KIMIA KIMIA ANALISA ANALISA 33 SKS SKS - Website Personalpersonal.its.ac.id/files/material/2852-heru-che-KA03.pdf · Graphite furnace atomic absorption spectroscopy Pb in Water Pb

KIMIA KIMIA ANALISAANALISA33 SKSSKS

Prof. Prof. DrDr. . HeruHeru SetyawanSetyawanJurusanJurusan TeknikTeknik Kimia FTI Kimia FTI –– ITSITS

The Language of Analytical The Language of Analytical ChemistryChemistry

Analysis, Determination, and MeasurementAnalysis, Determination, and Measurement

�� Analysis:Analysis:�� A process that provides chemical or physical information A process that provides chemical or physical information

about the constituents in the sample or the sample itself.about the constituents in the sample or the sample itself.

�� Analytes:Analytes:�� The constituents of interest in a sample.The constituents of interest in a sample.

�� Matrix:Matrix:All other constituents in a sample except for the All other constituents in a sample except for the

Dr. Heru Setyawan, Jurusan Teknik Kimia FTI Dr. Heru Setyawan, Jurusan Teknik Kimia FTI -- ITSITS

�� All other constituents in a sample except for the All other constituents in a sample except for the analytes.analytes.

�� Determination:Determination:�� An analysis of a sample to find the identity, An analysis of a sample to find the identity,

concentration, or properties of the analyte.concentration, or properties of the analyte.

�� Measurement:Measurement:�� An experimental determination of an analyte’s chemical An experimental determination of an analyte’s chemical

or physical properties.or physical properties.

Analysis, Determination, and MeasurementAnalysis, Determination, and MeasurementIn 1974, the federal government In 1974, the federal government enacted the Safe Drinking Water Act to enacted the Safe Drinking Water Act to ensure the safety of public drinking ensure the safety of public drinking water supplies. To comply this act water supplies. To comply this act municipalities regularly monitor their municipalities regularly monitor their drinking water supply for substantially drinking water supply for substantially harmful substances. One such harmful substances. One such substance is coliform bacteria. substance is coliform bacteria. Municipal water departments collect Municipal water departments collect and analyze samples from their water and analyze samples from their water supply. To determine the concentration supply. To determine the concentration of coliform bacteria, a portion of water of coliform bacteria, a portion of water


of coliform bacteria, a portion of water of coliform bacteria, a portion of water is passed through a membrane filter. is passed through a membrane filter. The filter is placed in a dish containing The filter is placed in a dish containing a nutrient broth and incubated. At the a nutrient broth and incubated. At the end of the incubation period the number end of the incubation period the number of coliform bacterial colonies is of coliform bacterial colonies is measured by counting (see figure). measured by counting (see figure). Thus, municipal water departments Thus, municipal water departments analyzeanalyze samples of water to samples of water to determinedeterminethe concentration of coliform bacteria the concentration of coliform bacteria by by measurinmeasuringg the number of bacterial the number of bacterial colonies that form during a specified colonies that form during a specified period of incubation. period of incubation.

• Upper left: typical dark blue fecal coliform colonies on a membrane filter after incubation on M-FC agar.

• Lower center (purple): M-FC agar before use.• Upper right (blue): a control plate streaked

with an E.coli culture.

Techniques, Methods, Procedures, and ProtocolsTechniques, Methods, Procedures, and Protocols

�� Technique:Technique:�� A chemical or physical principle that can be A chemical or physical principle that can be

used to analyze a sample.used to analyze a sample.

�� Method:Method:�� A means for analyzing a sample for a specific A means for analyzing a sample for a specific

analyte in a specific matrix.analyte in a specific matrix.


analyte in a specific matrix.analyte in a specific matrix.

�� Procedure:Procedure:�� Written directions outlining how to analyze a Written directions outlining how to analyze a

sample.sample.

�� Protocol:Protocol:�� A set of written guidelines for analyzing a A set of written guidelines for analyzing a

sample specified by an agency.sample specified by an agency.

