3种银杏黄酮苷类化合物定量分析方法比较研究

Comparative Study on Three Quantitative Methods for Determination of Flavonoid Glycosides in Ginkgo biloba Oral Preparations

目的为改进目前银杏叶类制剂黄酮醇苷成分的含量控制采用间接测定方式的现状,建立一测多评测定法以及标化对照提取物实现量值转移的测定方法,使得银杏黄酮苷类成分的控制更加准确可靠。方法采用YMC-Pack ODS-AQ (4.6 mm×250 mm,5μm)色谱柱;乙腈(A)-0.4%磷酸溶液(B),梯度洗脱,流速1.0mL·min^-1,检测波长360nm;以芦丁为内参物,建立银杏叶口服制剂中5种黄酮醇苷[山柰酚3-O-芸香糖苷(KGR)、异鼠李素3-O-芸香糖苷(IGR)、山柰酚3-O-鼠李糖-2-葡萄糖苷(KRG)、槲皮素3-O-鼠李糖-2-O-(6-O-对羟基反式桂皮酰)-葡萄糖苷(QRcG)及山柰酚3-O-鼠李糖-2-O-(6-O-对羟基反式桂皮酰)-葡萄糖苷(KRcG)]“一测多评”含量测定方法,确定了5种黄酮醇苷对于芦丁的相对校正因子;对中国食品药品检定研究院发放的对照提取物以及自制银杏黄酮苷对照提取物中5种黄酮醇苷的含量进行了标定。结果所建立的“一测多评”方法各检测成分的相对校正因子,在不同仪器、色谱柱、不同浓度及进样体积条件下重复性好,方法可行。标定了银杏叶对照提取物和制备的银杏黄酮苷对照提取物中黄酮苷的含量。3种方法含量测定结果与外标法测定结果进行统计分析,结果显示,4种测定方法间无明显差异(P>0.05)。结论 4种检测方法均可有效、准确地对银杏叶多剂型口服制剂进行含量测定,一测多评法和对照提取物法实现了量值转移,解决无对照物质问题,为评价银杏叶口服制剂的质量提供了可行的方法。

OBJECTIVE To establish a QAMS and a method for quantitative transfer of standardized control extracts to change the current status of controlling flavonoid glycosides in Ginkgo biloba preparations by means of indirect determination, thus making the control of flavonoid glycosides of Ginkgo biloba more accurate and reliable. METHODS The YMC-Pack ODS-AQ column (4.6 mm×250 mm, 5 μm) was adopted. The mobile phase consisted of acetonitrile-0.4% phosphoric acid and gradient elution was conducted at a flow rate of 1.0 mL·min-1. The detection wavelength was set at 360 nm. Five kinds of flavonoid glycosides in various oral preparations of Ginkgo biloba were determined by single marker(QAMS)method. Rutin was used as internal reference standard and the relative correction factors(RCFs)of KGR, IGR, KRG, QRcG and KRcG to rutin were calculated. The ginkgo flavonoid glycosides extract control was then prepared. Two batches of Ginkgo biloba extract control were demarcated,one was laboratory homemade, and the other was issued by the National Institutes for Food and Drug Control (NIFDC). RESULTS The RCFs determined by different instruments and columns at different concentrations and injection volumes had good reproducibility. The contents of flavonoid glycosides in the prepared ginkgo flavonoid glycosides extract control and Ginkgo biloba extract control were calibrated. No significant difference was found among the results of the above-mentioned three methods and the external standard method (P>0.05). CONCLUSION The four methods can all effectively and accurately determine the contents of flavonoid glycosides in various oral preparations of Ginkgo biloba. QAMS and standardized extract control method may achieve quantitative transfer. They can solve the problem of lack of reference substances and provide a feasible method for evaluating the quality of Ginkgo biloba oral preparations.