一测多评法测定黄蜀葵花中7个黄酮类成分

目的：建立以金丝桃苷为内标的一测多评法，用以评价黄蜀葵花药材质量的方法。方法：采用ACQUITY UPLC BEH C₁₈色谱柱（2.1 mm×100 mm，1.7 μm），0.05%甲酸水溶液（A）-乙腈（B）为流动相，流速为0.40 mL·min^-1，梯度洗脱（0~2 min，88%A→85%A；2~5 min，85%A→84%A；5~6 min，84%A→83%A；6~9 min，83%A→70%A；9~11 min，70%A→20%A；11~12 min，20%A→5%A；12~13 min，5%A→88%A），检测波长360 nm，柱温35 ℃，建立金丝桃苷与其他6个指标成分（芦丁、异槲皮苷、 棉皮素-8-O-葡萄糖醛酸苷、杨梅素、槲皮素-3’-O-葡萄糖苷、槲皮素）间的相对校正因子（RCF），并利用校正因子计算黄蜀葵花中7个主要黄酮成分含量，实现一测多评。结果：在一定的线性范围内，金丝桃苷与芦丁、异槲皮苷、棉皮素-8-O-葡萄糖醛酸苷、杨梅素、槲皮素-3’-O-葡萄糖苷、槲皮素间的RCF值分别为0.827、1.312、0.494、0.989、1.608、1.516，且在不同实验条件下重现性良好（RSD<5.0%），并与常规外标一点法比较，13批黄蜀葵花药材中7个黄酮成分一测多评法计算值与实测值间均无显著性差异（RSD<5%）。结论：以金丝桃苷建立的6个黄酮成分校正因子可用于黄蜀葵花药材的定量分析及质量评价。

Objective: To establish an objective and reasonable method evaluating the quality of the flowers of Abelmoschus manihot. Methods: The analysis was performed at 35 ℃,on an ACQUITY UPLC BEH C₁₈ column(2.1 mm×100 mm,1.7 μm),eluted with a gradient program(0-2 min,88%A→85%A;2-5 min,85%A→84%A;5-6 min,84%A→83%A;6-9 min,83%A→70%A;9-11 min,70%A→20%A;11-12 min,20%A→5%A;12-13 min,5%A→88%A)using 0.05% formic acid aqueous solution(A)-acetonitrile(B) as the mobile phase.The flow rate was 0.40 mL·min^-1,and the detection wavelength was 360 nm.Hyperoside was used as the internal reference substance,quantitative assay of multicomponents by single-marker(QAMS)was applied to calculate the relative correlation factors(RCFS)of rutin,isoquercetin,hibifolin,myricetin,quercetin-3'-O-glucoside,quercetin and determine the seven bioactive components in the flowers of Abelmoschus manihot by UPLC. Results: The RCFs of rutin,isoquercetin,hibifolin,myricetin,quercetin-3'-O-glucoside,quercetin with reference to hyperoide were 0.827,1.312,0.494,0.989,1.608,1.516 respectively,and the repeatability was good under different experimental conditions(RSDs<5%).No significant differences were found in the quantitative results of the seven flavonoids in 13 batches of Abelmoschi Corolla determined by external standard method and QAMS(RSD<5%). Conclusion: The QAMS method established in this research for simultaneous determination of the seven components is accurate and feasible to evaluate the quality of the flowers of Abelmoschus manihot.