Graphite furnaceatomic absorption spectroscopy

Pb inWater

Pb inSoil

Pb inBlood

APHA ASTM

Techniques

Methods

Procedures


EPAProtocols

Chart showing hierarchical relationship among a technique, methods using that technique, and procedures and protocols for one method.

Abbreviations:APHA = American Public Health AssociationASTM = American Society for Testing MaterialsEPA = Environmental Protection Agency

Classifying Analytical TechniquesClassifying Analytical Techniques• If a technique responds to the absolute absolute

amountamount of analyte in the sample, the signal due to the analyte SA

SA = knA

• Since cylinder 2 contains twice as many moles of Cu2+ as cylinder 1, analyzing the contents of cylinder 2 gives signal that is twice that of cylinder 1.


Graduated cylinders containing 0.01 M Cu(NO3)2. (a) Cylinder 1 contains 10 mL, or 0.0001 mol, of Cu2+. (b) Cylinder 2 contains 20 mL, or 0.0002 mol, of Cu2+.

cylinder 1.

• If a technique responds to the relative relative amountamount of analyte in the sample, the signal due to the analyte SA

SA = kCA

• Since the solutions in both cylinders have the same concentration of Cu2+, their analysis yields identical signals.

Classifying Analytical TechniquesClassifying Analytical Techniques

�� Total analysis techniquesTotal analysis techniques�� A technique in which the signal is proportional to the A technique in which the signal is proportional to the

absolute amount of analyte; also called “classical” absolute amount of analyte; also called “classical” techniques.techniques.

�� Mass, volume, and charge are the most common signals Mass, volume, and charge are the most common signals for total analysis techniques, and the responding for total analysis techniques, and the responding techniques are gravimetry, titrimetry, and coulometry.techniques are gravimetry, titrimetry, and coulometry.

�� Concentration techniquesConcentration techniques


�� Concentration techniquesConcentration techniques�� A technique in which the signal is proportional to the A technique in which the signal is proportional to the

analyte’s concentration; also called “instrumental” analyte’s concentration; also called “instrumental” technique, e.g., spectroscopy, potentiometry, and technique, e.g., spectroscopy, potentiometry, and voltammetry.voltammetry.

�� The relationship between the signal and the analyte is a The relationship between the signal and the analyte is a theoretical function that depends on experimental theoretical function that depends on experimental conditions and the instrumentation used to measure the conditions and the instrumentation used to measure the signal, so the value of signal, so the value of kk must be determined must be determined experimentally.experimentally.

Validation involves determining:

•selectivity

•linearity

•accuracy

•precision

•sensitivity

•range

•limit of detection


General process for evaluation/validation of methodology.

•limit of quantitation

•ruggedness/robustness

Standard reference materials (SRMs) best for determining accuracy.

Selecting an Analytical MethodSelecting an Analytical Method�� AccuracyAccuracy

�� A measure of the agreement between an experimental result and its A measure of the agreement between an experimental result and its expected value.expected value.

�� Analytical methods may be divided into three groups based on the Analytical methods may be divided into three groups based on the magnitude of their relative errors:magnitude of their relative errors:

�� < 1% : highly accuracy< 1% : highly accuracy

100result expected

result expected result obtained Error % ×−=


�� < 1% : highly accuracy< 1% : highly accuracy�� Between 1% and 5% : moderately accuratcyBetween 1% and 5% : moderately accuratcy�� > 5% : low accuracy> 5% : low accuracy

�� In general, total analysis methods produce results of high accuracy, In general, total analysis methods produce results of high accuracy, and concentration methods range from high to low accuracy.and concentration methods range from high to low accuracy.

�� PrecisionPrecision�� An indication of the reproducibility of a measurement or result.An indication of the reproducibility of a measurement or result.�� Depends on those factors affecting the relationship between the Depends on those factors affecting the relationship between the

signal and the analyte. Of particular importance are the uncertainty signal and the analyte. Of particular importance are the uncertainty in measuring the signal and the ease of handling sampling in measuring the signal and the ease of handling sampling reproducibly.reproducibly.

