Determination of Four Chemical Components in Qishen Yiqi Dripping Pills by UPLC-MS/MS Method

Abstract

Background In this study, the ultrahigh performance liquid chromatography mass spectrometry/mass spectrometry (UPLC-MS/MS) method was established to test the contents of protocatechuic acid (C7H6O4), salvianolic acid B (C36H30O16), rosmarinic acid (C19H14O3), and cryptotanshinone (C19H20O3) in Qishen Yiqi dripping pills. Materials and Methods Methanol was chosen as the solvent according to a certain material–liquid ratio as a solution, and the ultrasonic extraction method was used to prepare the sample solution. UPLC-MS/MS was used to complete a quantitative analysis of the contents of Qishen Yiqi dripping pills. Then, the chromatographic conditions were shown as follows. The stationary phase was Agilent ZORBAX SB-C18 (2.1 mm × 100 mm, 1.8 µm), and the mobile phase was acetonitrile–0.1% formic acid water. The column temperature was 40℃, the injection volume was 3 µL, and the flow rate was 0.2 mL⋅min–1. Finally, the determination method was built to test four chemical constituents under the multiple reaction monitoring (MRM) analysis mode. Results The retention times of protocatechuic acid, salvianolic acid B, rosmarinic acid, and cryptotanshinone were 1.26, 1.32, 1.35, and 6.76 min, respectively. In addition, the concentration ranges of four compounds were 0.25–2.0 µg⋅mL–1, 10.0–50.0 µg⋅mL–1, 5.0–50.0 µg⋅mL–1, and 1.0–10.0 µg⋅mL–1, respectively. And the standard curve showed a good linear relationship in a reasonable range ( r2 ≥ 0.9950). The average recovery rate was from 96% to 124%. The contents of protocatechuic acid, salvianolic acid B, rosmarinic acid, and cryptotanshinone were 0.12%–0.022%, 17%–8.4%, and 6.5%–8.6% in seven batches of Qishen Yiqi dripping pills, which were in the range of 23%–31%. The instrument ran stably, and the method had good accuracy and repeatability (relative standard deviation [RSD] ≤ 3%). Additionally, the test result was stable within 10 h. Conclusion The contents of the four constituents were different among different batches. The quantitative method was established to determine the four chemical components in Qishen Yiqi dripping pills, which could provide a reference for quality evaluation in vivo.