基于在线富集与原位衍生技术的植物组织中内源性油菜素甾醇的超高效液相色谱-串联质谱分析方法

doi:10.3724/SP.J.1123.2014.08010

色谱 ›› 2014, Vol. 32 ›› Issue (10): 1094-1103.DOI: 10.3724/SP.J.1123.2014.08010

基于在线富集与原位衍生技术的植物组织中内源性油菜素甾醇的超高效液相色谱-串联质谱分析方法

丁俊^1,2, 蒋丽¹, 冯钰锜¹

1. 生物医学分析化学教育部重点实验室, 武汉大学化学与分子科学学院, 湖北武汉 430072;
2. 中国科学院植物种质创新与特色农业重点实验室, 中国科学院武汉植物园, 湖北武汉 430074

收稿日期:2014-08-13 出版日期:2014-10-08 发布日期:2014-09-29
通讯作者: 冯钰锜，E-mail：yqfeng@whu.edu.cn
基金资助:
国家自然科学基金项目（91017013，91217309）.

An automatic and sensitive method for the determination of endogenous brassinosteroids in plant tissues by an online trapping-in situ derivatization-ultra performance liquid chromatography-tandem mass spectrometry system

DING Jun^1,2, JIANG Li¹, FENG Yuqi¹

1. Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, China;
2. Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture, Wuhan Botanical Garden of Chinese Academy of Sciences, Wuhan 430074, China

Received:2014-08-13 Online:2014-10-08 Published:2014-09-29

摘要/Abstract

摘要：

油菜素甾醇是一类重要的植物激素，对植物的生长发育过程起显著调节作用。已报道的油菜素甾醇分析方法存在样品前处理过程复杂和灵敏度低等问题。本研究采用C18 PEEK管填充柱为富集柱，以4-N，N-二甲氨基苯硼酸为衍生试剂，搭建了基于双泵-双六通阀的在线管内固相微萃取-超高效液相色谱-串联质谱（UPLC-MS/MS）联用系统，对油菜素甾醇进行在线富集和原位衍生，实现植物组织中内源性油菜素甾醇的自动化分析。该系统程序化地控制了油菜素甾醇的进样、萃取、衍生、分离和检测过程，有效简化前处理过程，节省人力；在线富集步骤实现了样品溶液大体积进样，提高样品利用率；原位衍生改善了油菜素甾醇电喷雾的离子化效率，使油菜素甾醇的电喷雾质谱检出限降低至pg/mL。该系统可在300 mg鲜重植物组织中检测到内源性油菜素甾醇，已成功用于水稻、油菜中多种油菜素甾醇的定量分析。

关键词: 超高效液相色谱-串联质谱, 油菜素甾醇, 原位衍生, 在线富集, 植物组织

Abstract:

Brassinosteroids (BRs) are a class of naturally occurring phytohormones with polyhydroxy steroid structure, which regulate general plant growth and many physiological processes. The reported methods for BR analysis were complicated, and the detection sensitivity was relatively low. To realize the automatic analysis of trace BRs in limited plant tissues, an in-tube solid phase microextraction-ultra performance liquid chromatography-tandem mass spectrometry (SPME-UPLC-MS/MS) system was constructed based on two valves-two pumps. Using C18 PEEK column as the trapping column and 4-(dimethylamino)phenylboronic acid (4-DMAPBA) as the derivatization reagent, an on line trapping and in situ derivatization assay method of BRs was developed. BRs could be programmed to fulfill the procedures of injection, extraction, derivatization, LC separation and MS detection in the system. The detection limits of BRs were improved more than one order of magnitude by the online trapping and in situ derivatization techniques, thus endogenous BRs could be quantified in only 300 mg plant tissues.

Key words: brassinosteroid, in situ derivatization, online trapping, plant tissues, ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)

中图分类号:

O658

丁俊, 蒋丽, 冯钰锜. 基于在线富集与原位衍生技术的植物组织中内源性油菜素甾醇的超高效液相色谱-串联质谱分析方法[J]. 色谱, 2014, 32(10): 1094-1103.

DING Jun, JIANG Li, FENG Yuqi. An automatic and sensitive method for the determination of endogenous brassinosteroids in plant tissues by an online trapping-in situ derivatization-ultra performance liquid chromatography-tandem mass spectrometry system[J]. Chinese Journal of Chromatography, 2014, 32(10): 1094-1103.

