Simultaneous quantification of 26 bile acids in Calculus Bovis Sativus and Calculus Bovis by HPLC-MS/MS / 中草药

ABSTRACT

Objective To develop an HPLC-MS/MS method for the simultaneous determination of 26 bile acids in Calculus Bovis Sativus (CBS) and Calculus Bovis (CB). Methods Chromatographic separation was performed on a Symmetry C18 column (150 mm × 2.1 mm, 3.5 μm) with linear gradient elution of water (0.1% formic acid + 10 mmol/L ammonium acetate) and methanol (0.1% formic acid + 10 mmol/L ammonium acetate) at a flow rate of 0.15 mL/min, and the injection volume was 10 μL. Switching electrospray ion source polarity in negative mode. The ion spray voltage was set at - 4 500 V, the turbo spray temperature was maintained at 350 ℃, and multiple-reaction monitoring (MRM) was employed. Results The 26 bile aicds showed good linearity within the linear ranges (r ≥ 0.991 4). The average recoveries of the bile acids ranged from 98.2% to 102.3%. The precisions (RSD) for the investigated components were less than 0.95%. There are some differences in variety and chemical components between CBS and CB. Conclusion The proposed method is reliable and has high sensitivity and good specificity, which allows the determination of various bile acids in CB.