Catecholamine interference with enzymatic determination of nonesterified fatty acids in two commercially available test kits.
We present evidence that catecholamines, which are commonly used to stimulate lipolysis in adipose tissue in vitro, interfere with the enzymatic determination of non-esterified fatty acid (NEFA) in two commercially available kits. Measurement of a 100 mumol/L standard with the Wako "NEFA C" test kit was 60% inhibited by 100 mumol/L norepinephrine and was completely inhibited by 100 mumol/L isoproterenol or by 1 mmol/L norepinephrine or epinephrine. Measurement with the Boehringer Mannheim "Free Fatty acids, Half-micro test" was completely inhibited by 100 mumol/L norepinephrine and was also affected by concentrations as low as 0.1 mumol/L. We propose that this effect is due to the catecholamines interfering with a step common to the two kits, the generation of hydrogen peroxide and oxidation of a chromagen; furthermore, this interference appears to be stoichiometric. We also give details of an alternative in-house method, which does not depend on the generation of hydrogen peroxide and is not affected by catecholamines.
Carr
R E
RE
Oxford Lipid Metabolism Group, Sheikh Rashid Laboratory, Radcliffe Infirmary, UK.
Humphreys
S M
SM
Frayn
K N
KN
D042962:Acyl-CoA Oxidase; D002395:Catecholamines / Q000097:blood*; D003066:Coenzyme A Ligases / Q000378:metabolism; D004837:Epinephrine / Q000494:pharmacology; D005230:Fatty Acids, Nonesterified / Q000097:blood*; D006801:Humans; D006861:Hydrogen Peroxide; D007545:Isoproterenol / Q000494:pharmacology; D009638:Norepinephrine / Q000494:pharmacology; D010088:Oxidoreductases / Q000378:metabolism; D011786:Quality Control; D011933:Reagent Kits, Diagnostic / Q000706:statistics & numerical data*; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D012680:Sensitivity and Specificity; D013053:Spectrophotometry; D000098859:Long-Chain-Fatty-Acid-CoA Ligase
D002395:Catecholamines; D005230:Fatty Acids, Nonesterified; D011933:Reagent Kits, Diagnostic; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D006861:Hydrogen Peroxide; D010088:Oxidoreductases; D042962:Acyl-CoA Oxidase; D003066:Coenzyme A Ligases; C090799:FAA2 protein, S cerevisiae; D000098859:Long-Chain-Fatty-Acid-CoA Ligase; D007545:Isoproterenol; D009638:Norepinephrine; D004837:Epinephrine