GC EIMS Research Articles

Differential heat stability of amphenicols characterized by structural degradation, mass spectrometry and antimicrobial activity

Heat stability of amphenicols and the relationship between structural degradation and antimicrobial activity after heating has not been well investigated. Florfenicol (FF), thiamphenicol (TAP), and chloramphenicol (CAP) were heated at 100 °C in water, salt water, soybean sauce and chicken meat for up to 2 h. Degradation and antimicrobial activity of the compounds was evaluated using capillary electrophoresis (CE) with UV-DAD spectrometry, minimum inhibitory concentration (MIC) assay, and gas chromatography with electron impact ionization mass spectrometry (GC–EI-MS). Heat stability of amphenicols in matrices was ranked as water ≥ salt water > soybean sauce > meat, suggesting that heat degradation of amphenicols was accelerated in soybean sauce and was not protected in meat. Heat stability by drug and matrices was ranked as FF > TAP = CAP in water, FF = TAP > CAP in salt water, TAP ≥ FF = CAP in soybean sauce, and TAP ≥ FF = CAP in meat, indicating differential heat stability of amphenicols among the 3 drugs and in different matrices. In accordance with the less than 20% degradation, the MIC against Escherichia coli and Staphylococcus aureus did not change after 2 h heating in water. A 5-min heating of amphenicols in water by microwave oven generated comparable percentage degradation to boiling in water bath for 30 min to 1 h. Both CE and GC–MS analysis showed that heating of FF produced TAP but not FF amine as one of its breakdown products. In conclusion, despite close similarity in structure; amphenicols exhibited differential behavior toward heating degradation in solutions and protein matrices. Although higher degradations of amphenicols were observed in soybean sauce and meat, heating treatment may generate product with antimicrobial activity (FF to TAP), therefore, heating of amphenicol residues in food cannot always be assumed safe.

ChemInform Abstract: A Novel Trisaccharide Glycolipid Biosurfactant Containing Trehalose Bears Ester-Linked Hexanoate, Succinate, and Acyloxyacyl Moieties: NMR and MS Characterization of the Underivatized Structure.

A Gram-positive actinomycete growing on n-hexadecane secreted a family of anionic glycolipid surfactant homologs. The major homolog, with a molecular weight of 1210.6347, had the formula C58H98O26. Following mild alkaline saponification, 1H and 13C NMR spectroscopy were used to characterize the non-reducing trisaccharide backbone: β-Glcp-(1→3)-α-Glcp-(1↔1)-α-Glcp (‘laminaratrehalose’). Hexanoate, succinate, 3-hydroxyoctanoate, and 3-hydroxydecanoate were found in 3:1:1:1 molar ratio using GC–EIMS analysis of fatty acid methyl esters (FAME) prepared by transesterification. We found that the β-hydroxy acids bore secondary hexanoate chains in 3-O-ester linkage, giving acyloxyacyl anions of appropriate m/z in FABMS and FABMS/MS spectra. COSY, HETCOR, HMBC, and HMQC NMR experiments established the acylation pattern: succinate at C-2 of the terminal α-glucopyranose ring; hexanoate at C-3′′ of the β-glucopyranose ring; 3-hexanoyloxyoctanoate and 3-hexanoyloxydecanoate at the 2′- and 4-positions. In FABMS spectra, the homologs flanked the molecular ion by ±14 and ±28 amu, suggesting heterogeneity in acyl chain length.

Spatial and temporal distribution of pesticide residues in surface waters in northeastern Greece

A monitoring study of 147 compounds in surface river waters from northeastern Greece near Greek/Bulgarian/Turkish borders was carried out during 1999–2007. Based on agricultural use eight sampling points along the rivers Ardas, Evros and Erythropotamos were set up, covering the distance from the Greek/Bulgarian borders down to the river's discharge (river's delta) in the Greek territory. In total, 88 sampling events were carried out from 1999 to 2007. Pesticides were extracted by solid-phase extraction (SPE) and analyzed by gas chromatography–mass spectrometry (GC–EI-MS) using a multiresidue in-house analytical method including pesticides belonging to different chemical classes. Aquatic risk concerning the detected pesticides was assessed on the basis of the risk quotient (RQ = PEC/PNEC). From the 28 compounds (pesticides, metabolites and caffeine) that were detected in surface waters of northeastern Greece the soil applied pesticides were the most frequently detected. High pesticide concentrations were detected within 2 months of their application. Extreme pesticide concentrations were detected in the beginning of the irrigation season or just after high rainfall events. Generally, low levels of pesticide residues were found in the first sampling point (Greek/Bulgarian borders) of all rivers, however o′, p′ DDT, o′, p′ DDE and γ-HCH were mainly detected in this sampling point regarded as cross-boundary contamination. The most commonly encountered compounds in the river waters were atrazine, DEA, alachlor, trifluralin, prometryne, molinate, carbofuran, carbaryl and diazinon. Increased loading (primary as well as secondary peaks) seemed to be a consequence of application (timing, rate, frequency) and intense rainfall during the application period. Aquatic risk assessment revealed that from the 28 compounds that were constantly detected 12 showed non-acceptable risk when median concentrations were used as PEC and 18 when extreme concentrations were used as PEC values.

