Development and validation of a detection method of alendronate in human plasma by high-performance liquid chromatography/tandem mass spectrometry (HPLC–MS/MS)

The aim of this study is to develop a method for detecting alendronate in human plasma. Due to its special chemical properties, alendronate is almost unretained on reverse-phase chromatography columns, and cannot be detected by UV or fluorescence detectors. Consequently, quantitative analysis and related research of alendronate in biological samples are extremely challenging. We developed a simple, sensitive and stable method for detecting alendronate in human plasma. In this method, plasma samples was treated with 10% trifluoroacetic acid to remove proteins and derivatized with trimethylsilyl diazomethane, then these samples were injected into the system for analysis using a PFP chromatographic column. The total run time was 10 min, and retention times of alendronate and Pamidronate disodium were 4.79 min and 4.00 min, respectively. The intra-batch and inter-batch accuracy values are between 90.88% and 104.50%, with CV% between 0.45% and 7.15%. And the <i>R</i>² value of the calibration curve was greater than 0.99 in the range of 2.015–793.333 ng/mL. The recovery of quality control samples at different concentrations were greater than 100%. This method had been validated to detect the concentration of alendronate in human plasma and can provide technical reference for clinical researchers to conduct pharmacokinetic studies of alendronate.