沙棘果高效薄层色谱分析及抗氧化能力研究

Study on high-performance thin-layer chromatography analysis and antioxidant ability of Hippophae rhamnoides L.fruits

该文建立了高效薄层色谱法定性定量分析沙棘果中苹果酸和奎宁酸,采用紫外法测定沙棘果对DPPH自由基、ABTS阳离子自由基、超氧阴离子自由基(·O_(2)^(-))的清除率。以V(二氯甲烷)∶V(乙酸乙酯)∶V(甲酸)=5∶6∶2.5溶液为展开剂,采用CamagTLC3扫描仪进行扫描并测定10个不同产地沙棘果中苹果酸和奎宁酸含量。采用DPPH法、ABTS法、·O_(2)^(-)法评价沙棘抗氧化能力。定性定量分析结果表明,苹果酸R_(f)值为0.59±0.05,奎宁酸R_(f)值为0.45±0.05,其方法学考察结果表明高效薄层色谱法分析方法良好。抗氧化试验结果表明,在试验浓度范围内,S7~S10沙棘果提取物DPPH自由基的IC_(50)为0.11~0.13 mg/mL,其中最大清除率为93.71%~96.27%;ABTS阳离子自由基的IC_(50)为0.11~0.61 mg/mL,其中最大清除率为92.18%~99.39%;·O_(2)^(-)的IC_(50)为1.80~2.90 mg/mL,其中最大清除率为81.92%~100.07%;DPPH自由基、ABTS阳离子自由基、·O_(2)^(-)清除率均仅次于维生素C。说明沙棘果提取物对DPPH自由基、ABTS阳离子自由基、·O_(2)^(-)氧化能力在一定浓度范围内具有较好的剂量依赖性。该文所建立的方法更易于广泛推广沙棘果的快速检测,在一定程度上丰富了沙棘果的鉴别方法,为沙棘果的质量控制提供一定参考,同时该研究表明沙棘果是具有高抗氧化性能的药食同源植物。

A high-performance thin-layer chromatography(HPTLC)method was established for the qualitative and quantitative analysis of malic acid and quinic acid in Hippophae rhamnoides L.fruits,and an ultraviolet method was established to determine the clearance rate of Hippophae rhamnoides L.fruits to DPPH free radicals,ABTS cationic free radicals and superoxide anion radicals(·O_(2)^(-)).Took V(dichloromethane)∶V(ethyl acetate)∶V(formic acid)=5∶6∶2.5 as the developing reagent,the CamagTLC3 scanner was used to determine the content of malic acid and quinic acid in Hippophae rhamnoides L.fruits from 10 different producing areas.The antioxidant ability of Hippophae rhamnoides L.fruits was evaluated by DPPH method,ABTS method,·O_(2)^(-) method.The qualitative and quantitative analysis results showed that the R_(f) value of malic acid was 0.59±0.05,and the R_(f) value of quinic acid was 0.45±0.05.The methodological investigation results showed that the analytical method of HPTLC was good.Furthermore,the antioxidant results showed that within the test concentration range,the IC_(50) of S7-S10 Hippophae rhamnoides L.fruits extract DPPH free radicals was 0.11-0.13 mg/mL and the maximum clearance rate was 93.71%-96.27%.The IC_(50) of ABTS cationic free radicals was 0.11-0.61 mg/mL and the maximum clearance rate was 92.18%-99.39%.And the IC_(50) of·O_(2)^(-) was 1.80-2.90 mg/mL and the maximum clearance rate was 81.92%-100.07%.The clearance rates of DPPH free radicals,ABTS cationic free radicals,and·O_(2)^(-) were all less than vitamin C.It was showed that the Hippophae rhamnoides L.fruit extract had a good dose-dependent DPPH free radical,ABTS cationic free radicals and·O_(2)^(-) antioxidation ability within a certain concentration range.The method established was simple and could be widely promote to the rapid detection of Hippophae rhamnoides L.fruits and provided a certain reference for the quality control of Hippophae rhamnoides L.fruits.This study showed that Hippophae rhamnoides L.fruits was medicinal and food plants with high antioxidant properties.