高效液相色谱法分离和测定小麦中的5种内源激素

引用本文: 张玉琼, 仲延龙, 高翠云, 董召荣, 陈娜, 王梅方. 高效液相色谱法分离和测定小麦中的5种内源激素[J]. 色谱, 2013, 31(8): 800-803. doi: 10.3724/SP.J.1123.2013.01011 shu

Citation: 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. doi: 10.3724/SP.J.1123.2013.01011 shu

高效液相色谱法分离和测定小麦中的5种内源激素

张玉琼 ^{1,
,} ,
仲延龙 ^1, ,
高翠云 ^1, ,
董召荣 ^{2,
,} ,
陈娜 ^1, ,
王梅方 ^1,

通讯作者: 张玉琼,董召荣; 张玉琼,董召荣

基金项目:
农业部科技项目国家公益性行业(农业)科研专项(201103001,201203032,201203096) (农业)科研专项(201103001,201203032,201203096)

安徽省自然科学基金面上项目(1208085mc48). (1208085mc48)

摘要: 建立了高效液相色谱法(HPLC)用于分离和测定小麦中的吲哚乙酸(IAA)、脱落酸(ABA)、赤霉素(GA₃)、玉米素(ZT)和水杨酸(SA)5种植物内源激素。经过条件优化,选用甲醇作为样品提取溶剂。然后,经石油醚和乙酸乙酯萃取,经Sep-Pak C18小柱纯化。液相色谱的分离采用Eclipse XDB-C₁₈ (250 mm×4.6 mm, 5 μm)反相色谱柱;流速为1 mL/min;进样量10 μL。检测器波长设置为254 nm; 14.5 min时切换到240 nm; SA洗脱后即18 min时切换回254 nm。流动相A为甲醇,B为乙酸溶液(pH 3.6)。梯度条件为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。结果表明,小麦中各激素的分离效果理想,加标回收率达96.9%~98%,相对标准偏差在1.54%~2.29%之间。因此,该方法的建立为快速、准确地分离和测定小麦内源激素提供了可靠的方法。

关键词:

English

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

1.
1. School of Life Sciences, Anhui Agricultural University, Hefei 230036, China;

Corresponding author: 张玉琼,董召荣; 张玉琼,董召荣

Received Date: 05 January 2013
Fund Project:
农业部科技项目国家公益性行业(农业)科研专项(201103001,201203032,201203096)

安徽省自然科学基金面上项目(1208085mc48).

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:

high performance liquid chromatography (HPLC)
/ endogenous hormones
/ wheat

1. [1] Maggio A, Barbieri G, Raimondi G, et al. J Plant Growth Regul, 2010, 29: 63 [1] Maggio A, Barbieri G, Raimondi G, et al. J Plant Growth Regul, 2010, 29: 63
2. [2] Liu H T, Li Y F, Luan T G, et al. Chromatographia, 2007, 66: 515 [2] Liu H T, Li Y F, Luan T G, et al. Chromatographia, 2007, 66: 515
3. [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[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. [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[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. [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[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. [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[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. [7] Tang Y, Wang L, Ma C, et al. Journal of Life Sciences, 2011, 5: 139[7] Tang Y, Wang L, Ma C, et al. Journal of Life Sciences, 2011, 5: 139
8. [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[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. [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[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. [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[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. [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[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. [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[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. [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[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. [14] Raskin I. Plant Physiol, 1992, 99: 799 [14] Raskin I. Plant Physiol, 1992, 99: 799
15. [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[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. [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[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. [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[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

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高效液相色谱法分离和测定小麦中的5种内源激素

通讯作者: 张玉琼,董召荣; 张玉琼,董召荣

English

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

Corresponding author: 张玉琼,董召荣; 张玉琼,董召荣

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

高效液相色谱法分离和测定小麦中的5种内源激素

通讯作者: 张玉琼,董召荣; 张玉琼,董召荣

English

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

Corresponding author: 张玉琼,董召荣; 张玉琼,董召荣

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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