Danton Malcolm, Lim Chang Kee
MRC Bioanalytical Science Group, School of Biological and Chemical Sciences, Birkbeck, University of London, UK.
Biomed Chromatogr. 2006 Jun-Jul;20(6-7):612-21. doi: 10.1002/bmc.656.
A high-resolution high-performance liquid chromatography/electrospray ionization quadrupole time-of-flight tandem mass spectrometry method is described for the analysis of porphyrins in blood, urine and faeces. The gradient elution reversed-phase HPLC system using acetonitrile-methanol-1 m ammonium acetate/acetic acid buffer (pH 5.16) as gradient solvent mixtures was able to separate all porphyrin metabolites, including the type I and type III isomers of uroporphyrin, hepta-, hexa- and penta-carboxylic acid porphyrins and coproporphyrin. The porphyrins were positively identified by the protonated molecules M+H and further characterized by tandem mass spectrometric analysis with each porphyrin giving a characteristic collisioninduced dissociation product ion spectrum. The mass chromatograms obtained by HPLC/ESI MS are useful for the differential diagnosis of the porphyrias, since each type of porphyria has a typical porphyrin excretion pattern.
描述了一种用于分析血液、尿液和粪便中卟啉的高分辨率高效液相色谱/电喷雾电离四极杆飞行时间串联质谱法。使用乙腈 - 甲醇 - 1 m醋酸铵/乙酸缓冲液（pH 5.16）作为梯度溶剂混合物的梯度洗脱反相高效液相色谱系统能够分离所有卟啉代谢物，包括尿卟啉的I型和III型异构体、七羧酸、六羧酸和五羧酸卟啉以及粪卟啉。通过质子化分子M + H对卟啉进行阳性鉴定，并通过串联质谱分析进一步表征，每种卟啉都给出特征性的碰撞诱导解离产物离子谱。通过HPLC/ESI MS获得的质量色谱图可用于卟啉症的鉴别诊断，因为每种类型的卟啉症都有典型的卟啉排泄模式。