参考文献

[1] Bajguz A, Tretyn A. Phytochem, 2003, 62(7): 1027
[2] Mitchell J W, Mandava N, Worley J F, et al. Nature, 1970, 225(5237): 1065
[3] Grove M D, Spencer G F, Rohwedder W K, et al. Nature, 1979, 281(5728): 216
[4] Ashraf M, Akram N A, Arteca R N, et al. Crit Rev Plant Sci, 2010, 29(3): 162
[5] Clouse S D, Sasse J M. Annu Rev Plant Physiol Plant Mol Biol, 1998, 49: 427
[6] Mertens R, Deus-Neumann B, Weiler E W. FEBS Lett, 1983, 160(1/2): 269
[7] Yokota T, Watanabe S, Ogino Y, et al. J Plant Growth Regul, 1990, 9(1/2/3/4): 151
[8] Swaczynová J, Novák O, Hauserová E, et al. J Plant Growth Regul, 2007, 26(1): 1
[9] Bajguz A. J Plant Physiol, 2009, 166(17): 1946
[10] Schrick K, Mayer U, Martin G, et al. Plant J, 2002, 31(1): 61
[11] Khripach V A, Zhabinskii V N, Litvinovskaya R P, et al. Immunoassays of Brassinosteroids//Hayat S, Ahamd A. Brassinosteroids: A Class of Plant Hormone. Netherlands: Springer, 2011: 375.[2014-08-13]. http://link.springer.com/chapter/10.1007/978-94-007-0189-2_14
[12] Yokota T, Watanabe S, Ogino Y, et al. J Plant Growth Regul, 1990, 9(1): 151
[13] Horgen P A, Nakagawa C H, Irvin R T. Can J Biochem Cell Biol, 1984, 62(8): 715
[14] Gupta D, Bhardwaj R, Nagar P K, et al. Plant Growth Regul, 2004, 43(2): 97
[15] Takatsuto S, Ying B, Morisaki M, et al. J Chromatogr A, 1982, 239: 233
[16] Sasse J M. Physiol Plant, 1997, 100(3): 696
[17] Winter J, Schneider B, Meyenburg S, et al. Phytochem, 1999, 51(2): 237
[18] Gamoh K, Takatsuto S. Anal Chim Acta, 1989, 222(1): 201
[19] Gamoh K, Sawamoto H, Kakatsuto S, et al. J Chromatogr A, 1990, 515: 227
[20] Svatos A, Antonchick A, Schneider B. Rapid Commun Mass Spectrom, 2004, 18(7): 816
[21] Huo F, Wang X, Han Y, et al. Talanta, 2012, 99: 420
[22] Wu Q, Wu D, Shen Z, et al. J Chromatogr A, 2013, 1297: 56
[23] Xin P, Yan J, Fan J, et al. Plant Physiol, 2013, 162(4): 2056
[24] Zhang H J, Wu J H, Feng Y Q. Journal of Analytical Science (张慧娟, 吴剑虹, 冯钰锜. 分析科学学报), 2008, 24(5): 589
[25] Atapattu S N, Rosenfeld J M. J Chromatogr A, 2013, 1296: 204
[26] Awan M A, Fleet I, Thomas C L P. Food Chem, 2008, 111(2): 462
[27] Ito R, Kawaguchi M, Honda H, et al. J Chromatogr B, 2008, 872(1/2): 63
[28] Magi E, Di Carro M, Liscio C. Anal Bioanal Chem, 397(3): 1335
[29] Zhang H J, Huang J F, Lin B, et al. J Chromatogr A, 2007, 1160(1/2): 114
[30] Park Y K, Choi K, Ahmed A Y B H, et al. J Chromatogr A, 1217(20): 3357
[31] Salvador A, Moretton C, Piram A, et al. J Chromatogr A, 2007, 1145(1/2): 102
[32] Risticevic S, Niri V, Vuckovic D, et al. Anal Bioanal Chem, 2009, 393(3): 781
[33] Fan Y, Feng Y Q, Da S L, et al. Anal Chim Acta, 2004, 523(2): 251
[34] Nie J, Zhang M, Fan Y, et al. J Chromatogr B, 2005, 828(1/2): 62
[35] Liang B H, Wen Y, Lü Y N, et al. Journal of Instrumental Analysis (梁炳焕, 文毅, 吕永宁, 等. 分析测试学报), 2008, 27(1): 18
[36] Huang J F, Lin X H. Chinese Pharmaceutical Journal (黄京芳, 林幸华. 中国药学杂志), 2009, 44(12): 941
[37] Fan Y, Feng Y Q, Zhang J T, et al. J Chromatogr A, 2005, 1074(1/2): 9
[38] Wei F, Fan Y, Zhang M, et al. Electrophoresis, 2005, 26(16): 3141
[39] Lin B, Zheng M M, Ng S C, et al. Electrophoresis, 2007, 28(15): 2771
[40] Ding J, Mao L J, Wang S T, et al. Phytochem Anal, 2013, 24(4): 386
[41] Springsteen G, Wang B H. Tetrahedron, 2002, 58(26): 5291

