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 analyses
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Cat no
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Quantity
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Reagents
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KR1391
|
1000 mlx1
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Mobile Phase
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KR1392
|
100 mlx1
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İnternal Standart
|
|
KR1393
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0.5 mlx5
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serum Calibrator
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ACCESSORIES
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KR1395
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1 adet
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HPLC column for the analysis of Serotonin in urine.
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|
KR1399
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100 adet
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Autosampler vial with teflon caps
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CONTROLS
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KR1397
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0.5 ml x5
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Urine Control, Level I
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|
KR1398
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0.5 ml x5
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Urine Control, Level II
|
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.
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.
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.
