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目的建立杜仲Eucommia ulmoides雄花6种活性成分的测定方法,分析杜仲雄花生长发育动态及活性成分的变化规律。方法采用超声法进行提取,采用Al Cl3比色法测定总黄酮,采用HPLC法测定其他6种成分,色谱柱为Thermo hypersil gold柱(250 mm×4.6 mm,5μm),以甲醇-0.5%磷酸水溶液为流动相,梯度洗脱;检测波长:206 nm(0~15 min)、236 nm(15~55 min)、255 nm(55~100 min);体积流量1.0 m L/min。结果花径、花高、鲜质量、干质量、雄蕊长度和雄蕊数均在盛花期达到最大值;总黄酮、绿原酸以及活性成分总量均以花蕾期最高,始花期最低;桃叶珊瑚苷量总体呈下降趋势;京尼平苷酸量花蕾期最低,至盛花期达到最高值,末花期有所下降;京尼平苷、异槲皮苷和紫云英苷量均以盛花期最高,始花期最低。结论花期对杜仲雄花形态、产量、活性成分量及质量均有很大影响,为杜仲雄花质量控制及相关产品开发提供了重要依据。
OBJECTIVE To establish a method for determination of 6 active constituents of male flowers of Eucommia ulmoides, and to analyze the growth and development of male flowers of Eucommia ulmoides and the variation of active ingredients. Methods The total flavonoids were determined by AlCl3 colorimetric method. The other six components were determined by HPLC. The column was Thermo hypersil gold column (250 mm × 4.6 mm, 5 μm) The mobile phase was gradient elution. The detection wavelength was 206 nm (0-15 min), 236 nm (15-55 min), 255 nm (55-100 min) and the volume flow rate was 1.0 mL / min. Results The flower diameter, flower height, fresh weight, dry weight, stamen length and number of stamens all reached their maximum at full flowering stage. The total flavonoids, chlorogenic acids and active ingredients were the highest in budding stage and the lowest in beginning flowering stage. The amount of geniposidic acid showed a decreasing trend in the whole, and the lowest in budding stage of geniposidic acid, reaching the peak value in full flowering stage and decreasing in late flowering stage. The amounts of geniposide, isoquercitrin and astragalin were all highest in full bloom , The lowest beginning flowering. Conclusions Florescence has great influence on the morphology, yield, active components and quality of male flowers of Eucommia ulmoides, providing an important basis for the quality control and related product development of male flowers.