Selecting an Analytical MethodSelecting an Analytical Method

5.8 5.9 6.0 6.1 6.2(a)


5.8 5.9 6.0 6.1 6.2

(b)

Two determinations of the concentration of K+ in serum, showing the effect of precision. The data in (a) are less scattered and, therefore, more precise than the data in (b).


�� SensitivitySensitivity�� A measure of a method’s ability to distinguish between A measure of a method’s ability to distinguish between

two samples; reported as the change in signal per unit two samples; reported as the change in signal per unit change in the amount of analyte (change in the amount of analyte (kk).).

�� If If ∆∆SSAA is the smallest increment in signal that can be is the smallest increment in signal that can be measured, then the smallest difference in the amount of measured, then the smallest difference in the amount of analyte that can be detected isanalyte that can be detected is


�� Detection limitDetection limit�� A statistical statement about the smallest amount of A statistical statement about the smallest amount of

analyte that can be determined with confidence.analyte that can be determined with confidence.

method) analysistion (concentra

method) analysis (total

k

SC

k

Sn

AA

AA

∆=∆

∆=∆


�� SelectivitySelectivity�� A measure of a method’s freedom from interferences as A measure of a method’s freedom from interferences as

defined by the method’s selectivity coefficient.defined by the method’s selectivity coefficient.SSsampsamp = = SSAA + + SSII = = kkAAnnAA + + kkIInnII (total analysis method)(total analysis method)SSsampsamp = = SSAA + + SSII = = kkAACCAA + + kkIInnII (concentration analysis method)(concentration analysis method)

�� Selectivity coefficientSelectivity coefficient


�� A measure of a method’s sensitivity for an interferent A measure of a method’s sensitivity for an interferent relative to that for the analyte (relative to that for the analyte (KKA,IA,I))

A

IIA, k

kK =

Selecting an Analytical MethodSelecting an Analytical Method1.1. A method for the analysis of CaA method for the analysis of Ca2+2+ in water suffers from an in water suffers from an

interference in the presence of Zninterference in the presence of Zn2+2+. When the . When the concentration of Caconcentration of Ca2+2+ is 100 times greater than that of is 100 times greater than that of ZnZn2+2+, an analysis for Ca, an analysis for Ca2+2+ gives a relative error +0.5%. gives a relative error +0.5%. What is the selectivity coefficient for this method?What is the selectivity coefficient for this method?

2.2. Barnett and colleagues developed a new method for Barnett and colleagues developed a new method for determining the concentration of codeine during its determining the concentration of codeine during its extraction from poppy plants. As part of their study they extraction from poppy plants. As part of their study they determined the method’s response to codeine relative to determined the method’s response to codeine relative to


determined the method’s response to codeine relative to determined the method’s response to codeine relative to that for several potential interferents. For example, the that for several potential interferents. For example, the authors found that the method’s signal for 6authors found that the method’s signal for 6--methoxycodeine was 6 (arbitrary units) when that for an methoxycodeine was 6 (arbitrary units) when that for an equimolar solution of codeine was 40.equimolar solution of codeine was 40.

a)a) What is the value for the selectivity coefficient What is the value for the selectivity coefficient KKA,IA,I when 6when 6--methoxycodeine is the interferent and codeine is the analyte?methoxycodeine is the interferent and codeine is the analyte?

b)b) If the concentration of codeine is to be determined with an If the concentration of codeine is to be determined with an accuracy of accuracy of ±±0.50%, what is the maximum relative concentration 0.50%, what is the maximum relative concentration of 6of 6--methoxycodeine (i.e., [6methoxycodeine (i.e., [6--methoxycodeine]/[codeine]) that can methoxycodeine]/[codeine]) that can be present?be present?


�� RobustRobust�� A method that can be applied to analytes in a A method that can be applied to analytes in a

wide variety of matrices is considered robust.wide variety of matrices is considered robust.

