Determination of five endogenous hormones in wheat by high performance liquid chromatography

doi:10.3724/SP.J.1123.2013.01011

Abstract

Abstract:

A high performance liquid chromatographic (HPLC) method was developed to determine the five endogenous hormones including indole-3-acetic acid (IAA), abscisic acid (ABA), gibberellic acid (GA₃), zeatin (ZT) and salicylic acid (SA) in wheat. The separation conditions were optimized, and methanol was chosen as the extraction solvent. Then the extract was extracted by petroleum ether and ethyl acetate, and purified with the Sep-Pak C18 column. The chromatographic conditions were as follows: Eclipse XDB-C₁₈ reversed phase column (250 mm×4.6 mm, 5 μm), the flow rate of 1 mL/min, the injection volume of 10 μL, and the detection wavelength of 240 nm were used for the separation of SA from 14.5 min to 18 min, while the detection at 254 nm used for the separation of the others. Methanol (A) and acetic acid aqueous solution (pH 3.6) (B) were used as the mobile phases with the linear gradient set as follows: 0-7 min 20%A, 7-10 min 20%A-28%A, 10-17 min 28%A, 17-19 min 28%A-40%A, 19-35 min 40%A. The results showed that: the hormones were separated well with the recoveries of 96.9%-98%, and the RSDs were in the range of 1.54% to 2.29%. It is a reliable method for rapid, accurate separation and determination of the endogenous hormones in wheat.

Key words: endogenous hormones, high performance liquid chromatography (HPLC), wheat

CLC Number:

O658

ZHANG Yuqiong, ZHONG Yanlong, GAO Cuiyun, DONG Zhaorong, CHEN Na, WANG Meifang. Determination of five endogenous hormones in wheat by high performance liquid chromatography[J]. Chinese Journal of Chromatography, 2013, 31(8): 800-803.

References

[1] Maggio A, Barbieri G, Raimondi G, et al. J Plant Growth Regul, 2010, 29: 63
[2] Liu H T, Li Y F, Luan T G, et al. Chromatographia, 2007, 66: 515
[3] Zhang S L, Li Z Q. Journal of Wheat Research (张酸亮, 李赈群. 小麦研究), 1996, 17(3): 33
[4] Zhao C J, Kang S J, Wang J H, et al. Acta Agriculturae Boreali-Sinica (赵春江, 康书江, 王纪华, 等. 华北农学报), 2000, 15(3): 51
[5] He J L, Liu H X. Journal of Tropical and Subtropical Botany (贺继临, 刘鸿先. 热带亚热带植物学报), 1998, 6(4): 341
[6] Liu C J, Liu Y Y, Li S Q, et al. Food Research and Development (刘长江, 刘旸旸, 李书倩, 等. 食品研究与开发), 2012, 33(2): 162
[7] Tang Y, Wang L, Ma C, et al. Journal of Life Sciences, 2011, 5: 139
[8] Yang T X, Wei A Z, Zheng Y, et al. Chinese Journal of Analytical Chemistry (杨途熙, 魏安智, 郑元, 等. 分析化学), 2007, 35: 1359
[9] Fu J H, Chu J F, Wang J D, et al. Chinese Journal of Analytical Chemistry (符继红, 褚金芳, 王吉德, 等. 分析化学), 2009, 37(9): 1324
[10] Yan S J, Guo L W, Wu C E, et al. Chinese Journal of Analytical Chemistry (闫师杰, 郭李维, 吴彩娥, 等. 分析化学), 2010, 38(6): 843
[11] Chen B L, Zheng C X, Sheng J D, et al. Journal of Xinjiang Agricultural University (陈波浪, 郑春霞, 盛建东, 等. 新疆农业大学学报), 2006, 29(1): 28
[12] Guo L J, Tan X Q, Lu P, et al. China Pharmacist (郭良君, 谭兴起, 陆萍, 等. 中国药师), 2009, 12(12): 1779
[13] Zhang Z, Zhang Q, Wang Z H, et al. Chinese Journal of Chromatography (张政, 张强, 王转花, 等. 色谱), 1994, 12(2): 140
[14] Raskin I. Plant Physiol, 1992, 99: 799
[15] Wang S L, Wang P, Wang C Y. Journal of Analytical Science (王水良, 王平, 王趁义. 分析科学学报), 2010, 5(26): 547
[16] Wang R Z, Xiao L T, Lin W H, et al. Chinese Journal of Chromatography (王若仲, 萧浪涛, 蔺万煌, 等. 色谱), 2002, 20(2): 148
[17] Bu J J, Hu Y L, Wei C B, et al. Northern Horticulture (步佳佳, 胡玉林, 魏长斌, 等. 北方园艺), 2012(8): 30