ELECTROCHEMICAL TECHNIQUES, OSMOMETRY AND THE PRINCIPLES OF RADIOACTIVITY

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1 ELECTROCHEMICAL TECHNIQUES, OSMOMETRY AND THE PRINCIPLES OF RADIOACTIVITY

2 ELECTROCHEMISTY ELECTROCHEMISTRY IS THE STUDY OF CHEMICAL REACTIONS THAT RESULT IN THE FLOW OF ELECTRONS (CURRENT) OR THE DEVELOPMENT OF ELECTROCHEMICAL POTENTIAL (VOLTAGE) DUE TO CHARGE SEPARATION AMPEROMETRY IS THE MEASUREMENT OF THE CURRENT AT A CONSTANT VOLTAGE POTENTIOMETRY IS THE MEASUREMENT OF THE VOLTAGE DEVELOPED DUE TO CHARGE SEPARATION WITH NO CURRENT FLOW; POTENTIOMETRIC SENSORS RESPOND TO ION ACTIVITY, NOT CONCENTRATION

3 POTENTIOMETRY A VOLTAIC OR GALVANIC CELL CONSISTS OT TWO HALF- CELLS CONNECTED BY A SALT BRIDGE (ELECTROLYTE SOLUTION) Fe +2 Fe +3 + e - THERE IS AN ELECTROCHEMICAL POTENTIAL OR VOLTAGE ASSOCIATED WITH THE FORCE DRIVING THE REACTION TO EQUILIBRIUM THIS ELECTROCHEMICAL POTENTIAL (VOLTAGE) IS RELATED TO THE FREE ENERGY OF THE SYSTEM THE MAGNITUDE OF THIS VOLTAGE IS GIVEN BY THE NERNST EQUATION

4 E NERNST EQUATION E RT nf log ared aox E = HALF-CELL POTENTIAL UNDER ACTUAL CONDITIONS E 0 = HALF-CELL POTENTIAL UNDER STANDARD CONDITIONS (ACTIVITIES OF OXIDIZED AND REDUCED SPECIES BOTH = 1 M) R = IDEAL GAS CONSTANT [8.314 J/(mole)( O K)] T = ABSOLUTE TEMPERATURE ( O K) n = NUMBER OF MOLES OF ELECTRONS PER MOLE OF REACTANT F = FARADAY S CONSTANT [96,500 C/eq] = NATURAL LOGARITHM OF 10 a = ACTIVITY OF THE ION

5 POTENTIOMETRY THE POTENTIAL OF A HALF-CELL ELECTRODE CAN NOT BE MEASURED BY ITSELF IT MUST BE CONNECTED TO ANOTHER HALF-CELL ELECTRODE, TO FORM A COMPLETE CIRCUIT TO WHICH A VOLTMETER CAN BE CONNECTED THE ELECTRODE (HALF-CELL) OF INTEREST IS CALLED THE INDICATOR ELECTRODE, AND THE ELECTRODE COMPLETING THE CIRCUIT IS CALLED THE REFERENCE ELECTRODE IN ORDER FOR THE MEASURED VOLTAGE IN AN ELECTRODE SYSTEM TO BE INDICATIVE OF THE ACTIVITY OF THE ION WE WANT TO MEASURE, ALL OTHER SOURCES OF POTENTIAL IN THE SYSTEM, INCLUDING THE REFERENCE ELECTRODE MUST BE KEPT CONSTANT, FROM SAMPLE TO SAMPLE DIFFERENCE IN POTENTIAL BETWEEN INDICATOR AND REFERENCE ELECTRODE IS MEASURED

6 REFERENCE ELECTRODES STANDARD HYDROGEN ELECTRODE INERT PLATINUM WIRE IMMERSED IN A PURE SOLUTION OF HCl WITH ah + = M ([H + ] =~1.2 M) HYDROGEN GAS BUBBLED THROUGH THE SOLUTION AT A PRESSURE OF ATMOSPHERE HALF-CELL REACTION: H + + e - 1/2H 2 E 0 = V THE REFERENCE ELECTRODE AGAINST WHICH ALL OTHER STANDARD HALF-CELL POTENTIALS ARE MEASURED NOT PRACTICAL FOR ROUTINE USE

7 REFERENCE ELECTRODES CALOMEL ELECTRODE Liquid Junction: Opening provides contact between reference and indicator electrodes Electrolyte solution slowly flows from this opening CONSISTS OF HG COVERED WITH CALOMEL (A HG 2 CL 2 PASTE ) IN CONTACT WITH A SOLUTION CONTAINING CL - acl- IS KEPT CONSTANT BY INTERNAL FILLING SOLUTION OF SATURATED KCl

