Determination of Free Tryptophan in Plasma and Its Clinical Applications

Abstract

Free tryptophan in plasma was separated by centrifugation through an Amicon ultrafiltration membrane cone. The value obtained without control of pH was found to be lower than that obtained with control of pH by an improved method ( Hijikata et al. (1981) Anal. Biochem. 118, 10-16). For determination of the total tryptophan concentration in the plasma, high performance liquid chromatography (HPLC) was better than the method of Denckla Dewey as modified by Bloxam & Warren ( (1974) Anal. Biochem. 60, 621-625), as judged on the basis of sensitivity, recovery rate and coefficient of variance. The total tryptophan concentration in the plasma determined by HPLC was lower than that determined by the Bloxam & Warren method. The total tryptophan concentration (t-Trp), free tryptophan concentration (f-Trp) and f-Trp/t-Trp ratio were 55.8 +/- 10.2 mumol/l, 11.6 +/- 1.5 mumol/l and 0.211 +/- 0.03 (mean +/- 1 SD) respectively, in healthy subjects (controls). No significant difference was observed between the values of controls and those of patients with liver cirrhosis, hepatocellular carcinoma, liver cirrhosis with hepatocellular carcinoma and hepatic encephalopathy of liver cirrhosis without bleeding. But in liver cirrhosis with bleeding, free tryptophan concentration (f-Trp, 48.0 +/- 23.3 mumol/l, p less than 0.001) and f-Trp/t-Trp ratio (0.645 +/- 0.289, p less than 0.001) were significantly higher than those of controls.(ABSTRACT TRUNCATED AT 250 WORDS)