47000 HPLC kit  t,t-muconic acid urine

trans,trans-Muconic Acid in Urine - HPLC

Order no.: 47000
100 Tests
  • t,t-Muconic acid

Occupational medicine
Low limit of quantification
Internal standard included
Easy, fast, reliable

Description

Clinical relevance

t,t-Muconic acid is an end product in the metabolism of benzene and is used as a biomarker for occupational monitoring of benzene exposure. Benzene is used in many different ways as a basic chemical for synthetic purposes and as a solvent. It is a toxic compound and has carcinogenic properties. It causes damage to the stem cells of bone marrow, which can induce leukaemia and lymphomas. Furthermore, t,t-Muconic acid is also important for biological monitoring of benzene because it also allows indirect verification of t,t-muconaldehyde, a metabolite that is also responsible for the toxicity of benzene.

Exposure equivalents for carcinogenic working materials (EKA values) can be used to assess exposure. These can be equated with the concentrations of the hazardous substance in the air and then compared with the technical standard concentration (TRK value).

 

Product advantages

  • Low limit of quantification
  • Custom-made internal standard
  • An easy, fast, and reliable method

 

This Chromsystems kit allows for the reliable and fast chromatographic determination of t,t-muconic acid using an isocratic HPLC system with UV detection. The sample preparation is based on efficient and selective purification with solid phase extraction. This includes the addition of a special internal standard to the sample with simultaneous pH adjustment and subsequent transfer to the SPE column. Sequenced washing steps are then performed to eliminate interfering substances. This ensures that the method maintains high levels of precision and reliability when quantifying the analyte. Finally, the t,t-muconic acid is eluted and stabilised simultaneously. This analysis method is very sensitive and allows the determination of concentrations upwards of 20 µg/l.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

100

Parameter

t,t-Muconic acid

Analysis Time

<17 min

Limit of quantification

0.02 mg/l

Linearity

0.02 up to 10 mg/l

Recovery

93-98 %

Intraassay

CV < 1.5 - 1.7 %

Interassay

CV < 2.9-3.4 %

Gradient

isocratic

Information

Exposition equivalents for carcinogenic substances

t,t-Muconic acid: 2 mg/l urine (correlates to an exposition of 1 ppm (3.25 mg/m3) benzene in the air)

Column temperature

35 °C (33-37°C)

Flow rate

1.2 ml/min (1.1-1.3 ml/min)

BEI values

1 ppm (3.25 mg/m3) Benzene (Occupational exposure limit
in air)

Specimen

urine

Sample Preparation

  • 250 µl urine +750 µl Internal Standard (vortex).
  • Apply to sample clean up column, centrifuge (1 min, ca. 250xg), draw through entirely.
  • Apply 1 ml Wash Buffer 1 on the sample clean up column, centrifuge (1 min, ca. 250xg).
  • Repeat the wash step using Wash Buffer  2 and Wash Buffer 3.
  • Add 1 ml Elution Buffer on the sample clean up column, centrifuge (1 min, ca. 250xg) and collect the eluate.
  • Mix eluate briefly, inject 20 µl into the HPLC system.

Sample Stability

Storage life is up to 3 days at ambient temperature, at +2 to +8 °C up to 2 weeks. For longer storage deep-freeze below –18 °C (maximum 3 months).

Injection Volume

20 µl

Flow

1.2 ml/min (1.1-1.3 ml/min)

Additional Info

HPLC system with UV detector.

Wavelength

264 nm

Autosampler rinsing solution

water/methanol = 90/10

Pressure

The backpressure of a new column at a flow rate of 1.2 ml/min and temperature of 35 °C is about 120 bar and may increase with column age and/or use

Please note

The freely available information on this website, in particular on the sample preparation, are not sufficient to work with our products. Please read instructions and warning notices on products and/or instruction manuals.

Kit Components

Components available separately

Mobile Phase


Order no.: 47001

Internal Standard


Order no.: 47004

Wash Buffer 1


Order no.: 47005

Wash Buffer 2


Order no.: 47006

Wash Buffer 3


Order no.: 47007

Sample Clean Up Columns


Order no.: 47008

Elution Buffer


Order no.: 47009

Reaction vials


Order no.: 33005

Calibrators

Calibrators available separately

Controls

Controls available separately

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