博士論文-AgⅢ配合物化學(xué)發(fā)光體系與發(fā)光機(jī)理及其在藥物分析中的應(yīng)用研究.pdf

1、 河北大學(xué)博士學(xué)位論文Ag（）配合物化學(xué)發(fā)光體系與發(fā)光機(jī)理及其在藥物分析中的應(yīng)用研究姓名：陳培云申請學(xué)位級別：博士專業(yè)：分析化學(xué)指導(dǎo)教師：孫漢文20100601 Ag(III)Cu(III)Ni( IV)( )()(-)()Ag(III)Ag(III)- H2SO41.310-85.410-6 g mL-12.510-83.310-6 g mL-13.110-9 g mL-12.210-84.510-6 g mL-1 1.910-84.910-6gmL-1,2.310-8 g mL-1 5.310-9 g mL-11.110-8 g mL-1I Ag(III)Ag(III)- H2SO49.1

2、10-9 g mL-1 3.110-9 g mL-1 4.410-9 g mL-13.010-83.710-6 g mL-1 4.010-73.010-6 g mL-1 1.510-72.810-6 gmL-14 Ag(III)-()()6.510-9gmL-12.010-88.010-6 g mL-197.0%-108.3%1.610 g-12.1%5-()()7.210-9, 1.710-8, 8.310-9 g mL1;91.3%112%(n=5)1.62.8%Ag(III)II AbstractAbstractChemiluminescence (CL) method is known

3、 to be a powerful analytical technique thatpossesses high sensitivity, wide linear range, rapidity, simple instrumentation, simpleoperation and so on. It has been widely applied in inorganic and organic species in differentfields such as biotechnology, pharmacology, molecular biology, clinical medic

4、ine andenvironmental detection. Chemiluminescent reagent is the basis of CL analysis. To reaseachand develop new chemiluminescent reagent or to find new chemiluminescent reagent from theknown commercialized reagent and to establish new CL system is very important to improveanalytical sensitivity and

5、 widen application range of CL analysis.The transition metal in highest oxidation state such as Ag(III) Cu(III) Ni(IV) can existstably in the alkaline medium by suitable multi-tooth ligand complexing. They are all strongoxidant with higher stability under the appropriate condition, its complex ion c

6、an produce freeradicals because of its unique structure. They have been widely used in the study of inorganic,organic reaction kinetics and oxidation mechanism. Unfortunately, they are less applied inanalytical chemistry, bis(hydrogenperiodato) argentate(III) complex anion is more lessresearch and i

7、ts application in chemiluminescence analysis.This study starts from the applications of transition metal in highest oxidation statecomplex in CL analysis, the chemiluminescence characteristics of them is explored. Thisresearch is made up of two aspects, one aspect is the establishment of the Ag (III

8、) complex -chemiluminescence system and the investigatation of preliminary mechanism of the reactions,the other aspect is its application in pharmaceutical analysis. The main content of this articleis as follows:1 Chemiluminescence characteristics and mechanism of transition metal in highestoxidatio

9、n stateThe chemiluminescence characteristics of the transition metals in highest oxidation statecomplex in acid medium is explored. For the first time the CL mechanism of the Ag (III)complex - Fluoroquinolones in acid medium is studied. At the same time the preliminaryIII Abstractmechanism of the re

10、actions is also investigated.2 Application of Ag (III) complex chemiluminescence system in pharmaceutical and bodyfluids analysisA novel chemiluminescence reaction system with bis(hydrogenperiodato) argentate(III)complex anion (Ag(III) complex, Ag(HIO6)25) with flow injection technique is developedf

11、orthedeterminationnorfloxacin(NFLX),enoxacin(ENX),ofloxacin(OFLX)andlofloxacin(LOFLX) in the pharmaceutical and body fluids. Under the optimize experimentalconditions, the CL intensity is proportional to the concentration of the drugs in the rang1.310-85.410-6 g mL-1 for NFLX, 2.510-83.310-6 g mL-1

12、for ENX, 2.210-84.510-6 gmL-1 for OFLX and 1.910-84.910-6 g mL-1 for LOFLX. The limit of detection (s/n = 3) is3.110-9 g mL-1 for NFLX, 2.310-8 g mL-1 for ENX 5.310-9 g mL-1 for OFLX and1.110-8 g mL1 for LOFLX. These new method can perform simple, sensitive, rapid andpractical.3 Application of Ag (I

