Urine analysis anabolic steroids

The same modified reagent has also been used with effect in the analysis of terpenoids and other steroids, and especially for the anabolic steroids. These applications are mentioned only as a passing reference because, like cholesterol, the analytes are more readily determined using zero and/or first order derivative spectrophotometry. If there is one distinct advantage associated with CD detection it is that the 17-ketosteroids, which are major interferences to absorbance detection in the analysis of urine for anabolic steroids, do react with the loss of all CD activity. 

Extensive drug screening is done at many athletic events, such as the Olympic Games. Usually, separate analyses, using different extraction procedures, are done for stimulants, narcotics, anabolic steroids, diuretics, and peptide hormones. In the analysis for stimulants, which are amines such as amphetamine and cocaine, a 5 rnL urine sample is first made basic with K.OH to ensure that the amines are present as the neutral molecules rather than as salts. The free amines are then extracted from the sample with diethyl ether. To save time and expense, the sample is first analyzed by gas chromatography only. If a peak appears with the retention time of one of the proscribed stimulants, then the sample is reanalyzed by GC/MS to confirm the identity of the suspected compound. 

With respect to the analysis of anabolic steroids, methods described are the analysis of 36 anabohc steroids in kidney fat, faeces, and urine [67], the analysis of the anabohc steroids 17P-19-nortestosterone, 17P-testosterone and progesterone and... 

Preparative LC fractionation of the kidney fat, faeces, and urine extracts into six fractions prior to the LC-MS analysis of 36 anabolic steroids [67],... 

In veterinary medicine, boldenone, a synthetic anabolic steroid (Figure 2.1), is commercially available, hence it is a concern in the horseracing industry. Pu et al. (2004) used ion-trap LC-MS analysis to detect boldenone conjugates (sulfate and glucuronide) and their 17-epimers in horse mine after intramuscular administration of boldenone undecylenate. Soon afterwards. Ho et al. (2004) reported the occurrence of endogenous boldenone sulfate in the urine of uncastrated male horses, and quantitated it by quadrupole time-of-flight (Q-TOF) LC-MS-MS. 

Draisci et al. (2000) used LC-MS-MS to quantitate T (Figure 2.2), 19-nortestosterone and their 17-epimer metabolites in bovine serum and urine, and subsequently stanozolol and its major metabolite (Draisci et al., 2001) and boldenone (Draisci et al., 2003) in bovine urine. Van de Wiele et al. (2000) also worked on stanozolol in cattle urine and feces, with or without derivatization before LC-MS-MS analysis. For trenbolone, Buiarelli et al. (2003) developed and characterized an LC-MS-MS method for bovine urine and serum. Van Poucke and co-workers extended the list of LC-MS-MS target analytes to four anabolic steroids (Van Poucke and Van Peteghem, 2002), then 21 anabolic steroid residues in bovine urine (Van Poucke et al., 2005). 

Buiarelli et al. (2004) extended the above analytical approach to many more related steroids when they published a method for the direct analysis of 15 urinary anabolic steroids in a single run, namely T, epitestosterone, dehydroepiandrosterone (DHEA), androsterone, etiocholanolone, their sulfates and their glucuronides (Figure 2,2), They extracted 2 mL of human urine by solid-phase extraction with methanol elution and reconstituted the residue in aqueous methanol in the presence of deuterated internal standards (da-epitestosterone glucuronide, [16,16,17-"H3 testosterone sulfate and [16,16,17-2H3]testosterone), then monitored, for example, mJz. 289-97 and 109 for T and epitestosterone, miz 367-97 for their sulfates, and m/z 463-113 and 287 for their glucuronides. The method does not achieve quantitation, but it allows the estimation of ratios, which makes it possible to monitor the urinary steroid profile, which is useful for monitoring the abuse of anabolic steroids. 

One anabolic steroid, the presence of which has proven difficult to analyze, is stanozolol [10418-03-8] C2 H22N20. A metaboHte of the parent drug, hydroxy stanozolol, detected in equine urine eight hours after ingestion of the parent drug is actually identified, usually at very low levels. Analysis was done by Ic/ms/ms which had a shortened analysis time advantage over gc/ms procedures because of the elimination of the need for a derivatization step (33). 

C/min), followed by an increase to 230°C (4°C/min), and finally to 300°C (30°C/min), which was held for 5 min. The MS analysis was performed on electron impact (El) mode. The analysis of residual anabolic steroids in meat was reported by Fuh, Huang, and Lin. The isolation was conducted by SPE. To derivatize the isolated steroids, MSTFA" "" " was used. The enzymatic hydrolysis was not conducted, as previous studies have discovered that there was no significant hormone liberation involved in muscle, fatty tissue, and meat. The derivatization products were then subjected to GC-MS/MS analysis. For this purpose, a DB-5 GC column (30 m X 0.25 mm, with 0.25 pm film thickness) was used, with a flow rate of 1.0 ml/min. The oven temperature was initially set at 180°C (1 min), increased to 240°C (6°C/min, for 2 min), and finally increased to 290°C (6°C/min, held for 10 min). The MS analysis was performed on El mode. Impens et al. " used a non-polar 5% phenyl-polysilphenylene-siloxane BPX-5 GC column to analyze anabolic steroids in bovine urine. The sample was derivatized with MSTFA" before being injected for the GC-MS/MS and GC-MS/MS/MS analyses. The oven temperature was initially set to 100°C (1 min), then increased to 250°C (30°C/min), followed by an increase to 290°C (2.5°C/min), and finally to 300 C (10°C/min), which was held for 1.5 min. Interested readers can consult Refs. 3, 8, and 18-25 for details. 

The use of HPLC-ED in the analysis of the anabolic steroids and metabolites, diethylstilbestrol, taleranol, zearalenol, zearalenone and zeranol (Figure 7.5), in mammalian tissue has been discussed.Diethylstilbestrol has been measured in animal tissue using an ODS-modified silica column with methanol-aq. phosphate buffer (50 mmol L pH 3.5) (67 + 33) as eluent and ED (GCE, +0.9 V vs Ag/ AgCl). Sample homogenisation was followed by LLE into MTBE, back-extraction into aqueous sodium hydroxide and SPE (ODS-modified silica). The LoD was approximately 0.5 pg kg wet weight (10 g sample). Clenbuterol assay in calf urine was discussed above (Chapter 6, Section 2). 

Kim SH, Cha EJ, Lee KM, Kim HJ, Kwon OS, Lee J. Simultaneous ionization and analysis of 84 anabolic androgenic steroids in human urine using liquid ehromatography-silver ion coordination ionspray/triple-quadrupole mass spectrometry. Drug Test Anal. 2014 6 1174-85. 

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