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目的:建立同时测定田基黄中4个黄酮类成分(异槲皮苷、槲皮苷、田基黄苷、槲皮素)含量的HPLC方法。方法:采用HPLC法,色谱柱为Agilent HC C18(250 mm×4.0 mm,5μm),流动相为甲醇-2.5%冰醋酸溶液(36∶64),检测波长为255 nm,流速为1.0 mL/min;分别采用外标测定法(以槲皮苷、异槲皮苷、田基黄苷、槲皮素对照品为对照)、一测多评法(以槲皮苷对照品为内标)测定含量。结果:外标测定法4个黄酮类成分在标准曲线范围内均呈良好线性关系(r>0.9990);平均加样回收率分别为:异槲皮苷101.66%、槲皮苷98.21%、田基黄苷100.98%、槲皮素99.00%,RSD分别为1.42%、2.61%、1.99%、2.29%。一测多评法异槲皮苷、田基黄苷、槲皮素相对槲皮苷对照品的校正因子f分别为1.155、0.993、0.737,RSD分别为0.52%、0.10%、1.77%。两种方法测得的含量相对偏差小于0.5%,无显著差异。结论:本实验建立的以槲皮苷对照品为内标的一测多评法具有可行性,只需一个对照品,经济实用,可替代外标测定法用于田基黄的质量分析,为全面评价田基黄药材的质量提供了依据。
OBJECTIVE: To establish an HPLC method for the simultaneous determination of four flavonoid constituents (isoquercetin, quercitrin, aglycones, quercetin) in Tianjihuang. Methods: The HPLC method was performed on an Agilent HC C18 column (250 mm × 4.0 mm, 5 μm) with a mobile phase of methanol-2.5% glacial acetic acid (36:64) at a detection wavelength of 255 nm and a flow rate of 1.0 mL / min ; Respectively using the external standard assay (quercitrin, isoquercitrin glycosides, glycosides of tinnitus glycosides, quercetin reference substance for the control), a test of multiple evaluation method (with quercitrin reference substance as an internal standard) determination of content . Results: The results showed that the four flavonoids in the external standard assay showed a good linearity (r> 0.9990) within the standard curve. The average recoveries were 101.66% for isoquercitrin, 98.21% for quercitrin, 100.98% of xanthosine, 99.00% of quercetin and RSD of 1.42%, 2.61%, 1.99% and 2.29% respectively. The results showed that the calibration factors f were 1.155,0.993,0.737, RSD 0.52%, 0.10%, 1.77%, respectively. The relative deviation of the two methods measured content is less than 0.5%, no significant difference. Conclusion: This test established quercetin reference substance as a standard test multiple evaluation method is feasible, only one reference substance, economical and practical, can be used to replace the external standard assay Tianji yellow mass analysis for a comprehensive Evaluation of the quality of Tianji Huang herbs provide the basis.