TLC展开剂选择及显色剂的汇总

薄层层析TLC通用显色剂薄层层析通用显色剂1 通用试剂(1) 重络酸钾-硫酸:检查一般有机物.喷洒剂:5克重络酸钾溶于100毫升40%硫酸中.薄层检查:喷洒后加热到150℃至班点出现(2) 荧光素-溴:检查不饱和化合物喷洒剂:0.1克荧光素溶于100毫升乙醇中溴试剂:5%的溴的四氯化碳溶液喷洒后处理:喷洒荧光素溶液后,放置存有溴溶液的缸内,可于紫外线分析灯下检查荧光,荧光素与溴化和成曙红(Eosin)(无萤光),而不饱和化合物则成溴加成物,保留了原有荧光;若点样较多,则呈黄色斑点,底板呈红色.(3) 碘:检查一般有机物.方法:a 层析谱放密闭缸内或瓷盘内，缸内预先放有碘结晶少许，大部分有机化合物呈棕色斑点。

b 层析谱放碘蒸气中5分钟（或喷5％碘的氯仿溶液）取出置空气中待过量的碘蒸气全部挥发后，喷1％淀粉的水溶液，斑点转成蓝色。

(4)硫酸：通用喷洒剂：5％的浓硫酸乙醇溶液，或15％浓硫酸正丁醇溶液，或浓硫酸－醋酸（1：1）喷洒后处理：空气中干燥15分钟，再热至110℃直至出现颜色或荧光。

(5)硝酸银－氢氧化铵（Tollen-Zaffaroni）试剂：检查还原性物质。

溶液I : 0.1%N硝酸银; 溶液II: 5N氢氧化铵喷洒剂: I和II以1:5混合(临用前混合)喷洒后处理: 105℃加热5~10分钟,至深黑色斑点出现.(6)磷钼酸或磷钨酸,硅钨酸:检查还原性物质,类脂体,生物碱,甾体喷洒剂: 5~10%磷钼酸或磷钨酸或硅钨酸乙醇溶液喷洒后处理: 120 ℃加热至斑点出现.沉淀试剂: 1克硅钨酸溶于20毫升水中,加10%盐酸至强碱性.2 生物碱(7)硫酸: 检查生物碱及含碘化合物喷洒剂: 0.1克硫酸?混悬于4毫升水中,加入1克三氯醋酸,加热至沸,逐滴加入浓硫酸至澄清.喷洒后处理: 110℃加热数分钟至斑点出现.(8)碘化铋钾(Drage ndorff)试剂:检查生物碱及其他含氟化合物.溶液I: 0.85克次硝酸铋溶于10毫升冰醋酸40毫升水中溶剂II: 8克碘化钾溶于20毫升水中.制备液I+II,等体积混合.可用于棕色瓶中保存较长时间,一般制备液可作沉淀试剂用.喷洒液: 制备液1毫升与2毫升醋酸,10毫升水混合即得(9).碘化汞钾(Mayer)试剂: 检查生物碱.制备液: 13.55克氯化汞和49.8克碘化钾各溶于20毫升水中,等体积混合并用水稀释至1000毫升.喷洒液: 制备液加1/10体积的17%盐酸.喷洒后处理: 观察斑点,并于紫外线荧光分析灯下检出.(10) ?酸纳-浓硫酸(Mandelin)试剂:检查生物碱.1%>酸纳的浓硫酸溶液.与多种生物碱呈不同颜色.(11)碘－碘化钾(Wagner)试剂:检查生物碱.1克碘及10克碘化钾,溶于50毫升水中,加热,加2毫升醋酸,再用水稀释至100毫升.可作纸层板显色剂,液可作沉淀试剂.3 酚类,鞣质(12)三氯化铁:检查酚类及??酸.喷洒剂:1~5%三氯化铁的水溶液或乙醇溶液.并加盐酸少许.??酸呈红色斑点,酚类称蓝色或绿色斑点.(13)铁氰化钾-三氯化钾:检查酚类,芳香胺类及还原性物质.喷洒剂: 1%铁氰化钾水溶液,2%三氯化铁水溶液.临用前等体积混合.喷洒后处理: 喷洒后酚性物质呈蓝色斑点.再喷2N盐酸,能使颜色加深,纸谱可用稀盐酸洗去喷洒液.(14) 4-胺基安替比林-铁氰化钾(Emerson反应): 检查酚类.喷洒剂: I . 2%4-氨基安替比林乙醇溶液;II. 8%铁氰化钾水溶液.或用0.9%4-氨基安替比林和5.4%铁氰化钾水溶液方法: 先喷洒I,再喷洒II,即显色,或再放入密闭缸中,缸内放25%氢氧化铵,即产生橙色至深红色.(15) 对氨基苯磺酸,重氮盐(Pauly试剂): 检查酚类,芳香胺类及能偶合的杂环化合物.喷洒剂: 4.5克对氨基苯磺酸,加热溶于45毫升12N盐酸中,用水稀释至500毫升,取10毫升稀释液用冰冷却,加10毫升冷4.5%亚硝酸钠水溶液,0℃放15分钟(此试剂于0℃可保存3天), 用前加等体积1%碳酸钠水溶液.一般重氮化试剂,也可用联苯胺,对硝基苯胺等.(16) 对甲苯磺酸: 检查甾体,黄酮,鞣质.喷洒剂: 20%对甲苯磺酸氯仿溶液.喷洒后处理: 100℃加热数分钟,紫外线分析灯下检查荧光斑点.4 含氧杂环及蒽醌类*(17) 三氯化铝: 检查黄酮体.喷洒1%三氯化铝乙醇液于紫外线荧光分析灯下检示,呈黄色荧光(18)碱式醋酸铅: 检查黄酮体.喷洒剂: 饱和碱式醋酸铅(或饱和醋酸铅)水溶液.于紫外线荧光分析灯下检查荧光斑点.(19)醋酸镁: 检查蒽醌甙,甙元及黄酮体喷洒剂: 0.5%醋酸镁甲醇溶液方法: 90℃加热5分钟,呈红色至紫色斑点.(20)氢氧化钾: 检查香豆素,蒽醌甙及甙元喷洒剂: 5~10%氢氧化钾的甲醇溶液.于日光及紫外线荧光分析灯下检示斑点.5 萜类,甾体(21) 三氯化锑(Carr-Price试剂): 检查甾体,萜类,皂类.喷洒剂: 25克三氯化锑溶于75克氯仿中(亦可以用氯仿或四氯化碳的饱和溶液). 喷洒后处理: 100℃加热5分钟,于紫外线荧光分析灯下检示荧光.(22) 五氯化锑: 检查甾体,萜类,皂甙.喷洒剂: 五氯化锑-氯仿或四氯化碳(1:4),用前新鲜配制.喷洒后处理: 120℃加热至斑点出现,并于紫外线荧光分析灯下检示.(23)香兰醛-硫酸: 检查高级醇类,酚类,甾体,萜类,芳香油.喷洒剂: 1克香兰素溶于100毫升浓硫酸,或0.5克香兰醛溶于100毫升硫酸-乙醇(4:1)中.喷洒后处理: 室温或120℃加热观察显色斑点.(24) 4-二甲氨基苯甲醛,醋酸,磷酸(E.P.试剂): 检查? Azulene 及?前体Proazulene.喷洒剂: 0.25克4-二甲氨基苯甲醛,溶于50毫升醋酸5克85％磷酸和20毫升水的混合液中(棕色瓶中保存数月):烃室温即成蓝紫色斑点,>前体于80℃加热10分钟出现蓝紫色斑点.(25) 氯胺T－三氯醋酸: 检查强心甙.喷洒剂: I. 3%氯仿T水溶液新鲜制备.II. 25%三氯醋酸乙醇溶液(能保存数天).10毫升I加40毫升II,用前混合.喷洒后处理: 110℃加热7分钟,紫外线荧光分析灯下检示呈蓝色或黄色荧光. (26) 亚硝酸基铁氰化钠-氢氧化钠(Legal试剂): 检查不饱和内酯;甲基酮或活性次甲基,常用于强心甙喷洒剂: 1克亚硝基铁氰化钠溶于100毫升2N氢氧化钠-乙醇(1:1)的水溶液.显红色或紫色斑点.(27) 3,5-二硝基苯甲酸(Legal试剂): 检查强心甙,α,β-不饱和内酯.喷洒剂: 1克3,5-二硝基苯甲酸溶于50毫升甲醇,加入1N氢氧化钾50毫升.强心甙呈紫红色斑点.6 糖类(28) 邻苯二甲基苯胺: 检查还原糖.喷洒剂: 0.93克苯氨,1.66克邻苯二甲酸溶于100毫升水饱和的正丁醇中.喷洒后处理: 105℃加热10分钟.(29) 2,3,5-Triphenyl-tetrazo lium chloride (T.T.C.)：检查还原糖及其他还原物质。

