Serotonin in Urine - HPLC

Serotonin in Urine - HPLC

Order no.: 4000
100 Tests
  • Serotonin

Now easier pH adjustment with pH indicator
Easy sample preparation
Short analysis 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 urine 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 serum/plasma and whole blood (# 3030).

  

Kit content

Mobile Phase, 1000 ml
Serotonin Calibration Standard, 10 ml
Internal Standard, 10 ml
Neutralisation Buffer, 300 ml
Wash Buffer, 300 ml
Elution Buffer, 500 ml
Sample Clean Up Columns, 100 pcs.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

100

Parameter

Serotonin

Analysis Time

aproxx. 8 min

Limit of quantification

5 µg/l

Linearity

up to 1000 µg/l

Recovery

~ 99 %

Intraassay

CV < 2 %

Interassay

CV < 3 %

Column temperature

ambient (~25 °C)

Flow rate

0.8-1.3 ml/min

Specimen

24 h urine (containing 10 ml 25 % HCl) or spontaneous urine (refer data to urine creatinine).

Pre-analytic Treatment

Note:

The following food and drugs can falsify the results of the determination: pineapples, eggplants, avocados, bananas, redcurrants, kiwis, melons, mirabelles, gooseberries, tomatoes, walnuts, plums, methocarbamol, mephenesin, guaifenesin, paracetamol, salicylic acid.

Sample Preparation

  • Mix 4 ml urine with 100 µl Internal Standard and 6 ml Neutralisation Buffer.
  • Add 2 N NaOH until the colour changes from yellow to green or green-grey (see figure).

 

 

  • Resuspend Sample Clean Up Column and allow buffer to flow out.
  • Apply entire urine dilution to the column, discard effluent.
  • Rinse the column with 1 volume of water, discard effluent.
  • Apply 3 ml Wash Buffer, discard effluent.
  • Apply 20 ml Elution Buffer, collect eluate, mix well.
  • Inject 20 to 50 µl eluate into the HPLC system.

Sample Stability

At +2 to +8 °C at least 5 days. Aliquot and freeze for longer storage.

Injection Volume

20-50 µl

Additional Info

For the HPLC analysis of serotonin in urine 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.: 4004

Neutralisation Buffer


Order no.: 4055

Wash Buffer


Order no.: 4006

Elution Buffer


Order no.: 4007

Controls

Controls available separately