�� RuggedRugged�� A method that is insensitive to changes in A method that is insensitive to changes in


experimental conditions is considered rugged.experimental conditions is considered rugged.


�� Scale of OperationScale of Operation�� Three limitations of particular importance:Three limitations of particular importance:

�� The amount of sample available for the analysis.The amount of sample available for the analysis.�� The concentration of analyte in the sample.The concentration of analyte in the sample.�� The absolute amount of analyte needed to obtain a The absolute amount of analyte needed to obtain a

measurable signal.measurable signal.


measurable signal.measurable signal.


-2

10-3%

10-4%

10-5%

10-6%

10-7%

10-8%

10-9%

10-10%

Trace

Ultratrace

log

(% a

nal

yte

as %

w/w

)

1 g sample, 1% analyte

0.1 g sample, 10% analyte


1 0.1 0.01100 10 1 0.1 0.01

100 10 1 0.1 0.01100 10 1

100%

10%

1%

0.1%

10-2%

MacroMeso Micro Ultramicro

gmgµgng

Major

Minor

-log(Weight of sample)

-lo

g(%

an

alyt

e as

%w

/w)



Scale of operation for analytical methods.

Selecting an Analytical MethodSelecting an Analytical MethodEquipment, Time, and CostEquipment, Time, and Cost

�� Analytical methods can be compared in terms ofAnalytical methods can be compared in terms of�� their need for equipmenttheir need for equipment�� the time required to complete an analysisthe time required to complete an analysis�� the cost per sample.the cost per sample.

�� Methods relying on instrumentation are equipmentMethods relying on instrumentation are equipment--intensive and may intensive and may require significant operator training.require significant operator training.�� The graphite furnace atomic absorption spectroscopic method for The graphite furnace atomic absorption spectroscopic method for

determining lead levels in water requires a significant capital investment in determining lead levels in water requires a significant capital investment in the instrument and experienced operator to obtain reliable results.the instrument and experienced operator to obtain reliable results.

�� Other methods such as titrimetry, require only simple equipment and Other methods such as titrimetry, require only simple equipment and


�� Other methods such as titrimetry, require only simple equipment and Other methods such as titrimetry, require only simple equipment and reagents and can be learned quickly.reagents and can be learned quickly.

�� The time needed to complete an analysis for a single sample is often The time needed to complete an analysis for a single sample is often fairly similar from method to method.fairly similar from method to method.�� This is somewhat misleading because much of this time is spent preparing This is somewhat misleading because much of this time is spent preparing

the solutions and equipment needed for the analysis. Once the solutions the solutions and equipment needed for the analysis. Once the solutions and equipment are in place, the number of samples that can be analyzed and equipment are in place, the number of samples that can be analyzed per hour differs substantially from method to method.per hour differs substantially from method to method.

�� The cost of an analysis is determined by many factors, including the The cost of an analysis is determined by many factors, including the cost of necessary equipment and reagents, the cost of hiring analysts, cost of necessary equipment and reagents, the cost of hiring analysts, and the number of samples that can be processed per hour.and the number of samples that can be processed per hour.�� In general, methods relying on instruments cost more per sample than other In general, methods relying on instruments cost more per sample than other

methods.methods.

Calibration and StandardizationCalibration and Standardization�� CalibrationCalibration

�� The process of ensuring that the signal measured by a piece of equipment The process of ensuring that the signal measured by a piece of equipment or an instrument is correct.or an instrument is correct.

�� For example: balances are calibrated using a standard weight whose mass For example: balances are calibrated using a standard weight whose mass can be traced to the internationally accepted platinumcan be traced to the internationally accepted platinum--iridium prototype iridium prototype kilogram.kilogram.

�� StandardizationStandardization�� The process of establishing the relationship between the amount of analyte The process of establishing the relationship between the amount of analyte

and a method’s signal.and a method’s signal.�� For a total analysis method, standardization is usually defined by the For a total analysis method, standardization is usually defined by the

stoichiometry of the chemical reactions responsible for the signal.stoichiometry of the chemical reactions responsible for the signal.


stoichiometry of the chemical reactions responsible for the signal.stoichiometry of the chemical reactions responsible for the signal.�� For a concentration method, the relationship between the signal and the For a concentration method, the relationship between the signal and the

analyte’s concentration is a theoretical function that cannot be calculated analyte’s concentration is a theoretical function that cannot be calculated without experimental measurements.without experimental measurements.

