植物组织中有机酸的提取方法比较

Study on the methods of extraction organic acids from different plant tissue by HPLC

以番茄果实和不结球白菜叶片为试验材料,用蒸馏水(水提)、80%乙醇旋转蒸发浓缩(醇提)、蒸馏水冷冻干燥浓缩(水提冻干)分别提取有机酸,采用高效液相色谱法测定。结果表明:3种提取方法下番茄果实中富马酸、丁二酸含量无显著性差异,草酸、酒石酸、苹果酸、草酰乙酸含量均以水提最高,而丙酮酸和柠檬酸以醇提含量最高;不同提取方法对不结球白菜叶片中草酸、酒石酸、富马酸含量无影响,其他几种有机酸(除草酰乙酸外)均以水提和水提冻干含量高于醇提,其中,水提法和水提冻干法提取丙酮酸、苹果酸、丁二酸的含量无显著差异,而柠檬酸含量以水提冻干法最高。综合考虑试验步骤和结果等各方面因素发现,蒸馏水直接提取法既能有效保证有机酸的提取量,又能大大简化试验步骤。

In this experiment, organic acids were extracted from tomato fruit and Chinese cabbage leaves by different extraction methods, such as by distilled water directly （W）, by 80% ethanol and then evaporation for concentration （E）, and by distilled water for extraction and then frozen-evaporation concentration （WF） and then determinated by HPLC. The results obtained are listed as follows. In terms of tomato fruits, the extracted amounts of fumarate and succinate were not influenced by extraction methods, while the amounts of oxalic acid, tartaric acid, malate and oxaloacetate extracted by W method were significantly higher than those extracted by the other two methods. E method could extract the highest amount of pyruvate and citrate. For Chinese cabbage leaves, no significant differences were found in oxalic acid, tartaric acid and fumarate extracted by the three extraction methods, whereas the amount of the other organic acids （ except for oxaloacetate） extracted by W and WF method were significantly higher than those extracted by E method. There were no significant difference of pyruvate, malate, and sueeinate extracted by W and WF method while citrate amount extracted by WF was higher than that by W. Therefore, distilled water extraction （ W） method was proposed to extract organic acids from tomato fruits and Chinese cabbage leaves when taking the extraction steps, working efficiency and extraction quality together into account.