一测多评法测定黑果腺肋花楸果中三种花色苷的含量

Quantitative analysis of multi-components with single-marker determine the content of three anthocyanins in black chokeberry

目的基于高效液相色谱外标法建立一测多评法测定新食品原料黑果腺肋花楸果中矢车菊素-3-O-半乳糖苷、矢车菊素-3-O-葡萄糖苷、矢车菊素-3-O-阿拉伯糖苷的含量。方法以AQ-C_(18)柱(250 mm×4.6 mm,5μm),乙腈(A)-0.5%甲酸(B)为流动相梯度洗脱,采用高效液相色谱外标法测定3种花色苷类成分含量,并进行方法学考察。在此基础上以矢车菊素-3-O-葡萄糖苷为内参,测定矢车菊素-3-O-半乳糖苷、矢车菊素-3-O-阿拉伯糖苷相对校正因子,采用相对校正因子计算含量,并与高效液相色谱外标法结果进行比较,验证一测多评法的准确性。结果外标法方法学考察均符合要求,以外标法测定6批黑果腺肋花楸果中矢车菊素-3-O-葡萄糖苷含量分别为0.139、0.148、0.131、0.204、0.088、0.159 mg/g;矢车菊素-3-O-半乳糖苷含量分别为3.899、4.184、3.698、5.855、2.334、4.537 mg/g;矢车菊素-3-O-阿拉伯糖苷含量分别为1.070、1.142、1.009、1.597、0.641、1.239 mg/g;一测多评法与外标法所得结果一致,两种方法所测6批黑果腺肋花楸果中3种花色苷含量结果相关性显著(P<0.05)。结论一测多评法可利用相对保留时间进行定性,可根据相对校正因子进行定量,实现以一种成分同时对多种成分进行定性定量的目的。

Objective To establish a quantitative analysis of multi-components with single-marker method for simultaneous determination of cyanidin-3-O-galactoside,cyanidin-3-O-glucoside and cyanidin-3-O-arabinoside in black chokeberry by HPLC.Methods Using AQ-C_(18)(250mm×4.6mm,5μm)chromatographic column in mobile phase comprising of acetonitrile-0.5%formic acid,gradient elution,external standard method were used to determine the content of three anthocyanin components and methodological investigation.Using cyanidin-3-O-glucoside as the internal marker to calculate relative correlation factors of cyanidin-3-O-galactoside and cyanidin-3-O-arabinoside,and the relative correction factor was used to determine the content of three components with one component.The results were compared between the calculated values of quantitative analysis of multi-components with the single-marker and the measured values with external standard method to validate the accuracy of quantitative analysis of multicomponents with single-marker.Results The external standard method methodological investigation meets the requirements.The contents of cyanidin-3-O-glucoside in six batches of black chokeberry were determined by external standard method that was 0.139,0.148,0.131,0.204,0.088 and 0.159 mg/g,and cyanidin-3-O-galactoside was 3.899,4.184,3.698,5.855,2.334 and 4.537 mg/g,respectively,and cyanidin-3-O-arabinoside were 1.070,1.142,1.009,1.597,0.641,1.239 mg/g,respectively;the results of the quantitative analysis of multi-components with singlemarker and external standard method of anthocyanins in six batches black chokeberry showed that the data were significantly correlated(P<0.05).Conclusion The quantitative analysis of multi-components with single-marker method can use relative retention time for qualitative,and it can be quantified,according to the relative correction factor,so as to achieve the purpose of qualitatively quantifying multiple components at the same time with one component.