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    5.0. Clinical Chemistry

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    1. Which formula correctly describes the relationship between absorbance and % 7? RA=2-log %T B.A=log 1/T CA=-log T D. All of these options Chemistry Identify basic principle(linstrumentation!2 2. A solution that has a transmittance of 1.0 %T would have an absorbance of A10 B20 €.1% D.99% Chemistry/CaleulatelBeer's lawl2 . In absorption spectrophotometry: A. Absorbance is ditectly proportional to ansmittance B, Percent transmittance is directly proportional to concentration €. Percent transmittance is direct the light path length D. Absorbance is directly proportional to portional to ChemistrylDefine fundamental characteristes! Beer's awit 4. Which wavelength would be absorbed strongly by a red-colored solution? 8.450 nm 8.585 am, €.600 nm, D. 650m Chemistry!Define fundamental characteristics! Spectrophotometry/2 5. A green-colored solution would show highest transmittance at A475 nm B.525 nm, €.585 nm, D.620 nm ChemistrylDefine fundamental characteristis! Spectrophotometry/2 5.1 Instrumentation Answers to Questions 1-5 1. D_Absorbanceis proportional to the inverse log of transmittance A=-logT Multiplying the numerator and denominator by 100 gives Az s00XT: A=log 104 A=I0g 100- log A=20-10g For example, iT = 100, then: A=20-fo9 100 9 100= 19 A=29 A=20- tog 7 A=20-log 1.0 The log of L0=0 A=20 3. D Beers law states that A absorptivity coeffi ent (a constant), isthe path length, and cis concentration. Absorbance is directly proportional to both b and « Dooling the path length results in incident light contac’ ‘the number of molecules in solution, This causes absorbance to double, the same effect as doubling the concentration of molecules. twice 4, A Asolution transmits ight corresponding in ‘wavelength to its colar, and usually absorbs light of wavelengths complementary to its color. A red solution transmits ight of 600-650 nm and strongly absorbs 400-800 nm light. 5. B Green light consists of wavelengths from S00-580 nm, A green-colored solution with a transmittance maximum of 525 nm and a S0-nmm bandpass transmis light of 525 nm and absores light below 475 nm and above 575 be quanttated using a wavelent strongly, such as 450 nm. 1m. A solution that is green would that it absores 173 174 Chapter 5 | Clinical Chemistry 6, SITUATION: A technologist is performing an enzyme assay at 340 nm using a visible-range spectrophotometer. After setting the wavelength and adjusting the readout to zero %T with the light path blocked, a cuvette with deionized water is inserted. With the light path fully open and the 100%T control at maximum, the instrument readout will not rise above 90%T. What is the ‘most appropriate first course of action? ‘A. Replace the source lamp B, Insert a wider cuvette into the light path . Measure the voltage across the lamp terminals D. Replace the instrument fuse ChemistrylSelect course of acion(Spectraphotometryl3 7. Which type of monochromator produces the purest monochromatic light in the UV range? A.A diffraction grating and a fixed exit slic B.A sharp cutoff filter and a variable exit slit G. Interference filters and a variable exit slit D.A prism and a variable exit slit ChemistrySelect componentSpectrophotometry!2 ‘Which monochromator specification is required in order to measure the true absorbance of a compound having a natural absorption bandwidth of 30 nm? ‘A.50-nm bandpass B. 25-nm bandpass . 15-nm bandpass D. S-nm bandpass ChemistrySelect componentiSpectrophotometryl2 9. Which photodetector is most sensitive to low levels of light? A. Barrier layer cell B. Photodiode ©. Diode array D. Photomultiplicr tube Chemistry/Define fundamental characteristics! Instrumentation! 10. Which condition is a common cause of stray light? A. Unstable source lamp voltage B, Improper wavelength calibration . Dispersion from second-order spectra D. Misaligned source lamp Chemistry densify sources of errorlSpectrophotometry!2 Answers to Questions 6-10, 6A 9D 10. ¢ Visible spectrophotometers are usualy supplied with a tungsten or quartz halogen source lamp. Tungsten lamps produce 2 continuous range of wavelengths ftom about 220-2,000 nm. Output Increases as wavelength becomes longer peaking at around 1,000 nm, and is poor below 400 nm. As the lamp envelope darkens with age, the amount of light reaching the photodetector at 240 nm becomes insufficient to set the blank reading to 100947. Quartz halogen lamps produce light from 300 nm through the infrared region. Deuterium or hydrogen lamps