Abstract

Circulating non-esterified fatty acids (NEFA) are toxic to mammalian cells. They increase in diseases such as diabetes and sepsis. Herein we propose a serum albumin-fatty acid saturation test.•We based our test on three methodologies: isoelectric focusing (IF) of human plasma albumin, staining proteins after isoelectric focusing in gels with Coomassie Brilliant Blue, and serum albumin measurement with bromocresol green.•The test consists in the determination of albumin IF and staining with bromocresol green. If albumin is saturated with NEFA, it focuses on lower pH, meaning it is the threshold to bind to them. Excessive NEFA is free and toxic. Many other tests are available for NEFA quantification as NEFA kit assay. All colorimetric assays are used for quantification of NEFA and other tests need expensive equipment to read out the results, and they do not measure albumin levels.•Our method focused on albumin-NEFA saturation instead of just NEFA quantification. Critically ill patients have an alteration in both albumin and NEFA. Therefore, our test undergoes less daytime variation compared to assays that measure absolute NEFA values, allowing a more reliable use as an indicator of albumin-fatty acid saturation and NEFA toxicity.

Highlights

  • Circulating non-esterified fatty acids (NEFA) are toxic to mammalian cells

  • Our method focused on albumin-NEFA saturation instead of just NEFA quantification

  • We adapted 3 methodologies, isoelectric focusing of human plasma albumin [1], staining proteins after isoelectric focusing in gels with Coomassie Brilliant Blue R 250 [2], and measurement of serum albumin with bromocresol green [3], in one

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Summary

Method Article

Cassiano Felippe Gonçalves-de-Albuquerqueb,d,*,1, Marcos Roberto Colombo Barnesea,, Mariana Alves Soaresc, Mauro Velho Castro Fariae, Adriana Ribeiro Silvab, Hugo Caire Castro Faria Netob, Patrícia Burthc, Mauricio Younes-Ibrahima,**.

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