Abstract
Flow injection method was developed for rapid, specific, and highly sensitive determination of histidine. The method is based on the fluorescence derivatization of histidine with orthophthalaldehyde in a carrier stream of neutral buffer. A linear calibration curve for histidine was obtained over the range of 0.02–1000 pmol per injection (10 μL) with the relative standard deviation of 0.58% at 1.0 pmol (n = 10) and with the detection limit (S/N = 8) of 15 fmol. Since the fluorescence derivatization is selective for histidine and the interfering substances are limited to glutathione and histamine, the flow injection method was applicable to the rapid and specific assay of histidine in deproteinized human serum, urine, and the extract of stratum corneum where the relative amounts of the interferents to histidine are considerably low. The present method can analyze forty samples per hour. Histidine values in the biological samples obtained by the present method correlated well with those determined by high-performance liquid chromatography. The present method is also applicable to biological samples containing large excess amounts of glutathione, by pretreatment of the samples with a thiol masking reagent such as N-ethylmaleimide.
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