铁—邻菲咯啉类金属配合物与DNA作用机理及DNA荧光光纤传感器的研究
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摘要
脱氧核糖核酸(DNA)是生命的最基本物质之一,广泛存在于各种生物体内,是储存、复制和传递遗传信息等生命过程的主要物质基础。金属离子及其配合物在许多生命过程中起着核心作用,是核酸等生物大分子研究的重要探针。本文以研究DNA传感器为目的,合成和表征了两种铁-邻菲咯啉类金属配合物;用荧光光谱法、紫外.可见光谱法、电化学法研究了它们与DNA的相互作用;采用共键合法对光纤表面进行了修饰,分别以这两种金属配合物为杂交指示剂制成了DNA荧光光纤传感器,具体工作如下:
(1)以邻菲咯啉及其中性衍生物咪唑并[f]邻菲咯啉和对羟基苯并咪哗并[f]邻菲咯啉为配体,合成了[Fe(phen)_2IP]·3ClO_4·2H_2O(phen=邻菲咯啉,IP=咪唑并[f]邻菲咯啉)和[Fe(phen)_2PHPIP]·3ClO_4·2H_2O(phen=邻菲咯啉,PHPIP=对羟基苯并咪哗[f]邻菲咯啉)两种金属配合物;通过配合物的红外光谱(IR)、元素分析(EA)测定,确定了配合物的分子组成;用3-D荧光光谱验证了它们的荧光特性。
(2)利用荧光光谱法、紫外-可见光谱法和电化学法研究了[Fe(phen)_2IP]~(3+)和[Fe(phen)_2PHPIP]~(3+)与鲑鱼精DNA的相互作用,确定了配合物与DNA的作用方,测定了结合比和结合常数。实验结果表明,[Fe(phen)_2IP]~(3+)与DNA的作用方式嵌插结合,结合比为1:1,结合常数为4.00×102L·mol~(-1)。[Fe(phen)_2PHPIP]~(3+) DNA的作用方式也为嵌插结合,结合比为1:2,结合常数为1.02×103L0.5·mol-0.5。
(3)对光纤表面进行了修饰,用共价键合法制备了DNA修饰石英光纤,分别以[Fe(phen)_2IP]~(3+)和[Fe(phen)_2PHPIP]~(3+)为杂交指示剂制成了DNA荧光光纤传器,分别用它们检测了与固定在光纤上的探针DNA互补的靶DNA。实验表明,以[Fe(phen)_2IP]~(3+)为杂交指示剂对互补DNA进行检测,检测的线性范围为1.25×10-8 mol-L-1～1.50×10-7 mol·L~(-1) ,检测限为1.35×10-9 mol·L~(-1)。以[Fe(phen)_2PHPIP]~(3+)为杂交指示剂对互补DNA进行检测,检测的线性范围为4.98×10-7 mol·L~(-1)～4.88×10-6 mol·L~(-1) ,检测限为1.08×10-7 mol·L~(-1)。
Deoxyribonucleic acid (DNA) is the basic material in the most of lives, which exits in all kinds of organism extensively, and it is the main material base of some lives course, such as storing, duplicating and transmitting hereditary information, etc.. Metal complexes play a key role in the course of a lot of lives, and they are important probes for studying some molecular such as nucleic acid. To study DNA biosensor, two 1,10-phenanthroline complexes of ferrum(III) were synthesized and characterized. The interactions between the complexes and salmon sperm DNA were studied by fluorescence spectrum, UV-Vis spectrum and electrochemical methods. With two 1,10-phenanthroline complexes of ferrum(III) as the hybridization indicators, the fluorescence fiber-optic DNA biosensors with covalently immobilized DNA modified optical fiber have been prepared.
(1) With 1,10-phenanthroline and 1,10-phenanthroline ramification using as ligands, two complexes imidazo [f]1,10-phenanthroline ferrum(III) ([Fe(phen)2 IP]·3ClO4·2H2O) and p-Hydroxyphenylimidazo [f] 1,10-phenanthroline ferrum(III) ([Fe(phen)2 PHPIP]·3ClO4·2H2O) have been synthesized successfully. The structures of these novel complexes have been determined by means of IR and EA, respectively. The fluorescent property of the two complexes were demonstrated by 3-D fluorescence spectrum.
(2) The interactions mechanism between two metal complexes and salmon sperm DNA have been studied by means of fluorescence spectrum, UV-Vis spectrum and electrochemical methods, respectively. The results show that the interaction mode between [Fe(phen)_2IP]~(3+) and DNA is intercalation, binding ratio is 1:1 and binding constant is 4.00×102 L·mol-1. The interaction mode between [Fe(phen)_2PHPIP]~(3+) and DNA is intercalation too, binding ratio is 1:2 and binding constant is 1.02×103 L0.5·mor-0.5。
(3) With the two complexes as the hybridization indicators respectively, the fluorescence fiber-optic DNA biosensors with covalently immobilized DNA modified optical fiber have been prepared. The fluorescence fiber-optic DNA biosensors can be used to the recognition of complementary ssDNA segment. Using [Fe(phen)_2IP]~(3+) as hybridization indicator, the complementary ssDNA could be quantified over the ranges from 1.25×10-8mol·L~(-1) to 1.50×l0-7 mol·L~(-1) and a detection limit of 1.35×10-9 mol·L~(-1) . Using [Fe(phen)_2PHPIP]~(3+) as hybridization indicator, the complementary ssDNA could be quantified over the ranges from 4.98×10-7 mol·L~(-1) to 4.88×10-6 mol·L~(-1) and a detection limit of 1.08x 10-7 mol·L~(-1) .
