Novel isolation procedure for short-, medium-, and long-chain acyl-coenzyme A esters from tissue.
A novel procedure for the quantitative isolation and purification of acyl-coenzyme A esters is presented. The procedure involves two steps: (1) tissue extraction using acetonitrile/2-propanol (3+1, v+v) followed by 0.1M potassium phosphate, pH 6.7, and (2) purification using 2-(2-pyridyl)ethyl-functionalized silica gel. Recoveries determined by adding radiolabeled acetyl-, malonyl-, octanoyl-, oleoyl-, palmitoyl-, or arachidonyl-coenzyme A to powdered rat liver varied 93-104% for tissue extraction and 83-90% for solid-phase extraction. The procedure described allows for isolation and purification, with high recoveries, of acyl-coenzyme A esters differing widely in chain length and saturation.
Minkler
Paul E
PE
Department of Pharmacology, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
Kerner
Janos
J
Ingalls
Stephen T
ST
Hoppel
Charles L
CL
D019840:2-Propanol / Q000737:chemistry; D000097:Acetonitriles / Q000737:chemistry; D000214:Acyl Coenzyme A / Q000737:chemistry / Q000302:isolation & purification*; D000818:Animals; D004952:Esters; D008099:Liver / Q000737:chemistry*; D008316:Malonyl Coenzyme A / Q000737:chemistry / Q000302:isolation & purification; D010171:Palmitoyl Coenzyme A / Q000737:chemistry / Q000302:isolation & purification; D010710:Phosphates / Q000737:chemistry; D017680:Potassium Compounds / Q000737:chemistry; D011522:Protons; D051381:Rats; D015203:Reproducibility of Results; D052616:Solid Phase Extraction / Q000379:methods*
D000097:Acetonitriles; D000214:Acyl Coenzyme A; D004952:Esters; D010710:Phosphates; D017680:Potassium Compounds; D011522:Protons; C026563:arachidonyl-coenzyme A; D010171:Palmitoyl Coenzyme A; D008316:Malonyl Coenzyme A; C013216:potassium phosphate; D019840:2-Propanol; C032159:acetonitrile