13、II) complex chemiluminescence system in determination offluoroquinolone in milk and body fluids.Based on the novel chemiluminescence reaction system-Ag(III)-H2SO4, A highsensitivity detection for enrofloxacin (ENLX), lomefloxacin (LMFX) and pefloxacin (PFLX)is achieved in food, pharmaceutical and hu

14、man body fluids. The detection limit is 9.1 10-9 gmL-1 for ENLX, 3.1 10 -9 g mL-1 for LMFX and 4.4 10-9 g mL-1 for PFLX. the CLintensity is proportional to the concentration of the drugs in the range 3.010-83.710-6 gmL-1 for LMFX, 4.010-73.010-6 g mL-1for ENLX and 1.510-72.810-6 g mL-1for PFLX.The p

15、roposed method offers the advantages of sensitivity, simplicity and rapidity for the threedrugs determination. The proposed methods are applied successfully to the determination ofthese compounds in real samples. The chemiluminescence characteristics of them is alsoexplored and compared .4 Enhancede

16、ffecton CL emission of Ag(HIO6)25-luminol system andHydroxycamptothecin analysis.A novel chemiluminescence method is developed for the determination ofIV Abstract10-Hydroxycamptothecin (HCPT) based on CL reaction between Ag(HIO6)25- and luminolin alkaline solution. The enhanced degree of CL emission

17、 is proportional to HCPTconcentration. The effect of the reaction conditions on CL emission is examined. Underoptimal conditions, the linear ranges for HCPT is 2.010-88.010-6 g mL-1 , the detectionlimits is 6.510-9 g mL1 with a relative standard deviation of 2.1% for the determination of1.610 -1(n=1

18、1).recoveries-108possible mechanism f o5uenchinganalysis.A(HIO6)25-luminolsulfonamidesTheCL(HIO6)25-(HIO6)25-luminol system isfound. The quenching degree of CL emission is proportional to sulfonamides concentration.The effects of the reaction conditions on CL emission and quenching are examined. Und

19、eroptimal conditions, the detection limits (s/n=3) are 7.210-9, 1.710-8, 8.310-9 g mL-1 forsulfadiazine, sulfameter, and sulfadimethoxine respectively. For spiked urine samples andspiked serum samples the recoveries of sulfanilamides are in the range of 91.3%112% withthe RSDs of 1.62.8% (n=5)and the

20、 results are satisfactory. A possible mechanism of CLemission and quenching effect was also suggested.Keywords Chemiluminescence Ag(III) complex mechanism luminolfluoroquinolones drug sulfanilamides drug 10-HydroxycamptothecinV 111.1(Chemiluminescence CL)ICLCL .dc/dt =EX EM dc/dtICLCLdc/dtEXEMCL(36)

21、(1).1 (2)SchmitzGleu19021928Albrecht1935Petsch(2 2-) (II)Ce( )1.21.2.1(5-2 3-1 4-3-)()1()21.12 11.1 LuminolFig. 1.1 CL mechanism of luminolLuminolLHluminolluminol(L) L(O2)(LO22) LO22LLO22(L), L(HO2-) O21.2.2luminolluminol3 31.2.2.14( )4( )1.31.2.2.21.2.2.2.1()(HRP)HRPHRP-I HRP-II, HRPHRP-I, HRP-IHRP

22、-IIHRP-IIluminolHRP1.2HRP-I HRP-IIluminol1.2luminol-H2O2Fig. 1.2 process diagram luminol-H2O2 system catalyzed HRP4 156-HRPJansen Van den Berg6Navas Diaz7pH=8.5Easton8luminolC1.2.2.2.2Fe( II) Fe(III) Co(II) Cu(II) Ni( II)luminolluminolK3Fe(CN)6Co(II)-EDTA Co(II)-Cu(II)Cu(II)MnCl42Cu(III)1.2.2.3H2O29

23、BaO-MgO105 1.31.3Fig. 1.3 CL mechanism for reaction of luminol and reactive oxygen speciesO2,C11,12O2O21336(DMSO)6 1-DMSO-OH-H2O2-Co2+,181420B615B1216-171819O2HO O2HO,luminol-H2O2-Co(II),20,6.42108 mol L-121luminol-H2O2O22223HOluminol-KIO4(1O2)O2HOluminolluminol1O224Khan Kasha252,1O226Luluminol-H2O2

