目的:建立一种同时测定黄芪建中汤中6个活性成分(毛蕊异黄酮-7-O-β-D-吡喃葡萄糖苷、桂皮酸、芍药苷、芍药内酯苷、甘草苷和甘草酸)含量的超快速液相色谱-串联质谱法。方法:采用Shim-Pack XR-ODS色谱柱(75 mm×3.0 mm, 2.2 μm),以0.1%甲酸水溶液(A)-乙腈(B)为流动相,梯度洗脱(0～6.0 min,12%B→25%B;6.0～7.0 min,25%B→45%B;7.0～11.0 min,45%B→60%B;11.0～13.0 min,60%B→70%B),平衡时间5 min,流速0.4 mL·min-1,柱温30 ℃,进样量5 μL;采用电喷雾电离源(ESI源),负离子模式检测,多反应监测(MRM)。结果:毛蕊异黄酮-7-O-β-D-吡喃葡萄糖苷、桂皮酸、芍药苷、芍药内酯苷、甘草苷和甘草酸质量浓度分别在0.2~10 μg·mL-1(r=0.999 6)、0.5~25 μg·mL-1(r=0.999 5)、2~100 μg·mL-1(r=0.999 6)、0.5~25 μg·mL-1(r=0.999 8)、0.5~25 μg·mL-1(r=0.999 7)和0.5~25 μg·mL-1(r=0.999 6)范围内与峰面积呈良好线性关系;平均回收率(n=9)分别为98.3%、98.4%、98.9%、98.0%、98.8%和99.2%。6批样品中上述6个成分的含量分别为0.114~0.128、0.118~0.144、4.19~4.52、1.03~1.12、0.277~0.345、0.679~0.798 mg·mL-1。结论:本方法灵敏度高,稳定性好,可用于黄芪建中汤的质量控制。
Objective: To establish an ultra-fast liquid chromatography with tandem mass spectrometry (UFLC-MS/MS) method for simultaneous determination of six active components (calycosin-7-O-β-D-glucopyranoside, cinnamic acid, paeoniflorin, albiflorin, liquiritin and glycyrrhizic acid) in Huangqi Jianzhong decoction Methods: Chromatographic separation was performed on a Shim-Pack XR-ODS column (75 mm×3.0 mm, 2.2 μm). The mobile phase consisted of 0.1% formic acid in water (A) and acetonitrile (B) at a flow rate of 0.4 mL·mL-1. The six analytes were determined with gradient elution (0-6 min, 12%B→25%B; 6-7 min, 25%B→45%B; 7-11 min, 45%B→60%B; 11-13 min, 60%B→70%B), and the equilibration time was 5 min. The column temperature was maintained at 30 ℃, and the injection volume was 5 μL. Detection was performed with electrospray ionization source in negative ionization mode. Quantification was performed using multiple reactions monitoring (MRM) mode. Results: The linear ranges of calycosin-7-O-β-D-glucopyranoside, cinnamic acid, paeoniflorin, albiflorin, liquiritin and glycyrrhizic acid were 0.2-10 μg·mL-1 (r=0.999 6), 0.5-25 μg·mL-1 (r=0.999 5), 2-100 μg·mL-1 (r=0.999 6), 0.5-25 μg·mL-1 (r=0.999 8), 0.5-25 μg·mL-1 (r=0.999 7) and 0.5-25 μg·mL-1 (r=0.999 6), respectively. The average recoveries (n=9) of the above-mentioned six components were 98.3%, 98.4%, 98.9%, 98.0%, 98.8% and 99.2%, respectively. The content ranges of six components in six batches were 0.114-0.128 mg·mL-1 for calycosin-7-O-β-D-glucopyranoside, 0.118-0.144 mg·mL-1 for cinnamic acid, 4.19-4.52 mg·mL-1 for paeoniflorin, 1.03-1.12 mg·mL-1 for albiflorin, 0.277-0.345 mg·mL-1 for liquiritin and 0.679-0.798 mg·mL-1 for glycyrrhizic acid. Conclusion: The established method is accurate, stable and reproducible,and can provide a research method for the quality control of Huangqi Jianzhong decoction.
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