Anabolic steroids, analysis

A. Leinonen, T. Kuuranne, R. Kostiainen, LC-MS in anabolic steroid analysis -optimization and comparison of three ionization techniques ESI, APCI and APPI-MS, J. Mass Spectrom., 37 (2002) 693. 

Leinonen, A. Kuuranne, T. Kostiainen, R. Liquid chromatography/mass spectrometry in anabolic steroid analysis—optimization and comparison of three ionization techniques Electrospray ionization, atmospheric pressure chemical ionization and atmospheric pressure photoionization. J. Mass Spectrom. 2002, 37 (7), 693-698. 

Foltin, R. W., The importance of drug self-administration studies in the analysis of abuse liability An analysis of caffeine, nicotine, anabolic steroids, and designer drugs. Annual Meeting of the American Academy of Psychiatrists in Alcoholism and Addictions (1990, Santa Monica, California). American Journal on Addictions Spr Vol 1(2), 139-149, 1992. 

Amundsen, L. K., Kokkonen, J. T., Rovio, S., and Siren, FI. (2004). Analysis of anabolic steroids by partial filling micellar electrokinetic capillary chromatography and electrospray mass spectrometry. J. Chromatogr. A 1040, 123—131. 

An issue for inhaled corticosteroid treatment is patient compliance. Analysis of prescription renewals shows that corticosteroids are taken regularly by a minority of patients. This may be a function of a general "steroid phobia" fostered by emphasis in the lay press over the hazards of long-term oral corticosteroid therapy and by ignorance over the difference between corticosteroids and anabolic steroids, taken to enhance muscle strength by now-infamous athletes. This fear of corticosteroid toxicity makes it hard to persuade patients whose symptoms have improved after starting the treatment that they should continue it for protection against attacks. 

This methodology traditionally used by biologists is now applied in the areas of environment and food analysis (with pesticides, aflatoxins, anabolic steroids, PAHs). However, it is only possible to measure a compound if the adapted antibodies and enzyme conjugates are available. 

M. J. Walters, R. J. Ayers, and D. J. Brown, Analysis of illegally distributed anabolic steroid productivity by LC with identity com-firmation by mass spectrometry or infrared spectrophotometry, JAOAC, 73 904(1990). 

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. 

In those cases where a suitable antibody for radioimmunoassay is not available, a full multi-component screening procedure is necessary, and also needs to be undertaken as a confirmatory test in cases where the nature of the anabolic steroid is not known. The full analysis involves the separation of the glucuronic acid and sulphate conjugate fractions and their separate hydrolysis followed by GC-MS of their silyl or methoxime-silyl derivatives. It should be noted that the sulphatase in Helix pomatia will not hydrolyse the sulphate conjugates of the 1713-hydroxy steroids, which require mild acid hydrolysis. 

A standard solution is prepared for the analysis of fluoxymesterone (C20H29FO3), an anabolic steroid. A stock solution is first prepared by dissolving 10.0 mg of fluoxymesterone in enough water to give a total volume of 500.0 mL. A 100.0-p.L aliquot (portion) of this solution is diluted to a final volume of 100.0 mL. Calculate the concentration of the final solution in terms of molarity. 

Huopalahti and Henion " reported anabolic steroid extraction from bovine tissues. SFE was performed with CO2 at 60°C and 405 bar, with analytes collected in precooled methanol and quantitated using LC-MS analysis. Residues of 2-thiouradl, 6-methyl-2-thiouradl, 6-propyl-2-thiouracil, and 6-phenyl-2-thiouracil in bovine muscle tissues were extracted with SC CO2 and analyzed using GC-MS. ... 

The group of Kostiainen [72-74] reported a number of comparative studies into the performance of APPI relative to APCI and ESI, e.g., in the LC-MS analysis of flavonoids [72], anabolic steroids [73], and naphthalenes [74], A variety of solvents were compared for the toluene-doped APPI of naphthalenes,... 

APPI is promoted as an alternative to APCI in the analysis of relatively nonpolar compounds. Leinonen et al. [25] compared ESI, APCI, and APPI in the analysis of three anabolic steroids, 3 -hydroxystanozolol, 6P-hydroxy-4-chlorodehydromethyl-testosterone, and oxandrolone. All three techniques proved suitable, but ESI was preferred. Optimum mobile phase was a methanol-water gradient containing 5 mmol/1 ammonium acetate and 0.01% acetic acid. For all three compounds, intact protonated molecules and some fragments due to consecutive water losses were observed. Significant better performance of APPI compared to ESI and APCI was also reported for corticosteroids by Greig et al. [26]. 

In this chapter, the role of LC-MS in various aspects of food safety analysis is discnssed. LC-MS is important in the quantification and confirmation of identity of food contaminants, e.g., residues of pesticides and related substances in frait and vegetables (Ch. 7.7), antibiotic and antibacterial compounds, other veterinary dmgs, anabolic steroids, and various toxins in feed and food. In addition, attention is paid to the analysis of heterocychc aromatic amines in food. 

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],... 

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