基于UHPLC的金银花和山银花的鉴别及测定研究

Study on identification and determination of Lonicerae Japonicae Flos and Lonicerae Flos by UHPLC

目的从定性、定量2个方面分别研究金银花和山银花中主要成分及其量的差异。方法采用UHPLC法对金银花和山银花中9种成分新绿原酸、绿原酸、隐绿原酸、木犀草苷、异绿原酸B、异绿原酸A、异绿原酸C、灰毡毛忍冬皂苷乙和川续断皂苷乙进行定性鉴别。定量测定绿原酸、木犀草苷、灰毡毛忍冬皂苷乙和川续断皂苷乙4种成分的量。采用Luna C_(18)-HST柱(100 mm×3.0 mm,2.5μm);柱温40℃;流动相为0.1%甲酸水溶液-0.1%甲酸乙腈溶液,梯度洗脱;体积流量1 mL/min;进样体积1μL;检测波长为327 nm(0~3 min)、350 nm(3~5 min),皂苷类化合物(5~8 min)用蒸发光散射检测器检测。结果各成分线性关系良好(r=0.997 6~0.999 9)。加样回收率为98.71%~100.8%,RSD为0.53%~1.87%。测定20批样品,并比较其色谱图。结论该方法操作简单、可靠、重现性好,可在8 min内通过检测皂苷类化合物快速鉴别金银花和山银花,同时可准确测定绿原酸、木犀草苷、灰毡毛忍冬皂苷乙和川续断皂苷乙的量。

Objective To explore the differences between chemical constituents of Lonicerae Japonicae Flos and Lonicerae Flos based on the quality and quantity analysis. Methods The chromatographic separation was carried out on a Luna C（18）-HST column（100 mm × 3.0 mm, 2.5 μm） with gradient elution kept at 40 ℃. The mobile phases consist of 0.1% formic acid aqueous solution and 0.1% formic acid acetonitrile solution at a flow rate of 1.0 mL/min. The samples were analyzed with a small volume（1 μL） per injection. DAD detection was performed at 327 nm from 0 to 3 min, 350 nm from 3 to 5 min and ELSD detection was performed from 5 to 8 min. Results Almost all the compounds had good linearities（r = 0.997 6—0.999 9）. The excellent recovery rates were 98.71%—100.8% with their RSD from 0.53%—1.87%. Twenty batches of samples obtained from different locations were examined and their chromatographic profiles were compared. Conclusion The results showed that this method was simple, reliable, and stable, and it could be used to rapidly distinguish Lonicerae Japonicae Flos and Lonicerae Flos with high resolution in a single run within 8 min. Furthermore, the method could accurately determinate chlorogenic acid, luteolin-7-O-glucoside, maranthoidin B, and dipsaccside B.