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High-performance Liquid Chromatographic Separation of Acylcarnitines Following Derivatization With 4'-bromophenacyl Trifluoromethanesulfonate

High-performance Liquid Chromatographic Separation of Acylcarnitines Following Derivatization With 4'-bromophenacyl Trifluoromethanesulfonate

P E Minkler, S T Ingalls, C L Hoppel

Anal Biochem. 1990 Feb 15;185(1):29-35.

PMID: 2344045

Abstract:

A high-performance liquid chromatographic method for the separation of acylcarnitines after derivatization with 4'-bromophenacyl trifluoromethanesulfonate is presented. Derivatization of acylcarnitines was achieved at room temperature within 10 min. Separation of the acylcarnitine 4'-bromophenacyl esters was accomplished by high-performance liquid chromatography using as the analytical column a Resolve-PAK 5-microns C18 radially compressed cartridge eluted with a tertiary gradient containing varying proportions of water, acetonitrile, tetrahydrofuran, triethylamine, potassium phosphate, and phosphoric acid. Acylcarnitine 4'-bromophenacyl esters were detected spectrophotometrically at 254 nm. Baseline separation was obtained for a standard mixture (5 nmol of each injected) containing carnitine, acetyl-, propionyl-, butyryl-, valeryl-, hexanoyl-, heptanoyl-, octanoyl-, nonanoyl-, decanoyl-, lauroyl-, myristroyl-, palmitoyl-, and stearoylcarnitine. Nearly complete separation was obtained for a standard mixture containing butyryl-, isobutyryl-, isovaleryl-, and 2-methylbutyrylcarnitine. The method was applied to a normal human urine and then to this same urine spiked with the acylcarnitine standards. Urinary acylcarnitine profiles from patients having propionic acidemia, isovaleric acidemia, and medium-chain acyl-CoA dehydrogenase deficiency were performed. Urinary isovalerylcarnitine was quantified in the patient with isovaleric acidemia using heptanoylcarnitine as an internal standard.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP93128042	4-Bromophenacyl trifluoromethanesulfonate		93128-04-2	Price

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