Impact of pesticides used in agriculture and vineyards to surface and groundwater quality (North Spain)

An environmental monitoring program was carried out to determine the impact of eight pesticides on the surface and groundwater quality of agricultural areas within the Ebro, Duero and Miño river basins. Three triazines and their desethyl degradation products, metolachlor and metalaxyl, were monitored during 18 months in 63 sites. Solid-phase extraction (SPE) using OASIS HLB 60 mg cartridges and gas chromatography–mass spectrometry (GC–EI-MS) provided good analytical quality parameters and limits of quantification of 0.01 μg/L. Environmental data were assessed using descriptive statistical analysis and multivariate data analysis with principal component analysis (PCA) to elucidate the relevant contamination patterns and provide a description of their seasonal trends, according to the pesticide application timing. Duero was the site with the highest frequency of detection and highest concentration levels, followed by the Ebro and Miño basins. The frequency of detection of the studied compounds, considering all surface and groundwater samples, was atrazine>desethylatrazine>simazine>desethylsimazine>metolachlor>desethylterbuthylazine>terbuthylazine>metalaxyl. Over all results, and taking into consideration the European Union (EU) maximum residual limit of pesticides in groundwater, only 12% of the results exceeded the 0.1 μg/L limit. However, sporadic high levels up to 2.46 μg/L in groundwater and 0.63 μg/L in surface water were detected. PCA permitted to state that Duero and Ebro river basins were especially affected by a contamination pattern dominated by atrazine, the Ebro river basin being occasionally affected by a contamination pattern dominated by simazine. Only trace levels were rarely detected in the Miño river basin. Groundwater levels were higher than surface water levels for the studied pesticides.

Solid-phase analytical derivatization of alkylphenols in fish bile for gas chromatography–mass spectrometry analysis

Solid-phase analytical derivatization (SPAD) with bis(trimethylsilyl)trifluoroacetamide (BSTFA) has successfully been used as a sample preparation method for determination of (APs) in fish bile treated with β-glucuronidase and sulfatase. Derivatized APs were analysed by gas chromatography–mass spectrometry in the electron ionization mode (GC–EI-MS). Overall limits of detection (LODs) ranged from 5 to 18 ng/g bile for 19 out of 21 investigated compounds. LODs were not determined for 4-methylphenol and 4- tert-octylphenol due to high background levels in control bile. Recoveries ranged from 83 to 109%. The analysed APs vary in degree of alkylation from methyl (C 1) to nonyl (C 9), and represent various pollution sources, including produced water (PW) discharge from the offshore oil industry. The applicability and sensitivity of the method has been demonstrated by analysis of bile taken from Atlantic cod ( Gadus morhua L.) exposed to two dilutions of PW (1:500 and 1:1500) in a continuous flow system.

Hyphenated gas chromatography–mass spectrometry analysis of 3-mercaptohexan-1-ol and 3-mercaptohexyl acetate in wine: Comparison with results of other sampling procedures via a robust regression

This work describes a new purge and trap gas chromatography electron impact mass spectrometry (PT–GC–EIMS) method for quantifying 3-mercaptohexan-1-ol (3-MH) and 3-mercaptohexyl acetate (3-MHA), two molecules able to characterize some wines with their tropical scents. Firstly the experimental conditions of the purge and trap extraction (sample temperature, extraction time, trap temperature, flow rate) following a multivariate approach were optimized. Then the method through the construction of the calibration curves and the establishment of the detection limits was validated. The purge and trap procedure appears faster and more sensitive than both the headspace solid phase microextraction (HS-SPME) and the solid phase extraction (SPE) procedures, reaching detection limits for the two thiols closer to their sensory thresholds. Evidence of similar performances of the three sampling methods considered was gained comparing the results relevant to same wine samples. The Theil's regression method was used for purpose of comparison.

