Simultaneous quantification of malonyl-CoA and several other short-chain acyl-CoAs in animal tissues by ion-pairing reversed-phase HPLC/MS.
Malonyl-CoA is a key intermediate involved in lipid synthesis and lipid oxidation. Here, we report on a novel method for the quantification of malonyl-CoA and seven other short-chain acyl-CoAs in various rat and mouse tissues using ion-pairing reversed-phase HPLC/MS. This method is capable of measuring malonyl-CoA, free coenzyme A (CoASH), acetyl-CoA, beta-hydroxyl-butyryl-CoA (HB-CoA), 3-hydroxy-3-methyl-glutaryl-CoA (HMG-CoA), propionyl-CoA, succinyl-CoA, and isobutyryl-CoA simultaneously with a dynamic linear range over two orders of magnitude in a 7.0 min HPLC gradient run. The lower limit of quantification (LLOQ) was 0.225 pmol for all acyl-CoAs studied, except for HMG-CoA which had a higher LLOQ of 0.90 pmol. The interference of HB-CoA on the quantification of malonyl-CoA in animal tissues was also explored for the first time.
Gao
Lan
L
Department of Biological Screening, Global Pharmaceutical Research and Development, Abbott Laboratories, 100 Abbott Park Road, Abbott Park, IL 60064, United States. lan.gao@abbott.com
Chiou
William
W
Tang
Hua
H
Cheng
Xueheng
X
Camp
Heidi S
HS
Burns
David J
DJ
D000105:Acetyl Coenzyme A / Q000032:analysis / Q000737:chemistry; D000214:Acyl Coenzyme A / Q000032:analysis* / Q000737:chemistry; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008099:Liver / Q000737:chemistry; D008316:Malonyl Coenzyme A / Q000032:analysis* / Q000737:chemistry; D051379:Mice; D015394:Molecular Structure; D018482:Muscle, Skeletal / Q000737:chemistry; D009206:Myocardium / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results
D000214:Acyl Coenzyme A; C024343:butyryl-coenzyme A; C009061:propionyl-coenzyme A; D008316:Malonyl Coenzyme A; D000105:Acetyl Coenzyme A; C012046:succinyl-coenzyme A