trans, trans -MUCONIC ACID (Urine) HPLC

trans, trans-MUCONIC ACID For HPLC

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INTENDED USE

KR148 Reagent kit for the HPLC determination of trans, trans-Muconıc Acıd in urine.

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KIT SPECIFICATION

Contents for 100 analyses

Cat no	Quantity	Reagents
KR1481	1000 mlx1	Mobile Phase
KR1482	15 mlx1	İnternal Standart
KR1483	0.5 mlx5	Urine Calibrator
KR1484	100 mlx1	Dilution Buffer
KR1485	100 mlx3	Washing Buffer
KR1486	100 mlx3	Elution Reagent

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ACCESSORIES

KR1487	1 adet	HPLC column for the analysis of trans, trans-Muconıc Acıd in urine.
KR1488	100 adet	Autosampler vial with teflon caps

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CONTROLS

KR1489	0.5 ml x5	Urine Control, Level I
KR1490	0.5 ml x5	Urine Control, Level II

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INTRODUCTION

The measuring of trans,trans-MUCONIC Acid is advised as a biological and reliable marker for observing workers subject to benzene.
Benzene is an industrial chemical product that often appears in mineral oils following combustion processes. Because of this reason, there is a high risk of environmental pollution . Its levels in the air can easily remain due to emissions from coal and oil burning, benzene leak and storage operations, exhaust fumes from cars, evaporation from gasoline service stations, and use of industrial solvents. Benzene may be produced by smoking in air since tobacco includes high level of benzene.
Breathing benzene for long periods may have different harmful effects. In fact, it is haematoxic and has been defined as a carcinogen, can bring to anaemia, excessive bleeding and it can have harm for the immune system and the reproductive organs.
Benzene is primarily metabolised by a cythocrome P-450 ( CYP-450 ) to phenolic compounds. The range of metabolic conversion of benzene to t,t-MA is about 2 – 25 % and dependent on the benzene exposure level.

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EQUIPMENT AND INSTRUMENTAL PARAMETERS

The analysis of trans,trans-MUCONIC ACID 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 10 min..

HPLC pump: Flow rate 1.0 ml/min is recommended

Column temperature: Ambient (~25 °C)

UV detector: 264 nm

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COLLECTION AND STORAGE OF PATIENT SAMPLES

The sample material is 24 hours urine. 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

1 ml urine, 150 µl Internal standart, 1 ml Dilution Buffer

Transfer the whole sample to a conditioned sample preparation column

Wash with 2ml Washing Buffer

Elute with1ml Elution Reagent

Inject 50 µl

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.