高效液相色谱法测定银杏叶提取物中4个酚酸类化合物的含量

HPLC determination of 4 hydroxybenzoic acids in extract of Ginkgo biloba leaves

目的:建立银杏叶提取物中4个酚酸类化合物(没食子酸,原儿茶酸,对羟基苯甲酸,间羟基苯甲酸)的HPLC含量测定方法。方法:采用Thermo Syncronis C18色谱柱(250 mm×4.6 mm,5μm),以0.1%磷酸-乙腈为流动相梯度洗脱(0～8 min,5.7%A;8～9 min,5.7%A→6.7%A;9～45 min,6.7%A),流速为1 mL.min-1,检测波长为切换波长(0～30 min,260 nm;30～45 min,234 nm),柱温35℃。结果:没食子酸、原儿茶酸、对羟基苯甲酸、间羟基苯甲酸进样量分别在30.53～457.9、391.2～5868、23.81～357.1、57.69～865.3 ng范围内线性关系良好(r=0.9998～0.9999),平均回收率(n=9)分别为99.8%、98.5%、100.2%、99.6%;检测限分别为0.654、0.559、0.425、1.03μg.g-1。结论:本研究所建立的含量测定方法可以用于银杏叶提取物中酚酸类成分的定量分析,各厂家中4个酚酸的测定结果差异大,本研究所建立的方法具有质控意义。

Objective： To establish an HPLC method for the determination of the contents of 4 hydroxybenzoic acids（ gallic acid,protocatechuic acid,p- hydroxybenzoic acid, mhydroxybenzoic acid） in Ginkgo biloba leafextract. Methods： The analyses were performed on a Thermo Syncronis C 18 column（250 mm×4.6 mm,5μm） , the mobile phase was water containing 0. 1% phosphoric acid - acetonitrile with gradient elution mode （0～8 min,5.7%A;8～9 min,5.7%A→6.7%A;9～45 min,6.7%A） at the flow rate of 1 mL·min^-1 ; The detection was set at 260 nm （0 - 30 min） and 234 nm（ 30 - 45 nm）, the column temperature was 30 ℃. Results： Good line-arity（r = 0. 9998 -0. 9999） was observed in the standard curves of gallic acid, protocatechuic acid,p -hydroxybenzoic acid and m - hydroxybenzoic acid in the ranges of 30.53 - 457.9,391.2 - 5868,23.81 - 357.1 and 57.69-865.3 ng, respectively; The average recoveries （ n = 9 ） of the method were 99.8%, 98.5%, 100. 2% and 99.6% ,respectively. The limits of detection were 0. 654,0. 559,0. 425 and 1.03 μg·g^-1, respectively. Conelu-sion： The HPLC method can be used for the contents determination of 4 hydroxybenzoic acids in Ginkgo biloba leaf extract. The assay results of 4 hydroxybenzoic acids in Ginkgo biloba leaf extracts supplied by 7 manufacturers arevery different, indicating that the content determination of hydroxybenzoic acids in Ginkgo biloba leaf extract should be controlled.