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.
A method has been developed for LC-EC quantitation of 5-hydroxyindoles (5-HT, 5-HTP, 5-HIAA and N-acetyl-5-HT) and the catecholamine metabolite HVA from brain tissue and biological fluids such as 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 um) and with the Waters C18 radial compression module with a 5 or 10 um C18 cartridge. The LC-EC quantitation of 5-HT, in brain tissue and of 5-HIAA and HVA in CSF has been validated by GC-MS-SIM. The LC-EC method is sensitive in the high pg range on a routine basis.
D006916:5-Hydroxytryptophan / Q000032:analysis*; D000818:Animals; D001792:Blood Platelets / Q000032:analysis; D001826:Body Fluids / Q000032:analysis; D001923:Brain Chemistry; D002853:Chromatography, Liquid / Q000379:methods*; D008401:Gas Chromatography-Mass Spectrometry; D006719:Homovanillic Acid / Q000032:analysis*; D006801:Humans; D006897:Hydroxyindoleacetic Acid / Q000032:analysis*; D008297:Male; D010648:Phenylacetates / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains; D012701:Serotonin / Q000032:analysis*
D010648:Phenylacetates; D012701:Serotonin; D006897:Hydroxyindoleacetic Acid; D006916:5-Hydroxytryptophan; D006719:Homovanillic Acid