一测多评法测定萹蓄中3个黄酮类成分的含量

目的：以第四次全国中药资源普查实地采集的76批萹蓄药材为样本，研究建立萹蓄一测多评的含量测定方法，考察同一类型黄酮类化合物之间采用相对校正因子进行准确定量的可行性。方法：采用UPLC进行含量测定，色谱柱为Phenomenex Kinetex C₁₈（2.1 mm×100 mm，1.7 μm）；流动相为乙腈-0.5%磷酸水溶液，线性梯度洗脱；柱温45℃；流速0.2 m L·min^-1；检测波长360 nm；以萹蓄为研究对象，以杨梅苷为参照物，测定杨梅苷、萹蓄苷与槲皮苷的相对校正因子，并利用该校正因子，计算萹蓄苷、槲皮苷的含量，实现一测多评；同时采用外标法测定萹蓄中杨梅苷、萹蓄苷及槲皮苷的含量，将其测定值与一测多评法计算值进行对比分析。结果：杨梅苷、萹蓄苷与槲皮苷分别在2.42~48.4 mg·L^-1（r=0.999 8），2.314~46.28 mg·L^-1（r=0.999 9）和5.523~50.46 mg·L^-1（r=0.999 9）范围内线性关系良好，回收率分别为101.2%、98.9%和97.6%。试验中采用夹角余弦法统计分析2种方法测得的76批萹蓄药材中萹蓄苷和槲皮苷含量数据，夹角余弦值分别为0.999 8、0.999 9、0.999 8，2组数据无显著性差异。结论：本研究建立的以杨梅苷对照品为参照物，采用相对校正因子实现对萹蓄苷和槲皮苷的含量测定方法是准确的、可行的，一测多评法可以实现对同类型化合物进行准确定量。

Objective:Using 76 batches of samples collected from the 4 th national Chinese Materia Medica Resource Inventory,to establish a method for simultaneous determination of three flavonoids in Polygoni Avicularis Herba by quantitative analysis of multi-components with single marker(QAMS)strategy and investigate the feasibility of using correction factors between the identical types of flavonoids.Methods:UPLC analysis was performed on a Phenomenex Kinetex C₁₈ column(2.1 mm×100 mm,1.7 μm).The mobile phase consisted of acetonitrile(solvent A)and 0.5% phosphoric acid water solution(solvent B)with linear gradient elution at a flow rate of 0.2 mL·min^-1.The detection wavelength and column temperature were 360 nm and 45℃,respectively.The relative correction factors(RCFs)of curcumin to the other two ingredients were calculated.The method was evaluated by comparison of the quantitative results between external standard method and QAMS method.Results:Good linear relationships were observed for myricitrin,quercitrin and avicularin within the ranges of 2.42-48.4 mg·L^-1(r=0.999 8),2.314-46.28 mg·L^-1(r=0.999 9)and 5.523-50.46 mg·L^-1(r=0.999 9),respectively.The average recoveries(n=6)were 101.2% for myricitrin,98.9% for quercitrin,97.6% for avicularin,respectively.No obvious differences were found in the quantitative results of myricetrin,avicularin and quercitrin in 76 batches of Polygoni Avicularis Herba determined by the two methods,according to the angle cosine value,the angle cosine values were 0.999 8 for myricitrin,0.999 9 for quercitrin and 0.999 8 for avicularin.Conclusion:It is feasible and suitable to determine myricetrin,avicularin and quercitrin in Polygoni Avicularis Herba by QAMS,and this method can be used for quality evaluation of the identical types of flavonoids.

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