Determination of free serotonin and its metabolite 5‐HIAA in blood human samples with consideration to pre‐analytical factors

Yubero-Lahoz, Samanta; Rodríguez, Joaquín; Faura, Anna; Pascual, Julio; Oliveras, Anna; Cao, Higini; Farré, Magı́; Torre, Rafael de la

doi:10.1002/bmc.3192

Cited by 12 publications

(9 citation statements)

References 25 publications

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“…Nonetheless, there is no reliable protocol to measure the free circulating 5HT (known as platelet-poor plasma (PPP) 5HT) [37]. Technical artefacts associated with the sample collection and processing, for example, the anticoagulant and antiaggregant agents used as well as the temperature and sheer forces, could dramatically affect the PPP 5HT values (due to platelet activation and releases of high amount of 5HT), and, thus, it is highly not recommended to make conclusions from PPP data [37, 38]. Those technical difficulties have been reflected in a wide range of PPP 5HT reference values reported for humans [37, 39].…”

Section: Discussionmentioning

confidence: 99%

Platelet Serotonin Levels Are Associated with Plasma Soluble Leptin Receptor Concentrations in Normoglycemic Women

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Suazo

Olmos

et al. 2019

Journal of Diabetes Research

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Most peripheral serotonin (5-hydroxytryptamine (5HT)) is synthetized in the gut with platelets being its main circulating reservoir. 5HT is acting as a hormone in key organs to regulate glucose and lipid metabolism. However, the relation between platelet 5HT levels and traits related to glucose homeostasis and lipid metabolism in humans remains poorly explored. The objectives of this study were (a) to assess the association between platelet 5HT levels and plasma concentration of nonesterified fatty acids (NEFAs) and some adipokines including leptin and its soluble leptin receptor (sOb-R), (b) to assess the association between platelet 5HT levels and anthropometric traits and indexes of insulin secretion/sensitivity derived from oral glucose tolerance test (OGTT), and (c) to evaluate changes in platelet 5HT levels in response to OGTT. In a cross-sectional study, 59 normoglycemic women underwent a standard 2-hour OGTT. Plasma leptin, sOb-R, total and high molecular weight adiponectin, TNFα, and MCP1 were determined by immunoassays. Platelet 5HT levels and NEFAs were measured before and after OGTT. The free leptin index was calculated from leptin and sOb-R measurements. Insulin sensitivity indexes derived from OGTT (HOMA-S and Matsuda ISICOMP) and plasma NEFAs (Adipose-IR, Revised QUICKI) were also calculated. Our data show that among metabolic traits, platelet 5HT levels were associated with plasma sOb-R (r=0.39, p=0.003, corrected p=0.018). Platelet 5HT levels were reduced in response to OGTT (779±237 vs.731±217 ng/109 platelets, p=0.005). In conclusion, platelet 5HT levels are positively associated with plasma sOb-R concentrations and reduced in response to glucose intake possibly indicating a role of peripheral 5HT in leptin-mediated appetite regulation.

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Section: Discussionmentioning

confidence: 99%

Platelet Serotonin Levels Are Associated with Plasma Soluble Leptin Receptor Concentrations in Normoglycemic Women

Cataldo

Suazo

Olmos

et al. 2019

Journal of Diabetes Research

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“…MEPH and metabolites were quantified in plasma and urine with a validated liquid chromatography tandem–mass spectrometry method . Free serotonin (5‐HT) concentrations in human blood were quantified by high‐performance liquid chromatography with electrochemical detection …”

Section: Methodsmentioning

confidence: 99%

Dose‐Response Pharmacological Study of Mephedrone and Its Metabolites: Pharmacokinetics, Serotoninergic Effects, and Impact of CYP2D6 Genetic Variation

Olesti

Farré

Carbó

et al. 2019

Clin Pharma and Therapeutics

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Mephedrone (MEPH), the most widely consumed synthetic cathinone, has been associated with acute toxicity episodes. The aim of this report was to study its metabolic disposition and the impact of genetic variation of CYP2D6 on MEPH metabolism, in a dose range compatible with its recreational use. A randomized, crossover, phase I clinical trial was performed. Subjects received 50 and 100 mg (n = 3) and 150 and 200 mg (n = 6) of mephedrone and were genetically and phenotypically characterized for the CYP2D6 allelic variation. Our results showed a linear kinetics of mephedrone at the dose range assayed: plasma concentrations, cardiovascular and subjective effects, and blood serotonin concentrations all correlated in a dose‐dependent manner. Mephedrone metabolic disposition is mediated by CYP2D6. Mephedrone pharmacology presented a linear dose‐dependence within the range of doses tested. The metabolism of mephedrone by CYP2D6 implies that recreational users with no or low CYP2D6 functionality are exposed to unwanted acute toxicity episodes.

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“…This is the first study to compare different centrifugation conditions on measured serotonin concentrations in PPP from healthy volunteers, without varying blood sampling techniques and type of anticoagulant added to identify the most suitable pre-analytical conditions for PPP serotonin quantification by ELISA. Three previously reported methods using double-stage centrifugation [[7], [8], [9]] and three alternative singel-stage centrifugation methods using separation gel or heat shock treatment for preparing PPP were compared. This work demonstrates that a combination of a milder first centrifugation of gel-free EDTA-tubes to separate blood cells (100 or 200× g ) followed by a stronger centrifugation of the supernatant (4500 or 14500× g ) resulted in the lowest measured serotonin concentration in PPP.…”

Section: Discussionmentioning

confidence: 99%

Comparison of pre-analytical conditions for quantification of serotonin in platelet-poor plasma

Volkmann

Brønstad

Fiskerstrand

et al. 2019

Practical Laboratory Medicine

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BackgroundReported concentrations of serotonin in platelet-poor plasma (PPP) in healthy subjects vary widely due to different pre-analytical procedures.AimTo examine how different pre-analytical conditions affect the measured concentration of serotonin in PPP.MethodSix pre-analytical protocols were compared for preparation of PPP from EDTA whole blood for quantification of serotonin from nine healthy individuals. Three combinations of centrifugation with a mild centrifugation of gel-free EDTA tubes followed by a stronger centrifugation were compared to single-stage centrifugation of EDTA tubes with separator gel and heat shock treatment of blood prior to centrifugation. All samples were analysed using the same enzyme linked immunosorbent assay (ELISA) method.ResultsFindings show that two consecutive centrifugations; first a mild centrifugation at 100 or 200×g followed by centrifugation at 4500 or 14500×g resulted in the lowest serotonin concentration in PPP.ConclusionTwo successive centrifugations to produce PPP for serotonin analysis; first a mild centrifugation to avoid mechanical stress on the platelets, and next a stronger centrifugation to remove platelets, is superior to the use of gel tubes and heat shock treatment.

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Determination of free serotonin and its metabolite 5‐HIAA in blood human samples with consideration to pre‐analytical factors

Cited by 12 publications

References 25 publications

Platelet Serotonin Levels Are Associated with Plasma Soluble Leptin Receptor Concentrations in Normoglycemic Women

Platelet Serotonin Levels Are Associated with Plasma Soluble Leptin Receptor Concentrations in Normoglycemic Women

Dose‐Response Pharmacological Study of Mephedrone and Its Metabolites: Pharmacokinetics, Serotoninergic Effects, and Impact of CYP2D6 Genetic Variation

Comparison of pre-analytical conditions for quantification of serotonin in platelet-poor plasma

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