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 ×4.6 mm，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 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.
参考文献
| [1] | Franceschi V R;Nakata P A.[J].Annu Rev Plant Biol,200556:41. |
| [2] | 陈峥;耿华春;王沙沙.[J].分子植物育种,20076(S):105. |
| [3] | Xu H W;Ji X M;He Z H.[J].J Exp Bot,200657(09):1899. |
| [4] | Yu L;Jiang J Z;Zhang C.[J].J Exp Bot,201061(06):1625. |
| [5] | Tuma P;Samcova E;Stulik K.[J].Anal Chim Acta,2011685(01):84. |
| [6] | 韩海峰;王庆;刘霞.[J].色谱,201230(05):538. |
| [7] | 关瑾;王慧泽;任丽艳.[J].色谱,201230(01):107. |
| [8] | 林晓婕;魏巍;何志刚.[J].色谱,201432(03):304. |
| [9] | 高海燕;廖小军;王善广.[J].分析化学,200432(12):1645. |
| [10] | 籍秀梅;杨崇;杨军.[J].分析化学,200533(04):527. |
| [11] | 郭瑛;肖朝萍;王红.[J].分析化学,200432(12):1624. |
| [12] | 金高娃;章飞芳;薛兴亚.[J].分析化学,200634(07):987. |
| [13] | 俞乐;彭新湘;杨崇.[J].分析化学,200230(09):1119. |
| [14] | Keates S E;Tarlyn N M;Loewus F A.[J].Phytochemis-try,200053(04):433. |
| [15] | 马瑞;欧阳嘉;李鑫.[J].色谱,201230(01):62. |
| [16] | 郭燕;梁俊;李敏敏.[J].食品科学,201233(02):227. |
| [17] | 陆敏;张绍岩;张文娜.[J].食品科学,201233(14):235. |
| [18] | Cawthray G R.[J].J Chromatogr A,20031011(1/2):233. |
| [19] | Zotou A;Loukou Z;Karava O.[J].Chromatographia,200460:39. |
| [20] | 林灵超.[J].应用化工,201342(04):756. |
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