53000 HPLC Kit 1-Hydroxypyrene in Urine

1-Hydroxypyrene in Urine - HPLC

Order no.: 53000
100 Tests
  • 1-Hydroxypyrene

Complete CE-IVD kit, including internal standard and mobile phase
Effective hydrolysis in only 2 hours – with integrated colour indicator for pH adjustment
Analysis time of less than 7 minutes

Description

Clinical relevance

Polycyclic aromatic hydrocarbons (PAHs) are hydrocarbons composed of multiple aromatic rings with potential carcinogenic properties. Physiological monitoring of these substances is therefore essential in occupational health and in particular in industries where workers are potentially exposed to high levels of PAHs, such as in aluminium, iron and steel manufacturing. One of the parent PAHs is pyrene, which undergoes a simple metabolism to 1-hydroxypyrene. 1-Hydroxypyrene and its glucuronide are excreted in urine, making it suitable for monitoring. Pyrene is always present in PAH mixtures and is therefore not only an indicator of pyrene accumulation, but also of an overall exposure to PAHs.

 

Product advantages

  • Complete CE-IVD kit, including internal standard and mobile phase
  • Effective hydrolysis in only 2 hours – with integrated colour indicator for pH adjustment
  • Analysis time of less than 7 minutes
  • Equilibrated, tested and ready to use HPLC column
  • Simple isocratic HPLC system sufficient

 

This Chromsystems kit enables the reliable determination of 1‑hydroxypyrene from urine using an isocratic HPLC run within 7 minutes. The sample preparation is fast and simple and requires only two steps: 1-hydroxypyrene is de-conjugated by an efficient and fast enzymatic hydrolysis, followed by solid phase extraction using Sample Clean Up Columns. The correct pH required for precise analysis is visually verified by a colour changing indicator. The inclusion of an internal standard ensures reproducible and reliable quantitative results. 1-Hydroxypyrene is measured using optimised chromatographic separation with highly sensitive fluorescence detection, ensuring elimination of most potential interferences. The human-matrix based calibration standard and controls ensure precision and accuracy of results.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

100

Parameter

1-Hydroxypyrene

Analysis Time

6.5 min

Limit of quantification

0.1 µg/l

Linearity

up to 20 µg/l

Recovery

92 %

Intraassay

CV = 0.9–2.7 %

Interassay

CV = 2.7–5.7 %

Column temperature

35 °C

Flow rate

1.2 ml/min

Reference ranges

For non-smokers aged 3 to 69 years, the following reference value applies:

1-Hydroxypyrene: 0.5 µg/l (0.3 µg/g creatinine)

Reference value, also known as Biological Reference Value (BRV) = 95th percentile of frequency distribution of urinary 1-hydroxypyrene concentration in the non-occupationally exposed population

Specimen

Urine

Pre-analytic Treatment

Patient specimens can be kept for up to 2 days at room temperature and at +2 to +8 °C. For longer periods of storage (up to 1 year), deep-freeze below –18 °C.

Sample Preparation

ENZYMATIC HYDROLYSIS

  1. Mix (vortex for 2 s) 1 ml urine (sample/control/calibrator) with 50 µl internal standard and 200 µl of the prepared enzyme solution in a reaction vial
  2. Adjust pH if necessary (prepared enzyme solution includes colour indicator)
  3. Close the vial tightly and incubate for at least 2 h at 37 °C

 

SOLID PHASE EXTRACTION

  1. Briefly mix (vortex for 2 s) the hydrolysed urine
  2. Add 1 ml of the hydrolysed urine to a Sample Clean Up Column and draw through by centrifugation (2 min at 700 x g) or suction. Discard effluent.
  3. Centrifuge (1 min at 700 x g) 1000 µl Wash Buffer through the Sample Clean Up Column. Discard effluent.
  4. Centrifuge (2 min at 700 x g) 300 µl Elution Buffer through the Sample Clean Up Column into a light-protected collection vessel (e.g. autosampler vial)
  5. Briefly mix (vortex for 2 s) the eluate
  6. Inject 10 µl eluate into the HPLC system

Sample Stability

The prepared samples can be kept for up to two weeks at +18 to +30 °C, at +2 to +8 °C or below –18 °C.

Injection Volume

10 µl

Additional Info

For the HPLC analysis of 1-hydroxypyrene in urine any isocratic system with fluorescence detector is suitable.

Wavelength

EX 242 nm
EM 388 nm

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.: 53001

Internal Standard


Order no.: 53004

Wash Buffer


Order no.: 53005

Hydrolysis Buffer


Order no.: 53006

Enzyme Solution


Order no.: 53007

Sample Clean Up Columns


Order no.: 53008

Elution Buffer


Order no.: 53009

Calibrators

Calibrators available separately

Controls

Controls available separately

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