Dogan Ozlem, Serdar Muhittin A, Murat Koza, Sonmez Cigdem, İspir Emre, Serteser Mustafa, Unsal Ibrahim
1Department of Medical Biochemistry, Ankara University Faculty of Medicine, Ankara, Turkey.
2Department of Medical Biochemistry, Acibadem University, Istanbul, Turkey.
Indian J Clin Biochem. 2019 Jan;34(1):82-88. doi: 10.1007/s12291-017-0716-8. Epub 2017 Nov 10.
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.
为了诊断和随访卟啉病，需要对生理液体中的卟啉、5-氨基酮戊酸（ALA）和卟胆原（PBG）进行分析。已经开发出具有更高特异性和灵敏度的高效液相色谱（HPLC）和液相色谱串联质谱（LC-MS）方法。这些方法的主要缺点是由于基质效应，它们需要更长的提取时间。本研究提出了一种简单、快速、灵敏且特异的检测方法，通过直接进样且无需样品预处理，使用LC-MS测定尿样中的粪卟啉、5-羧基卟啉、6-羧基卟啉、7-羧基卟啉、尿卟啉I以及ALA和PBG。为了本研究的目的，将LC-MS设备设置为多反应监测（MRM）和正离子模式。卟啉以及ALA和卟胆原通过它们的MS/MS产物离子光谱进行表征。同时检测到了ALA、PBG和5种卟啉。粪卟啉、5-羧基卟啉、6-羧基卟啉、7-羧基卟啉、尿卟啉I的检测限为2 nmol/L，其中ALA的检测限为5 μmol/L，卟胆原的检测限为2 μmol/L。本研究表明，与许多其他现有方法相比，本方法非常有效，因为它无需预处理，能在更短的时间内定量提供ALA、PBG和5种卟啉的同时检测结果，并且具有合适的灵敏度和选择性。LC-MS技术已用于临床尿卟啉水平的测定。
