Abstract:

In this study, we have developed a novel method for the simultaneous determination of calycosin 7-O-glucoside, hesperidin, and ammonium glycyrrhizinate in Buzhongyiqi Pills based on UPLC-Q/Orbitrap HRMS using Hypersil Gold C₁₈ chromatographic column. The gradient mobile phase comprised acetonile as well as water containing 0.1% formic acid. The flow rate, column temperature, and injection volume were 0.4 mL/min, 40 ℃, and 1 μL, respectively. Mass spectrometer was operated using an electrospray ionization source in the positive ion mode for detection. The scan range was 100~1 500 m/z. Quantification was performed by extracting the accurate mass of the target compounds.The linear ranges of calycosin 7-O-glucoside,hesperidin, ammonium glycyrrhizinate were 0.089 1~1.425 0 μg/mL,0.098 4~12.600 0 μg/mL, and 7.425 0~29.700 0 μg/mL (r≥0.999 3), respectively. Furthermore, the limits of detection were 5.57 ng/mL, 4.10 ng/mL, and 5.80 ng/mL, respectively, while the limits of quantitation were 22.26 ng/mL, 12.30 ng/mL, and 23.20 ng/mL, respectively. The results demonstrated good precision, repeatability, and sample stability. Spike recoveries were 91%~105%(δ_RSD≤5.0%,n=6). This method is simple, accurate, and highly sensitive, which is suitable for the simultaneous determination of calycosin 7-O-glucoside, hesperidin, and ammonium glycyrrhizinate in Buzhongyiqi Pills.

Key words: UPLC-Q/Orbitrap HRMS, Buzhongyiqi Pills, calycosin 7-O-glucoside, hesperidin, ammonium glycyrrhizinate