Simultaneous high-performance liquid chromatography determination of coenzyme A, dephospho-coenzyme A, and acetyl-coenzyme A in normal and pantothenic acid-deficient rats

We describe here a simultaneous high-performance liquid chromatography method for practical and rapid determination of coenzyme A (CoA), dephospho-CoA, and acetyl-CoA in tissues. These coenzymes are biosynthesized from the vitamin pantothenic acid (PaA), which is involved in the metabolism of fatty acids, amino acid catabolism, and several other nutrients. The method employed a Tosoh TSK-GEL ODS-100 V column (250 × 4.6 mm i.d., particle size 5 μm) eluted with 100 mmol/L NaH2PO4 and 75 mmol/L CH3COONa (pH was adjusted to 4.6 by the addition of concentrated H3PO4)–acetonitrile (94:6, v/v) at a flow rate of 1.0 ml/min. The ultraviolet detector was set at 259 nm. The limits of detection for CoA, dephospho-CoA, and acetyl-CoA all were 10 pmol. The method was applied to the analysis of several tissues of rats fed normal and PaA-free diets. The results clearly showed that the method was suitable for the simultaneous determination of CoA, dephospho-CoA, and acetyl-CoA in the liver, heart, kidney, spleen, testis, large colon, and muscle, but not for the small intestine, of rats.