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Lipids

, Volume 47, Issue 2, pp 185–193 | Cite as

Steryl Glucoside and Acyl Steryl Glucoside Analysis of Arabidopsis Seeds by Electrospray Ionization Tandem Mass Spectrometry

  • Kathrin SchrickEmail author
  • Sunitha Shiva
  • James C. Arpin
  • Nicole Delimont
  • Giorgis Isaac
  • Pamela Tamura
  • Ruth Welti
Original Article

Abstract

Establishment of sensitive methods for the detection of cellular sterols and their derivatives is a critical step in developing comprehensive lipidomics technology. We demonstrate that electrospray ionization tandem (triple quadrupole) mass spectrometry (ESI-MS/MS) is an efficient method for monitoring steryl glucosides (SG) and acyl steryl glucosides (ASG). Comparison of analysis of SG and ASG by ESI-MS/MS with analysis by gas chromatography with flame ionization detection (GC–FID) shows that the two methods yield similar molar compositions. These data demonstrate that ESI-MS/MS response per molar amount of sterol conjugate is similar among various molecular species of SG and ASG. Application of ESI-MS/MS to seed samples from wild-type Arabidopsis and a mutant deficient in two UDP-glucose:sterol glucosyltransferases, UGT80A2 and UGT80B1, revealed new details on the composition of sitosteryl, campesteryl and stigmasteryl glucosides and ASG. SG were decreased by 86% in the ugt80A2,B1 double mutant, compared to the wild-type, while ASG were reduced 96%. The results indicate that these glucosyltransferases account for much of the accumulation of the sterol conjugates in wild-type Arabidopsis seeds.

Keywords

Steryl glucosides Acyl steryl glucosides Sterols Electrospray ionization tandem mass spectrometry Gas chromatography Arabidopsis thaliana UDP-glucose:sterol glucosyltransferase ugt80A2,B1, UGT80A2, UGT80B1 

Abbreviations

SG

Steryl glucosides

ASG

Acyl steryl glucosides

ESI-MS/MS

Electrospray ionization tandem (triple quadrupole) mass spectrometry

GC

Gas chromatography

FID

Flame ionization detection

Notes

Acknowledgments

K.S. was supported by National Research Initiative Competitive Grants Program grant no. 2007-35304-18453 from the United States Department of Agriculture National Institute of Food and Agriculture and by the National Science Foundation (MCB 0517758). Equipment acquisition and method development at the Kansas Lipidomics Research Center were funded by the National Science Foundation (EPS 0236913, MCB 0455318 and 0920663, DBI 0521587), Kansas Technology Enterprise Corporation, Kansas IDeA Network of Biomedical Research Excellence (K-INBRE) of National Institutes of Health (P20RR16475), and Kansas State University. This is contribution no. 11-374-J from the Kansas Agricultural Experiment Station.

Supplementary material

11745_2011_3602_MOESM1_ESM.doc (82 kb)
Supplementary material 2 (DOC 83 kb)
11745_2011_3602_MOESM2_ESM.doc (38 kb)
Supplementary material 3 (DOC 39 kb)

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Copyright information

© AOCS 2011

Authors and Affiliations

  • Kathrin Schrick
    • 1
    Email author
  • Sunitha Shiva
    • 1
  • James C. Arpin
    • 1
  • Nicole Delimont
    • 1
  • Giorgis Isaac
    • 1
  • Pamela Tamura
    • 1
  • Ruth Welti
    • 1
  1. 1.Division of Biology116 Ackert Hall, Kansas State UniversityManhattanUSA

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