Determination of individual long-chain fatty acyl-CoA esters in heart and skeletal muscle.
A method has been developed for determination of individual long-chain fatty acyl-CoA esters from heart and skeletal muscle using high performance liquid chromatography (HPLC). The esters were extracted from freeze-clamped tissue of pig and rat hearts and rat skeletal muscle for analysis on a radially compressed C18 5mu reverse-phase column. Nine peaks in the extract with carbon chain lengths from C12 to C20 that subsequently disappeared on alkaline hydrolysis were identified. The major acyl-CoA peaks were 14:1, 18:2, 16:0 and 18:1 and additionally in rat heart 18:0. Total long-chain acyl-CoA esters obtained by summation of the individual molecular species was 11.34 +/- 1.48 nmol/g wet wt. pig heart; 14.51 +/- 2.11 nmol/g wet wt. in rat heart, and 4.35 +/- 0.71 nmol/g wet wt. in rat skeletal muscle. These values were approximately 132% of those obtained using a separate procedure that measured total CoA by HPLC after alkaline hydrolysis of the esters. The described method demonstrates the quantitation of individual acyl-CoA species in muscle tissue. Therefore, it has a number of advantages in that it permits information to be obtained on the individual molecular species under various nutritional and metabolic conditions.
Molaparast-Saless
F
F
Department of Medicine, University of Wisconsin, Madison 53706.
Shrago
E
E
Spennetta
T L
TL
Donatello
S
S
Kneeland
L M
LM
Nellis
S H
SH
Liedtke
A J
AJ
D000214:Acyl Coenzyme A / Q000032:analysis*; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D009132:Muscles / Q000032:analysis*; D009206:Myocardium / Q000032:analysis*; D013552:Swine
D000214:Acyl Coenzyme A