A fast and versatile method for extraction and quantitation of long-chain acyl-CoA esters from tissue: content of individual long-chain acyl-CoA esters in various tissues from fed rat.
A method for the extraction of acyl-CoA esters from tissue, and their subsequent analysis by HPLC is described. The lipids are removed by a two-phase extraction in a chloroform/methanol/water system. The long-chain acyl-CoA esters are extracted using methanol and a high salt concentration (2 M ammonium acetate). Reextraction of the dry residue after evaporation of extraction solvent results in low overall recoveries (20%). By adding 1 mg/ml acyl-CoA-binding protein to the extraction solvent the overall recovery was increased to 55%. The method is easy and fast to perform and is thereby suitable for analysis of a large number of samples. The advantages of the method over previously published methods are discussed.
Rosendal
J
J
Institute of Biochemistry, University of Odense, Denmark.
Knudsen
J
J
D000214:Acyl Coenzyme A / Q000302:isolation & purification*; D000818:Animals; D002250:Carbon Radioisotopes; D002352:Carrier Proteins / Q000494:pharmacology; D002623:Chemistry Techniques, Analytical / Q000379:methods; D002725:Chloroform; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004952:Esters / Q000302:isolation & purification*; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D008099:Liver / Q000737:chemistry; D008297:Male; D000432:Methanol; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000494:pharmacology; D015203:Reproducibility of Results; D013228:Stearates / Q000302:isolation & purification; D014867:Water
D000214:Acyl Coenzyme A; D002250:Carbon Radioisotopes; D002352:Carrier Proteins; D004952:Esters; C490174:Fabp7 protein, rat; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D011994:Recombinant Proteins; D013228:Stearates; D014867:Water; D002725:Chloroform; D000432:Methanol