Development and application of procedures for the highly sensitive quantification of cyclosarin enantiomers in hemolysed swine blood samples

The present study was initiated to develop a sensitive method for the analysis of cyclosarin ( O-cyclohexyl methylphosphonofluoridate, GF) enantiomers in biological samples utilizing classical configurations of GC–MS and automated solid phase extraction. To achieve this goal, a specific procedure had to be developed to extract cyclosarin from swine blood samples thereby stabilising and minimising the racemisation/deracemisation of its enantiomers. The chiral stationary phase was GAMMA DEX™ (gamma cyclodextrin), on which GF and deuterated GF enantiomers were baseline-resolved. The limit of detection was 1 pg for (−)-GF with GC–EI-MS and 5 pg for (+)-GF with GC–NCI-MS. The absolute recovery of the overall procedure for sample preparation was 85%. After an intravenous infusion of a supralethal dose of GF in anaesthetised swine only (−)-GF could be quantified, (+)-GF was not detected.

Insecticidal activity of 12- epi-hapalindole J isonitrile

12- epi-Hapalindole J isonitrile ( 1) and three previously described hapalindoles, 12- epi-hapalindole C isonitrile ( 2), hapalindole L ( 3) and 12- epi-hapalindole E isonitrile ( 4) were isolated and identified as insecticidal alkaloids of the biofilm-forming freshwater cyanobacterium Fischerella ATCC 43239 (Stigonematales). The structures of the purified compounds were elucidated by ESI–FTICR–MS, GC–EI–MS and various 2D NMR experiments. At 26 μM hapalindole 1 killed 100% of the larvae of the dipteran Chironomus riparius within 48 h. Insecticidal activities were also found at similar concentration for the hapalindoles 2–4. The bioactivity of hapalindoles demonstrates that cyanobacterial biofilms can be considered as promising sources of insecticidal metabolites which might be useful for the biocontrol of dipterans.

Qualitative analysis of phenols and alcohols in complex samples after derivatization to esters of ferrocene carboxylic acid by gas chromatography with mass spectrometric detection

Phenols and alcohols in complex petrochemical samples are derivatized to esters of ferrocene carboxylic acid in a rapid and simple reaction. These esters show several characteristic ions of high intensity in gas chromatography coupled with electron impact ionization mass spectrometry (GC–EI–MS) which can be used for the identification of the analyte. A differentiation between alcohols and phenols is possible due to the McLafferty rearrangement shown only by the alcohol esters. Selective ion monitoring of m/ z 213 yields a phenol-selective chromatogram and m/ z 230 an alcohol-selective chromatogram. For all iron containing fragments, the isotopic pattern of iron can be observed which enhances the reliability of the peak identification. The ferrocene esters of 11 alcohols, 20 alkylphenols (including octyl- and nonylphenol), phenylphenol, naphthol and hydroxyphenanthrene, several chloro- and all mononitrophenols were synthesized as well as the esters of pentadeutero- and two fluorinated phenols. Their fragmentation pattern under EI ionization is studied and a GC-ion trap-MS system was optimized for simultaneous use of the full scan mode and an MS/MS experiment in the same run. This provides for a very high selectivity in the detection of the esters and makes available the complete mass spectra without any additional measurements. The benefits of the simple and rapid derivatization procedure in combination with this powerful detection method are demonstrated for selected petrochemical samples. Several alkylphenols could be successfully identified in such samples and molecular information about unknown phenolic components could be easily obtained.

