Longas M O, Poh-Fitzpatrick M B
Clin Chim Acta. 1982 Feb 5;118(2-3):219-28. doi: 10.1016/0009-8981(82)90009-2.
Free acid porphyrins were isolated from plasma of a patient with variegate porphyria. Part of the total porphyrin content--which included protoporphyrin IX, harderoporphyrin and uroporphyrin in a molar ratio of 1.2:1:0.5 and traces of pentacarboxylic porphyrin--was extractable with ethyl acetate/acetic acid as described previously [1]. Unextractable porphyrins remained in the precipitate formed after mixing the lower liquid layer and precipitate from the extraction procedure (Fig. 1, [1]) with excess ethyl acetate/acetic acid. A portion of this precipitate was hydrolyzed in 8 mol/l HCl; its porphyrins were extracted with N-butanol and analyzed by high pressure liquid chromatography; another portion was chromatographed on Sephadex G-150 with 1 mol/l MgCl2, and the major porphyrin-protein pool was hydrolyzed in 8 mol/l HCl, reacted separately with AgNO3 and Ag2SO4, and subjected to cellulose acetate and polyacrylamide-gel electrophoresis. The results support the hypothesis that a dicarboxylic porphyrin, a major portion of which was unextractable by standard procedures [1] and which appeared to be covalently bound to a protein of approximately 68 000 mol. wt. that moved with human serum albumin during cellulose acetate electrophoresis, is the preponderant porphyrin in this plasma.
从一位患有混合型卟啉症的患者血浆中分离出游离酸卟啉。总卟啉含量的一部分（其中原卟啉IX、硬卟啉和尿卟啉的摩尔比为1.2:1:0.5，还有痕量的五羧基卟啉），可按先前所述方法[1]用乙酸乙酯/乙酸萃取。不可萃取的卟啉留在萃取过程中下层液体层与沉淀混合后形成的沉淀中（图1，[1]），将该沉淀与过量的乙酸乙酯/乙酸混合。将此沉淀的一部分在8mol/L盐酸中水解；其卟啉用正丁醇萃取并用高压液相色谱分析；另一部分在含1mol/L氯化镁的葡聚糖G - 150上进行色谱分离，主要的卟啉 - 蛋白质组分在8mol/L盐酸中水解，分别与硝酸银和硫酸银反应，然后进行醋酸纤维素和聚丙烯酰胺凝胶电泳。结果支持了这样的假设，即一种二羧酸卟啉是该血浆中的主要卟啉，其大部分不能用标准方法[1]萃取，并且似乎与一种分子量约为68000的蛋白质共价结合，在醋酸纤维素电泳过程中该蛋白质与人类血清白蛋白迁移情况相同。
