[Simple chromatographic determination of the neopterin marker in immunopathies]

Abstract

Estimation of neopterin in urine has become part of examination in phenylketonuria, malignities, immunodeficiency incl. AIDS and HIV infection. The authors describe a simple chromatographic method for estimation of neopterin which does not call for highly efficient liquid chromatography nor imported kits. The di- and tetrahydro- forms of neopterin are oxidized with MnO2 and stable neopterin is obtained. The specimen is purified on a Dowex 50WX4 (Fluka) column in a H+ cycle and on PRESEPC18 (TESSEK) columns. Then thin-layer chromatography on Merck Kieselgel 60F254 plates is used in a system of ethyl acetate--isopropanol--1 N NH4OH (35:45:25). Fluorescence is assessed on a densitometer (Shimadzu CS-920 at 367.5 nm). The RF of neopterin is 0.214. The fluorescence is linear within the range of 5-200 ng. The method is sufficiently sensitive also for the estimation of normal neopterin excretion. The authors submit the results of estimations in controls, immunopathies and AIDS.