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Comparison of Liquid Chromatography-Mass/Mass Spectrometry (MS) and Gas Chromatography-MS for Quantitative Analysis of Indole-3-acetic acid and Indole-3-butyric acid from the Concentrated Liquid Fertilizer

Liquid Chromatography-Mass/Mass Spectrometry (MS)와 Gas Chromatography-MS를 이용한 농축 액상 비료제품 중 Indole-3-acetic acid 및 Indole-3-butyric acid 정량분석능 비교

  • Kim, Jin Hyo (Chemical Safety Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Park, Jong Min (Chemical Safety Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Choi, Geun-Hyoung (Chemical Safety Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Park, Yun-Ki (Chemical Safety Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Im, Geon-Jae (Chemical Safety Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Kim, Doo-Ho (Chemical Safety Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Kwon, Oh-Kyung (Chemical Safety Division, National Academy of Agricultural Science, Rural Development Administration)
  • Received : 2012.10.04
  • Accepted : 2012.12.11
  • Published : 2013.03.31

Abstract

In here, we investigated the quantitative analysis method of indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) with liquid chromatography-mass/mass spectrometry (LC-MS/MS) or gas chromatography-MS. Two ways of clean-up process were investigated for LC-MS/MS instrumental analysis of IAA, but both a simple dilution and hydrophile-lipophile balance (HLB) solid phase extraction (SPE) were not met the optimal recovery rates for quantitative analysis. On the other hand, the clean-up method for GC-MS was finally optimized through HLB-SPE from 250-folds diluted sample and methylation with trimethylsilyl chloride in methanol for 4 h. The limit of detection for methyl ester of IAA and IBA were both 1.4 mg/L, and recovery rates showed 93-107% from the concentrated liquid fertilizer.

비료제품에는 의도적 혹은 비의도적으로 농약으로 등록된 indole-3-acetic acid (IAA) 및 indole-3-butyric acid (IBA)성분에 대한 잔류 가능성이 꾸준히 제기되고 있으나, 생장조정제가 아닌 비료제품에서 mg/L 수준의 옥신류 잔류분석법이 마련되어 있지 않았다. 따라서, 본 연구에서는 생체시료에 미량 잔류하는 IAA 및 IBA 분석에 사용되어 온 liquid chromatography-mass/mass spectrometry (LC-MS/MS)와 gas chromatography-MS 기기분석법을 활용하여 농축 액상비료제품에 적용할 수 있는지 조사하였으며, 액상 비료제품에 적용 가능한 정밀기기 분석법을 개발하고자 하였다. 수용액 상태의 시료에서 식물 생장호르몬인 IAA와 IBA를 가장 손쉽게 정제할 수 있는 방법으로 hydrophile-lipophile balance (HLB) solid phase extraction를 활용하였으나, 제한된 조건에서 LC-MS/MS를 통한 정량분석은 적합한 회수율을 확보하지 못하였고, 정성분석만 가능함을 확인하였다. 반면, 비료제품 250배 희석액을 사용하여 HLB 정제, trimethylsilyl chloride을 이용한 methylation을 통한 GC-MS 분석에서는 검출한계 1.4 mg/L과 93-107%의 회수율로 액상 비료제품에서 IAA와 IBA 정량분석법을 확립할 수 있었다.

