HPLC法测定大果山楂果实中八种酚酸类成分的含量

Determination of contents of eight phenolic acids in Malus doumeri fruit by HPLC

关键词:  HPLC, 大果山楂, 果实, 酚酸, 含量

Key words:  HPLC, Malus doumeri, fruits, phenolic acids, content

摘要:

建立HPLC法同时测定大果山楂果实中原儿茶酸、绿原酸、二氢咖啡酸、咖啡酸、根皮酸、p-香豆酸、阿魏酸和肉桂酸的方法,并通过HPLC法分析这八种酚酸在不同产地大果山楂果实中的含量。采用ZORBAX SB-C18色谱柱,以甲醇/1.5%甲酸水溶液(v/v)为流动相,梯度洗脱,柱温35 ℃,流速1.0 mL·min^-1,检测波长280、320 nm。结果表明:(1)在线性范围内八种酚酸质量浓度与色谱峰峰面积的线性关系良好,相关系数均>0.997,检出限0.08～0.20 μg·mL^-1,定量下限 0.27～0.67 μg·mL^-1,变异系数均<5.0%,加标回收率99.3%～103.3%;(2)10个不同产地的大果山楂果实中酚酸含量丰富,均检出原儿茶酸、绿原酸、二氢咖啡酸、咖啡酸、根皮酸、阿魏酸和肉桂酸七种酚酸,其中,二氢咖啡酸、根皮酸、阿魏酸均为首次检出,以绿原酸为主(8 410.2～13 826.7 μg·g^-1),占总酚酸的80%以上,总酚酸的质量分数在10 187.8～15 583.9 μg·g^-1之间,其中广西百色靖西和桂林恭城产的果实总酚酸质量分数相对较高,均大于15 000 μg·g^-1。综上结果表明,该研究的HPLC法适用于大果山楂果实中酚酸含量的测定,可为大果山楂优良品种的选育、产品质量控制及深度开发利用提供方法和科学参考。

Abstract:

The contents of phenolic acids in Malus doumeri fruits from different production areas were investigated, and at the same time, HPLC method was developed to detect eight phenolic acids, including protocatechuic acid, chlorogenic acid, dihydrocaffeic acid, caffeic acid, phloretic acid, p-coumaric acid, ferulic acid and cinnamic acid. ZORBAX SB-C18 column was adopted. The mobile phase was composed of methanol and 1.5% formic acid aqueous with a gradient elution. The column temperature was 35 ℃, the flow rate was 1.0 mL·min^-1, the detection wavelength was 280, 320 nm, and the injection volume was 20 μL. The results were as follows:(1)Linear relationship between the mass concentration and the peak area of the chromatogram of eight phenolic acids good, with the correlation coefficient > 0.997, the detection limit 0.08-0.20 μg·mL^-1, the lower limit of quantitation 0.27-0.67 μg·mL^-1, the coefficient of variation <5.0%, and the adding standard recovery 99.3%-103.3%.(2)Seven phenolic acids(protocatechuic acid, chlorogenic acid, dihydrocaffeic acid, caffeic acid, phloretic acid, ferulic acid and cinnamic acid)were detected over ten samples in different production areas. Among the seven phenolic acids, dihydrocaffeic acid, phloretic acid and ferulic acid were detected in M. doumeri fruits for the first time, and chlorogenic acid(8 410.2-13 826.7 μg·g^-1)was predominant, accounting for more than 80% of the total phenolic acids. The mass fraction of total phenolic acids ranged from 10 187.8 to 15 583.9 μg·g^-1. The mass fraction of total phenolic acids from fruits picked from Jingxi of Baise and Gongcheng of Guilin in Guangxi was relatively high, more than 15 000 μg·g^-1. The above results indicate that the HPLC method used in this study is suitable for the determination of phenolic acids contents in M. doumeri fruit, and can provide methods and scientific basis for screening superior variety, products quality control, and deep exploitation and utilization of M. doumeri fruits.