感冒清热颗粒中4种成分的LC-MS/MS定性分析及HPLC含量测定

Qualitative Analysis and Determination of Four Components in Ganmao Qingre Granules by LC-MS/MS and HPLC

目的建立感冒清热颗粒中苦杏仁苷、葛根素、升麻素苷和5-O甲基维斯阿米醇苷含量的LC-MS/MS定性分析方法和HPLC分析方法。方法采用LC-MS/MS对各成分进行确证。采用AgilentZorbaxSBC18(4.6mm×250mm,5μm)色谱柱;流动相为甲醇-水梯度洗脱;葛根素、升麻素苷和5-O-甲基维斯阿米醇苷检测波长为252nm,苦杏仁苷检测波长为207 nm;流速为1.0 mL·min^(-1);柱温为35℃。结果感冒清热颗粒中苦杏仁苷、葛根素、升麻素苷和5-O-甲基维斯阿米醇苷的一级、二级质谱图与其对照品溶液相同;各成分分别在5.278~105.558,12.868~257.351,0.938~18.761,0.493~9.861μg·mL^(-1)内呈良好线性关系(r≥0.999 9);平均回收率分别为100.86%,100.09%,100.06%和99.58%。结论该方法操作简便快速,分离效果好,灵敏度高,重复性好,为测定感冒清热颗粒主成分的含量提供了一种快速高效可靠的方法。

OBJECTIVE To establish a LC-MS/MS method and a HPLC method for qualitative analysis and determination of amygdalin, puerarin, prim-O-glucosylcimifugin and 5-O-methylvisammioside in Ganmao Qingre granules. METHODS The components were confirmed by LC-MS/MS. The analysis was performed on an Agilent Zorbax SB C18 column(4.6 mm×250 mm, 5 μm) under the gradient elution with methanol-water. The detection of puerarin, prim-O-glucosylcimifugin and 5-Omethylvisammioside was set as 252 nm, the detection of amygdalin was set as 207 nm. The flow rate was 1.0 mL·min^(-1)and the column temperature was 35 ℃. RESULTS The primary and secondary mass spectrograms of amygdalin, puerarin, prim-O-glucosylcimifugin and 5-O-methylvisammioside in Ganmao Qingre granules were consistent with the reference solution. Each component showed good linear relationship in the ranges of 5.278-105.558, 12.868-257.351, 0.938-18.761 and 0.493-9.861 μg·mL^(-1)respectively(r≥0.999 9). The average recoveries were 100.86%, 100.09%, 100.06% and 99.58%. CONCLUSION The established method is simple and rapid with the good separation and reproducibility, and high sensitivity. It provides a fast, efficient and reliable method for the determination of the main components of Ganmao Qingre granules.