1. 电分析应用 定量和定性分析 生物传感器

2. 电分析应用 定量和定性分析 生物传感器 AS : 电位传感器 恒电流 > 电位变化 AS: 电流传感器 恒电位 > 电流变化

3. 电分析应用 定量和定性分析 生物传感器 电位传感器： 玻璃电极 (pH) 固体膜 离子交换膜 气体微孔膜 生物酶电极 ISFET 涂层线电极

4. 电分析应用 定量和定性分析 1.ISE 阳离子剂： H +, Na +, K +, Ca +2, Ag +, 2.ISE 阴离子剂： Br -, Cl -, CN -, S -2, F -, 3.ISE 气体扩散： NH 3, SO 2, O 2, NO 2, 4.ISE 有机物：草酸，乙酸，可待因，罂粟碱， 安飞他命

5. 电分析应用 定量和定性分析 生物传感器 电流传感器： 简单电流传感器 膜和膜覆盖修饰电极

6. 电分析应用 定量和定性分析 生物传感器 1. 食物分析和药物分析 比如：生物酶和微生物

7. 电分析应用 定量和定性分析 1. 快速经济地分析食物的某些成分 2. 测量微生物侵袭氧化过程 3. 毒素，添加剂，保存剂 4. 作为色析法的指示剂

8. 电分析应用 定量和定性分析 生物传感器： 蔗糖， GLUCOSIO + O 2 ——OXIDASE  GLUCOLACTONE+ H 2 O 2 LACTATE + O 2 ——OXIDASE  PIRUVATE+ H 2 O 2

9. 电分析应用 定量和定性分析 生物传感器： 氨，缩氨酸，蛋白质 R-CHNH 2 COOH + O 2 H 2 O —OXIDASE  R-COCOOH +NH 3 +H 2 O 2 L-AMINOACID + O 2 —OXIDASE  CHETOACID+ H 2 O 2 NH 3 METHIONINE —LIASE  CHETOBUTIRRATE+ METHYLMERCAPTANE + NH 3

10. 电分析应用 定量和定性分析 BIOSENSORS FOR ASCORBIC ACID OXALIC ACID ALCHOLS (GLICEROLS, PHENOLS,) FATTY ACIDS (for control the quality of the food) PHOSPHOLIPIDS

11. ELECTROANALYSIS APPLICATIONS QUANTITATIVE AND QUALITATIVE ANALYSIS BIOSENSORS QUALITATIVE ANALYSIS: AMPEROMETRIC OR POTENTIOMETRIC SENSORS ? KINETICS OF REACTIONS STUDY OF BEST ENZYMES, TEMPERATURE, ELECTROLYTES, MEMBRANES MODIFICATION OF ELECTRODES

12. ELECTROANALYSIS APPLICATIONS QUANTITATIVE AND QUALITATIVE ANALYSIS IMUNOSENSORS: AMPEROMETRIC AND POTENTIOMETRIC SENSORS FOR DETERMINATION OF SPECIFIC ANTIBODY ANTIGENE REACTIONS STUDY OF KINETICS REACTIONS CROSS ANALYSIS WITH EQCM AND ESPR

13. APPLICATIONS ELECTROANALYSIS APPLICATIONS APPLICATIONS

14. ELECTROANALYSIS APPLICATIONS ELECTROCHEMICAL ENZYME IMMUNOARRAY FOR PCB

15. ELECTROANALYSIS APPLICATIONS Amperometric Detection of Uric Acid, Hypoxanthine, Inosine and Inosine Monophosphate with Xantine oxidase Immobilised and Carbon Screen-printed electrode. Application for fish freshness determination.

16. ELECTROANALYSIS APPLICATIONS EQCM  Surface Modification for the Development of Affinity Biosensors · Immobilisation of antibodies · Immobilisation of DNA probes · Development of Affinity Biosensors · Measuring affinity and rate costants · DNA biosensors · Polymorfism detection · Specific gene detection in bacterial DNA

17. ELECTROANALYSIS APPLICATIONS

19. Coupling PCR (polymerase Chain reaction) and piezoelectric DNA biosensors for detecting the pathogenicity in Aeromonas hydrophila species

20. ELECTROANALYSIS APPLICATIONS

23. QUANTITATIVE AND QUALITATIVE ANALYSIS BIOSENSORS FOR CONTROL THE BIOLOGICAL PROCESS WITH AN ELECTROCHEMICAL FUNDATION -TRASMISSION BETWEEN CELLULAR MEMBRANES -NERVE IMPULSE -CARDIOVASCULAR ELECTROCHEMISTRY -BIOENERGETICS AND BIOELECTROCATALYSIS -NEUROTRASMISSION (DOPAMINE GENERATION)

