A time efficient adaptation of GC-FID method for the analysis of PBMC lipid composition


Abstract

Peripheral blood mononuclear cells (PBMC) comprise of lymphocytes and monocytes which are involved in the pathogenesis of chronic inflammatory diseases. A method for estimation of PBMC lipid composition was modified from an existing method for plasma to support our fish oil intervention study in mild-moderate asthmatics. This modification involved an hour- long direct transesterification reaction using methanol and acetyl chloride followed by a modified GC-FID method. The identification of PBMC fatty acid composition was achieved by relative comparison with a 37 Fatty acid component standard. The validation demonstrated the method to be reproducible and accurate with a within batch and between batch CVs of <10%. The modified method was adapted for PBMC total lipid estimation and was considered suitable to detect changes in PBMC composition following fish oil supplementation. This method is quick, scalable, cheaper and cost effective for large scale population and intervention studies. (145 words)



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