Alka Beotra

Simultaneous identification of prednisolone and its ten metabolites in human urine by high performance liquid chromatography-tandem mass spectrometry

The use of prednisolone and prednisone is prohibited by the World Anti-Doping Agency (WADA) due to their performance-enhancing effect. The purpose of the present work was to explore the possibility of identification and detection of various metabolites of prednisolone by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in excretion study samples. Ten metabolites of prednisolone could be identified namely prednisone (11-oxo metabolite) [M-1], 6-β-OH-prednisolone [M-2], 20-β-OH-prednisolone [M-3], 20-α-OH-prednisolone [M-4], 20-α-OH-prednisone [M-5], 20-β-OH-prednisone [M-6], 2 tetrahydro epimers of 20-β-OH-prednisolone [M-7], 2 tetrahydro epimers of 20-α-OH-prednisolone [M-8], 2 tetrahydro epimers of 20-β-OH-prednisone [M-9], and 2 tetrahydro epimers of 20-α-OH-prednisone [M-10]. Prednisolone was administered in 10-, 20-, and 40-mg dosage to healthy volunteers to study detection of various metabolites. The parent, M-1, M-2, and M-3 could be detected up to 72 h while rest of the metabolites were detectable up to 24 h after drug administration. The detection of newer metabolites of the drug can further be used for confirmatory purposes.

Identification of Prednisolone, Methylprednisolone and Their Metabolites in Human urine using HPLC (+) ESI-MS/MS and Detection of Possible Adulteration in Indian Herbal Drug Preparations

Normal 0 false false false EN-IN X-NONE HI MicrosoftInternetExplorer4 Objective: To explore the possibility of identifying the maximum number of metabolites of prednisolone and methylprednisolone by LC-MS/MS and to further test the application of this developed method on six Indian herbal drug preparations. Method and Materials: The sample extraction procedure involves enzymatic hydrolysis and liquid-liquid extraction and further analysis using LC-MS/MS. The excretion profile was performed with four healthy male volunteers after administration of 40 mg and 8 mg of prednisolone and methylprednisolone, respectively. Six herbal drug preparations obtained from All India Institute of Medical Sciences, India were tested to detect the possible adulteration with prednisolone or methylprednisolone. Results: The Analytical method was validated as per the requirement of WADA International Standard of Laboratories (version 6.0). The parent compound and ten urinary metabolites of prednisolone could be identified. The parent, M-1, M-2 and M-3 could be detected up to 72 hours while rest of the metabolites were detectable up to 24 hours. In the case of methylprednisolone, the parent compound and six urinary metabolites were identified. M-1 and M-2 of methylprednisolone were detectable up to 48 hours while the parent drug methylprednisolone and other metabolites were detectable up to 24 hours. Out of the six herbal drugs tested, one showed the presence of prednisolone. Conclusion: The improved detection method developed for the identification and detection of prednisolone and methylprednisolone metabolites would prove highly beneficial in extending the time of detection of drug abuse in athletes and also detecting spiked ayurvedic, homeopathic and unani preparations. Keywords: Doping, Excretion study, LC-MS/MS, Anti-doping Rule Violation, Prednisolone, Methylprednisolone

Anti-doping programme in India: testing for major games

The control of drug abuse in athletes has become a highly specialised, complex task, which requires use of sophisticated testing procedures. National Dope Testing Laboratory (NDTL) is engaged in the fight against doping since 1990. The anti-doping programme in India is governed by its stringent testing protocol, which is regularly monitored for quality assurance by both National Accreditation Board of Testing and Calibration Laboratories and World Anti-Doping Agency. It is further strengthened by continuous research and training programmes. The year 2010 is very special for NDTL with a target of testing for two major international games. These are Singapore Youth Olympics (14–26 August 2010) and XIX Commonwealth Games (3–14 October 2010). The testing protocol for these major games will include both urine and blood samples. NDTL is ready with its new test methods viz. human growth hormone, erythropoietin, continuous erythropoietin receptor activator, haemoglobin-based oxygen carrier and blood transfusion. It is expected to test a total of 1100 (urine + blood) samples for Singapore Youth Olympic games and 1500 (urine + blood) samples for XIX Commonwealth Games. The aim of NDTL for these two major games is to provide fool-proof testing programmes that will achieve both deterrence and detection with athletes concerns in mind and a results management system that is both fair and time bound. The presentation will focus on the various strategies of the action plan so as to set a milestone of excellence.

Distribution of caffeine levels in urine in different sports in India

Caffeine (1,3,7-trimethylxanthine) is a xanthine alkaloid compound that acts as a stimulant and was banned for use in sports. Caffeine was removed from the World Anti-Doping Agency (WADA) prohibited list from January 2004. As per the requirement of WADA, the WADA-accredited labs are monitoring the presence of caffeine for signs of possible abuse. National Dope Testing Laboratory (NDTL), India, is monitoring caffeine levels in in-competition urine samples of sports persons from different sports disciplines. The aim of the present study is to find the distribution of caffeine levels in urine in different sport disciplines in Indian and other south-east Asian countries. Caffeine was measured using an alkaline extraction with tertiary butyl methyl ether followed by gas chromatographic nitrogen-phosphorus specific detection. A comparison of statistical data from the urine samples done from January 2008 to July 2010 in NDTL, India, for different discipline of sports since January 2008 to July 2010 was performed. The study proves beneficial in monitoring the abuse of caffeine in different disciplines of sports.