目的 运用超高效液相色谱串联三重四极杆质谱（HPLC-QQQ）建立一种能够同时测定银杏叶制剂中13个黄酮类成分的分析方法。方法 采用Agilent Poroshell 120 EC-C₁₈色谱柱（50 mm×3.0 mm，2.7 μm），以0.1%甲酸溶液-乙腈为流动相，梯度洗脱，体积流量0.3 mL/min，柱温30℃。在电喷雾离子化负离子模式下离子化，以多反应监测方式（MRM）检测。结果 13种黄酮类成分芦丁、异槲皮苷、槲皮素-3-O-β-D-葡萄糖基（1→2）-α-L-鼠李糖苷、山柰酚-3-O-芸香糖苷、水仙苷、槲皮苷、芹菜素-7-O-β-D-吡喃葡萄糖苷、木犀草素、槲皮素、芹菜素、山柰酚、异鼠李素、芫花素的分离度良好，在各自的质量浓度范围内的线性关系良好（r=0.999 0），且精密度试验、重复性试验的RSD值均小于3.5%，供试品溶液在12 h内稳定性好，各成分的平均回收率在94.5%~100.5%。不同批次样品之间，总黄酮含量相差不大；各批次的银杏黄酮类成分含量均不相同，差异较大，其中黄酮苷类化合物所占比例相对较大，芦丁、水仙苷含量均较单糖（异槲皮苷、槲皮苷）含量高，芫花素、木犀草素等黄酮苷元含量最低。结论 方法分离度好、操作简便、专属性强，能够明确反映出银杏叶提取物中13个黄酮类成分的分布和差异，可用于银杏叶提取物制剂的质量控制。

Objective To establish an High Performance Liquid Chromatography-Triple Quadrupole Mass Spectrometry (HPLC-QQQ) method for determination of thirteen flavonoids in Ginkgo leaves preparations. Methods The separation was performed on an Agilent Poroshell 120 EC-C₁₈ (50 mm×3.0 mm, 2.7 μm) with the mobile phase consisted of 0.1% formic acid solution acetonitrile for gradient elution. The column temperature was set at 30℃, and the flow rate was 0.3 mL/min. The flavonoids were detected by electrospray ionization mass spectrometry in negative mode with multiple reaction monitoring (MRM) mode. Results Thirteen flavonoids, such as rutin, isoquercetin, quercetin-3-O-β-D-glucosyl (1→2)-α-L-rhamnoside, kaempferol-3-O-rutinoside, narcissus, quercetin, apigenin-7-O-β-D-glucopyranoside, luteolin, quercetin, apigenin, kaempferol, isorhamnetin, and genkwanin had good resolution, and showed good linear relationships in their respective mass concentration ranges (r=0.999 0). The values of RSD of precision test and repeatability test were all less than 3.5%. The sample solution was stable within 12 h. The recoveries of thirteen flavonoids were within 94.5%-100.5%. The contents of total flavonoids in Ginkgo leaves preparations from different batches had little difference, and it was quite different in different batches of samples. The proportion of flavonoid glycosides was relatively large. The contents of rutin and narcissus were higher than those of monosaccharides (isoquercitrin and quercitrin), while the contents of genkwanin and luteolin were the lowest. Conclusion The method has good separation, simple operation, and strong specificity. It can clearly reflect the distribution and difference of flavonoids in Ginkgo leaves preparations, and can be used for the quality control of Ginkgo leaves preparations.

R927.2

上海市宝山区科学技术委员会科技创新专项资金项目（18-E-14）