Pavel S, Boverhof R, Wolthers B G
J Invest Dermatol. 1984 Jun;82(6):577-9. doi: 10.1111/1523-1747.ep12261318.
It has previously been shown that enzymatically hydrolyzed urine of patients with malignant melanoma contains 5,6-dihydroxyindole (5, 6DHI ). In this study we describe the elucidation of the entire structure of urinary 5, 6DHI -conjugate. Differential hydrolysis of melanotic urine revealed that, in contrast to beta-glucuronidase, sulfatase can liberate 5, 6DHI from its conjugated form. 5, 6DHI -sulfate was synthetized by reacting 5, 6DHI with sulfur trioxide trimethylamine complex. Thin-layer chromatography (TLC) documented its close similarity to the Thorm ahlen -positive compound usually entitled "C." Gas chromatographic-mass spectrometric (GC-MS) analysis of methylated and subsequently hydrolyzed synthetic 5, 6DHI -sulfate showed that the synthetic product consisted of a mixture of 5-hydroxy-6-indolyl-O-sulfate and 6-hydroxy-5-indolyl-O-sulfate (with a certain amount of 5, 6DHI -disulfate). 5, 6DHI -sulfate was purified with use of DEAE-cellulose column chromatography from melanotic urine. Methylation of this conjugate with deuterated dimethylsulfate and subsequent GC-MS analysis of the hydrolyzed product provided evidence that 5, 6DHI from melanotic urine was almost exclusively sulfated in position 6. It was concluded (1) that 5, 6DHI is excreted as a 6-O-sulfate, and (2) that this compound is consistent with Thorm ahlen -positive compound "C".
先前已有研究表明，恶性黑色素瘤患者经酶解的尿液中含有5,6 - 二羟基吲哚（5,6DHI）。在本研究中，我们描述了对尿中5,6DHI - 结合物完整结构的阐明。对黑色素尿进行差异水解显示，与β - 葡萄糖醛酸酶不同，硫酸酯酶可将5,6DHI从其结合形式中释放出来。通过使5,6DHI与三氧化硫三甲胺络合物反应合成了5,6DHI - 硫酸盐。薄层色谱（TLC）记录了它与通常称为“C”的托马伦阳性化合物非常相似。对甲基化并随后水解的合成5,6DHI - 硫酸盐进行气相色谱 - 质谱（GC - MS）分析表明，合成产物由5 - 羟基 - 6 - 吲哚基 - O - 硫酸盐和6 - 羟基 - 5 - 吲哚基 - O - 硫酸盐（以及一定量的5,6DHI - 二硫酸盐）的混合物组成。利用DEAE - 纤维素柱色谱法从黑色素尿中纯化了5,6DHI - 硫酸盐。用氘代硫酸二甲酯对该结合物进行甲基化，随后对水解产物进行GC - MS分析，结果表明黑色素尿中的5,6DHI几乎完全在6位硫酸化。得出的结论是：（1）5,6DHI以6 - O - 硫酸盐的形式排泄；（2）该化合物与托马伦阳性化合物“C”一致。
