化学发光体系的研究与应用
摘要
本文建立了几种化学发光体系并对其应用进行了研究,同时介绍了CdTe纳米晶在化学发光分析及荧光免疫分析中的应用。具体内容如下:
一、第一章对化学发光分析方法的研究进展进行了综述,介绍了化学发光分析方法的应用、现状及发展前景。
二、第二章研究了三价稀土离子与无机酸形成的络合物及三价稀土离子/强酸弱碱盐体系的化学发光现象。对实验参数和发光机理做了较详细的研究,建立了化学发光新体系。
三、第三章研究了几个发光体系在药物分析及检测中的应用:利用双[2,4,6-三氯苯基]草酸酯/H_2O_2/锌卟啉发光体系检测了牛奶中残留的氨苄青霉素;利用鲁米诺/溶解O_2/Tween20发光体系研究了部分药物的抗氧化活性;利用luminol/H_2O_2体系检测了蔬菜中残留的甲胺磷农药。
四、第四章详细地研究了CdTe纳米晶在化学发光分析及荧光免疫分析中的应用。
Chemiluminescence(CL)analytical method and related analytical technique have intrigued people extensive interest and have been developed to be an important and powerful tool in different fields such as food analysis,environmental monitoring,drag analysis,clinical diagnose,biomedical detection,and biological process,because of its high sensitivity,wide linear range,simple instrumentation,and no background scattering light interference.In the paper,some CL systems were studied,conditions of CL reaction were investigated,the application of systems was exploited.In addition,CL system of TCPO or luminol were used to detect residues of pesticide or antibiotics on the food and evaluate scavenging activity of same antioxidants.
In the work,the CL of complexes of trivalent lanthanide ion(Tb~(3+),Eu~(3+),Ce~(3+)and La~(3+))with simple inorganic acid or same metal ions was studied.It was found that the CL system can produce chemiluminescence only in alkaline medium,and CL intensity is the strongest when sodium hydroxide concentration is 0.1mol/L,strong CL signal was obtained when the molar ratio of Tb(Ⅲ)and HCl,HNO_3,H_2SO_4,H_3PO_4,H_3BO_3,HAc are 1:50、1:40、1:20、1:40、1:90 and 1:20,respectively.oxidants,especially hydrogen peroxide and ammonium persulfate could oxidize complexes to produce strong CL emission.Has a good function relationship between CL intensity and concentration of complexes of Tb~(3+),Eu~(3+),Ce~(3+)and La~(3+)with HCl.
Trivalent lanthanide ion solution containing FeCl_2,CuCl_2,CrCl_3 or Al_2(SO_4)_3 can react with oxidants to produce strong CL signal in alkaline condition.But trivalent lanthanide ion solution containing Ni(NO_3)_2、MnCl_2 or AgNO_3 only produce weak CL signal.The results showed that strong CL signal was obtained when the molar ratio of Tb(Ⅲ)and Fe~(2+)、Cu~(2+)、Cr~(3+)、Al~(3+)were 1:20,1:10,1:30 and1:500.The kinetics curve of Tb~(3+)/Cr~(3+)/Ce~(4+)is narrow and high,S_2O_8~(2-)can reacted with system Tb~(3+)/Fe~(2+)、Tb~(3+)/A13+to produce strong CL signal,The kinetics curve of system Tb~(3+)/Cu~(2+)/H_2O_2 and Tb~(3+)/Cr~(3+)/ H_2O_2 is very wide.But K_3Fe(CN)_6 did not react with the all system.
TCPO/H_2O_2/fluorophore system is very efficient,having a quantum yield higher than that observed for most of the chemiluminescent reaction discovered to date.When zinc porphyrin was added into TCPO/H_2O_2 system,strong CL signal was obtained.The system for the determanition of ampicillin combined with steady injection analysis is presented in this paper.The method is based on that the CL signal of the zinc porphyrin /TCPO/H_2O_2 system can be significantly enhanced in the presence of ampicillin.The calibration curve was linear over the range 0.18～270μg/mL with a detection limit of 40ng/mL(S/N=3).The relative standard deviation was 6.8%(n=7)for 3.6μg/mL of ampicillin.A liquid-liquid solvent extraction procedure was developed for eliminating the effect of a number of potential interferences.The proposed method was applied to the analysis of ampicillin in bovine milk sample with satisfactory results.
It was found that when surfactant Tween 20,Tween 80,sodium dodecyl sulfate orβ-cyclodextrin were added into alkaline luminol the strong chemiluminescence signal was obtained.The CL intensity of luminal/Tween 20 system is the strongest.The CL was produced when luminal react with oxygen studies Tween 20 as catalyzer.The scavenging activity of L-Ascorbic acid,Tocopherol and Traditional Chinese medicine was studied by luminol/Tween 20 CL system.It can be found that catechin and chlorogenic acid have a good scavenging activity than the other antioxidants,scavenging activity ofα-,δ-,γ-Tocopherol is equivalent than L-Ascorbic acid.And order of their scavenging activity wereα＞γ＞δ.Scavenging activity of Magnolol is poor.
