Serotonin in Serum/Plasma/Whole Blood - HPLC

Serotonin in Serum/Plasma/Whole Blood - HPLC

Order no.: 3030
100 Tests
  • Serotonin


Easy sample preparation
Short run time
Long column life time

Description

Clinical relevance

Serotonin (5-hydroxytryptamine) is a biogenic amine. In humans, serotonin is involved in a large number of physiological processes, e.g. it acts as a tissue hormone and neurotransmitter in the central nervous system, the enteric nervous system and the cardiovascular system. A large amount of serotonin is found in the chromaffin cells of the gastrointestinal tract where it is part of the gastrointestinal hormone system. When released into the capillary blood, serotonin is almost absorbed by the thrombocytes.

A number of diseases are associated with pathological changes in the serotonin metabolism. For the routine, the diagnosis of carcinoid tumours together with the analysis of 5-hydroxyindoleacetic acid (5-HIAA) is very important.

 

Product advantages

  • Simple and efficient sample preparation
  • Fast results due to short analysis times
  • Long column life times

 

This kit allows for reliable HPLC determination of serotonin in serum/plasma and whole blood as part of routine diagnostic laboratory testing. Easy and fast sample preparation leads to exact and reproducible results. The chromatographic determination requires an isocratic HPLC system with electrochemical detection. Calibrator and quality controls are available separately – they are produced from pure human matrices to prevent matrix-associated measurement variabilities.

Alternatively, Chromsystems offers a kit for determining serotonin in urine (# 4000).

 

Kit content

Mobile Phase, 1000 ml
Serotonin Calibration Standard, 10 ml
Internal Standard, 10 ml
Precipitation Reagent, 10 ml
Reaction vials, 100 pcs.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

100

Parameter

Serotonin

Analysis Time

< 8 min

Limit of quantification

1 µg/l

Linearity

up to 1000 µg/l

Recovery

> 95 %

Intraassay

CV < 2 %

Interassay

CV < 1.5 %

Column temperature

ambient (~25 °C)

Flow rate

1.0 to 1.2 ml/min

Specimen

Serum/Plasma/Whole Blood

Pre-analytic Treatment

Time of sample drawing

  • Fasting
  • Always at the same time of the day, preferably in the morning
  • Blood collection on ice

Serum samples

Care has to be taken to destroy all platelets completely. Recommendation: intensive clotting by the use of serum tubes with clot activator. Centrifuge at least 10 min at 1500 g [Reference: DG Klinische Chemie Mitteilungen 1998; 29(3)].

Plasma samples (platelet-poor plasma)

When measuring serotonin in plasma it is essential to obtain cell-free plasma without destruction of platelets. For this purpose the addition of an anticoagulant is necessary. Recommendation: Centrifuge minimum 15 min at 2000 to 3000 g [Reference: DG Klinische Chemie Mitteilungen 1998; 29(3)].

Platelet concentrate

Centrifuge EDTA-whole blood at low speed (1500 rpm, 15 min). The obtained platelet-rich plasma is centrifuged again at high speed (4000 to 5000 rpm, 5 to 10 min), and the platelet concentrate can be collected from the bottom of the tube.

 

Sample Preparation

Serum specimens

  • Vortex 100 µl serum + 100 µl Internal Standard + 100 µl Precipitation Reagent in a reaction vial for 30 s.
  • Incubate 10 min at +2 to +8 °C.
  • Centrifuge 10 min at approx. 13 000 rpm.
  • Inject 20 µl supernatant into the HPLC system.

Plasma specimens

  • For the determination of serotonin in plasma it is necessary to dilute the Internal Standard 1:10 with distilled water.

Whole blood specimens

  • Dilute whole blood samples 1:1 with distilled water before adding the Internal Standard.

Sample Stability

At +2 to +8 °C up to 12 hours. Freeze the samples if they are not analysed within a period of 12 hours after blood collection.

Injection Volume

20 µl

Additional Info

For the HPLC analysis of serotonin any isocratic HPLC system with electrochemical detector is suitable.

Potential

approx. 400 to 500 mV

Please note

The freely available information on this website, in particular on the sample preparation, are not sufficient to work with our products. Please read instructions and warning notices on products and/or instruction manuals.

Kit Components

Components available separately

Mobile Phase


Order no.: 3031

Internal Standard


Order no.: 3034

Precipitation Reagent


Order no.: 3035

Reaction vials


Order no.: 3006

Controls

Controls available separately