24、luminol27-NaIO4luminolluminolCLLan Mottola28CO2CO2O2Xiao29CO4 CO4CO2CLCO2Bonifacio Coichev30SO3 SO4 SO5 SO52O2luminol-Ni( II)21.2.2.47 3132luminol-H2O2337-K3Fe(CN)6,34,35luminol-H2O23610 nm375.65-10 nm SnO2,SnO2-O2,0.3 mg. L-11.2.2.5pH38-80- ( ),-( )-39-80-( )CTMAB-40Lin,41,42431.2.2.68-G25044,H2O2-

25、K2CO3CO2CO321.210-11 mol L-1,458 1luminol1.3Mn( ) Co( )Ag( ) Ag(III) Cu(III) Ni( IV)Fe( VI)H2O46Ag(III) Cu(III) Ni( IV)( )47(III)4849( )50( )51( )52(FeO42-)1.3.1 Cu(III)(III)(III)Cu(III)Cu(III)(III)(III)53Cu(III)PtKIO4-KOHCu(NO3)2Cu(HIO6)5-Cu(HIO6)5-5.3 10-8 g mL-154Cu(HIO6)5-Cu(HIO6)5-luminol9 Cu(H

26、IO6)5-luminol552-56-58Cu(HIO6)5-Cu(HIO6)5-4.110-11 mol L-17.210-11 mol L-1Cu(III)luminolluminol-Cu (III)3.510-9 g mL-1 1.210-9 g mL-1(III)3.110-9 g mL-1 3.210- 8 g mL-14.310- 8 g mL-11.3.259, 60Ag(II) Ag(III)Ag(I)Ag(II)1.710-7 mol L-1Ag(III)Ag(OH)4- Ag(H2TeO6)25- Ag(HIO6)25-6162Ag(HIO6)25-(III)6364(

27、III)-10 12.01011 mol L-1,65-67(III)(III)-1.468-707172-73( )( )(III)(III)11 2Ag(III)Ag(HIO6)25-O2(O2)2*Ag(III)-2.12.1.12.1.1.1 Ni(IV)Ni( IV)Baker743 g2 g NiS04 . 7H200.4 g50 mL25-3030-405 g403.59 mol L-1KOH50 mL40-501Ni( IV)2.1.1.2 Cu(III)75Cu( )Cu(III)KIO4(5.74 g)CuS04.5H20(3.12 g) K2S2O8(2.00 g) KO

28、H(18.0 g) 200 mL40Cu(III)0-4Cu(III)2.1.1.3 Ag(III)76Ag(I)Ag(III)KIO4(3.24 g) AgNO3(1.36 g)40Na2S2O8 (3.00 g) KOH(8.00 g) 200 mLAg(III)2.1.1.40-4Ag(III)BPCL(400 nm-640 nm10) IFFM-E()12 2TU-1900() RF-5301PC().2.12.1Fig. 2.1 Schematic diagram of flow injection chemiluminescence analysis system(peristal

29、tic pump) V (sampling inlet valve) C (flowing cell) PMT(photomultiplier tube) (high voltage)(recorder) W (waste) a (acid/ luminol alkaline solution)PAMP(amplifier)HVRb/cAg(HIO6)25-(solution)dAg(HIO6)25-(solution)solution)2.1.2 Ni(IV)Ni()2.1Ni ( )2.1 Ni()-Table 2.1 some drug test results of Ni ( ) -

30、sulfuric acid system(L-1)4.434.002.423.235.0013 4.883.763.125.001.601.605.005.005.005.004.345.005.005.005.002.1.3 Cu(III)(III)266nm415nmCu(III)2.2Cu(III)2.3420266nm415nm(1)(2)200300400500600Wavelength/ nm2.2()(III)UV-VisFig. 2.2 UV/visible spectra of Cu( HIO6)25-(1) Cu(HIO6)25-, (2) Cu(HIO6)25-H2SO4

31、14 2460nm3210-1400450500550600650Wavelength / nm2.3 Cu(HIO6)25-H2SO4Fig. 2.3 CLspectra (b) of Cu(HIO6)25- H2SO4 system2.4Fig. 2.4 kinetic characteristic of the CL reaction(a)(b)(III)2.4,0.3sCu (HIO6)25-H2SO4(III)15 (III)2.22.2 Cu(III)-H2SO4Table 2.2 some drug test results of Cu(III)- sulfuric acid s