8 REFERENCE ELECTRODES SILVER/SILVER CHLORIDE ELECTRODE CONSISTS OF AG WIRE COATED WITH AGCL IMMERSED IN A SOLUTION CONTAINING CL - acl- IS KEPT CONSTANT BY INTERNAL FILLING SOLUTION OF SATURATED KCl Ag/AgCl ELECTRODES ARE VERY COMMONLY USED AS REFERENCE ELECTRODES IN CLINICAL SYSTEMS

9 INDICATOR ELECTRODES MOST INDICATOR ELECTRODES ARE CONSTRUCTED WITH ION-SELECTIVE MEMBRANES ION-SELECTIVE MEMBRANES SELECTIVELY INTERACT WITH A PARTICULAR ION ON BOTH SIDES OF THE MEMBRANE IF THERE ARE UNEQUAL CONCENTRATIONS OF THAT ION ACROSS THE MEMBRANE, A MEMBRANE POTENTIAL WILL DEVELOP TYPES OF ION-SELECTIVE MEMBRANES GLASS MEMBRANE POLYMER MEMBRANE SOLID STATE MEMBRANE

10 INDICATOR ELECTRODES BASIC HYDROGEN-SELECTIVE MEMBRANE (ph) ELECTRODE Ag/AgCl electrode immersed in HCl contained in tube with a glass membrane tip; glass membrane composed of silicon dioxide with imbedded oxides of alkali metal ions When hydrated, movement of H + produces a potential difference between internal solution and test sample Calibration requires 2 buffers High concentrations of other cations can interfere Store electrode in liquid to maintain hydration Changes in glass membrane composition allows measurement of other ions such as Na, Lithium or Ammonium

11 ION-SELECTIVE MEMBRANE ELECTRODES ION-SELECTIVE POLYMER MEMBRANES ( Liquid Membrane) CONSISTS OF ION-EXCHANGE MATERIALS INCORPORATED INTO AN INERT MATRIX; MATRIX DISSOLVED IN WATER INSOLUBLE SOLVENT CONTACT BEWTEEN TEST SOLUTION ON ONE SIDE OF MEMBRANE AND REFERENCE SOLUTION ON THE OTHER SIDE CREATES A POTENTIAL EXAMPLE: VALINOMYCIN SELECTIVELY BINDS POTASSIUM CHANGING THE ION CARRIER ALLOWS MEASUREMENT OF OTHER IONS i.e. Ca, Mg, NH 4, Na

12 ION-SELECTIVE MEMBRANE ELECTRODES SOLID-STATE ELECTRODES MEMBRANE CONSISTS OF A SINGLE CRYSTAL OR A MIXTURE OF FINE CRYSTALS IMMOBILIZED IN IN AN INERT MATRIX MEMBRANE IS SELECTIVE FOR A PARTICLAR SIZE OR SHAPE OF ION IE. CHLORIDE SENSITIVE ELECTRODE COMPOSED OF AgCL/Ag 2 S

13 ION-SELECTIVE MEMBRANE ELECTRODES THE PRINCIPLES OF POTENTIOMETRY AND ION-SELECTIVE MEMBRANE ELECTRODES ARE EXTENDED TO MEASURE GASES IN WHOLE BLOOD IE. CO 2 AND AMMONIA pco 2 Electrode ph glass electrode in contact with sodium bicarbonate solution encased by outer silicon rubber membrane permeable to CO 2 gas; CO 2 from test sample diffuses into sodium bicarbonate solution changes ph Ammonia Electrode uses membrane permeable to ammonia gas

14 ION-SELECTIVE MEMBRANE ELECTRODES ENZYME ELECTRODES ISE COVERED BY GEL OR MEMBRANE CONTAINING IMMOBILIZED ENZYMES IE. UREASE CATALYZES HYDROLYSIS OF UREA TO NH 4+ WHICH IS THEN MEASURED BY THE ELECTRODE

