反相高效液相色谱法同时测定金莲花中4种黄酮类化合物的含量

RP-HPLC determination of flavonoids in Flos Trollii

目的:建立用于金莲花药材质量评价的实用有效的含量测定方法。方法:分析柱为 Thermo Hypersil BDS C_(18)柱(4.6 mm×250 mm,5μm),检测波长为340 nm,流速为0.6 mL·min^(-1),以乙腈-水-冰醋酸(14:86:0.05)为流动相等度洗脱反相高效液相色谱法同时测定金莲花药材中荭草苷-2″-O-β-L-半乳糖、荭草苷-2″-O-β-L-阿拉伯糖、荭草苷和牡荆苷4种黄酮类成分的含量。并采用所建立的方法测定了不同品种、部位和来源的金莲花药材中的4种黄酮类成分含量。结果:线性范围分别为47.7～318μg·mL^(-1)(荭草苷-2″-O-β-L-半乳糖)、3.82～255μg·mL^(-1)(荭草苷-2″-O-β-L-阿拉伯糖)、39.0～260μg·mL^(-1)(荭草苷)、7.20～72.0μg·mL^(-1)(牡荆苷);平均加样回收率(n=9)分别为99.8%,102%,100%,101%。不同金莲花药材中4种黄酮类成分含量差异较大。结论:本方法准确可靠,可用于金莲花药材的质量评价和控制。

Objective： To establish a new method for simultaneous quantitative analyzing flavonoids being applicable to the quality evaluation of Flos Trollii. Methods ： An RP - HPLC method was developed for quantitative determination of four flavonoids （2″-O-β-L- galactopyranosylorientin ,2″-O-β-arabinopyranosylorientin, orientin and vitexin）in Flos Trollii. The analysis was performed on a Thermo Hypersil BDS C18 column（4. 6 mm×250 mm,5 μm） using isocratic elution with the mobile phase consisting of acetonitrile - water - glacial acetic acid （ 14： 86： 0. 05） at 340 nm. The flow rate was 0. 6μg·mL^-1. The established method was applied to the determination of four flavonoids in different species and different parts of Flos Trollii from various sources. Results： A good linear relationship was obtained with correlation coefficient 〉0. 999 ,ranging from 47.7 to 318 μg·mL^-1 for2″-O-β-L-galactopyrano- sylorientin ,3.82 to 255μg·mL^-1 for 2″-O-β- arabinopyranosylorientin ,39.0 to 260 μg·mL^-1for orientin and 7.20 to 72. 0 μg·mL^-1 for vitexin. The recoveries were 99.8%, 102%, 100% and 101% ,respectively,with RSD below 1.3%. A significant variability in the contents of four flavonoids was found among the tested forty samples. Conclusion： The fully validated RP - HPLC method for quantitative analyzing four flavonoids is applicable to the quality evaluation of Flos Tmllii.