32、ystem(mg L-1)4.434.002.423.235.004.883.763.125.001.601.605.005.005.005.004.346-5.005.005.005.005-2.1.4 Ag(III)(III)Ag(III)16 2Ni( IV)Ag(III)Cu(III)Ag(III)Ag(III)Ag(HIO6)25-2.2 Ag(III)2.2.1 Ag(III)Ag(III)Ag(HIO6)25-Ag(HIO6)(OH)(H2O)2-5-2-OHOHOHOOOOOOOH 2AgOHK1+ 2H2OI+ H2IO63-IAgIOOOOOOOOOAg(HIO6)25-O

33、H-( )Ag(OH)2(H2IO6)25- Ag(OH)2(H3IO6)23-7778,79Ag(III)(III)(DPA)pH1.74 VAg(III)Ag(III)Ag(III)Ag(I)Ag(III)Ag(III)/L-/L-Ag(HIO6)25-Ag(III)Ag(HIO6)(OH)(H2O)2-Ag(HIO6)25-/Ag(III)Ag(HIO6)25- Ag(HIO6)(OH)(H2O)2-Ag(III)Ag(HIO6)25- Ag(HIO6)(OH)(H2O)2-Ag3+Ag+Ag3+ + H3O+ Ag+ + H5IO6 + O2 Ag3+Ag+17 81,822.2.2

34、Ag(III)(III)3571 nm 2471 nm2.5Ag(III)247nm210357nm12200300400500600Wavelength / nm2.5()(III)UV-VisFig. 2.5 UV/visible spectra of Ag(HIO6)25(1) Ag(HIO6)25, (2) Ag(HIO6)25H2SO4, Ag(HIO6)25,1.4104 M; H2SO4, 0.08M Ag(HIO6)25Ag(III)2.60.443ab0.30.20.10.021120-1350400450500400450500550600650/ nm/ nm2.6 Ag

35、(HIO6)25- H2SO4(a)(b)Fig. 2.6 Fluorescence emission spectra(a) and CLspectra (b) of Ag(HIO6)25-H2SO4 system(a) (1) Ag(HIO6)25-, (2) Ag(HIO6)25- H2SO4; ex = 290 nm; Ag(HIO6)25-, 2.8105 mol L1; H2SO4,4102 mol L1; (b) Ag(HIO6)25-, 1.4104 mol L1; H2SO4, 1.0 mol L1.18 22.6H2SO4490 nmAg(HIO6)25- Ag(HIO6)(

36、OH)(H2O)2-325-550 nmAg3+Ag(HIO6)25-Ag+Ag3+ + H3O+ Ag+ + H5IO6 + O2O2(O2)2* (O2)2*83,842.3Ag(III)(2.3)Ag(HIO6)25-2.3CompoundStructuresAbbreviationNoroxacin1-ethyl-6- uoro-1,4-dihydro-4-oxo-7-(1-pipera-zinyl)-3-quinoline-carboxylic acidEnrofloxacinENLXLMFX1-cyclopropyl- 7-(4-ethylpiperazin-1-yl)-6-flu

37、or-4-oxo- 1,4-dihydrochinolin-3-carbonateLomefloxacin1-ethyl- 6,8-difluoro-1,4-dihydro-7-(3-methyl-1-piperazinyl)-4-oxo-3-quinolinecarboxylic acidOfloxacinOFLX()9-fluoro-2,3-dihydro-3-methyl-10-(4-methyl-1-pieraziny)-7-oxo-7-H-pyrido(1,2,3-de)-1,4-benzoxazine-6-carboxylicacid19 EnoxacinENX1-ethyl-6-

38、 fluoro-1,4-dihydro-4-oxo-7-(1-piperazinyl)- 1,8-naphtyridine-3-carboxylic acid2.3.1 Ag(HIO6)25-NFLX 2SO4Ag(HIO6)25-NFLXH2SO4Ag(HIO6)25-NFLXH2SO42.724151230250300350400450500Wavelength/nm2.7-Fig.2.7 UV-vis spectra of different systems(1)NFLX, (2) Ag(HIO6)25, (3) Ag(HIO6)25H2SO4, (4) Ag(HIO6)25NFLX,

39、(5)Ag(HIO6)25NFLXH2SO4, NFLX, 1.44105 g mL1; Ag(HIO6)25, 1.4104 M; H2SO4, 0.08 MAg(HIO6)25-357 nmH2SO4Ag(HIO6)25-H2SO4 Ag(HIO6)25-NFLX271 nmAg(HIO6)25- Ag(HIO6)25- 2 SO4NFLXAg(HIO6)25-Ag(HIO6)25-H2SO4NFLXAg(HIO6)25-20 2350-550 nmAg(HIO6)25-NFLXH2SO42.84.54.03.53.02.52.01.51.00.50.01234350400450nm500

