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Biological Labeling Fluorescent Dye And Free Radical Chlorination Reaction

Posted on:2005-06-26Degree:MasterType:Thesis
Country:ChinaCandidate:Z H PengFull Text:PDF
GTID:2191360122997161Subject:Applied Chemistry
Abstract/Summary:PDF Full Text Request
In this thesis, four different fluorescent dyes for labeling biological molecules have been synthesized and the mechanism of free radical chlorination of nitrotoluenes have been investigated.Firstly: A practical and reliable method for the production of multigram quantities of carboxy fluorescein is described. The poor solubility of fluorescein derivatives and their existence as lactone or free acid tautomers complicate 5- and 6-isomeric separation.Both of the problems are circumvented by the reaction of pivaloyl chloride and carboxyfluorescein in pyridine to give a crude mixture of dipivalates. After resolved in ethanol and treated with diisopropylamine, pure 6-carboxy fluorescein Dipivalates diisopropylamine salt is obtained. After treated with dilute hydrochloric acid and N,N-disuccinimidyl carbonate, the corresponding 6-carboxyfluorescein dipivalate succinimidyl ester which is suitable for labeling protein is synthesized.Secondly:Two novel BODIPY dyes are designed and synthesized. The p-substituted benzaldehyde condense with two equivalent 2, 4~dimethylpyrrole give dipyrromethane. Subsequent treatment of dipyrromethane with p-chloranil affords a black reaction mixture. Treatment of the black mixture with triethylamine and BF.,-etherate affords the carboxy BODIPY dyes and the hydroxy BODIPY dye in 9.6% and 35.7% yield respectively. The NHS ester of the former can be used for labeling protein, and the latter is suitable for labeling oligonucletides.Thirdly: The relative reactivities of two kinds of radical chlorination have been investigated. The result shows that the reactivity of denitrochlorination is much less than the first a -chlorination of methyl group but more than the second a-chlorination.
Keywords/Search Tags:Fluorescent dyes, Radical chlorination, Carboxy fluorescein, Pivaloyl chloride, N,N-disuccinimidyl carbonate, p-chloranil, Label ing protein, Labeling oligonucletides, Denitrochlorination
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