Rapid method for the separation and detection of tissue short-chain coenzyme A esters by reversed-phase high-performance liquid chromatography.
A simple and rapid method for the separation and identification of tissue levels of short chain coenzyme A (CoA) esters by a reversed-phase high-performance liquid chromatography with ultraviolet-visible adsorbance detection is described. Samples of liver, heart and kidney tissues were homogenised in 5% sulfosalicylic acid containing 50 microM of dithioerythritol in 1:9 w/v proportion. Following centrifugation, 20 microliters of the supernatant were directly injected onto a 3-micron ODS C18 column (100 x 4.6 mm I.D.). The separation of acetyl-CoA, malonyl-CoA, methylmalonyl-CoA, succinyl-CoA, propionyl-CoA and free CoASH was achieved in less than 20 min using gradient elution with sodium phosphate, sodium acetate and methanol at a constant flow-rate of 1.5 ml/min. The lowest detection limit was 3 pmol.
Demoz
A
A
Department of Clinical Biology, Haukeland Hospital, University of Bergen, Norway.
Garras
A
A
Asiedu
D K
DK
Netteland
B
B
Berge
R K
RK
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003065:Coenzyme A / Q000737:chemistry / Q000378:metabolism*; D003080:Cold Temperature; D004952:Esters; D007668:Kidney / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008297:Male; D009206:Myocardium / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D004952:Esters; D003065:Coenzyme A