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Profiling of phytohormones and their major metabolites in rice using binary solid-phase extraction and liquid chromatography-triple quadrupole mass spectrometry.
J Chromatogr A. 2016 Jun 17; 1451:67-74.JC

Abstract

A high-throughput method was developed using liquid chromatography-triple quadrupole mass spectrometry (LC-MS/MS) for the profiling and quantification of 43 phytohormones and their major metabolites, including auxins, abscisic acid, jasmonic acid, salicylic acid, cytokinins and gibberellins in a single sample extract. Considerable matrix effects (MEs) were observed (with most ME values in the range of 29%-84%, but maximum MEs of more than 115%, even up to 206%, existed) in sample extracts for most of the compounds studied. The application of the proposed binary solid-phase extraction using polymer anion and polymer cation exchange resins, was performed to purify 25 acidic and 18 alkaline phytohormones and their major metabolites prior to the LC-MS/MS analysis, which markedly reduced the MEs to acceptable levels, with ME values in the range of ±15%. Moreover, all of the isomers of cytokinins and their metabolites were fully separated on a sub-2μm particle C18 reverse-phase column with the optimized mobile phase consisting of methanol and 5mM ammonium formate. The method showed good linearity for all 43 analytes with regression coefficients (R(2))>0.991. Limits of detection ranged from 0.19 to 7.57 fmol for auxin, gibberellins, abscisic acid and their metabolites, 29.7 fmol for jasmonic acid, 18.1 fmol for salicylic acid, and from 0.03 to 0.31 fmol for cytokinins and their metabolites. The mean recoveries for all of the analytes were from 70.7 to 118.5%, and the inter-day precisions (n=6) were less than 18.7%, with intra-day precisions (n=6) within 25.4%. Finally, 20 compounds were successfully quantified in rice sample profiles using the proposed method, which will greatly facilitate the understanding of hormone-related regulatory networks that influence rice growth and development. To our knowledge, there are limited reports that measure this level of phytohormone species in rice samples using a single analysis.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Rice Product Quality Inspection and Supervision Center of Ministry of Agriculture, China National Rice Research Institute, Hangzhou 310006, China.

Institute of Health Food, Zhejiang Academy of Medical Sciences, Hangzhou 310007, China.

Rice Product Quality Inspection and Supervision Center of Ministry of Agriculture, China National Rice Research Institute, Hangzhou 310006, China.

Rice Product Quality Inspection and Supervision Center of Ministry of Agriculture, China National Rice Research Institute, Hangzhou 310006, China.

Rice Product Quality Inspection and Supervision Center of Ministry of Agriculture, China National Rice Research Institute, Hangzhou 310006, China.

Rice Product Quality Inspection and Supervision Center of Ministry of Agriculture, China National Rice Research Institute, Hangzhou 310006, China.

Rice Product Quality Inspection and Supervision Center of Ministry of Agriculture, China National Rice Research Institute, Hangzhou 310006, China. Electronic address: cmingxue@live.com.

MeSH

Abscisic AcidChromatography, LiquidCyclopentanesCytokininsGibberellinsIndoleacetic AcidsIon Exchange ResinsOryzaOxylipinsPlant Growth RegulatorsSolid Phase ExtractionTandem Mass Spectrometry

Pub Type(s)

Journal Article

Language

eng

PubMed ID

27208989

Citation

Cao, Zhao-Yun, et al. "Profiling of Phytohormones and Their Major Metabolites in Rice Using Binary Solid-phase Extraction and Liquid Chromatography-triple Quadrupole Mass Spectrometry." Journal of Chromatography. A, vol. 1451, 2016, pp. 67-74.

Cao ZY, Sun LH, Mou RX, et al. Profiling of phytohormones and their major metabolites in rice using binary solid-phase extraction and liquid chromatography-triple quadrupole mass spectrometry. J Chromatogr A. 2016;1451:67-74.

Cao, Z. Y., Sun, L. H., Mou, R. X., Zhang, L. P., Lin, X. Y., Zhu, Z. W., & Chen, M. X. (2016). Profiling of phytohormones and their major metabolites in rice using binary solid-phase extraction and liquid chromatography-triple quadrupole mass spectrometry. Journal of Chromatography. A, 1451, 67-74. https://doi.org/10.1016/j.chroma.2016.05.011

Cao ZY, et al. Profiling of Phytohormones and Their Major Metabolites in Rice Using Binary Solid-phase Extraction and Liquid Chromatography-triple Quadrupole Mass Spectrometry. J Chromatogr A. 2016 Jun 17;1451:67-74. PubMed PMID: 27208989.

* Article titles in AMA citation format should be in sentence-case

TY - JOUR T1 - Profiling of phytohormones and their major metabolites in rice using binary solid-phase extraction and liquid chromatography-triple quadrupole mass spectrometry. AU - Cao,Zhao-Yun, AU - Sun,Li-Hua, AU - Mou,Ren-Xiang, AU - Zhang,Lin-Ping, AU - Lin,Xiao-Yan, AU - Zhu,Zhi-Wei, AU - Chen,Ming-Xue, Y1 - 2016/05/04/ PY - 2016/02/09/received PY - 2016/04/27/revised PY - 2016/05/03/accepted PY - 2016/5/23/entrez PY - 2016/5/23/pubmed PY - 2016/12/15/medline KW - Binary solid-phase extraction KW - LC–MS/MS KW - Metabolites KW - Phytohormones KW - Rice SP - 67 EP - 74 JF - Journal of chromatography. A JO - J Chromatogr A VL - 1451 N2 - A high-throughput method was developed using liquid chromatography-triple quadrupole mass spectrometry (LC-MS/MS) for the profiling and quantification of 43 phytohormones and their major metabolites, including auxins, abscisic acid, jasmonic acid, salicylic acid, cytokinins and gibberellins in a single sample extract. Considerable matrix effects (MEs) were observed (with most ME values in the range of 29%-84%, but maximum MEs of more than 115%, even up to 206%, existed) in sample extracts for most of the compounds studied. The application of the proposed binary solid-phase extraction using polymer anion and polymer cation exchange resins, was performed to purify 25 acidic and 18 alkaline phytohormones and their major metabolites prior to the LC-MS/MS analysis, which markedly reduced the MEs to acceptable levels, with ME values in the range of ±15%. Moreover, all of the isomers of cytokinins and their metabolites were fully separated on a sub-2μm particle C18 reverse-phase column with the optimized mobile phase consisting of methanol and 5mM ammonium formate. The method showed good linearity for all 43 analytes with regression coefficients (R(2))>0.991. Limits of detection ranged from 0.19 to 7.57 fmol for auxin, gibberellins, abscisic acid and their metabolites, 29.7 fmol for jasmonic acid, 18.1 fmol for salicylic acid, and from 0.03 to 0.31 fmol for cytokinins and their metabolites. The mean recoveries for all of the analytes were from 70.7 to 118.5%, and the inter-day precisions (n=6) were less than 18.7%, with intra-day precisions (n=6) within 25.4%. Finally, 20 compounds were successfully quantified in rice sample profiles using the proposed method, which will greatly facilitate the understanding of hormone-related regulatory networks that influence rice growth and development. To our knowledge, there are limited reports that measure this level of phytohormone species in rice samples using a single analysis. SN - 1873-3778 UR - https://www.unboundmedicine.com/medline/citation/27208989/Profiling_of_phytohormones_and_their_major_metabolites_in_rice_using_binary_solid_phase_extraction_and_liquid_chromatography_triple_quadrupole_mass_spectrometry_ DB - PRIME DP - Unbound Medicine ER -