15 ION-SELECTIVE MEMBRANE ELECTRODES FLOW-THROUGH DESIGN OF CURRENT ISE ANALYZERS

16 ION-SELECTIVE MEMBRANE ELECTRODES DRY SLIDE DESIGN

17 ION-SELECTIVE ELECTRODES PRACTICAL CONSIDERATIONS ELECTRODES MUST BE REGULARLY CONDITIONED TO STRIP AWAY ANY DEPOSITS ON MEMBRANE TO INSURE PROPER ION-EXCHANGE AT THE SURFACE NEVER WIPE AN ELECTRODE. IF EXCESS FLUID MUST BE REMOVED, BLOT IT DRY FOR SHORT TERM STORAGE, KEEP THE OUTER MEMBRANE SURFACE WET WITH A SOLUTION WITH CONCENTRATIONS SIMILAR TO WHAT YOU WILL BE MEASURING FREQUENT CALIBRATION OF ELECTRODES WITH TWO CALIBRATORS IS NEEDED TO VERIFY THE SLOPE MINIMIZE LIQUID JUNCTION POTENTIAL BY USING IONS WITH SAME MOBILITIES FOR THE REFERENCE FILLING SOLUTION OR CALIBRATORS WITH CHARACTERISTICS SIMILAR TO TEST SAMPLES MAINTAIN A CONSTANT TEMPERATURE

18 VOLATAMMETRY MEASUREMENT OF CURRENT FROM AN ELECTROCHEMICAL REACTION DIVIDED INTO AMPEROMETRY OR POLAROGRAPHY AMPEROMETRY A CONSTANT VOLTAGE OR POTENTIAL IS APPLIED BETWEEN TWO ELECTRODES, ONE SERVES AS AN ANODE (+), THE OTHER SERVES AS A CATHODE (-) IF A SUBSTANCE IS INTRODUCED THAT CAN BE OXIDIZED OR REDUCED BY THE APPLIED VOLTAGE, THEN CURRENT WILL FLOW BETWEEN THE TWO ELECTRODES THE CURRENT WILL BE PROPORTIONAL TO THE RATE AT WHICH THE SUBSTANCE CAN DIFFUSE TO THE ELECTRODES FROM THE SAMPLE, WHICH IS USUALLY PROPORTIONAL TO THE CONCENTRATION OF THE SUBSTANCE IN THE SAMPLE

19 AMPEROMETRY po2 (Clark) ELECTRODE Platinum cathode separated from test sample by membrane permeable to O 2 ; When voltage applied oxygen is reduced; Produces flow of electrons (current) AT THE CATHODE: AT THE ANODE: O 2 2H 2 O 4e 4OH o 4Cl 4Ag 4AgCl 4e CURRENT IS PROPORTIONAL TO po 2 IN THE SAMPLE

20 COULOMETRY MEASUREMENT OF THE # OF COULOMBS OF CHARGE INVOLVED IN A CHEMICAL REACTION CLINICAL APPLICATION: COULOMETRIC TITRATION OF CHLORIDE IONS BY SILVER IONS A OFF TIMER ON BECAUSE Ag + IS GENERATED AT A CONSTANT RATE, TIME TO REACH EQUIVALENCE POINT IS DIRECTLY PROPORTIONAL TO [Cl - ] Cl -

21 ANODIC STRIPPING VOLTAMMETRY CLINICAL APPLICATION: BLOOD LEAD TESTING Application of a negative potential reduces metal in test solution to the elemental state; plates onto mercury electrode Polarity is reversed and voltage increased; metals re-oxidize; flow back into solution producing current Amount of current proportional to conc of metal; Re-oxidation voltage specific for individual metals

22 BIOSENSORS REACTION BETWEEN ANALYTE AND THE BIODETECTION AGENTS (ANTIBODIES, NUCLEIC ACIDS, CELL RECEPTORS, ENZYMES) CONVERTED BY TRANSDUCER INTO AN ELECTRICAL SIGNAL

23 OSMOMETRY AN OSMOMETER IS AN INSTRUMENT THAT MEASURES THE OSMOLALITY OF A SOLUTION OSMOLALITY IS ONE OF FOUR COLLIGATIVE PROPERTIES OF SOLUTIONS A COLLIGATIVE PROPERY IS ONE THAT DEPENDS SOLELY ON THE CONCENTRATION OF IONS OR MOLECULES, AND NOT ON THE TYPE OF ION OR MOLECULE WHEN A SOLUTE IS DISSOLVED IN WATER, IT RAISES THE OSMOTIC PRESSURE OF THE SOLUTION RAISES THE BOILING POINT OF THE SOLUTION LOWERS THE FREEZING POINT OF THE SOLUTION LOWERS THE VAPOR PRESSURE OF THE SOLUTION

