Analysis of pipecolic acid in biological fluids using capillary gas chromatography with electron-capture detection and [2H11]pipecolic acid as internal standard.
A sensitive and accurate stable isotope dilution assay was developed for the measurement of pipecolic acid in body fluids using capillary gas chromatography with electron-capture detection. The method utilizes [2H11]pipecolic acid as the internal standard. Sample preparation consisted of derivatization in aqueous solution (pH 11.5) of the amine moiety with methyl chloroformate to the N-methylcarbamate, followed by acidic ethyl acetate extraction at pH less than or equal to 2 and further derivatization of the carboxyl moiety with pentafluorobenzyl bromide, the excess of which was removed by solid-phase extraction. Control values have been determined in the plasma of at-term infants, age greater than 1 week (n = 21, mean = 1.36 microM, range = 0.47-3.27 microM). The utility of the method was demonstrated by quantitating pipecolic acid in biological fluids derived from patients with peroxisomal disorders. The method was validated against an established electron-capture negative ion mass fragmentographic technique.
Zee
T
T
Department of Pediatrics, Free University Hospital, Amsterdam, Netherlands.
Stellaard
F
F
Jakobs
C
C
D001826:Body Fluids / Q000737:chemistry*; D002849:Chromatography, Gas; D004583:Electrons; D006801:Humans; D008661:Metabolism, Inborn Errors / Q000097:blood / Q000134:cerebrospinal fluid / Q000175:diagnosis* / Q000652:urine; D008830:Microbodies / Q000378:metabolism*; D010875:Pipecolic Acids / Q000097:blood / Q000134:cerebrospinal fluid / Q000378:metabolism* / Q000652:urine; D012015:Reference Standards
D010875:Pipecolic Acids; C031345:pipecolic acid