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Analysis of Membrane Lipids pp 83–101Cite as

Analysis of Sterols by Gas Chromatography–Mass Spectrometry

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Abstract

Sterols are a major component of cell membranes among all biological systems including bacteria, plant, fungi, and mammals. While the essential carbon skeleton found in all sterol-like structures is the sterane (cyclopentanoperhydrophenanthrenes) ring, there are minor variations which make each structure unique. These include hydroxylations, methylations, ketone groups, double bonds, etc. These structures play specific roles in the biological membranes. Earlier, sterols could only be detected using traditional methods like thin-layer chromatography or UV–vis spectrophotometry. However, these techniques are unable to accurately differentiate between these closely related sterol structures. Therefore, it becomes essential to develop new and sensitive methods for accurate quantification of sterols. In the last few decades, research on gas chromatography–mass spectrometry (GCMS)-based sterol structure determination and quantification has been on the rise. In this chapter, we have discussed some basic background of GCMS and its application in the absolute quantification of sterols using some examples.

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Abbreviations

BHT:

Butylated hydroxytoluene

BSTFA-TMCS:

(N,O-Bis(trimethylsilyl)trifluoroacetamide-trimethylchlorosilane)

CI:

Chemical ionization

EI:

Electron impact

EIC:

Extracted ion chromatogram

GC:

Gas chromatography

i.s.:

Internal standard

m/z :

Mass-to-charge ratio

MS:

Mass spectrometry

RT:

Retention time

TIC:

Total ion chromatogram

TLC:

Thin-layer chromatography

TMS:

Trimethylsilyl

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Acknowledgments

We thank grants to AS from ICMR No. 52/08/2019-BMS and University of Lucknow, Lucknow. NB thanks financial support from ICMR No. 56/2/Hae/BMS and his institution.

Financial and Competing Interest Disclosure

Author has no financial and competing interests with the subject matter or materials discussed in the manuscript.

Contribution to the Manuscript

AS, SAU, KA, and NB wrote the manuscript.

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Authors and Affiliations

  1. Department of Biochemistry, University of Lucknow, Lucknow, Uttar Pradesh, India

    Ashutosh Singh, Sana Akhtar Usmani & Khushboo Arya

  2. Department of Zoology and Environmental Science, Gurukul Kangri University, Haridwar, India

    Nitin Bhardwaj

Authors
  1. Ashutosh Singh
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  2. Sana Akhtar Usmani
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  3. Khushboo Arya
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  4. Nitin Bhardwaj
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Corresponding author

Correspondence to Ashutosh Singh .

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Editors and Affiliations

  1. Amity Institute of Biotechnology and Amity Institute of Integrative Sciences and Health, Amity University Haryana, Gurgaon, Haryana, India

    Rajendra Prasad

  2. Department of Biochemistry, University of Lucknow, Lucknow, Uttar Pradesh, India

    Ashutosh Singh

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Singh, A., Usmani, S.A., Arya, K., Bhardwaj, N. (2020). Analysis of Sterols by Gas Chromatography–Mass Spectrometry. In: Prasad, R., Singh, A. (eds) Analysis of Membrane Lipids. Springer Protocols Handbooks. Springer, New York, NY. https://doi.org/10.1007/978-1-0716-0631-5_6

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  • DOI: https://doi.org/10.1007/978-1-0716-0631-5_6

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-0716-0630-8

  • Online ISBN: 978-1-0716-0631-5

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