Abstract

Serotonin, an important neurotransmitter, is produced mainly in intestines, and serotonin levels in feces can be an indicator of the intestinal environment. Human feces, however, contain a large amount of contaminants, which vary widely owing to food contents and the intestinal environment, and these contaminants would be expected to interfere with the determination of serotonin levels in human feces. To remove these contaminants and determine serotonin levels, we developed a new method using solid phase extraction (SPE) and column-switching LC-MS/MS. Serotonin, labeled with a stable isotope, was added to human feces samples prior to SPE as an internal standard to correct for individual differences in matrix effects. The recovery rate for SPE was 55.9–81.0% (intraday) and 56.5–78.1% (interday) for feces from two subjects. We analyzed 220 fecal samples from 96 subjects including 76 pregnant and post-delivery women. The endogenous serotonin content per unit weight of dried feces was 0.09–14.13 ng/mg for pregnant and post-delivery women and 0.30–9.93 ng/mg for the remaining subjects.

Highlights

  • In recent years, the gut has attracted attention as a “second brain.” Investigations of bacterial ora in the human gut using the 16S rRNA-based approach to examine gut microbiota in feces1,2) have become popular in studies of relations between gut microbiota to diseases,3) diet,4) age and geography,5) cognition in infants,6) etc

  • A quantitative analytical method using a sandwich enzyme-linked immunosorbent assay for α-defensins, which are produced by Paneth cells and are thought to regulate the growth of gut microbiota, has been developed,10) and studies concerning the involvement of α-defensins in diseases and health have been reported.11,12)

  • FecaL SerotonIn AnaLYsIs is accompanied by an impaired release of serotonin.16) Serotonin has been implicated in depression, and it has been reported that blood serotonin levels are reduced in patients su ering from major depressive disorder with melancholia.17) It has been reported that blood serotonin levels are correlated with tension/anxiety in pregnant and post-delivery women who are at high risk for mood disorders18) and are generally prone to constipation.19)

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Summary

Original Article

Analysis of Serotonin in Human Feces Using Solid Phase Extraction and Column-Switching LC-MS/MS. Communicated by Yoshinao Wada, Osaka Women’s and Children’s Hospital, Japan (received January 27, 2020, accepted February 10, 2020). Contain a large amount of contaminants, which vary widely owing to food contents and the intestinal environment, and these contaminants would be expected to interfere with the determination of serotonin levels in human feces. To remove these contaminants and determine serotonin levels, we developed a new method using solid phase extraction (SPE) and column-switching LC-MS/MS. E endogenous serotonin content per unit weight of dried feces was 0.09–14.13 ng/mg for pregnant and post-delivery women and 0.30–9.93 ng/mg for the remaining subjects

INTRODUCTION
FecaL SerotonIn AnaLYsIs
Chemicals and materials
Fecal samples
Fecal sample preparation and solid phase extraction
LC condition
RESULTS AND DISCUSSION
Full Text
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