超高效液相色谱-四级杆/静电场轨道阱高分辨质谱法同时检测补中益气丸中毛蕊异黄酮葡萄糖苷、橙皮苷、甘草酸铵的含量

Simultaneous determination of calycosin 7-O-glucoside,hesperidin,and ammonium glycyrrhizinate in Buzhongyiqi Pills via UPLC-Q/Orbitrap HRMS

建立了一种超高效液相色谱-四级杆/静电场轨道阱高分辨质谱同时检测补中益气丸中毛蕊异黄酮葡萄糖苷、橙皮苷和甘草酸铵含量的方法。采用超高效液相色谱-四级杆/静电场轨道阱质谱,Hypersil Gold C_(18)型色谱柱,乙腈-0.1%甲酸水梯度洗脱,流速0.4 mL/min,柱温40℃,进样量1μL;采用电喷雾离子源,正离子模式下,在质荷比100~1500范围内全扫描,通过提取待测成分的精确质量数进行定量。结果表明,毛蕊异黄酮葡萄糖苷、橙皮苷、甘草酸铵线性范围分别为0.0891~1.4250μg/mL、0.0984~12.6000μg/mL、7.4250~29.7000μg/mL(r≥0.9993),检测限分别为5.57、4.10、5.80 ng/mL,定量限分别为22.26、12.30、23.20 ng/mL,精密度、重复性及样品稳定性(24 h)良好,加样回收率91%~105%(δ_(RSD)≤5.0%,n=6)。本方法简便快捷、特异性强、灵敏度高,可同时检测补中益气丸中毛蕊异黄酮葡萄糖苷、橙皮苷、甘草酸铵的含量。

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 C_(18) 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 1μL,respectively.Mass spectrometer was operated using an electrospray ionization source in the positive ion mode for detection.The scan range was 100~1500 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 0.0891~1.4250μg/mL,0.0984~12.6000μg/mL,and 7.4250~29.7000μg/mL(r≥0.9993),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.