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1、1、 生物碱（1）沉淀反应碘化汞钾试剂 白色或浅黄色沉淀碘化铋钾试剂 橘红色沉淀碘碘化钾试剂 浅棕或暗棕色沉淀硅钨酸试剂 浅黄或黄棕色沉淀磷钨酸试剂 浅黄色沉淀磷钼酸试剂 白色或淡黄色沉淀苦味酸试剂 黄色结晶或非结晶形沉淀鞣酸试剂 棕黄色沉淀氯化金试剂 黄色结晶氯化铂试剂 白色结晶雷氏铵盐 红色无定形沉淀（2）薄层层析检查：吸附剂碱性氧化铝（级，干法铺板）硅胶 G（稀碱湿法铺板）展开剂氯仿：甲醇显色UV；碘化铋钾2、 氨基酸、多肽、蛋白质（1）加热沉淀试验：加热煮沸 混浊或沉淀 （蛋白质）+5%H2SO4（不加热）混浊或沉淀（2）双缩脲反应：+40%NaOH，1%CuSO4 紫色、红色或紫红色
2、（多肽、蛋白质）（3）茚三酮反应：+0.2% 茚三酮试液 蓝或蓝紫色（氨基酸、多肽、蛋白质）（4）吲哚醌反应：+吲哚醌试液 各种颜色（氨基酸）（5）Millon 反应： +Hg，H2NO2 红色（蛋白质分子中有酪氨酸组成）（6）Hopkins-Cole 反应：+乙醛酸，浓硫酸 各色（蛋白质分子中有色氨酸组成）（7）氨基酸的薄层层析检查：吸附剂硅胶 G展开剂 n-BuOH，n-BuOH：HAc：H2O显色剂 0.25%茚三酮试液 紫红色斑点3、 有机酸（1）PH 试纸检查（2）溴酚兰试液：喷洒蓝色背景黄色斑点（3）薄层层析检查：吸附剂硅胶 G 或酸性氧化铝展开剂 C6H6：EtOH显色剂0.1%
3、 溴酚兰试液 黄色4、 酚类和鞣质（1）FeCl3 试剂：+1%FeCl3 试液 蓝、暗绿或蓝紫色（2）三氯化铁- 铁氰化钾试剂：喷洒 蓝色斑点（3）香草醛 -盐酸试剂：喷洒 红色（间苯二酚、间苯三酚）（4）重氮盐试剂：+对硝基苯胺、亚硝酸钠 红色（5）薄层层析检查：吸附剂硅胶 G 或纤维素展开剂 n-BuOH：HAc：H2O ；15%HAc显色剂1% FeCl3 试液1%三氯化铁-1% 铁氰化钾试液 蓝、绿或黑色鞣质与酚类的区别：+明胶 沉淀上清液 +1%FeCl3 试液 蓝、暗绿或蓝紫色5、 糖和苷（1）斐林试剂：+硫酸铜、酒石酸钾钠 砖红色沉淀（还原糖）（）+1%HCl +NaOH 沉淀
4、（苷元）30min 上清液（+ ） （多糖、苷）（2）Molish 反应：+-萘酚 -浓硫酸 紫红色环（3）银镜反应：+0.1N 硝酸银、5N 氨水 银褐色（还原糖）（4）薄层层析检查：吸附剂硅胶 G 或纤维素展开剂 n-BuOH：Pd：H2O；15%HAc显色剂 苯胺 -邻苯二甲酸6、 皂苷（1）泡末试验：振摇 大量持续性泡末+0.1M HCl 二管泡末高度相同（三萜皂苷）+0.1M NaOH 碱管高于酸管（甾体皂苷）（2）溶血试验：+2%红血球悬浮液 溶血（3）LiebermanBurchard 反应：+ 醋酐- 浓硫酸 紫红色（三萜皂苷）黄-红- 紫-污绿（甾体皂苷）7、甾体（1）Lie
5、bermanBurchard 反应：+ 醋酐- 浓硫酸 黄-红-紫-污绿（2）氯仿 -浓硫酸反应： +氯仿-浓硫酸 氯仿层红或青色硫酸层绿色荧光（3）五氯化锑或三氯化锑反应：+SbCl3 或 SbCl5 红色（4）薄层层析检查：吸附剂中性氧化铝或硅胶 G 展开剂 C6H6-MeOH；CHCl3-MeOH显色剂 10%磷钼酸 蓝-蓝紫色5%三氯化锑试液 红、棕红或绿色8、黄酮（1）盐酸 -镁粉反应： +HCl-Mg 红色（2）三氯化铝反应：+AlCl3 黄色（3）浓氨水反应：+NH3 亮黄或橙色（4）薄层层析检查：吸附剂聚酰胺或硅胶 G 展开剂 MeOH-H2O；EtOH-H2O显色剂 UV亮黄
6、或黄绿色荧光1%三氯化铝试液亮黄色9、香豆素、内酯（1）开闭环反应：+1%NaOH澄清 +2%HCl混浊（2）异羟污酸铁反应：+7%盐酸羟胺、10%KOH +稀 HCl、1%FeCl3 红色（3）重氮盐试剂：+对硝基苯胺、亚硝酸钠 红色（4）薄层层析检查：吸附剂酸性硅胶 G 或硅胶 G 或酸性氧化铝 展开剂 甲苯-乙酸乙酯 -甲酸（5：4：1 ）显色剂 UV蓝色荧光异羟污酸铁试液 红色10、强心苷（1）Kedde 试剂：+3 ，5- 二硝基苯甲酸试液 紫红色（2）Baljet 试剂：+碱性苦味酸试液 橙或橙红色（3）Legal 试剂：+亚硝酰铁氰化钠试液 紫红色（4）K-K 反应：+FeCl3
7、/冰 HAc、浓 H2SO4 上层绿蓝色 （2-去氧糖）界面红棕色（5）薄层层析检查：吸附剂硅胶 G 或中性氧化铝展开剂 n-BuOH：HAc：H2O （4 ：1：5）显色剂 碱性 3，5- 二硝基苯甲酸试液紫红色碱性苦味酸试液 橙红色11、蒽醌（1）碱液反应：+10%NaOH 红色 +H2O2 红色不褪 +H+ 红色褪去（2）醋酸镁反应：+1%MgAc2 红色（3）薄层层析检查：吸附剂硅胶 G展开剂Pet ：EtOAc显色剂 UV黄色荧光5%NaOH 红色12、挥发油、油脂（1）油斑检查：油斑挥发 挥发油； 油斑不消失油脂或类脂（2）磷钼酸反应：喷洒 5%磷钼酸试液 蓝色（油脂、三萜、甾醇）
8、TLC Visualization ReagentsThis is a brief selection of the many available TLC visualization reagents. Below each title is the type of compounds or structure which can be detected with the specific reagent. When beginning work with these reagents, acquired any MSDS (material safety data sheets) to se
