Revised sample preparation for the analysis of oxysterols by enzyme-assisted derivatisation for sterol analysis (EADSA)

Analytical and Bioanalytical Chemistry
Peter J CrickWilliam J Griffiths

Abstract

Sterols, and specifically oxysterols, play important roles in the biosynthesis of bile acids and steroid hormones as well as possessing biological activities in their own right. Analysis of oxysterols is complicated due to their low abundance in biological systems and poor ionisation characteristics in mass spectrometry. Over the past decade, we have developed a liquid chromatography-mass spectrometry method termed enzyme-assisted derivatisation for sterol analysis (EADSA). Our derivatisation procedure relies on two solid-phase extraction steps to (i) separate cholesterol from oxysterols and (ii) remove excess derivatisation reagents. Recent inter-batch variation in C18 reversed-phase cartridges has led us to experiment with alternative columns. Here, we present our findings and report an improved sample preparation procedure using polymeric hydrophilic-lipophilic balanced reversed-phase cartridges.

References

Apr 21, 2012·Journal of Lipid Research·Jeffrey G McDonaldDavid W Russell
Aug 1, 2012·Free Radical Biology & Medicine·William J GriffithsYuqin Wang
Feb 12, 2013·Biochemical Pharmacology·William J GriffithsYuqin Wang

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Citations

Feb 16, 2016·The Journal of Steroid Biochemistry and Molecular Biology·Hanne Roberg-LarsenSteven Ray Wilson
Feb 10, 2019·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Jonas Abdel-KhalikWilliam J Griffiths
Apr 23, 2019·Free Radical Biology & Medicine·William J GriffithsYuqin Wang

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