SEROTONIN (Serum) HPLC

SEROTONIN (Serum) For HPLC

INTENDED USE

KR139 Reagent kit for the HPLC determination of Serotonin in serum.

KIT SPECIFICATION

Contents for 100 analyzes

ACCESSORIES

CONTROLS

INTRODUCTION

Serotonin (MW=176,22) is a biological compound that initiates an indolic group as structural domain. It obtains from the bio-transformation (hydroxylation and decarboxylation) of tryptophan. The steps of the serotonin production are: formation of 5-hydroxytryptophan by tryptophan hydroxylase and manifacture of 5-hydroxytryptamine (or serotonin) by aromatic L-aminoacid decarboxylase. Monoamine oxidase causes Serotonin catabolism, forming 5- hydroxyindolacetic acid that is excreted by urine as free molecule or linked with glucuronic acid.

PATHOLOGY

Carcinoid tumors excrete increased quantities of serotonin causing the carcinoid syndrome. The major features of this syndrome are diarrhea, paroxysms of flushing, bronchospasm and cardiac disease. Thanks to measuring the urinary excretion of5- hydroxyindolacetin acid and/or the assay of plasma serotonin, the diagnosis can be established.

EQUIPMENT AND INSTRUMENTAL PARAMETERS

The analysis of Serotonin requires a HPLC including gradient system withHPLC pump, autosampler, autoinjector and FLD detector.

The use of a solvent degasser prevents air bubbles and ensure a stable baseline.

Sampler:Injection volume 25 μl, analytical run time 8 min..

HPLC pump:Flow rate 1.2 ml/min is recommended

Column temperature:Ambient (~25 °C)

FLD detector:Ex:285,Em:345

COLLECTION AND STORAGE OF PATIENT SAMPLES

The sample material is 24 hours serum. Storage life at room temperature is about 1 day, refried (+2 to +8 °C) up to 1 week and frozen below -20 °C up to 6 months.

SAMPLE PREPARATION PROCEDURE

200 µl Serum sample is added to vial with teflon cap

10 µl internal standard is added.

200 µl of preparation is added

Incubate in 10 min at 2-8 C.

Vortex the mix for 30s.

Centrifuge at 10 000 rpm for 10 min.

It is also possible to determine serotonin in serum (citrated, EDTA).

In this case it is necessary to centrifuge at.