Determination of bestatin in serum by high-performance liquid chromatography with fluorescence detection.
A simple method for the determination of bestatin and its major metabolite in man, p-hydroxybestatin, in human serum was investigated; the method employs high-performance liquid chromatography with fluorescence detection. Bestatin and p-hydroxybestatin are oxidized to phenylacetaldehyde and p-hydroxyphenylacetaldehyde, respectively, with periodate, which are then converted into fluorescent compounds with 4,5-dimethoxy-1,2-diaminobenzene. The compounds are separated by reversed-phase chromatography on LiChrosorb RP-18. The detection limits of bestatin and p-hydroxybestatin are 0.2 and 0.4 microgram/ml serum, respectively. This method permits the precise determination of bestatin in serum (20 microliter) from patients administered bestatin. p-Hydroxybestatin in serum can not be measured by this method because of its low concentration (less than the detection limit).
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D006900:Hydroxylation; D007700:Kinetics; D007930:Leucine / Q000031:analogs & derivatives* / Q000097:blood / Q000378:metabolism; D008297:Male; D010084:Oxidation-Reduction; D013050:Spectrometry, Fluorescence / Q000379:methods; D013997:Time Factors
D007930:Leucine; C012211:ubenimex