Pre-concentration of indolic compounds at a carbon paste electrode and indirect determination of L-tryptophan in serum by adsorptive stripping voltammetry

Abstract

The pre-concentration of indole and other indolic compounds of biochemical and pharmaceutical interest at a carbon paste electrode (Nujol/graphite) has been studied. All the compounds examined can be determined, by direct voltammetric measurements in their solutions, in the concentration range 1–8 µM. Indole, methylated indoles, harmaline and serotonin were accumulated at the electrode by a combined adsorption/extraction process. By applying the medium-exchange procedure, the accumulated compounds can be determined in the same concentration range, after a 60-s pre-concentration period, enhancing, therefore, the selectivity of the voltammetric determination. A procedure for the indirect determination of L-tryptophan in serum has been developed, which is based on these findings. L-Tryptophan was cleaved to indole by tryptophanase, and indole was subsequently determined voltammetrically after a 2-min pre-concentration period at the carbon paste electrode. From 0.3 to 1.2 µg of L-tryptophan in a total of 75 µl of serum sample (20–80 µM) can be determined with an average error of ca. 0.03 µg, whereas the recovery of added L-tryptophan in serum samples is in the range 105–115%.