(1)以邻菲咯啉及其中性衍生物咪唑并[f]邻菲咯啉和对羟基苯并咪哗并[f]邻菲咯啉为配体,合成了[Fe(phen)_2IP]·3ClO_4·2H_2O(phen=邻菲咯啉,IP=咪唑并[f]邻菲咯啉)和[Fe(phen)_2PHPIP]·3ClO_4·2H_2O(phen=邻菲咯啉,PHPIP=对羟基苯并咪哗[f]邻菲咯啉)两种金属配合物;通过配合物的红外光谱(IR)、元素分析(EA)测定,确定了配合物的分子组成;用3-D荧光光谱验证了它们的荧光特性。
(2)利用荧光光谱法、紫外-可见光谱法和电化学法研究了[Fe(phen)_2IP]~(3+)和[Fe(phen)_2PHPIP]~(3+)与鲑鱼精DNA的相互作用,确定了配合物与DNA的作用方,测定了结合比和结合常数。实验结果表明,[Fe(phen)_2IP]~(3+)与DNA的作用方式嵌插结合,结合比为1:1,结合常数为4.00×102L·mol~(-1)。[Fe(phen)_2PHPIP]~(3+) DNA的作用方式也为嵌插结合,结合比为1:2,结合常数为1.02×103L0.5·mol-0.5。
(3)对光纤表面进行了修饰,用共价键合法制备了DNA修饰石英光纤,分别以[Fe(phen)_2IP]~(3+)和[Fe(phen)_2PHPIP]~(3+)为杂交指示剂制成了DNA荧光光纤传器,分别用它们检测了与固定在光纤上的探针DNA互补的靶DNA。实验表明,以[Fe(phen)_2IP]~(3+)为杂交指示剂对互补DNA进行检测,检测的线性范围为1.25×10-8 mol-L-1～1.50×10-7 mol·L~(-1) ,检测限为1.35×10-9 mol·L~(-1)。以[Fe(phen)_2PHPIP]~(3+)为杂交指示剂对互补DNA进行检测,检测的线性范围为4.98×10-7 mol·L~(-1)～4.88×10-6 mol·L~(-1) ,检测限为1.08×10-7 mol·L~(-1)。
Deoxyribonucleic acid (DNA) is the basic material in the most of lives, which exits in all kinds of organism extensively, and it is the main material base of some lives course, such as storing, duplicating and transmitting hereditary information, etc.. Metal complexes play a key role in the course of a lot of lives, and they are important probes for studying some molecular such as nucleic acid. To study DNA biosensor, two 1,10-phenanthroline complexes of ferrum(III) were synthesized and characterized. The interactions between the complexes and salmon sperm DNA were studied by fluorescence spectrum, UV-Vis spectrum and electrochemical methods. With two 1,10-phenanthroline complexes of ferrum(III) as the hybridization indicators, the fluorescence fiber-optic DNA biosensors with covalently immobilized DNA modified optical fiber have been prepared.
(1) With 1,10-phenanthroline and 1,10-phenanthroline ramification using as ligands, two complexes imidazo [f]1,10-phenanthroline ferrum(III) ([Fe(phen)2 IP]·3ClO4·2H2O) and p-Hydroxyphenylimidazo [f] 1,10-phenanthroline ferrum(III) ([Fe(phen)2 PHPIP]·3ClO4·2H2O) have been synthesized successfully. The structures of these novel complexes have been determined by means of IR and EA, respectively. The fluorescent property of the two complexes were demonstrated by 3-D fluorescence spectrum.
(2) The interactions mechanism between two metal complexes and salmon sperm DNA have been studied by means of fluorescence spectrum, UV-Vis spectrum and electrochemical methods, respectively. The results show that the interaction mode between [Fe(phen)_2IP]~(3+) and DNA is intercalation, binding ratio is 1:1 and binding constant is 4.00×102 L·mol-1. The interaction mode between [Fe(phen)_2PHPIP]~(3+) and DNA is intercalation too, binding ratio is 1:2 and binding constant is 1.02×103 L0.5·mor-0.5。
(3) With the two complexes as the hybridization indicators respectively, the fluorescence fiber-optic DNA biosensors with covalently immobilized DNA modified optical fiber have been prepared. The fluorescence fiber-optic DNA biosensors can be used to the recognition of complementary ssDNA segment. Using [Fe(phen)_2IP]~(3+) as hybridization indicator, the complementary ssDNA could be quantified over the ranges from 1.25×10-8mol·L~(-1) to 1.50×l0-7 mol·L~(-1) and a detection limit of 1.35×10-9 mol·L~(-1) . Using [Fe(phen)_2PHPIP]~(3+) as hybridization indicator, the complementary ssDNA could be quantified over the ranges from 4.98×10-7 mol·L~(-1) to 4.88×10-6 mol·L~(-1) and a detection limit of 1.08x 10-7 mol·L~(-1) .
引文
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