TLC显色试剂及配方大全3(6)含氮化合物①FCNP(硝普钠／铁氰化物)试剂检出物：脂肪族含氮化物，如氨基氰、胍、脲与硫脲及其衍生物，肌酸及肌酐。

溶液：10％氢氧化钠溶液、10％硝普钠溶液、10％铁氰化钾溶液与水按1：1：1：3混合，在室温至少放置20min，冰箱保存数周，用前将混合液与丙酮等体积混合。

②Dragendorff(碘化铋钾试剂)试剂检出物：芳香族含氮化合物，如生物碱类、抗心律不齐药物。

溶液：I．碱式硝酸铋0.85g溶于10ml冰醋酸及40ml水中；Ⅱ．碘化钾8g溶于水20ml中。

将上述溶液I及Ⅱ等量混合，置棕色瓶中作为储备液，用前取储备液lml、冰醋酸2ml与水l0ml混合。

结果：呈橘红色斑点。

③4-甲基伞形酮检出物：含氮杂环化合物。

溶液：本品0.02g溶于乙醇35ml，加水至100ml。

方法：喷板后置25％氨水蒸气的容器中,取出后于紫外灯(365nm)下观察。

④碘铂酸钾检出物：生物碱类及有机含氮化物。

溶液：10％六氯铂酸溶液3ml与水97ml混合，加6％碘化钾溶液，混匀。

临用前配制。

⑤硫酸高铈铵／硫酸检出物：生物碱及含碘有机化物。

溶液：硫酸铈1g混悬于4ml水中，加三氯乙酸1g，煮沸，逐滴加入浓硫酸直至混浊消失。

方法：喷后薄层于1l0oC加热数分钟。

结果：阿朴吗啡、马钱子碱、秋水仙碱、罂粟碱、毒扁豆碱与有机碘化物均能检出。

⑥Ehrlich (对二甲氨基苯甲醛／盐酸)试剂检出物：吲哚衍生物及胺类。

溶液：1％本品的浓盐酸溶液与甲醇按1：1混合。

方法：喷后板于50oC加热20min。

结果：呈不同颜色的斑点。

(7)胺类①硝酸／乙醇检出物：脂肪族胺类。

溶液：50滴65％硝酸于乙醇100ml中。

方法：需要时120oC加热。

②2，6-二氯醌氯亚胺检出物：抗氧剂、酰胺(辣椒素)、伯、仲脂肪胺、仲、叔芳香胺、芳香碳氢化物、药物、苯氧基乙酸除草剂等。

溶液：新鲜制备的0.5％～2％本品乙醇溶液。

方法：喷后薄层于110oC加热10min，再用氨蒸气处理。

TLC展开剂选择及显色剂的总结(转自中国色谱网)★huyuchem(金币+1):谢谢选择适当的展开剂是首要任务.一般常用溶剂按照极性从小到大的顺序排列大概为：石油迷<己烷<苯<乙醚<THF<乙酸乙酯<丙酮<乙醇<甲醇使用单一溶剂，往往不能达到很好的分离效果，往往使用混合溶剂通常使用一个高极性和低级性溶剂组成的混合溶剂，高极性的溶剂还有增加区分度的作用，常用的溶剂组合有：Petroleumether/Ethylacetate,petroleumether/Acetone,Petroleumether/Ether,Petroleumether/CH2Cl2, ethylacetate/MeOH,CHCl3/ethylacetate展开剂的比例要靠尝试.一般根据文献中报道的该类化合物用什么样的展开剂，就首先尝试使用该类展开剂，然后不断尝试比例，直到找到一个分离效果好的展开剂。

展开剂的选择条件：①对的所需成分有良好的溶解性；②可使成分间分开；③待测组分的Rf在0.2~0.8之间，定量测定在0.3～0.5之间；④不与待测组分或吸附剂发生化学反应；⑤沸点适中，黏度较小；⑥展开后组分斑点圆且集中；⑦混合溶剂最好用新鲜配制。

一般来说，弱极性溶剂体系的基本两相由正己烷和水组成，再根据需要加入甲醇、乙醇，乙酸乙酯来调节溶剂系统的极性，以达到好的分离效果，适合于生物碱、黄酮、萜类等的分离；中等极性的溶剂体系由氯仿和水基本两相组成，由甲醇、乙醇，乙酸乙酯等来调节，适合于蒽醌、香豆素，以及一些极性较大的木脂素和萜类的分离；强极性溶剂，由正丁醇和水组成，也靠甲醇、乙醇，乙酸乙酯等来调节，适合于极性很大的生物碱类化合物的分离。

