【摘要】 目的采用超高效液相色谱—二极管阵列检测法(UPLC-DAD)建立健脾方中7种成分(尿囊素、橙皮苷、芸香柚皮苷、异橙黄酮、川陈皮素、3, 5, 6, 7, 8, 3’, 4’-七甲氧基黄酮、桔皮素)含量的测定方法,并开展其应用研究。方法采用Agilent 1290 series超高效液相色谱仪,进行紫外多波长检测(尿囊素:224 nm;橙皮苷、芸香柚皮苷:284 nm;异橙黄酮:342 nm;川陈皮素、3, 5, 6, 7, 8, 3’, 4’-七甲氧基黄酮、桔皮素:333 nm),并完成方法学验证。同时将此方法用于分析不同提取方法制备的健脾方提取液中7种化学成分的含量差异。结果以上7种成分在相应的浓度范围内,化合物浓度与色谱峰峰面积的线性关系良好(r>0.999 3)。方法学考察结果表明,该方法精密度、重复性、样品溶液的稳定性及准确度均良好。不同提取方法制备的健脾方提取液中7种成分的含量差异较大,水煎液中尿囊素浓度最高,乙醇回流液中橙皮苷、芸香柚皮苷、异橙黄酮、川陈皮素、3, 5, 6, 7, 8, 3’, 4’-七甲氧基黄酮和桔皮素浓度均高于水煎液。提示在健脾方提取纯化工艺中,应从多指标成分含量分析的角度评价提取制备工艺选择的合理性。结论本研究基于UPLC-DAD建立的健脾方中7种成分的含量测定方法稳定可靠,能够为其工艺研究和质量控制提供参考。更多还原

【Abstract】 Objective To establish a new method for determination of seven components(allantoin, hesperidin, narirutin, isosinensetin, nobiletin, 3, 5, 6, 7, 8, 3’, 4’-heptamethoxyflavone, tangeretin) in Jianpi Prescription by ultra performance liquid chromatography(UPLC) with diode array detector(DAD) and to carry out its application research. Methods Agilent 1290 series ultra high performance liquid chromatography was used to detect the seven components by UV multi-wavelength detection. The detection wavelength of allantoin was set at 224 nm, with narirutin and hesperidin at 284 nm, isosinensetin at 342 nm, and nobiletin, 3, 5, 6, 7, 8, 3’, 4’-heptamethoxyflavone and tangeretin at 333 nm. The method was used to analyze the contents of seven components in Jianpi Prescription extracts prepared by different extraction methods. Results In the appropriate concentration range for all above seven components, the concentrations showed quite good linear relationship with the peak area(r>0.999 3).All indexes of the methodological investigations including precision, repeatability, stability and accuracy met the requirements. The contents of seven components in Jianpi Prescription extracts prepared by various of extraction methods were significantly different. The concentration of allantoin in water decoction of Jianpi prescription was the highest, while the concentrations of hesperidin, narirutin, isosinensetin, nobiletin, 3, 5, 6, 7, 8, 3’, 4’-heptamethoxyflavone and tangeretin in ethanol reflux liquid were higher than those in water decoction of Jianpi Prescription. It was suggested that in the extraction and purification process of Jianpi Prescription, the rationality of the method should be evaluated from the point of view of multi-component content analysis. Conclusion The established method based on UPLC-DAD in this study is stable and reliable, and can provide reference for its process research and quality control.更多还原