Indian Journal of Clinical Biochemistry

, Volume 34, Issue 1, pp 82–88 | Cite as

A Simple Method for Quantification of Five Urinary Porphyrins, Porphobilinogen and 5-Aminolevulinic Acid, Using Liquid Chromatography Tandem Mass Spectrometry

  • Ozlem DoganEmail author
  • Muhittin A. Serdar
  • Koza Murat
  • Cigdem Sonmez
  • Emre İspir
  • Mustafa Serteser
  • Ibrahim Unsal
Original Research Article


Analysis of porphyrins and 5-aminolevulinic acid (ALA), porphobilinogen (PBG) in physiological liquids is required for diagnosis and follow-up of porphyrias. High performance liquid chromatography (HPLC) and liquid chromatography tandem mass spectrometry (LC–MS) methods with higher specificity and sensitivity have been developed. The major disadvantage of those methods is that they require longer extraction times due to their matrix effects. The present study suggests a simple, fast, sensitive, and specific assay for determination of Coproporphyrin, 5-carboxylporphyrin, 6-carboxylporphyrin, 7-carboxylporphyrin, Uroporphyrin I and ALA, PBG in urine sample by direct injection without sample pre-treatment using LC–MS. For the purposes of the present study LC–MS device was set to multiple reaction monitoring (MRM) and positive ion mode. Porphyrins and ALA, porphobilinogen were characterized by their MS/MS product ion, spectra. ALA, PBG and 5 porphyrins were detected simultaneously. Limit of detection for Coproporphyrin, 5-carboxylporphyrin, 6-carboxylporphyrin, 7-carboxylporphyrin, Uroporphyrin I were 2 nmol/L, where it was 5 μmol/L for ALA and 2 μmol/L for porphobilinogen. The present study suggests that the present method is very effective compared to many other available methods for it does not require pre-treatment, provides simultaneous results of ALA, PBG and 5 porphyrins quantitatively in a shorter span of time, and has suitable sensitivity and selectivity. LC–MS technique was used clinically for the determination of urine porphyrin levels.


Porphyrins LC–MS ALA Porphobilinogen 


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Copyright information

© Association of Clinical Biochemists of India 2017

Authors and Affiliations

  1. 1.Department of Medical BiochemistryAnkara University Faculty of MedicineAnkaraTurkey
  2. 2.Department of Medical BiochemistryAcibadem UniversityIstanbulTurkey
  3. 3.Ataturk Chest Disease and Thoracic Surgery Education and Research HospitalAnkaraTurkey
  4. 4.Department of BiochemistryDr Abdurrahman Yurtarslan Education and Research HospitalAnkaraTurkey
  5. 5.Department of BiochemistryGulhane Military Medical HospitalAnkaraTurkey

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