Combined high-performance liquid chromatographic-continuous-flow fast atom bombardment mass spectrometric analysis of acylcoenzyme A compounds.
A high-performance liquid chromatographic method for the analysis of coenzyme A thioesters which employs continuous-flow fast atom bombardment mass spectrometric detection is presented. The chromatographic system utilizes gradient elution with reversed-phase conditions using ammonium acetate-acetonitrile from both standard analytical (3.9 mm I.D.) and microbore (1 mm I.D.) columns. Applications to coenzyme A thioesters of various acyl group chain length (C2-C18) and functionality (-COOH, -OH, -C = C-) are described. The system is also applied to an in vitro enzyme reaction (crotonase) to directly follow the disappearance of substrate and appearance of product. The mass spectrometry of coenzyme A thioesters, their chromatographic behavior, system stability, and sensitivity of detection are discussed.
Norwood
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Division of Pediatric Genetics and Metabolism, Duke University Medical Center, Durham, NC 27710.
Bus
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Millington
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D000214:Acyl Coenzyme A / Q000032:analysis* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D004746:Enoyl-CoA Hydratase / Q000378:metabolism; D013058:Mass Spectrometry
D000214:Acyl Coenzyme A; C030372:3-hydroxybutyryl-coenzyme A; C010701:crotonyl-coenzyme A; D004746:Enoyl-CoA Hydratase