroxyoctadecanoate) chains. Molar mass distribution revealed range of 121–6900 kDa. The initial degradation temperature ranged from 181.83 to 287.50°C and the crystallinity ranged from 35.30 to 66.70%. The results obtained indicate that C. necator IPT 029 from CG 3 could produce copolymers with industrially applicable thermophysical properties. Copyright © 2015 John Wiley & Sons, Ltd.     

Determination of d‐limonene in mice plasma and tissues by a new GC–MS/MS method: Comparison of the pharmacokinetics and tissue distribution by oral and inhalation administration in mice

The aim of the present study was to develop a method based on gas chromatography–tandem mass spectrometry (GC–MS/MS) to determine and quantify the d ‐limonene in mouse plasma and tissue samples. This new method was validated for the quantification of d ‐limonene with the linearity ranges 1.0–1000.0 ng/mL (r² > 0.9952) for plasma samples and 5.0–5000.0 ng/g (r² > 0.9940) for tissue samples. The intra‐ and inter‐day assay of precisions in plasma and tissues were <13.4% and the accuracies were within 91.1–105.8%. In the oral/inhalation administration pharmacokinetics and tissue distribution studies, the main pharmacokinetic parameters were the peak concentration = (97.150 ± 34.450)/(4336.415 ± 1142.418) ng/mL, the area under the curve = (162.828± 27.447)/(2085.721 ± 547.787) h ng/mL and the half‐life = (3.196 ± 0.825)/(0.989 ± 0.095) h. The tissue distribution of d ‐limonene in mice after oral/inhalation administration demonstrated a decreasing tendency in different tissues (liver > kidney > heart > lung > spleen).     

Synchronous Extraction,Antioxidant Activity Evaluation,and Composition Analysis of Carbohydrates and Polyphenols Present in Artichoke Bud

This study investigated the optimization of ultrasonic-assisted aqueous two-phase synchronous extraction of carbohydrates and polyphenols present in artichoke bud, evaluated their antioxidant activities in vitro, and analyzed the composition of carbohydrates and polyphenols by high-performance liquid chromatography (HPLC). The powder mass, ultrasonic time, ammonium sulfate concentration, and alcohol–water ratio were considered the influencing factors based on the single-factor experiment results, and a dual-response surface model was designed to optimize the synchronous extraction process to extract carbohydrates and polyphenols. The antioxidant activity was evaluated by measuring the scavenging capacity of ABTS⁺· and DPPH· and the reducing capacity of Fe³⁺. The optimal process conditions in this study were as follows: the powder mass of 1.4 g, ammonium sulfate concentration of 0.34 g/mL, alcohol–water ratio of 0.4, and ultrasonic time of 43 min. The polyphenol content in artichoke bud was 5.32 ± 0.13 mg/g, and the polysaccharide content was 74.78 ± 0.11 mg/g. An experiment on in vitro antioxidant activity showed that both carbohydrates and polyphenols had strong antioxidant activities, and the antioxidant activity of polyphenols was stronger than that of carbohydrates. The HPLC analysis revealed that the carbohydrates in artichoke bud were mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, and arabinose, and the molar ratio was 10.77:25.22:2.37:15.74:125.39:48.62:34.70. The polyphenols comprised chlorogenic acid, 4-dicaffeoylquinic acid, caffeic acid, 1,3-dicaffeoylqunic acid, isochlorogenic acid B, isochlorogenic acid A, cynarin, and isochlorogenic acid C, and the contents were 0.503, 0.029, 0.022, 0.017, 0.008, 0.162, 1.621, 0.030 mg/g, respectively. This study also showed that the carbohydrates and polyphenols in artichoke bud could be important natural antioxidants, and the composition analysis of HPLC provided directions for their future research. Carbohydrates and polyphenols in artichoke buds can be separated and enriched using the optimized process technology, and it is an effective means of extracting ingredients from plants.     