�� To standardize method, the value of To standardize method, the value of kk is determined by measuring the is determined by measuring the signal for one or more standards, each containing a known concentration of signal for one or more standards, each containing a known concentration of analyte.analyte.

�� When several standards with different concentrations of analyte are used, When several standards with different concentrations of analyte are used, the result is best viewed visually by plotting the result is best viewed visually by plotting SSmeasmeas versus the concentration versus the concentration of analyte in the standards. Such plot is known as a of analyte in the standards. Such plot is known as a calibration curvecalibration curve..

ProblemsProblems1.1. When working with a solid sample, it often is necessary to bring the analyte When working with a solid sample, it often is necessary to bring the analyte

into solution by dissolving the sample in a suitable solvent. Any solid impurities into solution by dissolving the sample in a suitable solvent. Any solid impurities that remain are removed by filtration before continuing with the analysis. In a that remain are removed by filtration before continuing with the analysis. In a typical total analysis method, the procedure might readtypical total analysis method, the procedure might read

After dissolving the sample in a beaker, remove any solid impurities by passing After dissolving the sample in a beaker, remove any solid impurities by passing the solution containing the analyte through filter paper, collecting the solution in the solution containing the analyte through filter paper, collecting the solution in a clean Erlenmeyer flask. Rinse the beaker with several small portions of a clean Erlenmeyer flask. Rinse the beaker with several small portions of solvent, passing these rinsings through the filter paper, and collecting them in solvent, passing these rinsings through the filter paper, and collecting them in the same Erlenmeyer flask. Finally, rinse the filter paper with several portions the same Erlenmeyer flask. Finally, rinse the filter paper with several portions of solvent, collecting the rinsings in the same Erlenmeyer flask.of solvent, collecting the rinsings in the same Erlenmeyer flask.


For a typical concentration method, however, the procedure might stateFor a typical concentration method, however, the procedure might state

After dissolving the sample in a beaker, remove any solid impurities by filtering After dissolving the sample in a beaker, remove any solid impurities by filtering a portion of the solution containing the analyte. Collect and discard the first a portion of the solution containing the analyte. Collect and discard the first several milliliters of solution before collecting a sample of approximately 5 mL several milliliters of solution before collecting a sample of approximately 5 mL for further analysis.for further analysis.

Explain why these two procedures are different.Explain why these two procedures are different.

ProblemsProblems2.2. A certain concentration method works best when the analyte’s concentration A certain concentration method works best when the analyte’s concentration

is approximately 10 ppb.is approximately 10 ppb.a.a. If the sampling volume for the method is 0.5 mL, about what mass of If the sampling volume for the method is 0.5 mL, about what mass of

analyte is being measured?analyte is being measured?b.b. If the analyte is present at 10% w/v, how would you prepare the sample If the analyte is present at 10% w/v, how would you prepare the sample

for analysis?for analysis?c.c. Repeat for the case in which the analyte is present at 10% w/w.Repeat for the case in which the analyte is present at 10% w/w.d.d. Based on your results, comment on the suitability of this method for the Based on your results, comment on the suitability of this method for the

analysis of a major analyte.analysis of a major analyte.3.3. An analyst needs to evaluate the potential effect of an interferent, I, on the An analyst needs to evaluate the potential effect of an interferent, I, on the

quantitative analysis for an analyte, A. She begins by measuring the signal quantitative analysis for an analyte, A. She begins by measuring the signal


quantitative analysis for an analyte, A. She begins by measuring the signal quantitative analysis for an analyte, A. She begins by measuring the signal for a sample in which the interferent is absent and the analyte is present with for a sample in which the interferent is absent and the analyte is present with a concentration of 15 ppm, obtaining an average signal of 23.3 (arbitrary a concentration of 15 ppm, obtaining an average signal of 23.3 (arbitrary units). When analyzing a sample in which the analyte is absent and the units). When analyzing a sample in which the analyte is absent and the interferent is present with a concentration of 25 ppm, she obtains an average interferent is present with a concentration of 25 ppm, she obtains an average signal of 13.7.signal of 13.7.

