新型罗丹明类荧光标记探针的研究(Ⅱ)——罗丹明B-N-羟基琥珀酰亚胺酯的合成与标记应用  

A Study on Novel Fluorescent Probe Containing Rhodamine Skeleton (Ⅱ)——Synthesis of Rhodamine B Succinimidyl Ester and Its Application as a Labeling Reagent

作　　者：郁美娟[1] 张海凤[1] 赖奕坚[1] 孟庆华[1] 张万斌[1] 

机构地区：[1]上海交通大学化学化工学院,上海200240

出　　处：《染料与染色》2004年第5期249-251,共3页Dyestuffs and Coloration

摘　　要：Rhodamine B(R)在乙腈介质中加入N-羟基琥珀酰亚胺和二环己基碳二亚胺,在45℃时反应1小时,常温下搅拌20小时制得标题化合物(BR—SE),用H—NMR、IR进行了表征。选择甘氨酸进行标记得到产物乙氨酸Rhodamine B酰胺(RB—Gly);RB、RB—SE、RB—Gly的最大可见吸收分别为545nm、558nm、556nm;在HPLC的分离中(流动相:甲醇:水:醋酸=70:29.4:0.6;检测波长560nm),最大吸收波长不受色谱条件影响,其保留时间分别为1.811min(RB—Gly)、2.198min(RB-SE)、3.070min(BR);RB—SE的浓度、缓冲液pH、标记反应温度对标记效率有影响。Rhodamine B (C.I.BasicVioletlO)was reacted with N-hydroxysuccinimide and dicyclohexylcarbodiimide at 45℃ for 1 hour, agitated at ambient temperature for 20 hours, giving the title compound (RB-SE), which was characterized by 1H-NMR and IR. The compound was applied for labeling glycine, to give RB-Gly; the maximum absorption wavelengths of RB, RB-SE and RB-Gly were 545nm, 558nm and 556nm respectively; they were separated by HPLC, a mixture of CH3OH-H2O-CH3COOH(70: 29.4: 0.6, V/V/V) as a mobile phase and detected at 560nm, the maximum absorption wavelength was not affected by the separation conditions, their retention times were 1.811 min (RB-Gly) ,2.198min(RB-SE),3.070min(RB) respectively; the labeling effect of RB-SE was affected by concentration, pH value of buffer solution and labeling reaction temperature.

关 键 词：RHODAMINE B 荧光探针 标记化合物 氡基酸 HPLC 

分 类 号：O657.3[理学—分析化学]