Determination of histidine and 3-methylhistidine in physiological fluids by high-performance liquid chromatography.
High-performance liquid chromatography based on pre-column derivatization of histidine and 3-methylhistidine with o-phthalaldehyde-mercaptoethanol has been used to determine these amino acids in small volumes of plasma and urine. The elution is performed in 45 min on a 5-micron Resolve C18 column by a multi-step gradient. The eluted analytes are measured with a fluorescence detector which provides detection limits of less than 1 pmol per 20-microliter injection. The correlation between concentration and the integrated area of amino acids gives a linear relationship between 10 and 150 pmol per injection. Some preliminary results from patients with chronic renal failure under variation of diet are presented.
D000596:Amino Acids / Q000032:analysis; D001826:Body Fluids / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D004044:Dietary Proteins / Q000008:administration & dosage; D006639:Histidine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000097:blood / Q000652:urine; D006801:Humans; D007676:Kidney Failure, Chronic / Q000097:blood / Q000652:urine; D008762:Methylhistidines / Q000032:analysis* / Q000097:blood / Q000652:urine; D013050:Spectrometry, Fluorescence; D009764:o-Phthalaldehyde
D000596:Amino Acids; D004044:Dietary Proteins; D008762:Methylhistidines; D006639:Histidine; D009764:o-Phthalaldehyde; C028118:3-methylhistidine