produce ultravioletrich spectra optimal for ultraviolet (UV) work Mercury vapor lamps produce a discontinuous spectrum that includes a high output at around 365 nm that is Useful for fluorescent applications. Xenon lamps generate a continuous spectrum of fairly uniform intensity from 300-2,000 nr, making them useful for both visible and UY applications Diffraction gratings and prisms both produce a continuous range of wavelengths. A diffraction grating produces a uniform separation of ‘wavelengths. Aprism produces much better separation of high-frequency Ight because refraction is greater for higher-eneray wavelengths. Instruments using a prism and a variable exit sit can produce UV light of very narrow bandpass, The adjustable sit is required in order to allow sufficient ight to reach the detector to set 100%7 Bandpass refers tothe range of wavelengths passing through the sample. The narrower the bandpass, the greater the photometric resolution. Banapass can bbe made smaller by reducing the width ofthe exit sik, Accurate absorbance measurements require @ bandpass less than one-fith the natural bandpass ofthe chromophore, ‘The photomultiplier tube uses dynodes of increasing vokage to amplify the current produced by the photosensitive cathode, It's 10.000 times as sensitive as batter lyer cell, which has no amplification. A photomultiplier ube requires a OCregulated lame because it responds to ight fluctuations caused by the ACoycle Stray ight is caused by the presence of any light other than the wavelength of measurement reaching the detector It is most often caused by second-order spectra, deteriorated optics, ight dispersed by @ darkened lamp envelope, and extraneous room light oe 2 1B. 4 A linearity study is performed on a visible spectrophotometer at 650 nm and the following absorbance readings are obtained: [Concentration of Standai ‘Absorbance Yoomg/at 020, 200mg/al oat 300 moral 082 40.0 mala 079 Soomg/at 032 “The study was repeated using freshly prepared standards and reagents, but results were identical to those shown, What is the most likely cause of these results? A. Wrong wavelength used B, Insutficient chromophore concentration . Mattix interference D. Stray light Chemistrylldensify sources of errrlSpectrophotometr/3 Which type of filter is best for measuring stray lighe? A Wraten B. Didymium, ©. Sharp euro D. Neutral density ChemistrySelect methods/ReagentslMedial Spectrophotometry!2 ‘Which of the following materials is best suited for verifying the wavelength calibration of a speetrophotometer? A. Neutral density filters B, Potassium dichromate solutions traceable to the National Bureau of Standards reference ¢. Wratten filters D. Holmium oxide glass Chemistry tdemify standard operating procedure! Spectrophotometry/2 ‘Why do many optical systems in chemistry analyzers utilize a reference light path? ‘A. To increase the sensitivity of the measurement B. To minimize error caused by source lamp Actuation €. To obviate the need for wavelength adjustment D. To reduce stay light effe ChemistrylDefine fundamental characteristics! Spectrophotometry/2 5.1 | Instrumentation 175 Answers to Questions 11-14 11. D Stray lights the mast common cause of los of linearity athigh-analyte concentrations. Lig transmitted through the cuveties lowest when absorption is highest. Therefore stray light isa greater percentage of the detector response when sample concentration is high, Stray li significant when measurernents are made at the extremes of the visible spectrum because lamp output and detector 12, © Sharp cutoff fiters transmit almost all incident light Untl the cutoff wavelength is reached. At that point, they cease to transmit ight. Because they sive an ‘allor none effect only stay light reaches the detector when the selected wavelength is beyond the cutoff, 13. D Wavelength accuracy's verified by determining reading that gives the highest absorbance (or transmittance) when a substance with a narrow natural bandoass (sharp absorbance \ce peak) is scanned. For example dlidymium glass has a sharp absorbance peak at 585 nm. Therefore, an instrument should glve its highest absorbance reading when the wavelength dlalis set at: very narrow absoroance peak at 361 nm; tkewise, the hydrogen lam of 2 UV spectrophatometer produces a 656-nm emission line that can be used to verify wavelength, Neutral density filers and dichromate solutions are used to verify absorbance accuracy or linearity. A Wratten filter is a wide~ bandpass fiker made by placing a thin layer of lass plates and is tometriccalbration 14, B Ateference beam is used