A simple,rapid method has been described for the determination of organophosphorus pesticide.The method is based on the methamidaphos enhancing effect on the CL reaction of luminol with hydrogen peroxide in alkaline solution.The optimal experimental conditions were investigated.And under the optimal conditions, the CL intensity was linear to the methamidaphos concentration in the range of 0.2-13μg/mL(r=0.9992).The detection limit(3σ)was 0.047μg/mL.Relative standard deviation for analysis of three samples were 1.8%、2.5%and 3.7%(n=5)respectively. The recoveries for the method were 90%-109%.This method has been successfully applied to the determination of methamidaphos residue on the vegetable samples.
The study has been reported that CdTe nanocrystals reacted with H_2O_2 to can produce chemiluminecsence in alkaline condition.In the work,effects of metal ions, L-ascorbic acid and polyhydric phenol on the CdTe nanocrystals CL was studied.Ba~(2+), Ca~(2+),Fe~(2+),Pb~(2+)and Cu~(2+)enhanced the CL intensity.Cr~(3+),Ni~(2+),Zn~(2+)and Ag~+ ion inhibited the CL intensity,and reduced CL intensity is linear to logarithm of metal ion concentration.L-ascorbic acid can capture oxygen radical,it react with the produced superoxide radical and hydroxyl radical in the CL reaction process,inhibited the CL of the CdTe/H_2O_2 system.Reduced CL intensity is linear to logarithm of L-ascorbic acid concentration in a range of 0.5～100μg/mL.The synthetical sample was detected, veracity is nice.Same polyhydric phenol can react with active oxygen,so can inhibit the CL of the CdTe/H_2O_2 system,but the sensitivity is low.Tween 20 was chosen as a catalyzer of the CL system,the polyhydric phenol was sensitively detected with the CdTe/H_2O_2/Tween 20 system.The CL intensity was linear to the concentration of Catechol in a range 0.05-1μg/mL(R=0.996)with a detection limit of 0.038μg/mL.This method has been successfully used to determine the content of polyhydric phenol in tap water.The recovery percentage was 87-109%.
Thioglycolic-acid-stabilized CdTe nanocrystals were applied to immunoassay for the determination of human immunoglobulin G(HIgG).The immunoassay was based on a sandwich immunoreaction of primary antibody on the magnetic nanoparticles,HIgG, and the secondary antibody labeled with CdTe nanocrystals.The method proposed in the work was compared with a fluorescein isothiocyanate(FITC).This system is an order of magnitude more sensitive than the FITC.The serum samples were analysed by nephelometry and the proposed method.At the same time,the standard addition testswere taken.The results indicate that the accuracy of the method is satisfactory.
一、第一章对化学发光分析方法的研究进展进行了综述,介绍了化学发光分析方法的应用、现状及发展前景。
二、第二章研究了三价稀土离子与无机酸形成的络合物及三价稀土离子/强酸弱碱盐体系的化学发光现象。对实验参数和发光机理做了较详细的研究,建立了化学发光新体系。
三、第三章研究了几个发光体系在药物分析及检测中的应用:利用双[2,4,6-三氯苯基]草酸酯/H_2O_2/锌卟啉发光体系检测了牛奶中残留的氨苄青霉素;利用鲁米诺/溶解O_2/Tween20发光体系研究了部分药物的抗氧化活性;利用luminol/H_2O_2体系检测了蔬菜中残留的甲胺磷农药。
四、第四章详细地研究了CdTe纳米晶在化学发光分析及荧光免疫分析中的应用。
Chemiluminescence(CL)analytical method and related analytical technique have intrigued people extensive interest and have been developed to be an important and powerful tool in different fields such as food analysis,environmental monitoring,drag analysis,clinical diagnose,biomedical detection,and biological process,because of its high sensitivity,wide linear range,simple instrumentation,and no background scattering light interference.In the paper,some CL systems were studied,conditions of CL reaction were investigated,the application of systems was exploited.In addition,CL system of TCPO or luminol were used to detect residues of pesticide or antibiotics on the food and evaluate scavenging activity of same antioxidants.
In the work,the CL of complexes of trivalent lanthanide ion(Tb~(3+),Eu~(3+),Ce~(3+)and La~(3+))with simple inorganic acid or same metal ions was studied.It was found that the CL system can produce chemiluminescence only in alkaline medium,and CL intensity is the strongest when sodium hydroxide concentration is 0.1mol/L,strong CL signal was obtained when the molar ratio of Tb(Ⅲ)and HCl,HNO_3,H_2SO_4,H_3PO_4,H_3BO_3,HAc are 1:50、1:40、1:20、1:40、1:90 and 1:20,respectively.oxidants,especially hydrogen peroxide and ammonium persulfate could oxidize complexes to produce strong CL emission.Has a good function relationship between CL intensity and concentration of complexes of Tb~(3+),Eu~(3+),Ce~(3+)and La~(3+)with HCl.
