HPLC法同时测定金荞麦片中6个多酚类成分

目的: 建立金荞麦片中6个多酚类成分(原儿茶酸、儿茶素、表儿茶素及原花青素B₁、B₂、C₁)含量的同时测定方法。方法: 采用反相高效液相色谱法,使用Shim-pack VP-ODS C₁₈(250 mm×4.6 mm,5 μm)色谱柱,流动相为乙腈与磷酸水溶液(用磷酸调pH=3.0),梯度洗脱,流速1.0 mL·min^-1,检测波长220 nm。样品采用10%乙腈回流提取。结果: 6个多酚类成分分离良好,阴性无干扰,其平均加样回收率(n=6)为95.4%~104.8%(RSD为1.3%~4.6%);重复性RSD(n=5)为1.3%~3.5%;原儿茶酸、儿茶素、表儿茶素及原花青素B₁、B₂、C₁的线性范围分别为0.013~0.065、0.024~0.121、0.058~0.288、0.033~0.164、0.080~0.402、0.035~0.177 μg,r²分别为1.000 0、0.999 9、0.999 9、0.999 8、0.999 8、0.999 0;供试品溶液中各成分在24 h内稳定。3批样品中原儿茶酸、儿茶素、表儿茶素及原花青素B₁、B₂、C₁ 6个成分的总含量分别为2 451.85、1 965.32、2 570.28 μg·片^-1。结论: 新建立的方法经方法验证,可有效控制金荞麦片的质量,也可作为提高标准中新的含量测定方法。

Objective: To develop a method for simultaneous determination of six polyphenols(protocatechuic acid,catechin,epicatechin,and procyanidins B₁,B₂,C₁)in Jinqiaomai tablets. Methods: An HPLC method was adopted for the analysis,which was performed on a Shim-pack VP-ODS C₁₈ (250 mm×4.6 mm,5 μm)column with the mobile phase of acetonitrile and phosphoric acid solution(pH=3.0)in a gradient elution,the flow rate was 1.0 mL·min^-1,and the detection wavelength was 220 nm.The samples were pre-processed with 1 h reflux in 10% acetonitrile solution. Results: The six polyphenols were separated perfectly by chromatography without disturbing peaks from negative samples.Their average recoveries were 95.4%-104.8%(RSD 1.3%-4.6%,n=6),the repeatabilities were expressed by RSD with values of 1.3%-3.5%(n=6),and linearity ranges of protocatechuic acid,catechin,epicatechin and procyanidins B₁,B₂,C₁ were respectively 0.013-0.065 μg(r²=1.000 0),0.024-0.121 μg(r²=0.999 9),0.058-0.288 μg(r²=0.999 9),0.033-0.164 μg(r²=0.999 8),0.080-0.402 μg(r²=0.999 8),0.035-0.177 μg(r²=0.999 0).The six polyphenols in the sample solution were stable within 24 h.The total contents of protocatechuic acid,catechin,epicatechin,and procyanidins B₁,B₂,C₁ in 3 batches of Jinqiaomai tablets were respectively 2 451.85 μg per tablet,1 965.32 μg per tablet,2 570.28 μg per tablet. Conclusion: The methodology validation proved the developed method can effectively control the quality of Jinqiaomai tablets,and can be used as a new assay method in the quality standard as well.