Development and validation of a liquid chromatography–tandem mass spectrometric method for the quantification of 5-thio-d-glucose in rat and human plasma

A highly selective, sensitive, and robust liquid chromatography–tandem mass spectrometric method for the determination of 5-thio-d-glucose concentrations in rat and human plasma was developed and validated. The sample preparation procedure involved protein precipitation and solid phase extraction, which efficiently removed sources of interference present in the plasma. Chromatographic separation was obtained using an NH2-column with distilled water and acetonitrile as the mobile phase under gradient conditions. Detection was performed using tandem mass spectrometry equipped with an electrospray ionization interface in negative ion mode. The selected reaction monitoring (SRM) transitions for 5-thio-d-glucose and an internal standard (5-thio-d-glucose-13C6) were m/z 195 → m/z 105 and m/z 201 → m/z 108, respectively. The correlation coefficients of the calibration curves ranged from 0.9997 to 0.9999 over a concentration range from 10 to 3000 ng/mL plasma. The validated method was successfully applied to a pharmacokinetic study in rats.

► An LC–MS/MS method for the analysis of 5-thio-d-glucose in plasma was developed.
► The validated quantitation range was from 10 to 3000 ng/mL.
► This is the first method that does not require radioisotope-labelled compounds.
► We succeeded in the removal of interferences by unique sample preparation procedure.
► The method was successfully applied to evaluate pharmacokinetic studies in rats.