RP-HPLC法同时测定葡萄籽提取物中没食子酸、儿茶素和表儿茶素的含量

目的: 建立RP-HPLC法同时测定葡萄籽提取物中没食子酸、儿茶素、表儿茶素3种单体的含量。方法: 采用Agilent zobax SB-C₁₈色谱柱(4.6 mm×250 mm,5 μm),以乙腈(A)-水(1%甲醇+0.15%三氟乙酸)(B)为流动相梯度洗脱(0~10 min,0%A→7%A;10~30 min,7%A→16%A;30~40 min,16%A→90%A;40~50 min,90%A;50~55 min,90%A→0%A;55~65 min,0%A),流速1.0 mL·min^-1,检测波长280 nm,柱温40 ℃, 进样体积20 μL。结果: 没食子酸、儿茶素和表儿茶素3种单体质量浓度分别在1.0~54.0 μg·mL^-1(r=0.999 8)、2.7~136.0 μg·mL^-1(r=0.999 8)、2.7~136.0 μg·mL^-1(r=0.999 9) 范围内与峰面积呈良好的线性关系;平均回收率(n=6)分别为100.3%、99.15%和100.3%。3批GSE样品中没食子酸的含量分别为0.69%、0.69%、0.68%,儿茶素的含量分别为3.15%、3.12%、3.16%,表儿茶素的含量分别为2.48%、2.44%、2.42%。结论: 建立的HPLC法经方法学验证,可用于葡萄籽提取物中没食子酸、儿茶素、表儿茶素含量的同时测定。

Objective: To establish an RP-HPLC method for simultaneous determination of three monomers (gallic acid,catechin,epicatechin)in extracts of grape seeds. Methods: Chromatographic separation was performed on the Agilent zobax SB-C₁₈ column (4.6 mm×250 mm,5 μm).The mobile phase was acetonitrile (A)-water (1% methanol+0.15% TFA)(B)with the gradient elution (0-10 min,0%A→7%A;10-30 min,7%A→16%A;30-40 min,16%A→90%A;40-50 min,90%A;50-55 min,90%A→0%A;55-65 min,0%A),at the flow rate of 1.0 mL·min^-1. The detective wavelength was set at 280 nm,the column temperature was maintained at 40 ℃ and the injection volume was 20 μL. Results: The linear ranges were 1.0-54.0 μg·mL^-1 (r=0.999 8)for gallic acid,2.7-136.0 μg·mL^-1 (r=0.999 8)for catechin and 2.7-136.0 μg·mL^-1 (r=0.999 9)for epicatechin.The average recoveries (n=6)of the three monomers were 100.3%,99.15% and 100.3%.The contents of gallic acid,catechin and epicatechin in three lots of GSE samples were respectively 0.69%,0.69%,0.68%;3.15%,3.12%,3.16%;and 2.48%,2.44%,2.42%. Conclusion: Through method validation,the developed HPLC method is suitable for the simultaneous determination of gallic acid,catechin and epicatechin in extracts of grape seeds.

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