Complementary use of amperometric and spectrophotometric detection for concurrent monitoring of serum tryptophan metabolites by reversed-phase liquid chromatography.

We describe an improved reversed-phase liquid-chromatographic method for concurrent measurement of tryptophan and eight of its metabolites in serum. These were separated by gradient elution and detected both by ultraviolet absorption at 254 nm and amperometrically at an oxidation potential of +0.700 V vs Ag/Cl. Detection limits for the electrochemically-detected compounds were between 100 pg and 1.0 ng, and approximately 1-5 ng for the ultraviolet-absorbing compounds. Analytical recovery of the compounds after deproteinization (by ultrafiltration) varied between 65 and 95%. As little as 500 microL of serum is required. No interference(s) from other naturally occurring serum constituents were encountered. This highly sensitive method is relatively fast and offers a useful tool for diagnosis and treatment of diseases involving derangement in tryptophan metabolism.