很多时候,展开剂的选择要靠自己不断变换展开剂的组成来达到最佳效果。

我们在实验中，为了实现一个配体与其他杂质有效分离，曾经尝试了很多种的溶剂组合，最后才找到石油醚—EtOAc—HCOOH（5.5：3.5：0.1）混合溶剂。

1、生物碱(1 )沉淀反应一一碘化汞钾试剂T白色或浅黄色沉淀碘化铋钾试剂T橘红色沉淀碘一碘化钾试剂T浅棕或暗棕色沉淀硅钨酸试剂T浅黄或黄棕色沉淀磷钨酸试剂T浅黄色沉淀磷钼酸试剂T白色或淡黄色沉淀苦味酸试剂T黄色结晶或非结晶形沉淀鞣酸试剂T棕黄色沉淀氯化金试剂T黄色结晶氯化铂试剂T白色结晶雷氏铵盐T红色无定形沉淀(2 )薄层层析检查：吸附剂一一碱性氧化铝(川级，干法铺板)硅胶G (稀碱湿法铺板)展开剂一一氯仿：甲醇显色一一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T混浊或沉淀(蛋白质)+5%H2SO4 (不加热)T混浊或沉淀(2)双缩脲反应：+40%NaOH , 1%CuSO4 T紫色、红色或紫红色(多肽、蛋白质)(3)茚三酮反应：+0.2%茚三酮试液T蓝或蓝紫色(氨基酸、多肽、蛋白质)(4)吲哚醌反应：+吲哚醌试液T各种颜色(氨基酸)(5)Millon反应：+Hg , H2NO2 T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T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：吸附剂一一硅胶G展开剂——n-BuOH , n-BuOH : HAc: H2O显色剂一一0.25%茚三酮试液T紫红色斑点3、有机酸(1)PH试纸检查(2)溴酚兰试液：喷洒T蓝色背景黄色斑点(3 )薄层层析检查：吸附剂一一硅胶G或酸性氧化铝展开剂——C6H6 : EtOH显色剂一一0.1%溴酚兰试液T黄色4、酚类和鞣质(1)FeCI3试剂：+1%FeCI3试液T蓝、暗绿或蓝紫色(2 )三氯化铁-铁氰化钾试剂：喷洒T蓝色斑点(3)香草醛-盐酸试剂：喷洒T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T红色(5 )薄层层析检查：吸附剂一一硅胶G或纤维素展开剂——n-BuOH : HAc: H2O; 15%HAc显色剂一一1% FeCl3试液1%三氯化铁-1%铁氰化钾试液T蓝、绿或黑色鞣质与酚类的区别：+明胶一一沉淀上清液+1%FeCI3试液T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%HCI +NaOH 沉淀(苷元)△ 30min 上清液(+ )(多糖、苷)(2)Molish反应：+ a-萘酚-浓硫酸T紫红色环(3)银镜反应：+0.1N硝酸银、5N氨水T银褐色(还原糖)(4 )薄层层析检查：：吸附剂一一硅胶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T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T溶血(3) ---------------------------------------------------------------------- Lieberman —Burchard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)Lieberman —Burchard反应：+醋酐-浓硫酸T黄-红-紫-污绿(2)氯仿-浓硫酸反应：+氯仿-浓硫酸氯仿层T红或青色硫酸层T绿色荧光(3)五氯化锑或三氯化锑反应：+SbCI3或SbCI5 T红色(4 )薄层层析检查：吸附剂一一中性氧化铝或硅胶G展开剂——C6H6-MeOH ; CHCl3-MeOH显色剂一一10%磷钼酸T蓝-蓝紫色5%三氯化锑试液T红、棕红或绿色8、黄酮(1)盐酸-镁粉反应：+HCl-Mg T红色(2 )三氯化铝反应：+AlCl3 T黄色(3)浓氨水反应：+NH3 T亮黄或橙色(4 )薄层层析检查：吸附剂一一聚酰胺或硅胶G展开剂——MeOH-H2O ; EtOH-H2O显色剂一一UV T亮黄或黄绿色荧光1%三氯化铝试液T亮黄色9、香豆素、内酯(1)开闭环反应：+1%NaOH T澄清+2%HCl T混浊(2)异羟污酸铁反应：+7%盐酸羟胺、10%KOH △ +稀HCl、1%FeCl3 T红色(3)重氮盐试剂：+对硝基苯胺、亚硝酸钠T红色(4 )薄层层析检查：吸附剂一一酸性硅胶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 T蓝色荧光异羟污酸铁试液T红色10、强心苷(1)Kedde试剂：+3 , 5-二硝基苯甲酸试液T紫红色(2)Baljet试剂：+碱性苦味酸试液T橙或橙红色(3)Legal试剂：+亚硝酰铁氰化钠试液T紫红色(4)K-K反应：+FeC⑶冰HAc、浓H2SO4 T 上层绿~蓝色(2-去氧糖)界面红棕色(5)薄层层析检查：吸附剂一一硅胶G或中性氧化铝展开剂——n-BuOH : HAc: H2O (4 : 1 : 5)显色剂一一碱性3 , 5-二硝基苯甲酸试液T紫红色碱性苦味酸试液T橙红色11、蒽醌(1)碱液反应：+10%NaOH T红色+H2O2 T红色不褪+H+ T红色褪去(2)醋酸镁反应：+1%MgAc2 T红色(3 )薄层层析检查：吸附剂一一硅胶G展开剂—— Pet: EtOAc显色剂一一UV T黄色荧光5%NaOH T红色12、挥发油、油脂(1 )油斑检查：油斑挥发T挥发油；油斑不消失T油脂或类脂(2)磷钼酸反应：喷洒5%磷钼酸试液T蓝色(油脂、三萜、甾醇)TLC Visualizatio n Reage ntsThis is a brief selectio n of the many available TLC visualizati on reage nts. Below each title is the type of compo unds or structure which can be detected with the specific reage nt. When begi nning work with these reage nts, acquired any MSDS (material safety data sheets) to see if there are any extra precauti ons n eeded in safely using them.Before spraying, plates should be well dried in the hood of residual solvents and comp onen ts. Ami nes and orga nic acids used in the mobile phases may adversely affect the visualizati on react ion being attempted. If heat ing to remove these comp onents is done, con siderati on should be give n so that loss of comp onents or their decompositi on is avoided (by loweri ng the temperature or using a shorter t ime in the ove n).Always spray any of these reage nts onto plates in a well ven tilated hood while weari ng safety glasses. Also apply moderate amounts to the plate so it always appears dull and flat (if it looks wet, you have sprayed too much). You can alway s overspray to enhance thedetect ion.When in formati on about the results of using the visualizati on reage nts was available, this was put un dereach reage nt as ‘ Results If not. give, the user will have to do a few experime nts to see what the results might be.Always remember to look un der no rmal light and also short and long wavele ngth UV light so as not to miss any possibilities.Many of the reage nts for these visualizers can be found in the EMD Chemicals catalog or on the website ( ).Alumi nium chlorideFor flav ono idsSpray plate with a 1% etha nolic soluti on of alumi num chloride.Results: Yellow fluoresce nee in long wavele ngth UV light (36 Onm)4-Ami noan tipyri ne/potassium hexacya nferrate (III) (Emers on reacti on)(Emers on reage nt) for the detect ion of phe nols and arylam inesSoluti on I: 1g aminoan tipyri ne (4-am in ophe nazone) in 100ml 80% etha nolSoluti on II: 4g potassium hexacya no ferrate (III) in 20ml water , fill to 100ml with etha no I Procedure:Spray with soluti on IDry 5 minu tes with warm airPlace chromatogram in a chamber with vapor from 25% ammonium soluti on, making surethat the layer doesnot con tact the liquid.Results: Red-ora nge to salm on pink spots2-Am ino ethyl diphe ny Iborate, see Etha no lam ine diphe ny IborateAmmonium metava nadate, ammonium monovan adate, see Van adium(V) / sulfuric acidAmmon ium molydbateFor detect ion of phosphoric acid derivativesSoluti on I: 1M perchloric acid in water/acet one (1:1)Solution II: ammon ium molybdate sol n: 5g (NH4)6Mo7O24.4 H2O in 35ml semi-co nc.Nitric acid and 65ml water .Solution III: Ti n (II) chloride sol n: 0.5g Sn Cl2.2 H2O in 100ml 0.5M hydrochloric acid:Dry developed chromatogram and heat to 60 CHydrolyse di- and triphosphates by spra ying perchloric acid (soluti on I) onto the warmplate. After spra ying 2times, dry plate slowly at 50 C. Amidophosphates might not be decomposed.In any case, spray the still warm plate with ammon ium molybdate soluti on (soluti on II)Then spray the still wet plate with tin (II) chloride soluti on (soluti on III)Results: Phosphates appear as blue to blue-green spots. Polyphosphates can also be detected by dipp ing theplates in a soluti on of ammon ium molybdate (1g) dissolved in water (8ml) and perchloricacid （3ml, ca. 70%）,filled up to 100ml with acet one. Then phosphates appear as yellow-gree n spots on a blue backgro und. Also seeMolybde num blue reactio n accord ing to Dittmer and Lester .Ani li ne phthalate （邻苯二甲酸胺）For the detect ion of reduci ng sugarsDry the developed chromatogramSpray with 0.93g an ili ne and 1.66g o-phthalic acid dissolved in 100ml n-buta nol saturated with water.Briefly dry with hot air , then heat to 105 C for 10 min utesResults: Substa nee spots show differe nt colors on an almost colorless backgro und. Some spots give fluoresce nee at 365nm.p-Anisaldehyde -sulfuric acid （对甲氧基苯甲醛-硫酸）For detect ion of phe no Is, sugars, steroids, and terpe nesSpray with a soluti on of freshly prepared 0.5ml p-a ni saldehyde in 50ml glacial acetic acidand 1ml 97% sulfuric acid. and heat to 105 ° C until maximum visualization of spots. The backgro und might be brighte ned by water vapor .Results: Liche n con stitue nts, phe no Is, terpe nes, sugars, and steroids turn violet, blue, red,grey or gree n.For detect ion of