Measurement Uncertainty and Validation for Quantitation of Salbutamol in Human Urine by Gas Chromatography–Tandem Mass Spectrometry

Abstract

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%.