to produce an electical signal at ine dete which the measurement of light aosorption by the sample is compared. This safeguards against measurement errors caused power fluctuations that change the source lamp intensity. Although reference beams increase the accuracy of measurements, they do so at the expense of optical sensitivity since some of the incident light must be used to produce the reference beam 176 Chapter 5 | Clinical Chemistry 15. Which component is required in a spectrophotometer in order to produce a spectral absorbance curve? ‘A. Multiple monochromators B. A reference optical beam C. Photodiode array D. Laser light source ChemistrylDefine fundamental characteristiod Spectrophotometry/1 16. The half-band width of a monochromator is defined by: [A The range of wavelengths passed at 50% maximum transmittance B, One-half the lowest wavelength of optical purity . The wavelength of peak transmittance D. One-half the wavelength of peak absorbance ChemistrylDefine fundamental characteristic Spectrophotometry/t 17, The reagent blank corrects for absorbance caused by: A. The color of reagents B. Sample turbidity . Bilirubin and hemolysis D. All ofthese options Chemistrylldemtfy base principle@WSpectrophotomenryl2. 47, 18. A plasma sample is hemolyzed and turbid. What is required to perform a sample blank in order to correct the measurement for the intrinsic absorbance of the sample when performing a speetrophotometric assay? A. Substitute deionized water for the sample B, Dilute the sample 1:2-with a standard of 18. known concentration . Substituce saline for the reagent D. Use a larger volume of the sample Chemistry identify basic principle(y/Spectrophotometr/2 19, Which instrument requires a highly regulated DC power supply? A.A spectrophotometer with a barrier layer cell B. A colorimeter with multilayer interference filers A spectrophotometer with a photomultiplier ube D. A densitometer with a photodiode detector ChemistrySelect componentiSpectrophotometryl2 19. ¢ 16. A Answers to Questions 15-19 15, C There are wo ways to perform spectral scanning for compound identification. One sto use a stepping continuously turns the monochromator so avelength aligned with the ext sit changes ata constant ate. A more practical method isto use a diode array detector. This conssts ofa chip embedded vith as many as several hundred photodiodes. Each photodiode is aligned with a narrow part ofthe spectrum produced by a diffraction grating, and produces current proportional to the intensity of the band of lghtstking it (usually 1-2 nm in range). The diode signals are processed by a computer to create 2 spectral absorbance or transmittance curve. Hal-band width is 2 measure of bandpass made Using a soliton or fker having a narrow natural bandpass (transmittance peat), The wavelength ‘giving maximum transmittance is set to 100567 (r0.).Tmen, the wavelength dialis adjustes downward, untila readout of SORT (030 A) is obtained. Next, the wavelength is adjusted upward Lunt 50567 is obtained, The wavelength dference is the hal-band width. The nartower the hal"band ‘width, the better the photometric resolution of the instrument. When a spectrophotometer is set to 10037 with the reagent blank instead of water, the absorbance of reagents is automaticaly subtracred from each Unknown reading, The reagent blank does not cortect for absorbance caused by interfering chromagens in the sample such as bliin, hemolysis, or tury {sample blanks used to subzract the intrinsic absorbance ofthe sample usually caused by hemolysis, icterus, turbidity, or drug interference. (On automated analyzers this is accomelishea by measuring the absorbance after the addition of sample and a fist ceagent, usualy a diluent. For tests using a single reagent, sample blanking can be done prior to the incubation phase before any color develops, Substhuting deionized water for sample isdone to subtract the absorbance of the reagent (reagent blanking) Diluting the sample with a standard (standard addition) may be done when the absorbance is below the minimum detection limit for the assay Using a lager volume of sample wil make the interference worse, When AC voltage reaulators are used to isolate source lamp power, ight output fluctuates as the voltage changes. Because this occurs at 60 Hits not detected by eyesight or slow-responding detectors Photomukiplier tubes are sensitive ‘enough to respond to the AC equency and require a DC-regulated power supaly. 20. 22. 23. 