Application of high-performance liquid chromatography of plasma fatty acids as their phenacyl esters to evaluate splanchnic and renal fatty acid balance in vivo

J Chromatogr B Biomed Sci Appl. 1999 Apr 30;727(1-2):15-22. doi: 10.1016/s0378-4347(99)00051-1.

Abstract

Plasma fatty acids from renal and hepatic veins, and arterialized hand vein obtained in 20 subjects before and after insulin infusion were separated by reversed-phase high-performance liquid chromatography following phenacyl esterification. Separation and quantification over the range 1.0-100 nmol per injection of nine fatty acids was achieved within 60 min using [2H31]palmitic acid as internal standard. Analytical recoveries were greater than 90% and the intra- and inter-assay coefficients of variation were less than 2.5 and 4.0%, respectively. Following insulin infusion, net splanchnic uptake of total fatty acids decreased from 3.0+/-0.3 to 1.0+/-0.1 micromol/kg min (p<0.01), whereas net renal balance remained neutral (-0.04+/-0.04 vs. -0.06+/-0.03 micromol/kg min, p=N.S.). Individual fatty acid balance varied from a low of 0.012+/-0.005 (myristic acid) to a high of 0.95+/-0.08 (oleic acid) micromol/kg min across the splanchnic tissues and from 0.005+/-0.002 (stearic acid) to 0.21+/-0.1 (oleic acid) micromol/kg min across the kidney. There is a substantial diversity in changes in plasma concentration and regional balance of individual fatty acid during short-term fasting and hyperinsulinemia. This method is simple, accurate, and can be applied to assess individual fatty acid metabolism in vivo.

Publication types

  • Clinical Trial
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Chromatography, High Pressure Liquid / methods*
  • Esters
  • Evaluation Studies as Topic
  • Fatty Acids, Nonesterified / blood*
  • Fatty Acids, Nonesterified / chemistry
  • Humans
  • Insulin / administration & dosage
  • Reference Standards
  • Reference Values
  • Renal Artery / metabolism*
  • Spectrophotometry, Ultraviolet
  • Splanchnic Circulation*

Substances

  • Esters
  • Fatty Acids, Nonesterified
  • Insulin