2,5 HEXANDIONE (Urine)

2,5 -hexandione (Urine) For HPLC


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


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

 

ACCESSORIES

 

KR1490

1 adet

HPLC column for the analysis of         2,5 -hexanedıone in urine.

KR1499

100 adet

Autosampler vial with teflon caps


CONTROLS 

KR1497

0.5 ml x5

Urine Control, Level I

KR1498

0.5 ml x5

Urine Control, Level II

 
 

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. 
 

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.


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.

 
 

 

© KREON MEDİKAL SİSTEMLER SAN.VE TİC.A.Ş. 2017