High-Performance Liquid Chromatography-Tandem Mass Spectrometry for the Determination of Trospium Chloride in Human Plasma and Its Application in a Bioequivalence Study

Abstract

A selective, rapid, and sensitive high performance liquid chromatography-tandem mass spectrometry (HPLC-ESI-MS/MS) method was developed and validated for the quantification of trospium chloride in human plasma. With tramadol chloride as the internal standard, sample pretreatment involved a simple liquid-liquid extraction with chloroform-isopropyl alcohol (40/2, v/v) of 0.5 ml plasma. The analysis was carried out on a Hypsil C₁₈ column (150 mm × 2.1 mm, 5.0 μm) with a flow rate of 0.2 ml/min. The mobile phase was methanol-5% acetic acid-20 mM ammonium methylate (55/30/15, v/v/v). The detection was performed by a selected reaction monitoring (SRM) mode via electrospray ionization (ESI). Linear calibration curves were obtained in the concentration range of 0.2–50.0 ng/ml, with a lower limit of quantification of 0.2 ng/ml. The intra- and inter-day precision (RSD) values were below 15%. The method was successfully applied to a clinical bioequivalence study of trospium chloride in Chinese healthy volunteers following oral administration.

Keywords:

• Bioequivalence
• Determination
• HPLC-ESI-MS/MS
• Human plasma
• Trospium chloride

Notes

Test formulation ^a : 5 mg capsule and 20 mg tablet, from Shu Tai Shen Pharmaceutic Co., Lmt., Beijing, P.R. China.

Reference formulation ^b : Spasmo-lyt®-20 mg tablet, from Doctor Horse Pharmaceutic Co., Germany.