AAC  >> Vol. 7 No. 2 (May 2017)

    GPC-HPLC-MS/MS测定植物组织中IAA与ABA的方法
    Determination of IAA and ABA in Plant Tissue by the GPC-HPLC/MS/MS

  • 全文下载: PDF(790KB) HTML   XML   PP.131-138   DOI: 10.12677/AAC.2017.72018  
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作者:  

杨 柳,刘 帅,王建立,邢 宇,秦 岭:农业应用新技术北京市重点实验室•北京农学院植物科学技术学院,北京 ;
王宗义:农产品有害微生物及农药残留检测与控制北京市重点实验室,北京 ;
曹庆芹:北京农学院生物科学与工程学院•北京林果业生态环境功能提升协同创新中心,北京

关键词:
液相色谱-质谱联用仪(HPLC-MS/MS)净化-定量-浓缩多联机系统(GPC)内标法吲哚乙酸脱落酸Liquid Chromatography-Mass Spectrometry Purifying-Quantitative-Concentrated More Online System Internal Standard Method Indole-3-Acetic Abscisic Acid

摘要:

植物激素检测的有效性和高效性是开展植物研究的前提。本试验采用净化-定量-浓缩多联机系统(GPC)纯化样品、利用HPLC-MS/MS检测以及内标法定量测定植物组织内源IAA和ABA含量。通过正交试验设计优化适于激素提取和纯化的条件,结果表明:取样量0.5 g、80%甲醇提取液、浓缩温度35℃和C18 SPE柱为最优组合。IAA和ABA的含量分别为20.34 ng/g和789.93 ng/g;最低检出限分别为2.36 ng/g和31.95 ng/g;回收率为70.43%和80.17%;变异系数分别为1.87%和2.26%。

The effectiveness and efficiency of the plant hormone detection is a prerequisite for plant research. The sample was purified with the GPC system, and the method employed HPLC-MS/MS for multiple reaction monitoring of concentrations of IAA, ABA, and deuterated IAA and ABA analogs. The condition of extraction and purification of hormones were optimized by orthogonal design. The results show that the composite of 0.5 g samples, 80% methanol extraction solvent, concentration temperature 35˚C and C18 cartridge column for solid-phase extraction (SPE), were optimal extraction procedure for each type of plant tissue. The contents of IAA and ABA as high as 20.34 ng/g and 789.3 ng/g were achieved by this method, respectively; the detectable limits were 2.36 ng/g and 31.95 ng/g; the recoveries were 70.43% and 80.17%; the RSD were 1.87% and 2.26%.

文章引用:
杨柳, 刘帅, 王宗义, 曹庆芹, 王建立, 邢宇, 秦岭. GPC-HPLC-MS/MS测定植物组织中IAA与ABA的方法[J]. 分析化学进展, 2017, 7(2): 131-138. https://doi.org/10.12677/AAC.2017.72018

参考文献

[1] Barkawi, L.S.,Tam, Y.Y., Tillman, J.A.,et al. (2008) A High-Throughput Method for the Quantitative Analysis of Indole-3-Acetic Acid and Other Auxins from Plant Tissue. Analytical Biochemistry, 372, 177-188.
[2] 王耀, 张汉霞, 邹潍力, 等. ASE萃取/GPC-SPE净化/GC-MS法测定茶叶中的有机磷残留[J]. 食品研究与开发, 2011, 31(3): 128-131.
[3] 魏冬旭, 江连洲, 郭伟, 等. ASE-GPC-GC法测定大豆及豆制品中六六六、滴滴涕农药残留[J]. 食品科学, 2009, 30(24): 351-354.
[4] Ross, A.R.S., Ambrose, S.J., Cutler, A.J., et al. (2004) Determination of Endog-enous and Supplied Deuterated Abscisic Acid in Plant Tissues by High-Performance Liquid Chromatog-raphy-Electrospray Ionization Tandem Mass Spectrometry with Multiple Reaction Monitoring. Analytical Biochemistry, 329, 324-333.
[5] Vilaró, F., Canela-Xandri, A. and Canela, R. (2006) Quantification of Abscisic Acid in Grapevine Leaf (Vitis Vinifera) by Isotope-Dilution Liquid Chromatography-Mass Spectrometry. Analytical and Bioanalytical Chemistry, 386, 306- 312.
https://doi.org/10.1007/s00216-006-0664-2
[6] Zhang, F.J., Jin, Y.J., Xu, X.Y., et al. (2008) Study on the Extraction, Purification and Quantification of Jasmonic Acid,Abscisic Acid and Indole-3-Acetic Acid in Plants. Phytochemical Analysis, 19, 560-567.
https://doi.org/10.1002/pca.1085
[7] Cao, J., Murch, S.J., O’Brien, R., et al. (2006) Rapid Method for Accurate Analysis of Melatonin, Serotonin and Auxin in Plant Samples Using Liquid Chromatography-Tandem Mass Spectrom-etry. Journal of Chromatography A, 1134, 333-337.
[8] Vine, J.H., Noiton, D., Plummer, J.A., et al. (1987) Simulta-neous Quantitation of Indole 3-Acetic Acid and Abscisic Acid in Small Samples of Plant Tissue by Gas Chromatog-raphy-Mass Spectrometry/Selected Ion Monitoring. Plant Physiology, 85, 419-422.
https://doi.org/10.1104/pp.85.2.419