Ehmann A
J Chromatogr. 1977 Feb 11;132(2):267-76. doi: 10.1016/s0021-9673(00)89300-0.
The chromogenic reagent described has been tested with seventy-nine indole derivatives and found to be very sensitive and indole-specific. The lower limit of detection on silica gel thin-layer plates was between 25 and 50 ng for most indoles. Phenols and hydroxy-, and amino-benzoic acids, hydroxy-, and methoxy-cinnamic acids did not yield chromophores with the exception of p-amino-benzoic acid and p-hydroxy-cinnamic acid which gave yellow and pink chromophores at concentrations greater than 1 and 2 mug. Although many of the C-3 substituted indoles such as indole-3-acetic acid and tryptamine had colors in the reddish-violet-blue color region, most exhibited sufficient color differentiation to allow their identification by thin-layer chromatography. The procedure was simple and required only 10 min from the time of spraying the thin-layer plate until full color development was reached. The colors had a wide spectral range from yellow of the indole-3-glyoxylamide chromophore to blue of the melatonin chromophore, and were extremely stable.
所述显色剂已用79种吲哚衍生物进行了测试,发现其灵敏度很高且对吲哚具有特异性。在硅胶薄层层析板上,大多数吲哚的检测下限在25至50纳克之间。酚类、羟基苯甲酸、氨基苯甲酸、羟基肉桂酸和甲氧基肉桂酸不会产生发色团,但对氨基苯甲酸和对羟基肉桂酸除外,它们在浓度大于1微克和2微克时分别产生黄色和粉红色发色团。尽管许多C-3取代的吲哚,如吲哚-3-乙酸和色胺,在红紫蓝色区域有颜色,但大多数都表现出足够的颜色差异,以便通过薄层层析进行鉴定。该方法很简单,从喷洒薄层层析板到显色完全只需10分钟。这些颜色的光谱范围很广,从吲哚-3-乙醛酰胺发色团的黄色到褪黑素发色团的蓝色,而且极其稳定。