sugarsSpray with a soluti on of freshly prepared 1ml p-a ni saldehyde, 1ml 97% sulfuric acid in18ml ethanol and heat at110 ° C.Results: Sugar phe ny Ihydraz ones produce gree n-yellow spots in 3 min. Sugars will produce blue, gree n, violetspots in 10min. Also detects d igitalis glycosides.p-A nisidi ne HydrochlorideFor detect ion of carbohydrates / sugarsMix a solutio n of 3% p-a ni sid ine hydrochloride in n-buta nolSpray and heat at 100 ° C for 2-10min.Results: Aldohexoses are seen as green-brown spots, ketohexoses as yellow spots,aldope ntoses as gree nspots, and uronic acids as red spots.Ani sidi ne phthalateFor detect ion of carbohydrates and reduci ng sugarsSpray with a solution of 1.23 g p-a nisidi ne and 1.66g phthalic acid in 100ml 95% etha nol.Results: Hexoses, green; pentoses, red-violet - sensitiv ity 0.5ug; methylpentoses,yellow-gree n; uronic acids, brow n -se nsitivity 0.1-0.2ug.An timo ny (III) chloride (三氯化锑)For detect ion of flav ono idsSpray with a 10% solution of antimony (III) chloride in chloroform Results: Fluoresci ng spots in long wavele ngth light (36 Onm).Antimony (III) chlorideFor detect ion of vitam ins A & D, carote no ids, steroids, sapoge nins, steroid glycosides, terpenes Spray with a solution of 25g antimony (III) chloride in 75ml chloroform (generally asaturated soluti on ofan tio mony (III) chloride in chloroform or carb on tetrachloride is used).Heat 10min at 100C, view under long wavelength light (360nm).Bromi ne / Carbon tetrachloride (四氯化碳)For detect ion of orga no thiophosphorous pesticidesPlace chromatogram in a chamber with a 10% bromine and tetrachloride without con tact with the liquid.3Bromocresol gree nFor detect ion of orga nic acidsDip chromatogram in a soluti on of 0.1g bromocresol gree n in 500ml etha nol and 5ml 0.1M NaOHResults: Acids yield yellow spots on a blue backgro und.Bromthymol blueFor detecti on of lipids and phospholipidsReage nt: 0.1% bromthymol blue in 10% aqueous etha nol made just alkali ne with NH4OH Spray dried plate.Results: Compounds above produce blue-green colors; sensitiv ity 0.1- 1 卩g.Chlora nil reage nt For detecti on of phe nolsSpray with a soluti on of 1% tetrachloro-p-be nzoq uinone in tolue neChlori ne / o-tolidi neFor detecti on of of compo unds formi ng chloroam in es, e.g., urea derivatives, carbamated, an tibiotics Solution I: 160mg o-tolidine in 30ml glacial acetic acid, filled to 500ml with distilled water,plus 1g KI soluti onSolution II: saturated solution of o-tolidine in 2% acetic acid/0.85% KI solution (1:1, v/v) Procedure APlace chromatogram 15-20 min in a chlori ne atmosphere (e.g., Potassium perma ngan ate +10% Hydrochloricacid)Leave 5 minu tes at ambie nt temperature un til the chlori ne is evaporated completely(spray corner of plate to in sure no blue color is see n, show ing complete abse nee ofchlori ne).Spray with soluti on IProcedure BSpray with 2% potassium hypochlorite soluti on in waterLeave 1-1.5hr at ambie nt temperatureSpray with soluti on IICopper sulfate / phosphoric acid (硫酸铜-磷酸)Used as a charri ng reage nt for polymer bound TLC plates (the n ewer hard layer plates)Spray with a soluti on of 10% copper (II) sulfate in 10% phosphoric acidHeat 5-30min at 110 ° CResults: View freque ntly (every 5-10 min) to see if colored or fluoresce nt spots (at 254 and 360nm) can be seen.Charring can be continued until spots are brown, grey or black.Chromosulfuric acidSee un der Potassium dichromate / sulfuric acidDDQ Reage nt (Dichlorodicya noben zoq uinone)For detecti on of phe nolsSpray with a soluti on of 2% 2,3-dichloro-5,6-dicya no-1,4-be nzoq uinone in tolue neDichlorofluoresce in (二氯荧光黄)For the detect ion of sweete ners sacchari ne & cyclamateSpray with a 0.2% soluti on of dichlorofluoresce in in 96% etha nolDry with warm air; if n ecessary, spray with waterView under 360nm UV lightDichlorofluoresce in / fluoresce in sodium salt (荧光钠)For detect ion of N-substituted barbituratesSpray with a 0.1% etha no lie soluti on of dichlorofluoresce inThen spray with a 0.1% etha nolic soluti on of fluoresce in sodium salt2,6-Dichloro quinone -4- chloroimideFor detecti on of an tioxida nts, phe no Is, primary and sec on dary aliphatic amin es,sec on dary and tertiary aromaticamin es, aromatic hydrocarb ons, pharmaceuticals, phe no xyacetic acid herbicides, etcSpray with a freshly prepared 0.5-2% soluti on of 2,6-dichloro quinon e-4-chloroimide inetha nol (reage nt stablefor 3 weeks if refrigerated).Heat 10min at 110 C; treat with ammo nia vaporp-Dimethylam inoben zaldehydeFor detect ion of sulfo namidesSpray with a soluti on of 1% p-dimethylami noben zaldehyde in 5% hydrochloric acid; add 5% etha nolDetects sulfo namidesp- Dimethylam inoben zaldehyde / hydrochloric acid reage nt (Ehrlich For detect ion of amin es, in dolederivativesSpray with a soluti on of 1% p-dimethylami noben zaldehyde in cone, hydrochloricacid/metha nol (2:2)Heat plates for 20min at 50 C2,4-Di nitrophe nylhydraz ineFor detect ion of aldehydes and ket ones Spray plate with solution of 0.4 g 2,4-DNPH in100ml 2N hydrochloric acid, add 1ml etha nolResults: Yellow-red spots will be see n,Diphe ny lam ine (二苯胺)For detect ion of glycosides, glycolipidsReage nt: 10ml 10% diphe nylami ne in etha nol, 100ml HCl and 80ml glacial acetic acid Spray lightly, cover plate with ano ther glass plate, heat 30-40min at 110areas appearResults: Glycolipids produce blue spots,s-Diphe ny Icarbaz oneFor detect ion of barbituratesSpary with a solution of 0,1% s-diphenyIcarbazone in 95% ethanolResults: Barbiturates will produce purple spots2,2 -'iphenylpicrylhydrazylFor detect ion of aldehydes and ket onesReage nt: dissolve 15mg of 2,2 -DPPH in '25ml chloroformSpray, heat 5-10min at 110 ° C;Results: Yellow spots on a purple backgro und will be see n,Dithizone (双硫腙)For detect ion of heavy metal ionsDissolve 20mg dithiz one in 100ml acet one, store in a brow n bottle in a refrigeratorProcedure: s reage nt)C untilSpray with dithiz one soluti onSpray with 25% ammonia soluti onDittmer and LesterSee Molybde num blueDrage ndorff reage ntFor detect ion of n itroge n compo un ds, alkaloids, an tiarrhythmic drugs, surfacta ntsSolution 1) 1.7g basic bismuth nitrate and 20g tartaric acid in 80ml waterSolution 2) 16 g potassium iodide in 40ml waterStock soluti on (stable for several weeks in a refrigerator):Mix equal volumes of solutio ns 1 and 2Procedure:Spray with a solution of 10g tartaric acid, 50ml water and 5ml stock solutionEtha no lam ine diphe ny Iborate (flav one reage nt accord ing to Neu)For detect ion of flav ono idsSpray with a 1% soluti on of etha no lam ine diphe ny Iborate in metha nolSpray with a 5% ethanolic solution of polyyethylene glycol for fluorescence stabilization Irradiate 2 minutes with intense 365nm UV lightView under 