A liquid chromatographic-tandem mass spectrometric method for the analysis of serotonin and related indoles in human whole blood.
This paper describes a high-performance liquid chromatographic-tandem mass spectrometric (HPLC-MS-MS) method for the determination of serotonin (5-HT) and the related indoles tryptophan (TRP), 5-hydroxyindoleacetic acid (5-HIAA), indole-3-acetic acid (IAA), and indole-3-proprionic acid (IPA) in whole blood. Ionization was achieved by atmospheric pressure chemical ionization (APCI). The method uses a solvent gradient to achieve baseline separation of all analytes. Linearity was established for all components from 10 to 500 ng/mL, with the exception of TRP, which was calibrated from 0.4 to 20 microg/mL. Limits of quantitation were established that were well below the normal endogenous concentrations of 5-HT and its related indoles. Recoveries of 5-HT, TRP, 5-HIAA, IAA, and IPA were 95%, 91%, 98%, 79%, and 65%, respectively, from whole blood matrix and varied less than 2%. Serotonin, TRP, and IAA results were verified by cross-validation with an external laboratory using a previously validated HPLC method with fluorometric detection. A regression analysis of the two data sets resulted in correlation coefficients of 0.981, 0.964, and 0.987 for 5-HT, TRP, and IAA, respectively.
Danaceau
Jonathan P
JP
Center for Human Toxicology, Department of Pharmacology and Toxicology, University of Utah, 20 South 2030 East, Room 490, Salt Lake City, Utah 84112, USA.
Anderson
George M
GM
McMahon
William M
WM
Crouch
Dennis J
DJ
D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards; D006801:Humans; D007211:Indoles / Q000097:blood; D013058:Mass Spectrometry / Q000379:methods* / Q000592:standards; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012701:Serotonin / Q000097:blood*
D007211:Indoles; D012701:Serotonin