Identification of Phenolic Dermal Sensitizers in a Wound Closure Tape

A latex-allergic patient presented with a severe local reaction to a non-latex wound closure bandage following surgery. Extracts of the bandage were analyzed by gas chromatograph-electron impact-mass spectrometry (GC EI-MS) in the total ion monitoring mode. Components were identified by their ion mass fingerprint and elution time as a corresponding standard from the GC column. The chemicals identified were 4,4′-thiobis-(6-tert-butyl-m-cresol) (TBBC), 6-tert-Butyl-m-cresol (BC), 2,4-di-tert-butylphenol (BP) and erucamide (EA). Sensitization potential of these chemicals was evaluated using two quantitative structure-activity relationship (QSAR) programs. The phenol 2,6-di-tert-butyl-4-(hydroxymethyl)phenol (BHP) was also included in the test series. It was initially thought to be present in the bandage but detectable levels could not be confirmed. The potential for TBBC to induce a sensitization response was predicted by both Derek for Windows and TOPKAT 6.2. The potential for BC and BP to induce a sensitization response was predicted by Derek for Windows, but not TOPKAT. BHP and EA were not predicted to be sensitizers by either QSAR program. Local lymph node assay (LLNA) analysis of the chemicals identified TBBC, BP, and BC as potential sensitizers with EC3 values between 0.2 and 4.5%. None of the animals exhibited body weight loss or skin irritation at the concentrations tested. In agreement with the toxicological modeling, BHP did not induce a sensitization response in the LLNA. Following a positive LLNA response, TBBC, BP, and BC were further characterized by phenotypic analysis of the draining lymph nodes. A positive LLNA result coupled with a lack of increase in B220+IgE+ cell and serum IgE characterize these chemicals as Type IV sensitizers. These studies used a multidisciplinary approach combining clinical observation, GC-EI-MS for chemical identification, QSAR modeling of chemicals prior to animal testing, and the LLNA for determination of the sensitization potential of chemicals in a manufactured product.

Detection of cholesteryl ester hydroperoxide isomers using gas chromatography–mass spectrometry combined with thin-layer chromatography blotting

Oxidative modification of low-density lipoprotein (LDL) has been implicated in the pathogenesis of atherosclerosis. During the oxidation of LDL, cholesteryl esters, the major lipid components in LDL, are oxidized to cholesteryl ester hydroperoxides (CEOOH). The isomers of CEOOH may reflect the reactive species that initiate the peroxidation reaction. In the current study, a novel analytical method for the determination of CEOOH isomers, especially cholesteryl linoleate hydroperoxide isomers, was developed using the combination of two chromatographic techniques: (i) thin-layer chromatography blotting with diphenyl-1-pyrenylphosphine (DPPP) fluorescent detection (DPPP–TLC blotting) and (ii) gas chromatography–electron ionization–mass spectrometry (GC–EI–MS). CEOOH was applied to DPPP–TLC blotting, the obtained DPPP-derived fluorescent spots containing cholesteryl ester hydroxides were extracted and derivatized (hydrogenation, transmethylation, and trimethylsilylation), and the formed methyl ester/trimethylsilylether derivatives of hydroxyoctadecenoic acid were then analyzed by GC–EI–MS. The CEOOH isomers were determined by selected ion monitoring of isomer-specific fragment ions originated from the alpha-cleavage of the trimethylsilyloxyl group. Using these two chromatographic techniques, we were able to detect isomeric CEOOH in the oxidized human LDL. Our results indicated that GC–EI–MS analysis combined with DPPP–TLC blot is a specific method for analyzing cholesteryl ester hydroperoxide isomers in biological samples such as oxidized LDL.

Chemotaxonomy of Portuguese Ulex: Quinolizidine alkaloids as taxonomical markers

Six species of Portuguese Ulex L. in a total of nineteen populations were studied by GC–EIMS as to their content in quinolizidine alkaloids. Sparteine, β-isosparteine, jussiaeiine A, N-methylcytisine, cytisine, 5,6-dehydrolupanine, rhombifoline, lupanine, jussiaeiine B, N-formylcytisine, N-acetylcytisine, anagyrine, jussiaeiine C, jussiaeiine D, pohakuline, baptifoline, and epibaptifoline were detected. Analysis of the chromatograms showed that the chemical profile of all species was mainly composed of N-methylcytisine, cytisine, anagyrine, and jussiaeiines A, B, C and D. Therefore a quantification study of these alkaloids in all the populations studied was done by GC. These data were then submitted to cluster analysis and principal component analysis, which allowed the definition of five chemotypes and the recognition of hybrids. N-methylcytisine, cytisine, and jussiaeiines A, C and D are recognized as markers of this genus in Portugal.

Determination of ethephon residues in water by gas chromatography with cubic mass spectrometry after ion-exchange purification and derivatisation with N-( tert-butyldimethylsilyl)- N-methyltrifluoroacetamide

An analytical method has been developed for the determination of residues of ethephon (2-chloroethyl phosphonic acid) in drinking and surface water. The procedure is based on de-ionisation with an anion/cation-exchange resin, solid phase extraction by means of anion-exchange polystyrene-divinylbenzene extraction disks, elution with a mixture of methanol and 10 M hydrochloric acid (98/2, v/v), redisolution into acetonitrile after evaporation and silylation with N-( tert-butyldimethylsilyl)- N-methyltrifluoroacetamide (MTBSTFA). Quantification is performed by gas chromatography with ion-trap cubic mass spectrometric detection in the electron impact mode (GC–EI–MS 3). Method validation was conducted using samples of mineral, tap, and river water that were fortified with ethephon at concentration levels ranging from 0.1 to 1.0 μg/L. The mean recovery from all the fortified samples ( n = 36) amounted to 88% with a relative standard deviation of 17%. The method, therefore, was shown to allow accurate determination of ethephon residues in drinking and surface water with a limit of quantification of 0.1 μg/L.