Keywords

References

  1. Ali B, Sabri AN, Ljung K, and Hasnain S (2009) Auxin production by plant associated bacteria: impact on endogenous IAA content and growth of Triticum aestivum L. Lett Appl Microbiol 48, 542-7. https://doi.org/10.1111/j.1472-765X.2009.02565.x
  2. Barkawi LS, Tam Y-Y, Tillman JA, Normanly J, and Cohen JD (2010) A high-throughput method for the quantitative analysis of auxins. Nat Protoc 5, 1609-18. https://doi.org/10.1038/nprot.2010.118
  3. Bottcher C, Keyzers RA, Boss PK, and Davies C (2010) Sequestration of auxin by the indole-3-acetic acid-amido synthetase GH3-1 in grape berry (Vitis vinifera L.) and the proposed role of auxin conjugation during ripening. J Exp Bot 61, 3615-25. https://doi.org/10.1093/jxb/erq174
  4. Chen KH, Miller AN, Patterson GW, and Cohen JD (1988) A rapid and simple procedure for purification of indole-3-acetic acid prior to GCSIM-MS analysis. Plant Physiol 86, 822-5. https://doi.org/10.1104/pp.86.3.822
  5. Davies PJ (2004) Plant hormones: biosynthesis, signal transduction, action. In The Final Action of Hormones, Davies PJ (ed.), pp. 204-20. Kluwer Academic Publishers, Dordrecht, The Netherlands.
  6. Esmon CA, Pedmale UV, and Liscum E (2005) Plant tropisms: providing the power of movement to a sessile organism. Int J Dev Biol 49, 665-74. https://doi.org/10.1387/ijdb.052028ce
  7. Finet C and Jaillais Y (2012) Auxology: When auxin meets evo-devo. Develop Biol 369, 19-31. https://doi.org/10.1016/j.ydbio.2012.05.039
  8. Hager A (2003) Role of the plasma membrane H+-ATPase in auxin-induced elongation growth: historical and new aspects. J Plant Res 116, 483-505. https://doi.org/10.1007/s10265-003-0110-x
  9. Koo YJ, Yoon E, Song JT, Seo HS, Kim J-H, Lee Y-W et al. (2008) An advanced method for the determination of carboxyl methyl esterase activity using gas chromatography-chemical ionization-mass spectrometry. J Chromatogr B 863, 80-7. https://doi.org/10.1016/j.jchromb.2008.01.015
  10. Kurepin L, Haslam T, Lopez-Villalobos A, Oinam G, and Yeung E (2011) Advantitious root formation in ornamental plants: II. The role of plant growth regulators. Propag Ornam Plants 11, 161-71.
  11. Movac M, Muller A, Jarh DM, Milavec M, Duchting P, and Ravnikar M (2009) Multiple hormone analysis indicates involvement of jasmonate signalling in the early defence of potato to potato virus Y-NTN. Biologia Plantarum 53, 195-9. https://doi.org/10.1007/s10535-009-0034-y
  12. NAAS (2010) Pesticide official analysis method. pp. 598-9, National Academy of Agricultural Science (NAAS), RDA, Gyeonggi, Korea.
  13. Nakhooda M, Watt MP, and Mycock D (2011) Auxin stability and accumulation during in vitro shoot morphogenesis influences subsequent root induction and development in Eucalyptus grandis. Plant Growth Regul 65, 263-71. https://doi.org/10.1007/s10725-011-9597-7
  14. Pencik A, Rolcik J, Novak O, Magnus V, Bartak P, Buchtik R et al. (2009) Isolation of novel indole-3-acetic acid conjugates by immunoaffinity extraction. Talanta 80, 651-5. https://doi.org/10.1016/j.talanta.2009.07.043
  15. Piccoli P, Travaglia C, Cohen A, Sosa L, Cornejo P, Masuelli R et al. (2011) An endophytic bacterium isolated from roots of the halophyte Prosopis strombulifera produces ABA, IAA, gibberellins A1 and A3 and jasmonic acid in chemically-defined culture medium. Plant Growth Regul 64, 207-10. https://doi.org/10.1007/s10725-010-9536-z
  16. Whang TE, Lim HO, and Lee JW (1991) Analysis of plant hormones by immunoassay. J Crop Sci 36, 506-12.
  17. You OJ (2011) Fertilizer registration and quality management status, In Fast Detection Method for Formaldehyde in Wood. pp. 3-17, National Academy of Agricultural Science (NAAS), RDA, Gyeonggi, Korea.
  18. Zhang R, Wang Y, Wang C, Wei Z, Xia D, Wang Y et al. (2011) Time-course analysis of level of indole-3-acetic acid and expression of auxinresponsive GH3 genes in Betula platyphylla. Plant Mol Biol Rep 29, 898-905. https://doi.org/10.1007/s11105-011-0306-5

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