A CRITICAL-EVALUATION OF LC-EC FOR THE SIMULTANEOUS DETERMINATION OF 5-HYDROXYTRYPTOPHAN, 5-HT, 5-HIAA AND HVA IN BIOLOGICAL SAMPLES USING GC-MS FOR VALIDATION

  • 1 January 1982
    • journal article
    • research article
    • Vol. 37  (3) , 413-430
Abstract
A method was developed for LC-EC [liquid chromatography-electrochemistry] quantitation of 5-hydroxyindoles (5-HT [5-hydroxytryptamine], 5-HTP [5-hydroxytryptophan], 5-HIAA [5-hydroyindoleacetic acid] and N-acetyl-5-HT) and the catecholamine metabolites HVA [homovanillic acid] from rat brain tissue and biological fluids such as human CSF. It involves tissue homogenization in chromatographic buffer containing a non-biological internal standard, 5-hydroxyindole, that can be used to quantitate all the indoles of interest as well as HVA. Comparable chromatographic separations were obtained with conventional C18 stainless steel columns (5 and 10 .mu.m) and with the Waters C18 radial compression module with a 5 or 10 .mu.m C18 cartridge. The LC-EC quantitation of 5-HT, in brain tissue and of 5-HIAA and HVA in CSF was validated by GC-MS-SIM [gas chromatography-mass spectrometry-selected ion monitoring]. The LC-ME method was sensitive in the high pg range on a routine basis.