ESI-MS/MS Analysis of Phenolic Compounds from Aeonium arboreum Leaf Extracts and Evaluation of their Antioxidant and Antimicrobial Activities

Aeonium is a genus of succulents belonging to the Crassulaceae family. Their importance in traditional medicine has stimulated both pharmacological and chemical research. In this study, we optimized extraction, separation, and analytical conditions using a high performance liquid chromatographic method coupled with electrospray ionization mass spectrometry by the negative mode (HPLC-ESI-MS) in order to, for the first time, determine thirty-four compounds from Aeonium arboreum leaves. Twenty-one of them are assigned among which are sixteen flavonoids and five phenolic acids. FRAP, TAC, DPPH, and ABTS^•+ radical scavenging were used to evaluate antioxidant activity. The obtained IC₅₀ values ranged from 0.031 to 0.043 mg.mL⁻¹ for DPPH and between 0.048 and 0.09 mg·mL⁻¹ for ABTS^•+. Antimicrobial activity was also assessed. The obtained minimum inhibitory concentrations (MIC) of these extracts ranged from 12.5 to 50 µg·mL⁻¹ against Micrococcus luteus, Listeria ivanovii, Staphylococcus aureus, Salmonella enterica, Escherichia coli, Pseudomonas aeruginosa, Aspergillus niger, and Fusarium oxysporum, and from 25 to 50 µg·mL⁻¹ against Candida albicans. Therefore, these extracts can be considered as a potential source of biological active compounds.     

HESI-MS/MS Analysis of Phenolic Compounds from Calendula aegyptiaca Fruits Extracts and Evaluation of Their Antioxidant Activities

Considering medicinal plants as an inexhaustible source of active ingredients that may be easily isolated using simple and inexpensive techniques, phytotherapy is becoming increasingly popular. Various experimental approaches and analytical methods have been used to demonstrate that the genus Calendula (Asteraceae) has a particular richness in active ingredients, especially phenolic compounds, which justifies the growing interest in scientific studies on this genus’ species. From a chemical and biological viewpoint, Calendula aegyptiaca is a little-studied plant. For the first time, high-performance liquid chromatography combined with negative electrospray ionization mass spectrometry (HPLC-HESI-MS) was used to analyze methanolic extracts of Calendula aegyptiaca (C. aegyptiaca) fruits. Thirty-five molecules were identified. Flavonoids (47.87%), phenolic acids (5.18%), and saponins (6.47%) formed the majority of these chemicals. Rutin, caffeic acid hexoside, and Soyasaponin βg’ were the most abundant molecules in the fruit methanolic extract, accounting for 17.49% of total flavonoids, 2.32 % of total phenolic acids, and 0.95% of total saponins, respectively. The antioxidant activity of the fruit extracts of C. aegyptiaca was investigated using FRAP, TAC, and DPPH as well as flavonoids and total phenols content. Because the phenolic components were more extractable using polar solvents, the antioxidant activity of the methanolic extract was found to be higher than that of the dichloromethane and hexane extracts. The IC50 value for DPPH of methanolic extract was found to be 0.041 mg·mL⁻¹. Our findings showed that C. aegyptiaca is an important source of physiologically active compounds.     

Liquid-Solid Extraction of Vitamin D3 Metabolites from Plasma for Analysis by HPLC,GC/MS and Protein Binding Techniques

Abstract

Liquid-solid extraction of vitamin D₃ metabolites from human plasma using octadecylsilane bonded silica has been studied. Steroid-protein interactions were minimized by diluting the plasma (or serum) with two volumes of saline and passing the solution through the sorbent at 64°C. Highly purified secosteroid fractions were obtained by washing with aqueous methanol, drying the sorbent in situ with a stream of nitrogen for one minute and eluting with mixtures of hexane/chloroform. Recoveries of vitamin D₃ metabolites were essentially quantitative. Applications to the rapid analysis of 25-hydroxy- and 1α, 25-dihydroxy-vitamin D in plasma by high-performance liquid chromatography, gas chromatography-mass spectrometry or by a receptor protein assay are reported.     

