SEROTONIN (Serum) HPLC

SEROTONIN (Serum) For HPLC

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INTENDED USE

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

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KIT SPECIFICATION

Contents for 100 analyses

Cat no	Quantity	Reagents
KR1391	1000 mlx1	Mobile Phase
KR1392	100 mlx1	İnternal Standart
KR1393	0.5 mlx5	serum Calibrator

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ACCESSORIES

KR1395	1 adet	HPLC column for the analysis of Serotonin in urine.
KR1399	100 adet	Autosampler vial with teflon caps

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CONTROLS

KR1397	0.5 ml x5	Urine Control, Level I
KR1398	0.5 ml x5	Urine Control, Level II

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INTRODUCTION

Serotonin (M.W.=176,22) is a biological compound that initiate 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.

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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, refrigerated (+2 to +8 °C) up to 1 week and frozen below -20 °C up to 6 months.

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SAMPLE PREPARATION PROCEDURE

200 µl Serum sample is added to vial with teflon cap

10 µl internal standard is added.

200 µl 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.