2,5 HEXANDIONE (Urine)

2,5 -hexandione (Urine) For HPLC

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INTENDED USE

KR149 Reagent kit for the HPLC determination of 2,5 -hexanedione in urine.

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KIT SPECIFICATION

Contents for 100 analyses

Cat no	Quantity	Reagents
KR1491	1000 mlx1	Mobile Phase
KR1492	100 mlx1	İnternal Standart
KR1493	0.5 mlx5	Urine Calibrator
KR1494	5.0mlx4	Derivatization solution
KR1495	2.5mlx1	Hydrolysis Reagent

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ACCESSORIES

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KR1490	1 adet	HPLC column for the analysis of 2,5 -hexanedıone in urine.
KR1499	100 adet	Autosampler vial with teflon caps

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CONTROLS

KR1497	0.5 ml x5	Urine Control, Level I
KR1498	0.5 ml x5	Urine Control, Level II

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INTRODUCTION

2,5 Hexandione is composed of an aliphatic diketone. The product of 2,5 Hexandione is transformation of the hexane. Glues which is used in the production of shoes or leathers contains hexane. Major way of absorption, even if ingestion is inhalation. Eye and skin contact can also happen. It can be metabolised so, 2,5 hexandione determination is decisive to determine the risk owing to the exposition to hexane.

From a clinical point of view 2,5 hexandione is regarded one of the most significant metabolites of the hexane in the improvement of polyneuropathies in leather and shoemakers.

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CHEMICAL ANALYSIS BY HPLC

2,5 Hexandione (P.M.=114.14) is an aliphatic volatile diketone. Because of its molecular structure, it is not directly revealed by UV determination.

This method allows to perform the HPLC urinary determination of 2,5 hexandione avoiding the extraction and concentration steps.

After acid hydrolysis, 2,5 hexandione is pullulated with a certain reagent for ketones and then directly injected into the HPLC.

EQUIPMENT AND INSTRUMENTAL PARAMETERS

The analysis of 2,5 -hexandione requires a HPLC system including isocratic pump, autosampler and UV detector.

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

Sampler: Injection volume 50 μl, analytical run time 20 min..

HPLC pump: Flow rate 1.2 ml/min is recommended

Column temperature: Ambient (~25 °C)

UV detector: 334 nm

COLLECTION AND STORAGE OF PATIENT SAMPLES

Before and soon after work shift, collect 10 ml of urine in a suitable test tube.

The sample is stable up to 4 weeks at - 20 °C.

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SAMPLE PREPARATION PROCEDURE

200 µl urine sample is added to vial with teflon cap

20 µl hydrolysis reagent is added.

Incubation is carried out at 100°C for 60 min.

Vortex the mix for 10s and then Centrifuge at 4 000 rpm for 5 min.

200 µl hydrolysis reagent is added

10 µl internal standard is added.

Incubation is carried

Samples prepared as above can be stored at ambient temperature for about 24 hours, at +2 to +8 °C up to 36 h, frozen below -18 °C up to 3 months.