Method development and validation for detection of procaterol in human urine using gas chromatography-tandem mass spectrometry

Abstract

Background: A simple, sensitive and accurate method for the determination of procaterol in human urine was developed and validated using Gas Chromatography coupled to tandem mass spectrometry (GC-MS/MS) as per the requirements of World Anti-Doping Agency (WADA) and ICH guidelines. Methodology: The sample processing includes deconjugation with enzymatic hydrolysis, Solid Phase Extraction (SPE) procedure using XAD2 column and Liquid-liquid Extraction (LLE) followed by the derivatisation using N-methyl trimethylsilysl trifluoroacetamide (MSTFA). Results: The method included a chromatographic run of 17 min and the calibration curve was linear over the concentration range of 5-40 ng/mL with a correlation coefficient (r) of 0.98. The intra-and inter-day assay precision ranged from 0.75 to 0.94% and 1.2 to 1.70%, respectively. The absolute recovery percentage for procaterol was found to be 83%. The recovery was sufficient to reliable identify the procaterol at or below the level prescribed by WADA ie MRPL 20ng/ ml. The LOQ and LOD was found to be 5 and 3 ng/ml respectively. Conclusion: A simple and sensitive GC-MS/MS method for quantitative estimation of Procaterol was developed and validated as per the WADA ISL and ICH guidelines. This developed method could be also used for therapeutic drug monitoring purposes.