24. ‘Which statement regarding reflectometry is truc? A. The relation between reflectance density and concentration is linear B. Single-point calibration can be used to determine concentration C. 100% reflectance is set with an opaque film called a white reference D. The diode array is the photodetector of choice ChemistrylApply principles of special procedures! Instrumentation/2 Bichromatic measurement of absorbance can correct for interfering substances if: ‘A. The contribution of the interferent to absorbance is the same at both wavelengths B, Both wavelengths pass through the sample simultaneously . The side band is a harmonic of the primary wavelength D. The chromogen has the same absorbance at both wavelengths ChemistrlApply principles of special procedures! Instrumentation?2 Which instrument requires a primary and secondary monochromator? A Spectrophotometer B. Atomic absorption speettophotometer €. Fluorometer D. Nephelomecer Chemistry/Apply principles of special procedures! Instrumentation/I ‘Which of the following statements about fluorometry is accurate? A. Fluorometry is less sensitive than, spectrophotometry B, Fluorometty is less specific than spectrophotometry . Unsaturated cyclic molecules are often Auorescent D. Fluorescence is directly proportional to temperature ChemistrlApply principles of special procedures! Instrumentation?2 ‘Which of the following components is not needed in a chemiluminescent immunoassay analyzer? A Source lamp B. Monochromator €. Photodetector D. Wash station Chemistry!Define fundamental characteristics! Instrumentation! 21. 22. € 23. € 24. 5.1 | Instrumentation 177 Answers to Questions 20-24 20. C Reflectometty does not follow Beer’ aw, but the relationship between concentration and reflectance can be desctbed by a logistic formuls or algorithm that can be solved for concentration, For example, K/S=(1~ RP/AR, where K=Kubelka-Munk absorptivity constant, coefficient, = reflectance density, K/Sis proportional to concentration, The hie ferences analogous to the 10087 set spectrophotomety and serves a a reference signal = log RyfR, where D isthe reflectance density, Ris the white reference signal, and 8 isthe photodetector signal forthe test sample. In bichromatic ahotomety, she absorbance of sample is measured at two diferent wavelengths The primary wavelength is ator near the absorbance maximum. An interfering substance having the same absorbance at both primary and secondary (side banal wavelengths does not affect the absorbance difference (a) A fluorometer uses a primary monochromator to Isolate the wavelength for excitation, and a secondary monachromator to solate the wavelength emitted by the fluorochrome, Increasing temperature results in more random collsion between molecules by increasing their motion. This causes eneray to be cissipated as heat instead of fluorescence, Temperature is inversely proportional te uorescence Fluorescence is more Sensitive than spectrophotometty because the detector signal can be amplified when lute solutions are measuted Its also more specific than spectrophotometry because both the exctation and emission wavelengths ate characterstics of the compound being measured. ‘Chemiluminescence is te production of ight following a cherical reaction, Inmuroassays based upon chemiluminescence generate light when the chemiluminescent molecule becomes excited; ‘therefore, alignt source is not used. In immunoassay platforms, chemiluminescent molecules such as ‘acriainium can be use to label antigens or antibodies. Alternatively, chernluminescent substrates such as luminal or dioxetane phosphate ‘may be used, Light willbe emitted when the enzyme-labeled molecule reacts with the substrate. In such assays, free and bound antigen separation is requited and s usually accomplished using paramagnetic particles bound te elther antibody or reagent antigen 178 Chapter 5 | Clinical Chemistry 25. 26. 27. 28. 29. ‘Which substance is used to generate the light signal in electrochemiluminescence? A. Acridinium B. Luminol €. Dioxetane phosphate D. Ruthenium ChemistrylApply principles of special procedures Iritrumentationl2 25. Light scattering when the wavelength is greater than 10 times the particle diameter is described by: A Rayleigh’s law B. The BeetLambert law ©. Mie's lw D. ‘the Rayleigh-Debye law ChemistrylApply principles of special procedures! Instrumentations2 26. A ‘Which statement regarding nephelometry is true? A. Nephelometry is less sensitive than absorption, spectrophotometry B, Nephelometry follows Beer's law . The optical desiga is identical to a curbidimeter except that a HeNe laser light source is used D. The detector response is directly proportional to concentration ChemistrylApply principles of special procedures! Instrumentation?2 27. “The purpose of the nebulizer in an atomic absorption spectrophotometer that uses a flame isto: ‘A. Convert ions to atoms B. Cause ejection of an outer shell electron C. Reduce evaporation of the sample D. Burn off organic imputities ChemistylApply principles of basic procedures! Instrumentation!2 28. A flameless atomic absorption spectrophotometer dehydrates and atomizes a sample using: A.A graphite capillary furnace B. An clectron gun, C.A thermoelectric semiconductor D. A thermospray platform ChemistrylApply principles of special procedures! Instrumentation! 