Citation: HUANG Tianzhi, WANG Shijie, LIU Xiuming, LIU Hong, WU Yanyou, LUO Xuqiang. Rapid determination of eight organic acids in plant tissue by sequential extraction and high performance liquid chromatography[J]. Chinese Journal of Chromatography, ;2014, 32(12): 1356-1361. doi: 10.3724/SP.J.1123.2014.09016 shu

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

HUANG Tianzhi ^1,2,3 ,
WANG Shijie ^1,2,, ,
LIU Xiuming ^1,2 ,
LIU Hong ¹ ,
WU Yanyou ^1,2 ,
LUO Xuqiang ⁴

1.
1. State Key Laboratory of Environmental Geochemistry, Institute of Geochemistry, Chinese Academy of Sciences, Guiyang 550002, China;

Corresponding author: WANG Shijie,
Received Date: 10 September 2014
Available Online: 10 October 2014
Fund Project: 国家重大科学研究计划项目(2013CB956700) (2013CB956700)中国科学院战略性先导科技专项(XDA05070400) (XDA05070400)国家自然科学基金项目(31100187). (31100187)

A sequential extraction method was developed to determine different forms of oxalate and seven oxalate-metabolism-related organic acids (glyoxylic acid, tartaric acid, glycolic 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 seven water-soluble organic acids. After the extraction of the water-soluble organic acids, the residues 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 1-2000 mg/L for oxalic acid, 25-2000 mg/L for acetic acid, and 10-2000 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 ≥ 0.9996. 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 ranged from 1 to 10 ng (S/N=3). The results indicated that the method is accurate, rapid and reproducible for the determination of organic acids in plant samples.
- sequential extraction,
- high performance liquid chromatography (HPLC),
- organic acids,
- plant
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