365nm UV lightErhlich ' s reage ntSee p-Dimethylam inoben zaldehydeEmers on reage ntSee 4-am inoan tipyri ne/potassium hexa-cya no ferrate (III)Fast Blue B reage ntFor detect ion of cannabino ids, phe no Is, tanning age nts, amines with can be coupled Spray with a soluti on of 0.5g Fast Blue B (tetraazotized di-o-a ni sidi ne) in acet on e/water (9:1, v/v), always prepared freshThe n overspray with 0.1M sodium hydroxide soluti onResults: Cannabino ids tur n dark red/purple in colorFerric Chloride / sulfuric acidUsed as a charri ng reage nt for polymer bound TLC plates (the n ewer hard layer plates) Spray with a solution of 2g FeCl3 in 83ml n-butanol and 15ml cone, sulfuric acid.Heat 5-30min at 110 ° CResults: View freque ntly (every 5-10 min) to see if colored or fluoresce nt spots (at 254 and 360nm) can be see n.Charri ng can be con ti nued un til spots are brow n, grey or black.Flav one reage nt accord ing to NeuSee etha no lam ine diphe nyl borateFluorescam ineFor detecti on of primary and sec on dary amin es, peptides, sulfo namides, e.g.,n itrosoam ines after photolysisSpray plate with a solution of 0.1mg/ml 4-phe nyl-spiro[fura n-2(3H),1-phthala n]-3,3-dio nein acet one preparedfresh dailyFor stabilizati on of fluoresce nee at 366nm spray with 10g triethylam ine, brought to 100ml with dichlorometha ne.Fluoresce nt In dicatorFor detect ion of compo unds which absorb UV lightSome TLC plates whe n manu factured have an in orga nic fluoresce nt in dicator added to the slurry poured to make the final plates. This type of in dicator will not dissolve or elute off. They are activated at 254nm or 360nm (see recomme ndati ons of the manu facturer for that type of plate).Results: When activated the fluoresce nt in dicator will turn a gree n or white (depe nding on the in dicator added)and the compo unds appear as dark spots or shadows aga inst this backgro un d. If view ing at other tha n the activati on wavele ngth, the compo unds might also have some fluoresce nee of their own, so various colors aga inst a dark backgro und would be see n.Formaldehyde / sulfuric acidFor detect ion of alkaloids, aromatic hydrocarb ons, e.g., an tihyperte nsive drugsSpray with a soluti on of 37% formaldehyde in cone, sulfuric acid (1:10) immediately after taking the plate from the develop ing chamber . Heati ng is not n ecessary.Results: various colored spots.Formaldehyde / phosphoric acidFor detect ion of steroid alkaloids, steroid sapoge nins and phe no thiaz ine derivatives6Spray with a soluti on of 0.03g formaldehyde in 100ml of 85% phosphoric acid with stirri ng at room temperature.The reage nt is stable for several weeks.Furfural / sulfuric acidFor detect ion of carbamate estersSpray soluti on I: 1% soluti on of furfural in acet oneSpray soluti on II: 10% soluti on of sulfuric acid in acet oneSpray plate with I, the n II.Gen tia n Violet -Bro mineFor detect ion of lipidsSpray 0.1% gentian violet (crystal violet) in methanol onto plate and place in a tankcontaining bromine vapor .Results: lipids produce blue spots on a yellow backgro und.Gibb ' s reage ntFor detect ion of phe no Is. For further applicati ons see 2,6-dichloro quinon e-4-chloroimideSpray with a solution of 3% 2,6-dibromo-N-chloro-p-benzoquinone imine in toluene ormetha nol.Hydroxylam ine / ir on (III) chlorideFor detect ion of amides, lact on es, carboxylic acid esters and an hydridesSoluti on 1) Mix 1 vol part of 7g hydroxylam monium chloride in 100ml metha nol w 1 vol partof a soluti on of 7.2 gpotassium hydroxide in 100ml metha nol. Filter from precipitated potassium chloride.Soluti on 2) 2% soluti on of iron (HI) chloride in 1% aqueous hydrochloride acidSpray air dried plate first with soluti on 1, the n with soluti on 2Iodi ne containing compo undsDetect ion by decompositi on un der UVDry plates at 100 CAfter cooli ng spray with a small amount of 50% acetic acidIrradiate some minu tes with un filtered UV light.Results: Iodi ne compo unds show weakly violet to brow n spots. The color can be enhanced by spray ing with 10% acetic acid and irradiatio n with UV light (sudde n appeara nee of blue spots).Iodi ne vaporRelatively un specific uni versal reage nt for many orga nic compo undsCharge chamber with some crystals of iodi nePlace developed, dried chromatogram in iodi ne vaporResults: spots tur n tan-brow n in colorIodoplati nateFor detect ion of orga nic n itroge n compo un ds, alkaloids, e.g., coca ine metabolitesSpray with a freshly prepared mixture of 3ml hexachloroplati nic (IV) acid soluti on (10%) in97ml/m in water and100ml aqueous potassium iodide soluti on.Note - use 5% ethanol or methanol in water to prepare these solutions for the new polymer bound TLC plates.Iron (HI) chloride / potassium hexacya no ferrate / sodium arse nate (accord ing to Patters on& Cleme nts)For detect ion of iodi ne compo un ds, e..g., thyroid gla nd horm onesSolution I: 2.7% iron (HI) chloride hexahydrate in 2N hydrochloric acidSoluti on II: 3.5% potassium hexacya no ferrate in waterSoluti on III: dissolve 3.8g arse nic trioxide in 25ml 2N sodium hydroxide soluti on heat ingslightly, cool to 5 C, andadd 50ml 2N sulfuric acid, fill to 200ml with waterImmediately before use mix 5ml soluti on I, 5ml soluti on II and 1ml soluti on III.Spray onto the dry layer and dry carefully (temp below 50 C)Cover with glass plate and leave 15min in the darkResults: Iodi ne containing compo und show light blue spots on a yellowish backgro und.Lead tetraacetate / 2,7-dichlorofluoresce inFor detect ion of vicinal diols, glycosides and phe no Is, e.g., sugar acidsSoluti on I: 2% (w/v) lead tetraacetate in glacial acetic acidSolution II: 1% (w/v) 2,7-dichlorofluorescein in etha nolMix 5ml of each soluti on 1 and 2, fill to 200ml with dry tolue ne. This reage nt soluti on is stable for only about 2 hours.Mangan ese / salicylaldehydeFor detect ion of orga no thiophosphorus pesticidesSolution I: dissolve 100mg manganese chloride (Mn CI2.4H2O) in 100ml 80% alcoholSolution II: dissolve 1.3g 2-hydrozine quinoline in the lowest possible volume of hot etha nol. Dissolve 1 g salicyaldehyde in 5ml etha nol and add 1-2 drops glacial acetic acid. Comb ine both soluti onsand reflux 30minu tes. The crystals of salicyl-2-aldehyde -2-quin oli nehydraz one precipitated duri ng thecooli ng arerecrystallized from etha nol. For soluti on dissolve 50mg of the salicylate derivative in 100mletha nolSpray with a mixture of equal volumes of soluti ons 1 and 2.Man deli n s 'age ntSee Van adium(V) / sulfuric acidMercury (II) chloride / diphe ny Icarbaz oneFor detect ion of barbituratesSoluti on I: 2% etha nolic mercury (II) chlorideSoluti on II: 0.2% etha nolic diphe nl ycarbaz oneMix freshly before use in equal partsResults: Pink spots on a violet backgro undMercury (II) chloride / dithiz oneFor detect ion of barbituratesSpray with a freshly prepared 1:1 mixture of 1-2% mercury (II) chloride in etha nol and0.1-0.2% dithizo ne inetha nol.View under 360nm UV light4-Methoxybe nzaldehyde / sulfuric acid / etha nolFor detect ion erythromyci n and metabolitesSpray with 4-methocybe nzaldehyde/sulfuric acid/etha no I (1:1:9)Heat 1 mi nute at 100 CMethyl yellowFor detect ion of chlori nated in secticides and an timicrobial compo undsSpray dried plate with a soluti on of 0.1g methyl yellow (N,N-dimethyl-4-phe ny lazoa nili ne) in 70ml etha nol, add25ml water and fill to 100ml with etha nol.Dry at ambie nt temperatureIrradiate 5 min with UV light without a filterResults: Red spots on a yellow backgro undMolybdatophosphoric acidSee un der Phosphomolybdic acidMolybde num blue reactio n accord ing to Dittmer and LesterFor detect ion of phospholipids and phosphoric acid derivativesSolution I: Boil 40.11g MoO3 in 1 liter 25N sulfuric acid for 3-4 hours until the molybde num oxide is completely dissolved. Let the light yellow soluti on slowly cool to ambie nt temperature overni ght. The soluti on will tur n light blue.Soluti on II: Boil 1.78 g molybde num powder and 500ml of soluti on I for 15m in, cool and deca nt from the rema ining residue.8For preparati on of the spray reage nt, add equal volumes of solutio ns I and II to 4.5 volume parts water. A darkgree n soluti on is formed.Soluti ons I and II are stable for several mon ths whe n stored in the dark. The spray reage nthas to be prepared weekly.Nin hydri nFor detect ion of amino acids, amin es, amino sugars.Spray with a soluti on of 0.2g nin hydri n in 100ml etha nol and heat to 110 C un til spots appear.Results: reddish spots appearNin hydri n / cadmium acetateFor detect ion of amino acids and heterocyclic aminesDissolve 1g ninhydrin and 2.5g cadmium acetate in 10ml glacial acetic acid and fill to500ml with etha nol.Spray and heat 20min at 120 CResults: Red, pink, or purple spots are see n.Nin hydri n / pyrid ine / glacial acetic acidFor detect ion of peptidesSpray with a 1% nin hydri n in pyrid in e/glacial acetic acid (5:1, v/v)Heat 5 min at 100 CNitric acid / etha nolFor detect ion of amines and alkaloidsSpray with a solution of 50 drops 65% nitric acid in 100ml ethanol (higher acidconcen trati ons are also possible).If n ecessary, heat to 120 C for some time.Orcino I (Bials reage nt)For detect ion of glycosides, glycolipidsReage nt: dissolve 0.1g orci nol in 40.7ml c one. HCl, add 1ml 1% ferric (111) chloride, and dilute to 10ml Spray and heat at 80 ° C for 90 minutes.Results: Glycolipids produce violet spots.Patters on and Cleme ntsSee un der Iron (III) chloride/potassium hexacya no ferrate/sodium arse nateParaffi n oilFor enhan ceme nt of fluoresce nee spots —more stable and greater in te nsity1% paraffi n oil in hexa neSpray eve nly over the TLC plateResults: spots should be more stable (no fadi ng with time) for sca nning and are of greater inten sitym-Phe nylen ediam ineFor detect ion of reduci ng sugarsSpray with a soluti on of 3.6g m-phe nylen ediam ine dihydrochloride in 100ml 70% etha nol and heat briefly at 105 ° CResults: Inten sely fluoresce nee colors in UV light (wavele ngth not specified, so check 254and 366nm).o-Phe nylen ediam ine -trichloroacetic acidFor detect ion of alpha-keto acidsSpray with a soluti on of 0.05g 1,2-phe nylen ediam ine in 100ml 10% aqueous trichloroacticacid and heat plate at100C for no more tha n 2 minu tes.Results: Gree n fluoresce nee spots in long wavele ngth UV light.p-Phe nylen ediam ine -phthalic acidFor detect ion of conjugated 3-ketosteroids9Spray with a soluti on of 0.9 p-phe nylen ediam ine and 1.6g phthalic acid in 100ml 1-buta nol saturated with waterand heat plate at 100-110 ° CResults: Yellow to orange spotsPheny Ihydraz ine sulfo nateFor detect ion of some an timicrobial compo undsSoluti on I: dissolve 3.5g phe ny Ihydraz ine 4-sulfo nic acid hemihydrate in 10ml water and20ml 1N NaOH solutionSoluti on II: mix 30ml 1N sodium hydroxide soluti on with 40ml acet oneThe spray reage nts have to be prepared fresh each time.Procedure:Wet chromatogram eve nly with spray soluti on 1After air drying the plate, shake spray soluti on 2 and spray plate.Phosphoric acidFor detect ion of sterols, steroids, and bile acidsSpray heavily un til the layer appears tran spare nt with a solutio n of 85% phosphoric acidwith water (1:1, v/v)Then heat 10-15minutes at 120 ° CResults: Sterols, steroids and bile acids and bile acids produce various colors un der visible and UV light. Phosphoric acid - bromineFor detect ion of digitalis glycosidesSpray soluti on I: 10% aqueous phosphoric acid soluti onSpray soluti on II: Mix 2ml saturated aqueous potassium bromide, 2ml saturated soluti on aqueous potassium bromate and 2ml 25% hydrochloric acid.Procedure: Spray plate with I and heat 12 min at 120 ° C.Results: Digitalis glycosides of the series B, D, and E show blue fluoresce nee in long wavele ngth UV light Procedure continued: Heat the plate again at 120 ° C and spray lightly with II Results: Glycosides of the series A show orange, of the series C show grey-green to grey-blue fluoresce nee in UV light.Phosphomolydbic acidFor detect ion of reduci ng substa nces, e.g, alcohols, bile acids, lipids, fatty acids, steroidsAlso used as a charri ng reage nt for polymer bound TLC plates (the n ewer hard layer plates)Spray with a soluti on of 250mg molybdatophosphoric acid in 50ml etha nolHeat to 120 C un til spots appear (ove n or heat gun)If n ecessary, treat with ammonia vapors to remove some backgro und colorati on.The reage nt soluti on is stable for only 10 days even in the dark.Results whe n using as a charri ng reage nt: View freque ntly (every 5-10 min) to see if colored or fluoresce nt spots(at 254 and 360nm) can be see n. Charri ng can be con ti nued un til spots are brow n, grey or black. Phosphot un gstic acidFor detection of cholesterol and its esters, reducing compounds, lipids, sterols, and steroidsSpray with 20% phosphotungstic acid in ethanol, heat at 110 ° Cfor 5-15min or until maximum visualizati on of thespots occursResults: Cholesterol, esters will produce red spots.Pin acryptol yellowFor detect ion of sweet ners, surfacta nts, alkyl- and ary lsulfo nic acidsDissolve 100mg pin acryptol yellow in 100ml hot water or etha nol (or some comb in ati on forpolymer boundplates)Spray with reage nt soluti onResults: Yellow to orange fluoresce nee spots un der long wavele ngth UV light (366nm)Potassium dichromate / sulfuric acid (chromosulfuric acid) - see below10Potassium perma ngan ate / sulfuric acid -see belowRhodam ine BFor detect ion of a wide variety of compo undsDry the developed chromatogram and spray with 0.025-0.25% etha nolic rhodam ine B reage nt.。