Trivalent lanthanide ion solution containing FeCl_2,CuCl_2,CrCl_3 or Al_2(SO_4)_3 can react with oxidants to produce strong CL signal in alkaline condition.But trivalent lanthanide ion solution containing Ni(NO_3)_2、MnCl_2 or AgNO_3 only produce weak CL signal.The results showed that strong CL signal was obtained when the molar ratio of Tb(Ⅲ)and Fe~(2+)、Cu~(2+)、Cr~(3+)、Al~(3+)were 1:20,1:10,1:30 and1:500.The kinetics curve of Tb~(3+)/Cr~(3+)/Ce~(4+)is narrow and high,S_2O_8~(2-)can reacted with system Tb~(3+)/Fe~(2+)、Tb~(3+)/A13+to produce strong CL signal,The kinetics curve of system Tb~(3+)/Cu~(2+)/H_2O_2 and Tb~(3+)/Cr~(3+)/ H_2O_2 is very wide.But K_3Fe(CN)_6 did not react with the all system.
TCPO/H_2O_2/fluorophore system is very efficient,having a quantum yield higher than that observed for most of the chemiluminescent reaction discovered to date.When zinc porphyrin was added into TCPO/H_2O_2 system,strong CL signal was obtained.The system for the determanition of ampicillin combined with steady injection analysis is presented in this paper.The method is based on that the CL signal of the zinc porphyrin /TCPO/H_2O_2 system can be significantly enhanced in the presence of ampicillin.The calibration curve was linear over the range 0.18～270μg/mL with a detection limit of 40ng/mL(S/N=3).The relative standard deviation was 6.8%(n=7)for 3.6μg/mL of ampicillin.A liquid-liquid solvent extraction procedure was developed for eliminating the effect of a number of potential interferences.The proposed method was applied to the analysis of ampicillin in bovine milk sample with satisfactory results.
It was found that when surfactant Tween 20,Tween 80,sodium dodecyl sulfate orβ-cyclodextrin were added into alkaline luminol the strong chemiluminescence signal was obtained.The CL intensity of luminal/Tween 20 system is the strongest.The CL was produced when luminal react with oxygen studies Tween 20 as catalyzer.The scavenging activity of L-Ascorbic acid,Tocopherol and Traditional Chinese medicine was studied by luminol/Tween 20 CL system.It can be found that catechin and chlorogenic acid have a good scavenging activity than the other antioxidants,scavenging activity ofα-,δ-,γ-Tocopherol is equivalent than L-Ascorbic acid.And order of their scavenging activity wereα＞γ＞δ.Scavenging activity of Magnolol is poor.
A simple,rapid method has been described for the determination of organophosphorus pesticide.The method is based on the methamidaphos enhancing effect on the CL reaction of luminol with hydrogen peroxide in alkaline solution.The optimal experimental conditions were investigated.And under the optimal conditions, the CL intensity was linear to the methamidaphos concentration in the range of 0.2-13μg/mL(r=0.9992).The detection limit(3σ)was 0.047μg/mL.Relative standard deviation for analysis of three samples were 1.8%、2.5%and 3.7%(n=5)respectively. The recoveries for the method were 90%-109%.This method has been successfully applied to the determination of methamidaphos residue on the vegetable samples.
The study has been reported that CdTe nanocrystals reacted with H_2O_2 to can produce chemiluminecsence in alkaline condition.In the work,effects of metal ions, L-ascorbic acid and polyhydric phenol on the CdTe nanocrystals CL was studied.Ba~(2+), Ca~(2+),Fe~(2+),Pb~(2+)and Cu~(2+)enhanced the CL intensity.Cr~(3+),Ni~(2+),Zn~(2+)and Ag~+ ion inhibited the CL intensity,and reduced CL intensity is linear to logarithm of metal ion concentration.L-ascorbic acid can capture oxygen radical,it react with the produced superoxide radical and hydroxyl radical in the CL reaction process,inhibited the CL of the CdTe/H_2O_2 system.Reduced CL intensity is linear to logarithm of L-ascorbic acid concentration in a range of 0.5～100μg/mL.The synthetical sample was detected, veracity is nice.Same polyhydric phenol can react with active oxygen,so can inhibit the CL of the CdTe/H_2O_2 system,but the sensitivity is low.Tween 20 was chosen as a catalyzer of the CL system,the polyhydric phenol was sensitively detected with the CdTe/H_2O_2/Tween 20 system.The CL intensity was linear to the concentration of Catechol in a range 0.05-1μg/mL(R=0.996)with a detection limit of 0.038μg/mL.This method has been successfully used to determine the content of polyhydric phenol in tap water.The recovery percentage was 87-109%.
Thioglycolic-acid-stabilized CdTe nanocrystals were applied to immunoassay for the determination of human immunoglobulin G(HIgG).The immunoassay was based on a sandwich immunoreaction of primary antibody on the magnetic nanoparticles,HIgG, and the secondary antibody labeled with CdTe nanocrystals.The method proposed in the work was compared with a fluorescein isothiocyanate(FITC).This system is an order of magnitude more sensitive than the FITC.The serum samples were analysed by nephelometry and the proposed method.At the same time,the standard addition testswere taken.The results indicate that the accuracy of the method is satisfactory.
引文
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