29.4 Answers to Questions 25-29 All of these substances are chemiluminescent, Dioxetane phosphate is excited by alkaline phosphatase. Acrcinium and luminol are excited. by hydrogen peroxide. In electrochemiluminesence, ruthenium is used to label antibody or antigen, Antigen-antibody complexes containing the ruthenium label are bound to paramagnetic particles via @ strepavidin-biotin reaction, The paramagnetic particles are attracted to an electrode surface, The flowcells washed with a solution containing tripropylamine (TPA) to remove unbound rutheniam Iabel Ac the electrode surface, the TPA is oxdized and the electrons excite the uthenium, causing production of 620-nm light Rayleigh’s law states that when the incident wavelength is much longer than the particle ameter, there is maximum backscatter and ‘minimum right-angle scatter. The Rayleigh-Debye law predicts maximum right-angle scatter when wavelength and particle diameter approach equality In nephelomety, the relationship between wavelength and diameter deterines the angle at which the detector is located D Innephelometry, the detector output is proportional to.concentration (as opposed to turbidimenry where the detectors behind the cuvette). The detectors) 1s (ae) usualy placed at an angle between 25° and 90° to the incident light, depending upon te application. Neghelometets, tke fluorometers, are calorated to read zero with the ight path blocked, and sensitivity can be incteased up to 1,000 times by amalification ofthe detector output or increasing the photomultiplier tube dynode vokage, ‘The atomizer of the atomic absorption spectrophotometer consists of either a nebulizer and flame or a graphite furnace. The nebulizer disperses the sample into a fine aerosol distributing it evenly into the flame. Heat from the flame is used to evaporate water and break the ionic bonds of salts, forming ground state atoms. The flame also excites a small percentage ofthe atoms, which release a characteristic emission line. Flameless atomic absorption uses a hollow tube ‘of graphite with quartz ends. The tube is heated in stages by an electric curtent to successively dry, ash, and atomize the sample. During the ash and atomization steps, argon is injected into the tube todistroute the atoms, The furnace is more sensitive than a lame atomizer and more efficient in atomizing thermostable salts. However, its prone to greater rmatix interference and is slower than the flame atomizer because it must cool down before introduction ofthe next sample, 30. When measuring lead in whole blood using atomic 32, 33. 34. absorption spectrophotometry, what reagent is required to obtain the needed sensitivity and precision? A. Lanthanum B. Lithium €. Triton X-100 D. Chloride ChemistrylApply principles of special procedures! Instrumentation/T 30. ¢ Interference in atomic absorption speetrophotometry caused by differences in viscosity is called: A. Absorption interference B, Matrix effect €.Tonization interference D. Quenching. ChemistrylBvaluate sources of errrllnstramentation!2 All of the following are required when ‘measuring magnesium by atomic absorption spectrophotometty except: A. Ahollow cathode lamp with a magnesium cathode B. A chopper to prevent optical interference from magnesium emission €.A monochromator to isolate the magnesium ‘emission line at 285 nm. D. A.285-nm reference beam to corsect for background absorption ChemistrySelect methods/Reagentsl Medial Electrolytes 2 ‘When measuring calcium by atomic absorption spectrophotometry, which is required? A. An organic extraction reagent to deconjugate calcium from protein B. An internal standard A magnesium chelator D. Lanthanum oxide to chelate phosphates ChemistrySeleet meshodadReagents/Medial Electrolyte!2 31.8 32. D 33.0 Ton selective analyzers using undiluted samples 34.8 have what advantage over analyzers that use a diluted sample? A. Can measure over a wider range of B. Are not subject to pseudohyponatremia