最全的TLC经验薄层层析显色剂TLC（Thhin-layer chromatography）是一种常用的分析技术，利用固定相上的物质分离样品中的化合物。

在TLC分析中，选择合适的显色剂是非常重要的，它可以使分离后的化合物形成明显的色斑，便于后续的观察和分析。

以下是一些常用的TLC显色剂及其应用经验：1.碘气（I2）显色剂：碘气广泛应用于氨基酸、激素、脂肪类化合物等的分析。

在整个TLC显色的过程中，容器内需维持相对湿润的环境，这有助于碘分子与分离出的化合物结合并形成色斑。

2.磷钼酸（H3PO4-MoO3）显色剂：磷钼酸显色剂对一些有机物具有较高的选择性，适用于血红素、古菌素等类似化合物的分析。

但是，过多的样品可能会导致背景的显色。

3.臭氧-有机酸显色剂：臭氧-有机酸显色剂可用于许多有机物的分析，如甾体激素、有机酸等。

显色过程中需要在高浓度臭氧气体下进行，非常危险，必须小心操作。

4.特殊显色剂：一些特殊化合物需要使用特殊的显色剂才能形成色斑，例如卡尔曼反应、尼特罗蒽酮反应等。

这些显色剂需要额外的处理步骤，并需要小心操作以避免有毒物质的接触。

此外，除了选择合适的显色剂，还有一些TLC操作经验值得注意：1.选用合适的固定相：根据分析样品的理化性质选择合适的固定相。

非极性固定相适用于极性化合物的分离，疏水性固定相适用于疏水化合物的分离等。

2.控制溶剂系统：在TLC中，正确的溶剂系统对于样品的分离非常重要。

使用不同比例的溶剂混合物，根据样品的极性和亲水性来优化分离效果。

3.控制样品施加的量：在施加样品前，要先进行样品的预处理，并确保样品施加均匀。

过多的样品可能会导致色斑扩散，影响分离效果。

4.控制板子的浸渍时间：浸渍时间的控制也非常重要，如果时间过短，样品可能没有完全被吸附;如果时间过长，可能会扩散过多的色斑，导致分离效果不佳。

5.注意保护眼睛和皮肤：在整个TLC显色的过程中，一些显色剂具有刺激性和毒性。

在操作时，必须佩戴适当的防护设备，如手套、护目镜等，以避免对人体健康的影响。

TLC常用显色剂适用范围显色剂名称配制方法使用方法显色通用碘蒸气将碘结晶置于密闭容器产生饱和碘蒸气放入碘蒸汽数分钟黄棕色硫酸浓H2SO4:MeOH(1:1)或５％H2SO4乙醇溶液喷雾110℃加热5min各种颜色磷钼酸试液5%5%磷钼酸乙醇溶液磷钼酸乙醇溶液喷雾120℃加热蓝色具还原性物质中性KMnO40.05%KMnO4水溶液喷雾淡黄色背景黄色斑点生物碱改良碘化铋钾试剂Dragendoff’sreagentⅠ.碱式硝酸铋0.85g溶于10ml冰醋酸和40mlH2O Ⅱ.8g碘化钾溶于20mlH2O贮存液：Ⅰ与Ⅱ等体积混合喷雾剂：1ml贮存液、2ml冰醋酸和10mlH2O混合喷雾桔红色碘－碘化钾试剂(wagner’s reagent) 1gI2和10gKI加50mlH2O溶解，再加2ml冰醋酸，用H2O稀释至100ml 喷雾棕褐色硫酸铈－硫酸试剂将0.1g硫酸铈悬浮与4mlH2O,加1g三氯乙酸煮沸，后滴加浓H2SO4至浑浊消失喷雾110℃加热８min各种颜色还原糖草酸苯胺试剂0.9g苯胺溶于0.1N草酸水溶液100ml中喷雾1０0℃加热粉红－棕红糖茴香醛－硫酸试剂1ml浓H2SO4加至含0.5ml茴香醛的50ml乙醇溶液中（用时配）喷雾105℃加热颜色不一a-萘酚-硫酸试剂(Molish’reagent)21ml 15%-萘酚乙醇溶液，13ml浓H2SO4,87mlEtOH,8mlH2O混合喷雾100℃加热３~6min蓝色（鼠李糖橙色）黄酮碱性试剂氨气，10%NaOH或KOH甲醇溶液喷雾前后在日光和紫外灯下观察本身颜色或荧光加强或改变三氯化铝试剂1%或5%AlCl3乙醇溶液醋酸镁试剂2%MgAc2甲醇溶液三氯化铁试剂1-2%FeCl3乙醇溶液蒽醌酸性试剂氨气，10% KOH甲醇，3%NaOH或NaCO3溶液喷雾日光和紫外灯下观察同上醋酸铝试剂０.5%AlCl3溶液醋酸镁试剂０.５%MgOAc2甲醇溶液喷雾90℃显色5min 紫色或红色等1适用范围适用范围 显色剂名称显色剂名称配置方法配置方法使用方法使用方法显色显色香豆素，酚芳香胺和能偶合的杂环化合物重氮化氨基苯磺酸试剂酸试剂0.9g 对氨基苯磺酸加热溶解于9ml 12N HCl 中，用水稀释至100ml,100ml,取取10ml,10ml,冷却，冷却，加热冰冷的4.5%4.5%亚硝酸钠亚硝酸钠10ml,010ml,0℃放置℃放置15min,用前加等体积１%NaCO 3喷雾喷雾不同颜色不同颜色酚 三氯化铁－铁氰化钾试剂化钾试剂 Ⅰ. 2%FeCl 3乙醇溶液乙醇溶液Ⅱ. 2%铁氰化钾水溶液铁氰化钾水溶液 使用前Ⅰ、Ⅱ等量混合使用前Ⅰ、Ⅱ等量混合喷雾喷雾蓝－紫色蓝－紫色 挥发油挥发油香草醛－硫酸试剂5%5%香草醛乙醇溶液香草醛乙醇溶液100ml,100ml,临用临用加4ml 浓H 2SO 4喷雾喷雾110℃加热℃加热各种颜色各种颜色 茴香醛－硫酸试剂1ml 浓H 2SO 4加入50ml 冰醋酸中冷却加0.5ml 茴香醛茴香醛 喷雾喷雾 105℃加热℃加热各种颜色各种颜色 有机酸有机酸甲红－溴酚兰指示剂示剂1g 甲红、3g 溴酚兰溶于95%EtOH100ml 如展开剂有HOAc,HOAc,先于先于先于 120℃挥去喷雾喷雾 黄色背景红色斑点红色斑点 溴甲酚绿指示剂溴甲酚绿指示剂1ml 浓H 2SO 4加至含0.5ml 茴香醛的50ml 乙醇溶液中（用时配） 蓝色背景黄色斑点黄色斑点皂甙皂甙血球试液血球试液取羊血或兔血1份，加玻璃球不断振摇除去玻璃球和凝集的血蛋白，加pH7.4磷酸缓冲液7份稀释而成份稀释而成喷雾喷雾 红色背景白色斑点白色斑点皂甙、萜皂甙、萜磷钼酸试剂磷钼酸试剂5%5%磷钼酸乙醇溶液磷钼酸乙醇溶液磷钼酸乙醇溶液喷雾喷雾140℃加热℃加热5~10min深蓝色深蓝色皂甙、强心苷、甾、萜三氯化锑试剂三氯化锑试剂 25%25%或饱和三氯化锑氯仿溶液或饱和三氯化锑氯仿溶液或饱和三氯化锑氯仿溶液 喷雾喷雾 100℃加热℃加热 5min同上同上五氯化锑试剂五氯化锑试剂25%25%五氯化锑氯仿溶液五氯化锑氯仿溶液五氯化锑氯仿溶液 同上同上 各种颜色各种颜色 甾体、三萜三萜 氯磺酸－乙酸试剂 氯磺酸－乙酸氯磺酸－乙酸(1:2) (1:2)喷雾喷雾 130℃加热℃加热 5min 各种颜色紫外灯下观察观察 皂甙皂甙 三氯乙酸试剂三氯乙酸试剂 三氯乙酸：乙酸三氯乙酸：乙酸(1:2) (1:2) 喷雾喷雾 120℃加热℃加热黄色黄色 强心苷、甾体三氯乙酸试剂三氯乙酸试剂Ⅰ.25%.25%三氯乙酸的乙醇或氯仿三氯乙酸的乙醇或氯仿溶液Ⅱ．取Ⅰ取Ⅰ10ml 10ml 加过氧去氢4滴或与新配置的3%3%氯胺氯胺T 水溶液4:1混合混合喷雾喷雾110℃加热℃加热7~10min 紫外灯下观察荧光观察荧光皂甙及甾醇 醋酐－浓硫酸试剂(Liebermann-Bur chard chard’’s reagent)冷却下，小心将5ml 醋酐与5ml 浓硫酸混合，并将其加入50ml 无水乙醇中（用前配置）无水乙醇中（用前配置） 喷雾喷雾100℃加热℃加热 10min 长波紫外灯下观察灯下观察强心甙强心甙Kedde Kedde’’sreagent 2%3,5-2%3,5-二硝基苯甲酸甲醇溶液与二硝基苯甲酸甲醇溶液与2N 氢氧化钾溶液用前1:1混合混合喷雾喷雾 紫红色紫红色 碱性三硝基苯试剂Ⅰ．间三硝基苯试剂100mg 溶于二甲基甲酰氨40ml 40ml，再加浓硫，再加浓硫酸3~4滴加水至100ml Ⅱ．Ⅱ．5%5%5%碳酸钠溶液碳酸钠溶液碳酸钠溶液先喷雾Ⅰ再喷雾Ⅱ90~100℃ 加热5min 浅橙色背景橙色红斑点斑点氨基酸氨基酸茚三酮试剂茚三酮试剂 茚三酮0.3g 溶于正丁醇100ml,加冰醋酸3ml 或0.2%0.2%乙醇溶液乙醇溶液乙醇溶液 喷雾喷雾110℃加热℃加热 蓝紫色蓝紫色 1,2-1,2-萘醌萘醌萘醌-4--4--4-磺酸磺酸试剂试剂1,2-1,2-萘醌萘醌萘醌-4--4--4-磺酸磺酸0.02g 溶于5%碳酸钠溶液100ml 中（用前配置）置） 喷雾喷雾 室温干燥室温干燥 各种颜色各种颜色。

薄层色谱的显色反应显色试剂显色剂可以分成两大类：一类是检查一般有机化合物的通用显色剂；另一类是根据化合物分类或特殊官能团设计的专属性显色剂。

显色剂种类繁多，本章只能列举一些常用的显色剂。

l．通用显色剂①硫酸常用的有四种溶液：硫酸-水(1：1)溶液；硫酸-甲醇或乙醇(1：1)溶液；1.5mol／L硫酸溶液与0.5-1.5mol／L硫酸铵溶液，喷后110oC烤15min，不同有机化合物显不同颜色。

②0.5％碘的氯仿溶液对很多化合物显黄棕色。

③中性0.05％高锰酸钾溶液易还原性化合物在淡红背景上显黄色。

④碱性高锰酸钾试剂还原性化合物在淡红色背景上显黄色。

溶液I：1％高锰酸钾溶液；溶液Ⅱ：5％碳酸钠溶液；溶液I和溶液Ⅱ等量混合应用。

⑤酸性高锰酸钾试剂喷1.6％高锰酸钾浓硫酸溶液(溶解时注意防止爆炸)，喷后薄层于180oC加热15～20min。

⑥酸性重铬酸钾试剂喷5％重铬酸钾浓硫酸溶液，必要时150oC烤薄层。

⑦5％磷钼酸乙醇溶液喷后120oC烘烤，还原性化合物显蓝色，再用氨气薰，则背景变为无色。

⑧铁氰化钾-三氯化铁试剂还原性物质显蓝色，再喷2mol／L盐酸溶液，则蓝色加深。

溶液I：1％铁氰化钾溶液；溶液Ⅱ：2％三氯化铁溶液；临用前将溶液I和溶液Ⅱ等量混合。

2．专属性显色剂由于化合物种类繁多，因此专属性显色剂也是很多的，现将在各类化合物中最常用的显色剂列举如下：(1)烃类①硝酸银／过氧化氢检出物：卤代烃类。

溶液：硝酸银O.1g溶于水lml，加2-苯氧基乙醇lOOml，用丙酮稀释至200ml，再加30％过氧化氢1滴。

方法：喷后置未过滤的紫外光下照射；结果：斑点呈暗黑色。

②荧光素／溴检出物：不饱和烃。

溶液：I．荧光素0.1g溶于乙醇lOOml；Ⅱ．5％溴的四氯化碳溶液。

方法：先喷(I)，然后置含溴蒸气容器内，荧光素转变为四溴荧光素(曙红)，荧光消失，不饱和烃斑点由于溴的加成，阻止生成曙红而保留荧光，多数不饱和烃在粉红色背景上呈黄色。