24. ELECTROANALYSIS APPLICATIONS QUANTITATIVE AND QUALITATIVE ANALYSIS BIOSENSORS FOR CONTROL THE BIOLOGICAL PROCESS WITH AN ELECTROCHEMICAL FUNDATION -BIOSENSORS FOR IN VIVO OR FLOW ANALYSIS -POTENTIOMETRIC SENSORS (ENZYME AND ISFET) -AMPEROMETRIC SENSORS -MICROBIAL SENSORS (contact between an electrode and living cells, conversion of biochemical signal from the microorganism in variation of V)

25. Different sensors Conductivity Thermal Spectroscopic Mass spectroscopy Electrochemical »Lack of selectivity »Lack of sensitivity »Expensive

26. Biosensor Transducer Bio- molecule Measurable, quantifiable signal

27. Bio-molecules Antibodies Enzymes Cells Synzymes - artifical enzymes

28. Redox enzymes Contains a co-factor that is reduced or oxidised by a substrate The enzyme needs to be regenerated with a second substrate or directly with an electrode

29. Sugar catalysis by oxidoreductases FADH 2 FAD O CH 2 OH HO OH O OHOH O CH 2 OH HO OH H Glucose Gluconolactone 2H + + 2e -

30. Amperometric biosensors S P O2O2 H2O2H2O2 e-e- 1st generation S P M ox M red e-e- 2nd generation S P e-e- 3rd generation

31. Redox polymer based amperometric biosensor S red S ox e-e- e-e- e-e- e-e- Enzyme Redox polymer Crosslinker

32. Ping-pong mechanism 1.E + SESE* + P 2.E* + AE*AE + A* 3.A*A Rate = d[S]/dt = k’[S] K M + [S]

33. Rate vs. concentration

34. Enzyme theory Michaelis-Menten kinetics

35. Detection of 2nd substrate FAD heme e-e- e-e- e-e- e-e- e-e- Electrode

36. Size exclusion Chromatography In sugar analysis the retention time depends on the size of the sugar molecule and not on the racemic form With a biosensor, sugars with the same size but of different form can be detected selectively. Applications: Glucose in blood, ethanol in beer, lactose in food etc.

37. Simultaneous amperometric determination of sugars Tid Spektrophotometer Detects everything (big window), but not very selective. Biosensors Narrow window, but very selective Tid Size exclusion separation Sample with many different kind of sugar molecules

38. IUPAC definition of a biosensor A biosensor can be regenerated PS EE*E* S + E P + E * S can be detected with an irreversible reaction Another kind of bio- sensing principle

39. SPR as a tool for biorecognition Analyte is ”washed” away!

40. The electrochemistry The redox active group of an enzyme is usually buried inside the enzyme Enzyme is often denatured at an metal electrode

41. Electrochemical methods General CV, SWV, CM to investigate the rate between active site and electrode Spectroelectrochemistry as a tool to proove the electrochemistry of active site

42. Formal potential for simpler molecules and ions. e-e- Electrode Ox/Red

43. The complexity of enzymes and isolation of active site Electrode e-e- e-e- e-e- e-e- e-e-

44. Spectra for oxidised and reduced enzyme Absorbance Wavelength / nm Reduced Oxidised

45. Coulometry Extinction coefficient Concentration Important to have a closed system! Charge Absorbance

46. Absorbance vs. potential Absorbance Potential vs. Ag/AgCl / mV

47. Cellobiose dehydrogenase adsorbed on to an electrode S P 2H + 2e - FAD heme (a) (b) I

48. Enzyme fragments adsorbed on to an electrode III SP FAD 2e - II heme S

49. Commercial biosensors Roche ”Accu-Chek” Medisense Bayer ”Glucometer”. LifeScan ”EuroFlash” and ”FastTake” Therasense ”FreeStyle” Chemel ”SIRE Biosensor” and more...

50. The future Development of tools for basic research and investigation of new biosensors: Spectroelectrochemistry, surface modification (FRA), ESPR, EQCM Production of more redox enzymes Site directed mutagenesis Development of applications with already existing biosensors

51. Biosensor with pre-oxidation BiosensorPre-oxidation membrane +0.4 V +0.7 V Analyte with interferences

52. Cascade systems Diacyl glycerol Monoacyl glycerol Glycerol Triacylglycerol lipas Diacylglycerol lipas Monoglycerol lipas Glycerol oxidase e-e- Electrode Membrane Aceton Fat