基于在线富集与原位衍生技术的植物组织中内源性油菜素甾醇的超高效液相色谱-串联质谱分析方法

An automatic and sensitive method for the determination of endogenous brassinosteroids in plant tissues by an online trapping-in situ derivatization-ultra performance liquid chromatography-tandem mass spectrometry system

PDF

可视化

摘要/Abstract

引用本文

使用本文

扫码分享

参考文献

相关文章 15

编辑推荐

Metrics

[1]	赵芮, 黄晴雯, 余智颖, 韩铮, 范楷, 赵志辉, 聂冬霞. QuEChERS-超高效液相色谱-串联质谱法同时测定水果中36种真菌毒素[J]. 色谱, 2023, 41(9): 760-770.
[2]	岳超, 赵超群, 毛思浩, 王展华, 施贝, 徐欣丰, 梁晶晶. 通过式固相萃取净化-超高效液相色谱-串联质谱法测定液体乳中10种氨基甲酸酯类农药残留[J]. 色谱, 2023, 41(9): 807-813.
[3]	孟二琼, 念琪循, 李峰, 张秋萍, 许茜, 王春民. 磺酸化磁性氮化碳固相萃取-超高液相色谱-串联质谱筛检淡水鱼中孔雀石绿和隐色孔雀石绿[J]. 色谱, 2023, 41(8): 673-682.
[4]	王秋旭, 冯启言, 朱雪强. 加速溶剂萃取-固相萃取净化结合超高效液相色谱-串联质谱法测定沉积物中双酚类化合物[J]. 色谱, 2023, 41(7): 582-590.
[5]	夏宝林, 汪仕韬, 殷晶晶, 张维益, 杨娜, 刘强, 吴海晶. 自动上样固相萃取-超高效液相色谱-串联质谱法同时测定水中9类43种抗菌药物残留[J]. 色谱, 2023, 41(7): 591-601.
[6]	陈东洋, 张昊, 张磊, 王一红, 王小丹, 冯家力, 梁静, 钟旋. 固相萃取-超高效液相色谱-串联质谱法测定发酵食品中的曲酸[J]. 色谱, 2023, 41(7): 632-639.
[7]	童学智, 陈东洋, 冯家力, 范翔, 张昊, 杨胜园. QuEChERS-超高效液相色谱-串联质谱法快速测定水产品中5种卤代苯醌[J]. 色谱, 2023, 41(6): 490-496.
[8]	曾永福, 陈美芳, 邵雨, 闫永欢, 张海超, 王敬, 艾连峰, 康维钧. 植物中27种典型药品及个人护理品多残留检测方法的建立及其在芽苗菜中迁移规律的分析[J]. 色谱, 2023, 41(5): 386-396.
[9]	李振环, 胡小键, 陆一夫, 谢琳娜, 朱英. 基于高通量全自动固相萃取的超高效液相色谱-串联质谱法测定人尿中16种抗生素和4种β-受体激动剂[J]. 色谱, 2023, 41(5): 397-408.
[10]	吴少明, 欧阳立群, 孟鹏, 何孟杭, 林钦, 陈言凯, 刘文菁, 苏晓明, 戴明. 固相萃取净化-超高效液相色谱-串联质谱法测定畜肉中18种卡因类麻醉剂[J]. 色谱, 2023, 41(5): 434-442.
[11]	孙佳林, 牛宇敏, 高群, 张晶, 邵兵. 超高效液相色谱-串联质谱法测定灰尘中26种双酚类化合物[J]. 色谱, 2023, 41(5): 417-425.
[12]	朱卫红, 王超, 张霖琳, 袁懋. 超高效液相色谱-串联质谱法直接测定水中5种黄原酸[J]. 色谱, 2023, 41(4): 339-347.
[13]	韩林学, 张续, 胡小键, 张海婧, 邱天, 林潇, 朱英. 超高效液相色谱-串联质谱法测定人尿中12种典型个人护理品[J]. 色谱, 2023, 41(4): 312-322.
[14]	邱天, 张续, 杨艳伟, 胡小键, 罗嵩, 朱英. 超高效液相色谱-串联质谱法测定尿液中7种苯系物代谢物[J]. 色谱, 2023, 41(4): 366-375.
[15]	刘柏林, 赵紫微, 詹子悦, 戴雁羽, 丁刚, 谢继安, 许相川, 孔洪涛. 改进QuEChERS法结合超高效液相色谱-串联质谱法同时测定冷冻食品中7种季铵盐化合物[J]. 色谱, 2023, 41(3): 233-240.