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Measurement of synthesis rate of long-chain acyl-coenzyme a ester in bovine liver by high-performance liquid chromatography |
T. Mitsuhashi, M. Mitsumoto, Y. Yamashita, S. Ozawa |
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Abstract |
A high performance liquid chromatographic procedure is described for the direct determination of the picomole amount of palmitoyl-Coenzyme A and stearoyl-Coenzyme A, using a stainless steel column packed with C-18 derivatized porous silica (5關m), an isocratic elution with a mixture of 33 mM KH2PO4/acetonitrile as a mobile phase and a UV detector. The long-chain acyl-Coenzyme A esters were determined in incubated microsomal fractions of a bovine liver to demonstrate the utility of this method for monitoring acyl-CoA synthesis in biological samples. The reaction rate of palmitate was higher than that of stearate. After a 60 minute incubation period, the generated amount of palmitoyl-Coenzyme A and stearoyl-Coenzyme A were approximately 70 and 20 n mol/mg micresomal protein, respectively. The advantage of this method are in that no decomposition of the CoA esters is involved, while the constituent molecular species is detected. |
Keywords:
HPLC; Acyl CoA; Bovine Liver |
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