No 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.

A simple, rapid, sensitive and specific gas chromatography–tandem mass spectrometry (GC–MS/MS) method was developed for quantitation of salbutamol in human urine using salbutamol-d3 as the internal standard. The processing of urines samples includes deconjugation with enzymatic hydrolysis, solid phase extraction procedure utilizing XAD2 column and liquid–liquid extraction accompanied by the derivatization by means of MSTFA/IODO–TMS/DTE mixture. The GC column was a HP Ultra-1 (17 m × 0.22 mm × 0.11 μm) used to separate the peak of interest. The data for GC–MS/MS were acquired and processed utilizing GC Labs Solution and Insight GCMS Software. The detection of spectra was performed on TQ 8050. This method included a chromatographic run of 13.67 min and the linearity was found over the concentration range of 250–2000 ng/mL with a regression coefficient (r2) of 0.99. The coefficient of variation for intra and interday assay precision was between 1.85 and 2.85% and the accuracy was between 95.50 and 107.04% for low quality control (QC), medium QC and high QC. The recovery was adequate to reliable detect the analyte at or below the level recommended by the World Anti-Doping Agency i.e., threshold 1000 ng/mL. The limit of detection and limit of quantification were found to be 10 and 100 ng/mL, respectively. The expanded measurement uncertainty (Uexp%) was found to be 8.28%.