trans, trans -MUCONIC ACID (Urine) HPLC

trans, trans-MUCONIC ACID For HPLC

INTENDED USE

KR148 Reagent kit for the HPLC determination of trans, trans-Muconic Acid in urine.

KIT SPECIFICATION

Contents for 100 analyzes

ACCESSORIES

CONTROLS

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 metabolized 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.

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

COLLECTION AND STORAGE OF PATIENT SAMPLES

The sample material is 24 hours urine. Storage life at room temperature is about 1 day, refried (+2 to +8 °C) up to 1 week and frozen below -20 °C up to 6 months.

SAMPLE PREPARATION PROCEDURE

1 ml urine, 150 µl Internal standard, 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.