高效液相色谱法测定大果山楂果实中6种三萜酸类成分的含量

Determination of Content of 6 Triterpenic Acids in Malus doumeri Fruit by High-performance Liquid Chromatography

建立同时测定大果山楂果实中蔷薇酸、2α,19α-二羟基-3-羰基-12-烯-28-乌苏酸、山楂酸、科罗索酸、齐墩果酸和熊果酸的高效液相色谱法(high performance liquid chromatography,HPLC),并对这6种三萜酸在广西6个产地大果山楂果实中的分布及含量进行分析。采用Diamonsil C_(18)(2)色谱柱,以0.1%甲酸水溶液/甲醇(v/v)为流动相,梯度洗脱,流速0.8 mL/min,检测波长210 nm,柱温30℃。结果表明:6种三萜酸在线性范围内质量浓度与色谱峰峰面积具有良好的线性关系,相关系数均>0.9990,检出限0.46~2.00μg/mL,定量下限1.53~6.67μg/mL,变异系数均<5.0%,加标回收率99.2%~101.3%;6个产地的大果山楂果实中三萜酸含量丰富,均检出蔷薇酸、2α,19α-二羟基-3-羰基-12-烯-28-乌苏酸、山楂酸、科罗索酸、齐墩果酸和熊果酸6种三萜酸,均为五环三萜酸化合物,其中蔷薇酸、2α,19α-二羟基-3-羰基-12-烯-28-乌苏酸首次检出,以熊果酸、蔷薇酸、2α,19α-二羟基-3-羰基-12-烯-28-乌苏酸为主,三者含量之和占总三萜酸的71.82%~81.38%,总三萜酸含量在808.74~1090.75μg/g FM之间,其中柳州柳江区和桂林平乐县的果实总三萜酸含量相对较高,分别达到1090.75μg/g FM和1045.20μg/g FM。本研究的HPLC法可作为大果山楂果实中三萜酸含量的检测方法,为大果山楂果原料和产品的品质评估提供科学参考。

A HPLC method was developed for the simultaneous determination of euscaphic acid,2α,19α-dihydroxy-3-oxours-12-en-28-oic acid,maslinic acid,corosolic acid,oleanic acid,and ursolic acid in Malus doumeri fruits.Analyses were conducted on the distribution and content of these six triterpenic acids in Malus doumeri fruits from six producing areas in Guangxi.Using gradient elution at a flow rate of 0.8 mL/min,a Diamonsil C_(18)(2)column was used with a mobile phase of 0.1%formic acid aqueous solution/methanol(v/v).The detection wavelength was 210 nm and the column temperature was 30℃.The results showed that the mass concentrations of the six triterpenic acids had a good linear relationship with the chromatographic peak areas in the linear range.The correlation coefficients were greater than 0.9990,the detection limits were within 0.46~2.00 μg/mL, the lower quantitation limits were within 1.53~6.67 μg/mL, the coefficients of variation were less than 5.0%, and the recovery percentages were within 99.2%~101.3%. Six samples from different production areas contained six triterpenic acids (euscaphic acid, 2α,19α-dihydroxy-3-oxo-urs-12-en-28-oic acid, maslinic acid, corosolic acid, oleanic acid, and ursolic acid). Each of the six triterpenic acids was a pentacyclic triterpenic acid. Among the six triterpenic acids, euscaphic acid and 2α,19α-dihydroxy-3-oxo-urs-12-en-28-oic acid were first detected in Malus doumeri fruits. Ursolic acid, euscaphic acid, and 2α,19α-dihydroxy-3-oxo-urs-12-en-28-oic acid were predominant, accounting for 71.82%~81.38% of the total triterpenic acids. The total triterpenic acids content ranged from 808.74 to 1090.75 μg/g FM. The total triterpenic acids content of fruits from Liujiang district of Liuzhou and Pingle of Guilin were relatively high, reaching 1090.75 μg/g FM and 1045.20 μg/g FM, respectively. Overall, this study's HPLC method can detect the triterpenic acids content of Malus doumeri fruits and provide scientific references for the quality evaluation of Malus doumeri fruit's raw materials and products.