24 OSMOMETRY WHEN 1 OSMOLE OF SOLUTE DISSOLVES IN 1 kg OF WATER, IT LOWERS THE FREEZING POINT OF WATER TO o C METHOD Supercool solution; vibrate probe to initiate freezing; measure temperature at which solution freezes Osmolality is calculated from temperature of the FP

25 OSMOMETRY FREEZING PT. OF WATER (0 o C) FREEZING PT. DEPRESSION IS DIRECTLY PROPORTIONAL TO THE OSMOLALITY A STIRRING WIRE INDUCES CRYSTALLIZATION IN THE SUPERCOOLED SAMPLE CURRENT IN THE THERMISTOR PROBE CIRCUIT IS CALIBRATED TO DISPLAY IN UNITS OF mosm/kg MEASURES ALL PARTICLES INCLUDING ANY CONTAMINANTS

26 OSMOMETRY VAPOR PRESSURE OSMOMETER SAMPLE IS INTRODUCED TO A SEALED CHAMBER CONTAINING A THERMOCOUPLE PROBE ELECTRICAL CURRENT COOLS THE CHAMBER BELOW DEW POINT ALLOWING WATER TO CONDENSE ON THE PROBE CURRENT SHUT OFF; HEAT OF CONDENSATION CAUSES TEMPERATURE TO RISE MEASURE THE TEMPERATURE AT WHICH EQUILIBRIUM OCCURS BETWEEN LIQUID (CONDENSATION) AND VAPOR (EVAPORATION) PHASE; INVERSELY RELATED TO THE VAPOR PRESSURE OF WATER FROM THE SAMPLE VAPOR PRESSURE OSMOMETERS GENERALLY REQUIRE LESS SPECIMEN VOLUME SPECIMENS CONTAINING VOLATILE ORGANICS SUCH AS ETHANOL, METHANOL, AND ACETONE YIELD FALSE RESULTS

27 PRINICPLES OF RADIATION NATURE OF RADIOACTIVITY ATOMIC NUMBER (z) = THE NUMBER OF PROTONS ATOMIC MASS (A) = THE SUM OF THE NUMBER OF PROTONS AND NEUTRONS WRITTEN FORM FOR FOR NUCLIDE ISOTOPES ARE NUCLIDES WITH THE SAME ATOMIC NUMBER BUT DIFFERENT MASSES NUCLEAR STABILITY DEPENDS ON THE n-p RATIO RADIOACTIVE DECAY IS A PROCESS OF SPONTANEOUS CHANGE WITHIN THE NUCLEUS TO INCREASE STABILITY; INVOLVES LOSS OF MASS AND EMISSION OF RADIATION 14 6 C

28 PRINICPLES OF RADIATION TYPES OF RADIOACTIVITY ALPHA DECAY EMISSION OF A PARTICLE WITH 2 PROTONS AND 2 NEUTRONS (HELIUM NUCLEUS) BETA DECAY EMISSION OF EITHER AN NEGATRON OR A POSITRON; RESULTS IN CONVERSION OF A NEUTRON TO A PROTON OR VICE VERSA ELECTRON CAPTURE NUCLEUS CAPTURES AN ELECTRON FROM AN INNER SHELL ORBITAL; RESULTS IN CONVERSION OF A PROTON TO A NEUTRON; AN ELECTRON FROM ANOTHER ORBITAL FILLS INNER SHELL VACANCY; ENERGY RELEASED AS AN X-RAY GAMMA RADIATION EMISSION OF ELECTROMAGNETIC RADIATION OF VERY SHORT WAVELENGTH (GAMMA RAYS)

29 PRINICPLES OF RADIATION RATE OF RADIOACTIVITY DECAY RATE OF DECAY CHARACTERISTIC OF EACH RADIONUCLIDE HALF-LIFE (t 1/2 ): TIME REQUIRED FOR RADIOACTIVITY IN SAMPLE TO DECREASE TO HALF OF ITS INITIAL VALUE UNIT OF RADIOACTIVITY DECAY CONVENTIONAL UNIT: 1 CURIE (Ci) = 3.7 X dps SI UNIT: 1 BECQUEREL (Bq) = ONE DECAY PER SECOND (dps)

30 PRINICPLES OF RADIATION MEASUREMENT RADIOACTIVITY SCINTILLATION COUNTERS MEASURE FLASHES OF LIGHT PRODUCED FROM IONIZATION OF A SCINTILLANT BY RADIATION GAMMA RAYS ARE COUNTED WITH A CRYSTAL SCINTILLATION COUNTER BETA RADIATION IS COUNTED WITH A LIQUID SCINTILLATION COUNTER

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