逐级提取-高效液相色谱法快速测定植物组织中8种有机酸  

Rapid determination of eight organic acids in plant tissue by sequential extraction and high performance liquid chromatography

作　　者：黄天志[1,2,3] 王世杰[1,2] 刘秀明[1,2] 刘虹[1] 吴沿友[1,2] 罗绪强[4] 

机构地区：[1]中国科学院地球化学研究所环境地球化学国家重点实验室,贵州贵阳550002 [2]中国科学院普定喀斯特生态系统观测研究站,贵州安顺562100 [3]中国科学院大学,北京100049 [4]贵州师范学院,贵州贵阳550018

出　　处：《色谱》2014年第12期1356-1361,共6页Chinese Journal of Chromatography

基　　金：国家重大科学研究计划项目(2013CB956700);中国科学院战略性先导科技专项(XDA05070400);国家自然科学基金项目(31100187)

摘　　要：针对植物组织中草酸存在的不同形态,建立了水和稀盐酸作为提取介质的逐级提取方法,获得了水溶态和酸溶态草酸及乙醇酸、乙醛酸、酒石酸、苹果酸、乙酸、柠檬酸、琥珀酸等有机酸。采用Hypersil ODS(200 mm×4.6mm,5μm)色谱柱,以5 mmol/L磷酸二氢钾水溶液(pH 2.8)作为流动相,在进样量5μL、检测波长210 nm、柱温30℃的条件下,通过分时段控制流速实现了8种有机酸的快速分离,同时去除了盐酸对酸溶态草酸测定的干扰。本方法精确灵敏、回收率高、重复性好,可应用于实际样品的测定分析。A sequential extraction method was developed to determine different forms of oxalate and seven oxalate metabolism related organic acids（glyoxylic acid,tartaric acid,gly colic acid,malic acid,acetic acid,citric acid,succinic acid）in plant tissue. The ultra pure water was used as the extraction medium to obtain water soluble oxalic acid and the other sev en water soluble organic acids. After the extraction of the water soluble organic acids,the resi dues were extracted by dilute hydrochloric acid successively to get the acid soluble oxalate which entered the liquid phase. A Hypersil ODS column was used with 5 mmol / L potassium dihydrogen phosphate buffer solution（pH 2. 8）as the mobile phase. The diode array detector was set at 210 nm and the column temperature at 30 ℃ with the injection volume of 5 μL. The flow rate was controlled at different times which allowed a good and rapid separation of the organic acids and hydrochloric acid. Under these conditions,the linear ranges of the method were 1- 2 000 mg / L for oxalic acid,25- 2 000 mg / L for acetic acid,and 10- 2 000 mg / L for glyoxylic acid,tartaric acid,glycolic acid,malic acid,citric acid and succinic acid,with the correlation coefficients of the eight organic acids ≥ 0. 999 6. The average recoveries of the eight organic acids in leaves and roots were 93. 5%- 104. 4% and 85. 3%- 105. 4% with RSDs of 0. 15%- 2. 43% and 0. 31%- 2. 9%（n = 7）,respectively. The limits of detection rangedfrom 1 to 10 ng（S / N = 3）. The results indicated that the method is accurate,rapid and repro ducible for the determination of organic acids in plant samples.

关 键 词：逐级提取 高效液相色谱 有机酸 植物 

分 类 号：O658[理学—分析化学]