探索利用活血生物效价检测方法和化学指纹图谱方法评价大黄的品质。采用血小板聚集仪测定不同大黄药材体外抗血小板聚集率,根据简化概率单位法计算生物效价;采用超高效液相色谱仪建立不同大黄药材的指纹图谱;利用SPSS 22.0软件的双变量(bivariate)分析方法进行谱效相关分析,最后用化学单体进行了验证。活血效价测定结果显示掌叶大黄的活血效价普遍高于唐古特大黄和药用大黄;酒大黄的活血效价高于生大黄,大黄炭活血效价显著降低。指纹图谱中23个共有色谱峰,指认了其中10个成分。谱效相关分析得到3个相关性较好的化学成分,包括大黄酸-8-O-β-D-葡萄糖苷、大黄素-8-O-β-D-葡萄糖苷和大黄酸,活血效价验证结果表明三者均具有较强的抑制血小板聚集的作用。本文建立的基于测定血小板聚集率模型的大黄活血生物效价检测方法对于大黄药材,尤其是酒大黄炮制过程中的质量评控具有一定的参考价值,同时初步证明大黄中活血化瘀药效物质为大黄酸等蒽醌类衍生物。
The biological potency assay and chemical fingerprint chromatogram were applied to quality evaluation of rhubarb. Using the biological potency as indicators, we evaluated the differences in quality of multiple batches of rhubarbs and related products. Using the platelet aggregation analyzer, we determined platelet aggregation rate in the different rhubarbs preparations, and calculated the biological potency based on the simplified probit principle. UPLC was adopted to establish the fingerprint spectra for rhubarbs. The spectral efficiency correlation analysis between chromatograms and biological potencies were conducted using the double variables of SPSS 22.0 software. We used three chemical composition to verify the potency. The biological potency results suggest that Rheum palmatum has a more potent activity than Rheum tanguticum, and wine-treated rhubarb had a higher potentcy than charred. We identified 10 elements in the Fingerprint Spectrum. The relevant elements including rhein-8-O-β-D-glucoside, emodin-8-O-β-D-glucoside and rhein have the strongest activity in the inhibition of platelet aggregation. In conclusion, this study provides a analytical method for rhubarb biological potency based on determination of the maximum antagonism rate model. The rhein may be the effective substance. It may serve as a reference in the quality control of wine processed rhubarb products.