Validation of various extraction techniques for the quantitative analysis of polycyclic aromatic hydrocarbons in sewage sludges using gas chromatography-ion trap mass spectrometry

A new GC-ion trap MS method has been developed for the determination of polycyclic aromatic hydrocarbons (PAHs) in sewage sludge samples. The sludge samples were extracted with Soxhlet, Soxtec, and pressurized liquid extraction (PLE) using 1:1 (v/v) dichloromethane (DCM): n-hexane solvent mixture. A multi-layer clean-up (silica/Al 2O 3) column were used, followed by gel permeation chromatography (GPC) to eliminate the interfering organic compound as well as the lipids. The extracts were quantified with GC–EI-SIS and GC–EI-MS–MS. The method was successfully applied to determine the concentration of PAHs present in sewage sludge samples collected from four waste water treatment plants (WWTPs). The method recovery values varied from 61.5 to 90.5%, 65.0 to 91.8% and 60.0 to 93.4% for Soxtec extraction, Soxhlet extraction and PLE extraction, respectively. The total concentrations of the 16 PAHs in the sewage sludge samples were found to vary from 1.56 to 6.18 mg/kg. The concentration of PAHs in WWTPs did not significantly vary.

Determination of polycyclic musks in sewage sludge from Guangdong, China using GC–EI-MS

Polycyclic musks [Cashmeran (DPMI), Celestolide (ADBI), Phantolide (AHMI), Traseolide (ATII), Tonalide (AHTN) and Galaxolide (HHCB)] were Soxhlet extracted, separated and analyzed in sewage sludge from six different wastewater treatment plants from Guangdong, China, using GC and GC–EI-MS. DPMI, ADBI, HHCB, AHTN were found in all samples, and ATII was not found in any sample. AHMI was detected in five out of six samples. HHCB and AHTN were the two major polycyclic musks found in highest concentrations in sludge. The sludge from municipal wastewater treatment plants contained HHCB, AHTN and DPMI at concentrations between 5.416 and 21.214, 0.715 and 6.195 and 0.599 and 2.870 mg/kg (dry), respectively. The highest concentration was found in sludge from one cosmetic plant at 703.681 mg HHCB/kg (dry). The results indicate that there are two sources of polycyclic musks: domestic sewage and industrial wastewater.

Determination of 14 benzodiazepines and hydroxy metabolites, zaleplon and zolpidem as tert-butyldimethylsilyl derivatives compared with other common silylating reagents in whole blood by gas chromatography–mass spectrometry

The most common commercially available silylating reagents, N-methyl- N-(trimethylsilyl)trifluoroacetamide (MSTFA), N, O-bis-(trimethylsilyl)trifluoroacetamide + 1% trimethylchlorosilane (BSTFA + 1% TMCS) and N-methyl- N-( tert-butyldimethylsilyl)trifluoroacetamide (MTBSTFA) were evaluated to achieve optimal derivatization conditions for analyzing various benzodiazepines based on gas chromatography–electron impact ionization–mass spectrometry (GC–EI–MS). Sensitivity, repeatability, retention times and stability of the derivatives, as well as specificity of mass fragmentation, were studied in detail. Also other parameters affecting the derivatization chemistry of benzodiazepines are discussed. tert-Butyldimethylsilyl (TBDMS) derivatives proved to be more stable, reproducible and sensitive than corresponding trimethylsilyl (TMS) derivatives for the GC–EI–MS analysis of benzodiazepines. Based on the TBDMS derivatives, a rapid, reliable, sensitive and quantitative GC–MS method was developed for the determination of 14 benzodiazepines and two hydroxy metabolites, as well as two non-benzodiazepine hypnotic agents, zolpidem and zaleplon, using 500 μl of whole blood. The method was completely validated in terms of accuracy, intra- and interday precision, limit of detection (LOD), limit of quantitation (LOQ), linearity, selectivity and extraction efficiency; these were all within the required limits, except for the accuracy of nitrazepam at a medium concentration level.

