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


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.


Technical Data

Method of Analysis


Number of Tests



t,t-Muconic acid

Analysis Time

< 17 min

Limit of quantification

0.02 mg/l


0.02 up to 10 mg/l


93–98 %


CV < 1.5–1.7 %


CV < 2.9–3.4 %


Exposition equivalents for carcinogenic substances = EKA

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



Pre-analytic Treatment

Patient samples are stable up to 3 days at ambient temperature, at +2 to +8 °C up to 2 weeks and below -18 °C up to 3 months.

Sample Preparation

  • Pipette 250 µl urine +750 µl Internal Standard into a reaction vial, mix briefly (vortex).
  • Apply to the Sample Clean Up Column, centrifuge (1 min, ca. 250 x g), draw through entirely.
  • Apply 1 ml Wash Buffer 1 on the Sample Clean Up Column, centrifuge (1 min, ca. 250 x g). 
  • 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. 250 x g) and collect the eluate.
  • Mix eluate briefly, inject 20 µl into the HPLC system.

Sample Stability

The prepared samples are stable at ambient temperature for 4 days, at +2 to +8 °C or below -18 °C up to 2 weeks.

Additional Info

Any HPLC system with UV detector is suitable.

Injection Volume

20 µl

Flow rate

1.1–1.3 ml/min



Column temperature



264 nm

Autosampler rinsing solution

water/methanol – 90/10


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 available separately


Controls available separately