Determination of urinary amino acids by liquid chromatography with "dabsyl chloride".

Abstract

We describe a "high-performance" liquid-chromatographic procedure for measuring amino acids in 0.5 mL of urine. The procedure includes direct derivatization of amino acids in urine samples with "dabsyl chloride" (4-dimethylaminoazobenzene-4'-sulfonyl chloride). An aliquot of this dabsylated amino acid solution is analyzed on a muBondapak C18 column with ethanol/sodium acetate (20 mmol/L, pH 4.0) 4/6 (by vol.), as mobile phase. Dabsylated amino acids are detected by their absorbance at 425 nm and quantitated by measuring peak heights. The procedure allows for the reliable analysis of amino acids in urine at concentrations near 16 mg/L. The sensitivity of this analysis on column approaches 5 ng/sample. Higher urinary tryptophan concentrations were found in the urines of some cancer patients, whereas we saw no significant difference in urinary glycine between cancer patients and control subjects. The present method was shown to be a straightforward procedure for detecting phenylalanine in phenylketonuric urine. Extension of this procedure to screening for other inborn errors of aminoaciduria is recommended.