caused by high lipids €. Do not require temperature equilibration D. Requite less maintenance ChemistrylApply knowledge to identify sources of errorlElectroytes/2 5.1 | Instrumentation 179 Answers to Questions 30-34 A graphite furnace is preferred over aflame for measuring lead because itis sufficiently sensitive to detect levels below 5 ug/L, the cutoff needed for lead screening of children. The mavrx modifier consists of Trton X-100, ammonium pnasphate and ritric acid. This allows for release of Pb from the RBCs, and solubilization of cel stroma, The matrix modifier also prevents loss of Pe caused by forration of lead halides and promotes interaction between Pb and the tube wall, preventing its loss during the ashing oye, Significant differences in aspiration anc atomization result when the matrx of sample and unknowns differ. Differences in viscosty and protein content are ‘major causes of matrix error Matrix effects can be reduced by using protein-based callraters and diluting both standards and samples prior to assay. Atomic absorption requires a lamp with a cathode made ftom the metal tobe assayed. The lamp emits the ne spectrum of the metal, providing the wavelength that the atoms can absorb, The chopper pulses the source light, allowing it to be discriminated from light emitted by excited atoms, A monochromator eliminates light emitted by the ideal gas in the lamp, Deuterium (wide bandpass light} or Zeeman correction {splitting the incident light into side bands by a magnetic field) may be used to correct for background absorption. An acide diluent such as hydrochloric acid (HCD il cisplace calcium bound to albumin, However, calcium forms a thermostable band with phosphate ‘that causes chemical interference in atomic ‘absorption. Lanthanum disolaces calcium, forming lanthanum chosphate, and eliminates interference from phosphates. Unlike in some colorimetric methods for calcium (e.g, o-ctesolphthalein complexonel, magnesium does notincerfere because it does not absorb the 422.7 nm line fom the cakcium-nollow cathode lamp. lon-selective analyzers measure the electrolyte dissolved in the Fuld phase of the sample in milimoles perlite of plasma water. When undiluted blood is assayed, the measurement is independent of colloids such as protein and lpia, Hyperlipemic samples cause falsely low sodium measurements ‘when assayed by flame photometry ang ion-select've analyzers requiring dilution because Ipids displace plasma water containing the electrobtes, One ‘trawback to undiluted or direct measuring systems Isthat the electrodes require more frequent ddeproteinzation and usually have a shorter duty oye. 180 Chapter 5 | Clinical Chemistry 35. 36. 37, 38, 39. Select the equation describing the potential that develops at the surface of an ion-selective electrode. A. van Deemter equation B. van Slyke equation €. Nernst equation D. Henderson-Hasselbalch equation ChemistrylDefine fundamental characteristiod Instrumentation! “The reference potential ofa silver-silver chloride cleetrode is determined by the: ‘A. Concentration of the potassium chloride filling solution B, Surface area of the electrode . Activity of total anion in the paste covering the electrode D. The concentration of silver in the paste covering the electrode. ChemistrylDefine fundamental characteristics! Instrumentation! ‘The term RT/nF in the Nernst equation defines the: A. Potential at the ion-selective membrane B. Slope of the electrode C. Decomposition potential D. Isopotential point of the electrode ChemistrylDefine fundamental characteristic! Instrumentation! ‘The ion-selective membrane used to measure potassium is made of: A. High-borosilicate glass membrane B, Polyvinyl chloride dioctylphenyl phosphonate jon exchanger €. Valinomycin gel D. Calomel Chemistry/Apply principles of basic laboratory procedures Electrolytes] ‘The response of a sodium electrode to a 10-fold increase in sodium concentration should be: A.A 10-fold drop in potential B. An increase in potential of approximately 60 mV. An increase in potential of approximately 10 mV. D. A decrease in potential of approximately 10 mV Chemistry!CaleulatelElectroytes2 Answers to Questions 35-39 35. C The van Deemter equation describes the 36. 