常用试剂配制及显色方法（一）通用显色剂1．重铬酸钾——硫酸一般有机物均能显色，不同化合物显示不同颜色。

喷洒剂：5克重铬酸钾溶于100ml，40%硫酸中。

喷洒后加热至150o C斑点出现。

2．碘：检查一般有机物（1）碘蒸汽：在一个密闭玻璃皿先放入碘片，使缸内空气被碘蒸气饱和将薄层或纸层放入缸内数分钟即显色，有时在缸内放一盛水的小杯，增加缸内的湿度，可提高显色的灵敏度。

（2）0.5%碘的氯仿溶液，取出挥发散过量的碘再喷1%淀粉的水溶液，斑点转成蓝色。

3．碘——碘化钾溶液：很我有机物虽黄色（配法见后）。

4．5%——磷钼酸乙醇溶液：喷后120o C烤，还原性物质显兰色，再用氨气熏，则背景变为无色。

5．20%磷酸乙醇溶液：喷后120o C，还原性物质显兰色。

6．碱性高锰酸钾试剂：还原性物质在淡红背景上显黄色。

溶液I:1%高锰酸钾溶液。

溶液II：5%碳酸钠溶液。

溶液I和溶液II等量混合使用。

7．中性0.05%高锰酸钾溶液：易还原性物质在淡红背景上显黄色。

8．硝酸银——氢氧化铵（Tollen’s--zoffaronl）试剂，喷后105o C烤5~10分钟，还原性物质显黑色。

溶液I：0.1N硝酸银溶液。

溶液II：氢氧化铵溶液。

临用前溶液I和溶液II以1：5混合。

注意：放久则形成爆炸性的叠氦化银。

9．硝酸银——高锰酸钾试剂：还原性物质在兰绿色背景上立即显黄色。

溶液I：0.1N硝酸银溶液：2N氢氧化铵溶液：2N氢氧化钠（1：1：2）。

临用前配制。

溶液II：高锰酸钾0.5g，碳酸钠1g，加水成100ml溶液,临用前溶液I和溶液II等量混合。

10．四唑试剂：还原性物质，在室温或微加热时显紫色。

溶液I：0.5%四唑兰甲醇溶液。

溶液II：6N氢氧化钠溶液。

临用前溶液I和溶液II等量混合。

11．铁氰化钾：三氯化铁试剂：还原性物质显兰色，再喷射2N/L盐酸溶液，则兰色加深。

溶液I：1%铁氰化钾溶液。

溶液II：2%三氯化铁溶液。

甘氨酸H2 N - CH2 - COOH缬氨酸CH3 - CH(CH3 ) - CH(NH2) - COOHTLC展开条件：（水:乙醇:乙酸= 1:6:0.5）显色剂：茚三酮显色丙氨酸赖氨酸赖氨酸50×150mm层析滤纸层析纸在展缸中饱和过夜纸色谱条件：正丁醇:乙酸:水(4:1:5)显色剂：茚三酮显色核苷酸测定通则，tlc条件硅胶GF板，展开剂：（正丁醇：丙酮：冰乙酸：5%氨水：水=7：5：3：3：2）体积比。

紫外254nm显色赖氨酸、丝氨酸、丙氨酸、缬氨酸、亮氨酸，天门冬氨酸等六种氨基酸以正丁醇-乙酸-水（4∶1∶1）作展开剂，0．5％茚三酮作显色剂进行的薄层层析，效果较好。

列出溶剂极性参数表，方便以下比较展开剂。

环已烷:-0.2、石油醚（Ⅰ类，30~60℃）、石油醚（Ⅱ类，60~90℃）、正已烷:0.0、甲苯:2.4、二甲苯:2.5、苯:2.7、二氯甲烷:3.1、异丙醇:3.9、正丁醇:3.9、四氢呋喃:4.0、氯仿:4.1、乙醇:4.3、乙酸乙酯:4.4、甲醇:5.1、丙酮:5.1、乙腈:5.8、乙酸:6.0、水:10.2。

首先是极性较小的挥发性物质。

比如：冰片：石油醚(30～60℃)—醋酸乙酯(17:3)、厚朴酚：苯－醋酸乙酯(9:1.5)、α-香附酮：苯－醋酯乙酯－冰醋酸(92:5:5)、丹皮酚：环己烷－醋酸乙酯(3:1)，这类化合物，以石油醚、正构烷和苯为体积百分数比较大的溶剂，通常起溶解和分离化合物的作用，而用醋酸乙酯为调节Rf （比移值）的溶剂。

为了减少拖尾之类其他相似相溶原则以外的影响，适当加入添加剂，如有机酸或者有机碱。

极性较小的不挥发性物质。

比如：β -谷甾醇：环己烷－醋酸乙酯－甲醇(6:2.5:1)或者环己烷－丙酮(5:2) 、熊果酸：甲苯－醋酸乙酯－冰醋酸(12:4:0.5)、齐墩果酸：氯仿－甲醇(40:1)、猪去氧胆酸：氯仿－乙醚－冰醋酸(2:2:1)、大黄素：苯—醋酸乙酯—甲醇(15:2:0.2)或者苯—乙醇(8:1)、丹参酮ⅡA：苯-醋酸乙酯-甲酸(40:25:4) 、穿心莲内酯：氯仿-无水乙醇(9:1)、靛玉红、靛蓝氯仿－乙醇(9:1)或者苯－氯仿－丙酮(5:4:1)。

选择适当‎的展开剂是‎首要任务.‎一般常用溶‎剂按照极性‎从小到大的‎顺序排列大‎概为：石油‎迷<己烷<‎苯<乙醚<‎T HF<乙‎酸乙酯<丙‎酮<乙醇<‎甲醇使用单‎一溶剂，往‎往不能达到‎很好的分离‎效果，往往‎使用混合溶‎剂通常使用‎一个高极性‎和低级性溶‎剂组成的混‎合溶剂，高‎极性的溶剂‎还有增加区‎分度的作用‎，常用的溶‎剂组合有：‎Pet‎r oleu‎m ethe‎r/Eth‎y lace‎t ate,‎p etro‎l eume‎t her/‎A ceto‎n e,Pe‎t role‎u meth‎e r/Et‎h er, ‎P etro‎l eume‎t h er/‎C H2Cl‎2, et‎h ylac‎e tate‎/MeOH‎,CHCl‎3/eth‎y lace‎t ate‎展开剂‎的比例要靠‎尝试.一般‎根据文献中‎报道的该类‎化合物用什‎么样的展开‎剂，就首先‎尝试使用该‎类展开剂，‎然后不断尝‎试比例，直‎到找到一个‎分离效果好‎的展开剂。

‎展开剂的选‎择条件：①‎对的所需成‎分有良好的‎溶解性；②‎可使成分间‎分开；③待‎测组分的R‎f在0.2‎~0.8之‎间，定量测‎定在0.3‎～0.5之‎间；④不与‎待测组分或‎吸附剂发生‎化学反应；‎⑤沸点适中‎，黏度较小‎；⑥展开后‎组分斑点圆‎且集中；⑦‎混合溶剂最‎好用新鲜配‎制。

一般‎来说，弱极‎性溶剂体系‎的基本两相‎由正己烷和‎水组成，再‎根据需要加‎入甲醇、乙‎醇，乙酸乙‎酯来调节溶‎剂系统的极‎性，以达到‎好的分离效‎果，适合于‎生物碱、黄‎酮、萜类等‎的分离；中‎等极性的溶‎剂体系由氯‎仿和水基本‎两相组成，‎由甲醇、乙‎醇，乙酸乙‎酯等来调节‎，适合于蒽‎醌、香豆素‎，以及一些‎极性较大的‎木脂素和萜‎类的分离；‎强极性溶剂‎，由正丁醇‎和水组成，‎也靠甲醇、‎乙醇，乙酸‎乙酯等来调‎节，适合于‎极性很大的‎生物碱类化‎合物的分离‎。