44000 HPLC kit porphyrins urine

Porphyrins in Urine - HPLC

Order no.: 44000
100 Tests
  • Coproporphyrin I
  • Coproporphyrin III
  • Heptacarboxyporphyrin
  • Hexacarboxyporphyrin
  • Pentacarboxyporphyrin
  • Uroporphyrin

Easy sample preparation
Specifically developed internal standard
Stable standards and controls

Description

Clinical relevance

Porphyrins are precursors of haem and usually only occur in urine in negligible amounts. However, disorders in haem biosynthesis lead to the syndrome of porphyria with increased formation and excretion of porphyrins. Porphyrias can be hereditary-related or acquired through exogenous influences such as alcohol consumption or lead poisoning. Different enzymes of the haem biosynthesis can be deficient, thus, several different porphyrias can be distinguished. Since these share some common symptoms, chromatographic measurement of the porphyrins is the only way to enable an exact diagnosis.

 

Product advantages

  • Easy sample preparation
  • Custom-made internal standard
  • Stable standards and controls

 

This kit is designed for the fast and reliable determination of all porphyrins in urine that are relevant to differential diagnosis. Sample preparation requires only the addition of an internal standard and a precipitation reagent, followed by the centrifugation of the urine. The internal standard elutes in the middle of the chromatogram, making it easier to interpret the results of the measurement.

 

Kit content

Mobile Phase A, 1000 ml
Mobile Phase B, 1000 ml
Porphyrins Urine Calibration Standard (lyoph.), 5 x 2 ml
Internal Standard, 5 ml
Stabilisation Reagent, 5 ml
Priming Solution, 5 ml
Reaction vials, amber coloured (light protection), 100 pcs.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

100

Parameter

Coproporphyrin I, Coproporphyrin III, Heptacarboxyporphyrin, Hexacarboxyporphyrin, Pentacarboxyporphyrin, Uroporphyrin

Analysis Time

approx. 15 min + 5 min equilibration

Limit of quantification

1 µg/l

Linearity

up to 5000 µg/l

Recovery

101-112 %

Intraassay

CV < 3 %

Interassay

CV < 5 %

Column temperature

ambient (~25 °C)

Flow rate

1.2 ml/min

Specimen

Urine

Sample Preparation

  • Pipette 500 µl urine into a light-protected reaction vial.
  • Add 50 µl Stabilisation Reagent and 50 µl Priming Solution, vortex briefly; incubate 10 min at ambient temperature.
  • Add 50 µl Internal Standard, vortex briefly.
  • Centrifuge 10 min at 13000 rpm.
  • Inject 25 µl supernatant into the HPLC system.

Sample Stability

At ambient temperature: approx. 24 h, 2 days at +2 to +8 °C and pH 6 to 7, at -20 °C up to 1 month. Keep specimens cool and light protected during transport. Stabilize acidic urines by sodium hydrogen carbonate (attention: foaming).

Injection Volume

25 µl

Additional Info

For the Chromsystems HPLC analysis of porphyrins in urine any binary HPLC gradient system with fluorescence detector is suitable.

Wavelength

EX 405 nm
EM 620 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 A


Order no.: 44001

Mobile Phase B


Order no.: 44002

Internal Standard


Order no.: 44004

Stabilisation Reagent


Order no.: 44005

Priming Solution


Order no.: 44006

Reaction vials


Order no.: 33005

Calibrators

Calibrators available separately

Controls

Controls available separately

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