The GC–MS detection and characterization of reticuline as a marker of opium use

Reticuline (a precursor of opium alkaloids) was detected and characterised as its trimethylsilyl ethers, acetyl esters and methyl ethers by GC–EIMS and GC–CIMS in opium and the urine of opium users after hydrolysis by acid or β-glucuronidase as coextractive of morphine. Because this compound cannot be detected in heroin and poppy seeds, it is suggested as a differentiating marker between opium and heroin use, opium and poppy seeds use, or opium and “pharmaceutical” codeine use in cases when opiate use has been confirmed by detection of morphine and codeine in the urine. As well as being a constituent of opium, reticuline in the urine of opium users may also result from the metabolic demethylation of the three other benzyltetrahydroisoquinoline opium alkaloids: codamine, laudanosine and laudanine.

The polysaccharides of red wine: total fractionation and characterization

Ethanol-precipitated red wine polysaccharides were fractionated by a combination of anion-exchange, size-exclusion and affinity chromatography steps. This comprehensive fractionation allowed us to prepare a collection of wine polysaccharides in sufficient amount to permit the determination of their intrinsic properties. Glycosyl-residue composition of each polysaccharide fraction was determined by GC–EI–MS of the per- O-trimethylsilylated methyl glycoside derivatives (TMS), a method that has been recently developed and adapted to suit simultaneous determination of neutral and acidic glycosyl-residue compositions of polysaccharides present in plant-derived products. The results showed that mannoproteins released by yeast during fermentation, and grape derived arabinogalactan-proteins, rhamnogalacturonans I and II are the main wine polysaccharides and accounted for 35, 42, 4 and 19%, respectively, of the total polysaccharides. Structural characterization revealed that rhamnogalacturonan I fractions were linked with xyloglucan-like polysaccharides. This finding represents compelling evidence of the existence of cross-linking between pectin and hemicellulose domains in plant primary cell walls.

Distribution of seed fatty acids and the taxonomy of Vochysiaceae

The distribution of seed fatty acids of 16 species of Vochysiaceae native to Brazil, corresponding to the genera Callisthene, Qualea, Salvertia and Vochysia, and amounting to 57 samples, were studied. The methyl esters of the fatty acids were analyzed by GC–EIMS. For most species, fatty acid patterns are seemingly useful for species characterization. Intraspecific variability of samples of Salvertia grandiflora seems to have geographic significance. With one exception, palmitic and oleic acids predominate in the fatty acids distribution of Qualea and Callisthene. Q. grandiflora possesses exceptional amounts of lauric acid. Relatively high amounts of either C 20–C 22 (saturated or monounsaturated) or stearic acids characterize Salvertia and Vochysia. UPGMA analysis of fatty acids distribution based on Euclidean distances resulted in a cluster combining Callisthene and all samples of Qualea, except Q. grandiflora. Samples of Vochysia are combined into several clusters. The dendrogram is suggestive of taxonomic relationships, allowing the distinction of taxa down to the sub-sectional level. V. pyramidalis, from section Ciliantha, subsection Ferrugineae, has closer chemical affinities with species of section Vochysiella, subsection Decorticantes. Comparing with monomers of polysaccharide seed cell walls of Vochysiaceae, fatty acid patterns revealed lower intraspecific variability and higher taxonomic resolution.

Determination of nandrolone and metabolites in urine samples from sedentary persons and sportsmen

Metabolites of nandrolone were determined in the urine of several sportsmen, sedentary and post-menopausal women by capillary gas chromatography–mass spectrometry quadrupole (GC–MS) and capillary gas chromatography mass–mass spectrometry ion trap (GC–MS–MS) methods. The method employed was GC–EI-MS with 17α-methyltestosterone as internal standard with ethyl ether extraction prior to selected ion monitoring of the bis(trimethylsilyl) ethers at ion masses m/ z 405 and 420 for the nandrolone metabolites, and 418 and 403 for nandrolone derivative. Recovery for nandrolone, 19-norandrosterone (19-NA) and 19-noretiocholanolone (19-NE) was 97.20, 94.17 and 95.54%, respectively. Detection limits for nandrolone, 19-NA and 19-NE were 0.03, 0.01 and 0.06 ng/ml. Metabolites of nandrolone (19-NA and 19-NE) were found in 12.5% ( n=40) of sportsmen and 40% ( n=10) of post-menopausal women.