37. A 38. C 39. relation between the velocity of mobile phase to column efficiency in gas chromatography. The Henderson-Hasselbalch equation is used to determine the pH of a solution containing # weak acid and its salt. van Slyke developed an apparatus to measure CO, and 0, content using a manometer. ‘The activity of any solid orion ina saturated solution isunity Fora siher electrode covered with silver chloride paste, the Nemst equation is E= £°—ATinF x 23 logyp Aa x CFIIAGCU, Because sive and silver chloride have an activity of 10, and all components except chloride are constants the potential of the reference electrode is determined by the chloride concenttation of the filing solution, E=E-RY/nE x23 logyfCI]= £592 Vx log( CF fat room temperature) In the term RT/nF, R= the molar gas constant, T=temperature in degrees Kelvin, F= Faraday’'s “constant, and n = the number of electrons donated per atom of reductant. The slope depends upon the temperature ofthe solution and the valence of the reductant. At room temperature, the slope is 582.mV fora univalent on and 29.6 mv fora divalent on Valinomycinisan antibiotic with a highly selective reversble-binging affinty for potassuen ns. Sodium ‘electrodes ate usualy composed af a lass membrane witha high content of aluminum silicate. Calcium and Itium ion-selective electrodes are made rom organic lquidion exchangers called neutral carierionophares Galorelis made of mercury covered with a paste of mercurous chloride g"/g,C) ands used 2s 2 reference electrode fr oH. Te Nemst equation predicts an increase of approximately 69 mV per 10d increase in sodium actnty. For sodium E=E"+RT/MF x23 logyNo] Therefore: E=°+ 60 mV x/og,gINarl Hf sodium concentration is 19 mmeV, then: E=E"+ 60 mV 0g,(10)=E* + 60 mV. If sodium concentration increases from 10 mmol/l. to 100 mmol/L ther: E=E°+ 60 mV x09, 9{100] = Ey 60mVx2=E"+ 120m, 40. a 43. 4a, 45. Which of the electrodes below is a current- ‘producing (amperomettic) rather than a voltage-producing (potentiometric) electrode? A. Clark electrode B. Severinghaus electrode €. pH electrode D. Ionized calcium electrode ChemistrylDefine fundamental characteristics! Instrumentation! ‘Which of the following would cause a “response” error from an ion-selective electrode for sodium when measuring serum but not the calibrator? A Incerference from other electrolytes B, Protein coating the ion-selective membrane . An overrange in sodium concentration D. Protein binding to sodium ions Chemistry identify sources of eror/Blectrolytes/2 In polarography, the voltage needed to cause depolarization of the cathode is called the: A. Half-wave potential B. Isoporential point €. Decomposition potential D. Polarization potential Chemistry Define fundamental characteristics! Instrumentation/i Persistent noise from an ion-selective electrode is most often caused by: ‘A. Contamination of sample B. Blocked junction at the sale bridge €. Overrange from high concentration D. Improper calibration Chemistrylidentify sources of erorlBlectrolytes 2 ‘Which clement is reduced at the cathode of a Clark polarographic electrode? A Silver B. Oxygen ©. Chloride D. Potassium hemistrylDefine fundamental characteristic! Instrumentation/i ‘Which of the following statements accurately characterizes the coulomettic titration of chloride? [A The indicator electrodes generate voltage B. Constant current must be present across the generator electrodes C. Silver ions are formed atthe generator cathode D. Chloride concentration is inversely proportional co titration time ChemistrylDefine fundamental characteristid Instrumentation!2 5.1 | Instrumentation 181 Answers to Questions 40-45 40. A The Clark electrode is composed of two half cells a. 4s. that generate current, not voltage. is used to measure partial pressure of oxygen (Po), and is based upon an amperometric method called polarography. When ~0.8 V's aoplied so tne cathode, 0, s reduced, causing current to flow. Current is proportional o the Po, ofthe sample. Response is the time required for an electrode to reach maximum potential. lor-selectve analyzers Use a micropracessor to moniter electiode response, slope, drt, ang noise. When an electrode gives an acceptable response time when measuring an aqueous calibrator, but not when measuring serum, the cause is often protein buildup on the membrane, In polarography, a minimum negative vokage must bbe applied to the cathode to cause reduction of rmetalions (or 0.) in solution. This s called the decomposition potential. It's concentration dependent (dilute solutions requite greater negative voltage), and can be determined using the Nernst equat'on. Electrode noise most ten resus fom an unstable junction potential, Most reference electrodes contain ahigh concentration of KC internal soluton used to produce the eference potential. This forms a salt bridge with the measuring half cel by contacting sample, bts

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