Porphyrins in Urine - HPLC

Order No.: 44000, for 100 tests
Parameters:
Coproporphyrin I, Coproporphyrin III, Heptacarboxyporphyrin, Hexacarboxyporphyrin, Pentacarboxyporphyrin, Uroporphyrin

Easy sample preparation
Specifically developed internal standard
Stable standards and controls

Coproporphyrin I

Coproporphyrin III

Heptacarboxyporphyrin

Hexacarboxyporphyrin

Pentacarboxyporphyrin

Uroporphyrin

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.

More Information
Number of Tests 100
Limit of quantification 3 µg/l
Linearity up to 5000 µg/l
Recovery 100–106 %
Intraassay CV ≤ 3.9 %
Interassay CV ≤ 6 %
Analysis Time 15 min + approx. 5 min reequilibration
Sample Preparation
  • Pipette 500 µl urine into a light-protected reaction vial.
  • Add 50 µl Stabilisation Reagent and 50 µl Priming Solution, mix briefly (vortex); incubate 10 min at ambient temperature.
  • Add 50 µl Internal Standard, mix briefly (vortex).
  • Centrifuge 10 min at 9000 x g.
  • Inject 25 µl supernatant into the HPLC system.
Sample Stability The prepared samples are stable at ambient temperature at least for 24 h, at +2 to +8 °C or below -18 °C up to 6 days.
Specimen Urine
Pre-analytic Treatment Keep patient specimens cool and light protected during transport. At a pH value of 6 to 7 storage life is up to 2 days at +2 to +8 °C. For longer storage (up to 1 month) keep samples below -18 °C. Stabilize acidic urines by sodium hydrogen carbonate (attention: foaming). Avoid repeated freeze/thaw cycles!
Injection Volume 25 µl
Flow rate 1.2 ml/min
Column temperature ambient (+18 to +30 °C)
Wavelength

EX 405 nm
EM 620 nm

Gradient binary
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.
Method of Analysis HPLC
Parameter Coproporphyrin I, Coproporphyrin III, Heptacarboxyporphyrin, Hexacarboxyporphyrin, Pentacarboxyporphyrin, Uroporphyrin
Complete Specification
Limit of quantification:
Linearity: up to 5000 µg/l
Intra-assay:
Inter-assay:
Recovery:
Run time: approx. 15 min + approx. 5 min equilibration
Stability of specimens: : approx. 24 h (ambient temperature)

HPLC parameters

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

Injection volume: 25 µl
Flow rate: 1.2 ml/min
Wavelengths: EX 405 nm
EM 620 nm
Column temperature: ambient (~25 °C)
Additional Info

Any binary HPLC gradient system with fluorescence detector is suitable.

  1. 44001 HPLC mobile phase A porphyrins urine
    Mobile Phase A
    Order no.: 44001
    Porphyrins in Urine - HPLC
    Details
  2. 44002 HPLC mobile phase B porphyrins urine
    Mobile Phase B
    Order no.: 44002
    Porphyrins in Urine - HPLC
    Details
  3. 44003 HPLC urine calibration standard porphyrins
    Porphyrins Urine Calibration Standard
    Order no.: 44003
    Porphyrins in Urine - HPLC
    Details
  4. 44004 HPLC internal standard porphyrins urine
    Internal Standard
    Order no.: 44004
    Porphyrins in Urine - HPLC
    Details
  5. 44005 HPLC stabilisation reagent porphyrins urine
    Stabilisation Reagent
    Order no.: 44005
    Porphyrins in Urine - HPLC
    Details
  6. 44006 HPLC priming solution porphyrins urine
    Priming Solution
    Order no.: 44006
    Porphyrins in Urine - HPLC
    Details
  7. Reaction Vials, 1.5 ml
    Reaction Vials, 1.5 ml
    Order no.: 33005
    Amber colour, light protected
    Details
  1. 44003 HPLC urine calibration standard porphyrins
    Porphyrins Urine Calibration Standard
    Order no.: 44003
    Porphyrins in Urine - HPLC
    Details
  1. 0144 0145 0146 HPLC porphyrins urine controls
    Porphyrins Urine Controls
    Order no.: 0144/0145/0146
    Details
  1. PEEK-encased Prefilter, 2 µm
    Details
  2. PEEK Prefilter Housing
    PEEK Prefilter Housing
    Order no.: 15010
    Details
  3. 44100 HPLC column porphyrins urine
    HPLC Column Porphyrins in Urine
    Order no.: 44100
    Porphyrins in Urine - HPLC
    Details
  4. PEEK-encased Prefilter, 5 µm
    Details
  5. Precolumn Cartridge Holder 4/10
    Details
  6. Precolumn Cartridge 4/10
    Precolumn Cartridge 4/10
    Order no.: 18044
    Porphyrins in Urine - HPLC
    Details
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