40、2.8Fig. 2.8 Fluorescence spectra of different systems.(1) NFLX, (2) NFLXH2SO4, (3) Ag(HIO6)25NFLX, (4)Ag(HIO6)25 2SO4NFLX;ex = 275 nm; NFLX, 2107 g mL1; Ag(HIO6)25, 1.4104 M; H2SO4, 1.0 M2.8NFLX 434 nm367 nm444 nm367 nmNFLXAg(HIO6)25-390 nmNFLXAg(HIO6)25-H2SO42.9543210-1400450500550600650/ nm2.9 Ag(

41、HIO6)25- H2SO4- NFLXFig. 2.9 CL spectra of Ag(HIO6)25- H2SO4- NFLX systemAg(HIO6)25-1.410-4 mol L-1 H2SO4 1.0 mol L-1NFLX210-5 g mL-121 Ag(III)-NFLX-H2SO4Ag(III)-NFLX-H2SO4Ag(HIO6)25-440 nm 460 nm 490 nmH2SO4NFLXAg(HIO6)25-H2SO4NFLXAg(HIO6)25- H2SO4(O2)2* 83,84 (O2)2*490 nmO2O2O2490 nmNFLX 440 nmNFL

42、X(O2)2*440 nmAg(HIO6)25-H2SO4-NFLX460 nm(O2)2*NFLX/Ag(HIO6)25-+ H2O Ag(HIO6)(OH)(H2O)2-+H2IO63-Ag(HIO6)(OH)(H2O)2- + Analyte Analyte oxide + H2OAg(HIO6)(OH)(H2O)2- + H3O+ Ag+ + H5IO6 + O2Ag3+/Ag+ + Analyte Analyte complex + H2O2O2-(O2)2*(O2)2* 2O2 + hv (490 nm)(O2)2*+ Analyte/Analyte oxide Analyte*/

43、Analyte oxide* + O2Analyte*/Analyte oxide* Analyte/ Analyte oxide + hv (440 nm)(O2)2* + Analyte O2 + Analyte complex *Analyte complex*Analyte complex + hv (460 nm)2.3.2 Ag(HIO6)25-H2SO4ENLX/LMLXAg(HIO6)25-H2SO4ENLX/LMLX2.102.1122 2543210a32b121304-1400350400450Wavelength/nm500550450500550600650Wavel

44、ength/nm2.10 Ag(HIO6)25- H2SO4- LMLX(a)(b)Fig. 2.10 Fluorescence emission spectra (a) and CL spectra(b) of Ag(HIO6)25-H2SO4LMFX system(a): (1) LMFX, (2) LMFXH2SO4, (3) Ag(HIO6)25LMFX, (4) Ag(HIO6)25- H2SO4 LMFX. ex =290 nm; LMFX,1.26108 g mL1; Ag(HIO6)25-, 2.8105 mol L1; H2SO4, 4102 mol L1;(b):Ag(HI

45、O6)25-, 1.4104 mol L1; H2SO4, 1.0mol L1; LMFX, 1.57105 g mL1.121076154321086420234350400450500400450500550600650Wavelength/nmWavelength/nm2.11 Ag(HIO6)25- H2SO4- ENLX(a)(b)Fig. 2.11 Fluorescence emission spectra (a) and CL spectra(b) of Ag(HIO6)25-H2SO4ENLX(a): (1)ENLX, (2) ENLXH2SO4 , (3) Ag(HIO6)2

46、5-ENLX , (4) Ag(HIO6)25- H2SO4 ENLX. ex =270nm; ENLX,1.26108 g mL1; Ag(HIO6)25-, 2.8105 mol L1; H2SO4, 4102 mol L1; (b):Ag(HIO6)25-, 1.4104 mol L1; H2SO4, 1.0 mol L1; ENLX, 1.0105 g mL-1.23 350-550 nmAg(III)457 nm 452 nm460 nmLMLX ENLXAg(III)-H2SO4490nmAg(III)-H2SO4-LMFX/ENLXAg(HIO6)25-H2SO4-ENLX/LMLX(O2)2*490 nm 460 nm490 nm(O2)2*ENLX/LMLX460 nmAg(HIO6)25-+ H2O Ag(HIO6)(OH)(H2O)2-+H2IO63-Ag(HIO6)(OH)(H2O)2- + Ana