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Published:
Journal of Analytical Toxicology,
ISSN 0146-4760,
Volume 27, Number 7, October 2003,
pp. 440-444
A Liquid Chromatographic-Tandem Mass Spectrometric
Method for the Analysis of Serotonin and Related Indoles in Human Whole Blood
Jonathan P. Danaceau[1], George M. Anderson[2],
William M. McMahon[3], and Dennis J. Crouch[1]
[1]Center for Human Toxicology, Department of Pharmacology and Toxicology,
University of Utah, 20 South 2030 East, Room 490, Salt Lake City, Utah 84112;
[2]Departments
of Laboratory Medicine and Child Psychiatry, Yale University School of Medicine,
New Haven, Connecticut 06510; and
[3]Department of Psychiatry, University of
Utah, School of Medicine, 30 North 1900 East, Room 5R110, Salt Lake City, Utah
84112
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 µg/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. Reproduction
of editorial content of this journal is prohibited without publisher’s
permission.
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