9、e if there are any extra precautions needed in safely using them.Before spraying, plates should be well dried in the hood of residual solvents and components. Amines and organic acids used in the mobile phases may adversely affect the visualization reaction being attempted. If heating to remove thes
10、e components is done, consideration should be given so that loss of components or their decomposition is avoided (by lowering the temperature or using a shorter time in the oven).Always spray any of these reagents onto plates in a well ventilated hood while wearing safety glasses. Also apply moderat
11、e amounts to the plate so it always appears dull and flat (if it looks wet, you have sprayed too much). You can always overspray to enhance the detection.When information about the results of using the visualization reagents was available, this was put under each reagent as Results. If not give, the
12、 user will have to do a few experiments to see what the results might be.Always remember to look under normal light and also short and long wavelength UV light so as not to miss any possibilities.Many of the reagents for these visualizers can be found in the EMD Chemicals catalog or on the website()
13、.Aluminium chlorideFor flavonoidsSpray plate with a 1% ethanolic solution of aluminum chloride.Results: Yellow fluorescence in long wavelength UV light (360nm)4-Aminoantipyrine/potassium hexacyanferrate (III) (Emerson reaction)(Emerson reagent) for the detection of phenols and arylaminesSolution I:
14、1g aminoantipyrine (4-aminophenazone) in 100ml 80% ethanolSolution II: 4g potassium hexacyanoferrate (III) in 20ml water, fill to 100ml with ethanolProcedure:Spray with solution IDry 5 minutes with warm airPlace chromatogram in a chamber with vapor from 25% ammonium solution, making sure that the la
15、yer doesnot contact the liquid.Results: Red-orange to salmon pink spots2-Aminoethyl diphenylborate, see Ethanolamine diphenylborateAmmonium metavanadate, ammonium monovanadate, see Vanadium(V) / sulfuric acidAmmonium molydbateFor detection of phosphoric acid derivativesSolution I: 1M perchloric acid in water/acetone (1:1)Solution II: ammonium molybdate soln: 5g (NH4)6Mo7O24.4 H2O in 35ml semi-conc. Nitric acid and 65ml water.Solution III: Tin (II) chloride soln: 0.5g SnCl2.2 H2O in 100ml 0.5M hydrochloric acid:Dry developed chromato