GC-MS联用技术结合化学计量学方法分析厚朴叶挥发油成分

利用水蒸气蒸馏法提取湖南产厚朴叶的挥发油成分,采用GC-MS联用技术结合化学计量学方法(直观推导式演进特征投影法和选择性离子法)进行分析,同时结合程序升温保留指数辅助定性。结果共鉴定出54种化合物,占挥发油总量的84.95%。主要化学成分为α-、β-和γ-桉油醇,含量分别为13.10%、28.21%和14.67%。此外,含量较高的化合物还有α-蒎烯(2.96%)、芳樟醇(1.71%)、丹皮酚(1.88%)、石竹烯(2.04%)、佛术烯(3.60%)、α-瑟林烯(3.84%)和[1AR-(1Aα,4α,4Aβ,7Bα)]-1A,2,3,4,4A,5,6,7B-八氢化-1,1,4,7-四甲基-1H-环丙烯并[E]奥(1.34%)。研究表明,将化学计量学方法用于中药挥发油成分的分析可提高定性分析的准确性。所得结果可为厚朴药植物资源的开发利用提供依据。     

Quality criteria of the isotope dilution method for the determination of polycyclic aromatic hydrocarbons, petroleum-derived hydrocarbons and phenol in leachate of large-scale lysimeter experiments

A comprehensive analytical procedure for polycyclic aromatic hydrocarbons, petroleum-derived hydrocarbons and phenol using the isotope dilution method and employing high-resolution gas chromatography and mass spectrometry was developed and validated for leachates from source-term experiments and the different sampling sites of lysimeters. The use of glassware and other materials is efficiently limited to minimize the risk of contamination. The relative standard deviation elaborated allows precise reliable measurements. Limit of detection and recovery data are useful to judge the quality of each single measurement.     

Measurement of the malondialdehyde-2'-deoxyguanosine adduct in human urine by immuno-extraction and liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry

The major adduct of malondialdehyde with guanine, M(1)G, was measured in human urine from non-smoking healthy individuals. M1G is a mutagenic DNA lesion and a terminal product of lipid peroxidation in vivo that may be implicated in cancer related to lifestyle and diet. On the basis of a recently developed method for the quantification of M1G as an excreted deoxynucleoside using immuno-extraction purification, chemical NaBH4 reduction and liquid chromatography combined with atmospheric pressure chemical ionization tandem mass spectrometry, we demonstrate that the average 24 h excretion rate of M1G-dR is about 12 +/- 3.8 fmol kg(-1) (n = 5).     

Liquid chromatography mass spectrometry for the determination of salvianolic acid B,a natural compound from the herb Danshen in rat plasma and application to pharmacokinetic study

A sensitive and specific liquid chromatographic–electrospray ionization mass spectrometric method was developed for quantification of salvianolic acid B in rat plasma with resveratrol as the internal standard. The analytes were separated on a reversed‐phase column with acetonitrile (40%) and water (60%) containing 0.75% formic acid as mobile phase at a flow rate of 1 mL/min. Liquid–liquid extraction was adopted for the sample preparation, and the analytes were determined using electrospray negative ionization mass spectrometry in the selective monitoring mode. The method was validated over the concentration range 0.1–40 µg/mL using 0.1 mL of plasma with coefficients of correlation >0.999. The intra‐ and inter‐day precisions of analysis were <10%, and accuracy ranged from 94 to 101%. This method was successfully applied to a pharmacokinetics of salvianolic acid B in rats. Copyright © 2009 John Wiley & Sons, Ltd.     

Rethinking the Process of Animal Mummification in Ancient Egypt: Molecular Characterization of Embalming Material and the Use of Brassicaceae Seed Oil in the Mummification of Gazelle Mummies from Kom Mereh,Egypt

The study of animal mummification in ancient Egypt has recently received increasing attention from a number of modern scholars given the fact that this part of ancient Egyptian funerary and religious history is a practice yet to be fully understood. In this study, nine samples of embalming matter were extracted from six gazelle mummies from the archaeological site of Kom Mereh (modern village of Komir), dated to the Roman period of dominance in ancient Egypt. All samples were analyzed for the presence of inorganic and organic matter applying a multi-analytical approach based on Fourier transform infrared spectroscopy (FT-IR) and gas chromatography–mass spectrometry (GC-MS). Furthermore, in order to identify more specific compounds such as bitumen and beeswax in studied balms, each sample was subjected to a solid phase extraction (SPE) and saponification separation process, respectively. The results of this study revealed that the majority of the analyzed embalming substances sampled from six gazelle mummies from Kom Mereh were complex mixtures of plant oils, animal fats, conifer resin, and beeswax. In this regard, this study was able to report a practice until now unmentioned in the scientific literature, namely, the use of cruciferous oil, derived from seeds of Brassicaceae plants, in animal mummification.