高效液相色谱法测定植物叶片中乙醇酸及几种酮酸含量  

Determination of Glycolic Acid and Several α-Keto Acidsin Plant Leaves by High Performance Liquid Chromatography

作　　者：籍秀梅[1] 杨崇[2] 杨军[2] 彭新湘[1] 

机构地区：[1]华南农业大学生命科学学院分子植物生理研究室,广州510642 [2]华南农业大学生命科学学院测试中心,广州510642

出　　处：《分析化学》2005年第4期527-530,共4页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(No. 30270799);广东省自然科学基金(No. 010300)资助项目

摘　　要：植物组织中的乙醇酸经乙醇酸氧化酶催化生成乙醛酸,乙醛酸及其他酮酸类有机酸用苯肼衍生化反应生成苯腙。采用C18柱、324nm紫外检测, 5%甲醇: 95%磷酸缓冲液( 13mmol/LKH2PO4; 1mmol/LK2HPO4,pH6. 0)为流动相,可有效分离几种酮酸的苯腙衍生物。利用该方法可定量测定植物组织中的乙醇酸、乙醛酸、草酰乙酸、羟基丙酮酸、α酮戊二酸及丙酮酸。经多方面检验表明,此方法精确灵敏,回收率高,重复性好,并且成本低、操作简便。可应用于各种生物材料、食品、饮料等产品中的上述有机酸含量检测。Several α-keto acids were first derivatized by phenylhydrazine to form phenylhydrazones, then the latters were separated and determined by reversed-phase high performance liquid chromatography. Hypersil C 18 column, a mobile phase containing 5% methanol and 95% potassium phosphate buffer (13 mmol/L potassium biphosphate;1 mmol/L potassium phosphate dibasic,pH 6.0), detection at 324 nm were applied in the system . By this system, glycolic acid could be also determined by its conversion into glyoxylic acid with glycolic acid oxidase. The method was repeatedly tested for the analysis of different plant leaves, showing it is precise, sensitive and highly reproducible. The detection limits are 0.119, 0.144, 1.651, 0.913 and 0.908 ng for glycolic acid, glyoxylic acid, oxaloacetic acid, α-keto-glutaric acid, pyruvic acid, respectively. Relative standard deviation is less than 3.9% and recovery is close to 100%.

关 键 词：高效液相色谱法 乙醇酸 植物叶片 酸含量 KH2PO4 Α-酮戊二酸 植物组织 磷酸缓冲液 乙醛酸 反应生成 C18柱 紫外检测 定量测定 草酰乙酸 生物材料 含量检测 有机酸 丙酮酸 酶催化 衍生化 流动相 衍生物 回收率 重复性 成本低 

分 类 号：O657.7[理学—分析化学]