a.a. What is the analyte’s sensitivity?What is the analyte’s sensitivity?b.b. What is the interferent’s sensitivity?What is the interferent’s sensitivity?c.c. What is the value of the selectivity coefficient?What is the value of the selectivity coefficient?d.d. Is the method more selective for the analyte or the interferent?Is the method more selective for the analyte or the interferent?e.e. What is the maximum concentration of interferent relative to that of the What is the maximum concentration of interferent relative to that of the

analyte (i.e., [interferent]/[analyte]), if the error in the analysis is to be analyte (i.e., [interferent]/[analyte]), if the error in the analysis is to be less than 1%?less than 1%?

ProblemsProblems4.4. A sample was analyzed to determine the concentration of an analyte. Under A sample was analyzed to determine the concentration of an analyte. Under

the conditions of the analysis, the sensitivity is 17.2 ppmthe conditions of the analysis, the sensitivity is 17.2 ppm--11. What is the . What is the analyte’s concentration if analyte’s concentration if SSmeasmeas is 35.2 and is 35.2 and SSreagreag is 0.6?is 0.6?

5.5. A method for the analysis of Ca2+ in water suffers from an interference in the A method for the analysis of Ca2+ in water suffers from an interference in the presence of Zn2+. When the concentration of Ca2+ is 50 times greater than presence of Zn2+. When the concentration of Ca2+ is 50 times greater than that of Zn2+, an analysis for Ca2+ gives a relative error of that of Zn2+, an analysis for Ca2+ gives a relative error of --2.0%. What is the 2.0%. What is the value of the selectivity coefficient for this method?value of the selectivity coefficient for this method?

6.6. The quantitative analysis for reduced glutathione in blood is complicated by The quantitative analysis for reduced glutathione in blood is complicated by the presence of many potential interferents. In one study, when analyzing a the presence of many potential interferents. In one study, when analyzing a solution of 10solution of 10--ppb glutathione and 1.5ppb glutathione and 1.5--ppb ascorbic acid, the signal was 5.43 ppb ascorbic acid, the signal was 5.43 times greater than that obtained for the analysis of 10times greater than that obtained for the analysis of 10--ppb glutathione. What ppb glutathione. What is the selectivity coefficient for this analysis? The same study found that is the selectivity coefficient for this analysis? The same study found that


is the selectivity coefficient for this analysis? The same study found that is the selectivity coefficient for this analysis? The same study found that when analyzing a solution of 350when analyzing a solution of 350--ppb methionine and 10ppb methionine and 10--ppb glutathione the ppb glutathione the signal was 0.906 times less than that obtained for the analysis of 10 ppbsignal was 0.906 times less than that obtained for the analysis of 10 ppb--glutathione. What is the selectivity coefficient for this analysis? In what way glutathione. What is the selectivity coefficient for this analysis? In what way do these interferents behave differently?do these interferents behave differently?

7.7. Oungpipat and Alexander described a new method for determining the Oungpipat and Alexander described a new method for determining the concentration of glycolic acid (GA) in a variety of samples, including concentration of glycolic acid (GA) in a variety of samples, including physiological fluids such as urine. In the presence of only GA, the signal is physiological fluids such as urine. In the presence of only GA, the signal is given asgiven as

SSsamp,1samp,1 = = kkGAGA CCGAGAand in the presence of both glycolic acid and ascorbic acid (AA), the signal isand in the presence of both glycolic acid and ascorbic acid (AA), the signal is

SSsamp,2samp,2 = = kkGAGA CCGAGA + + kkAAAA CCAAAA

ProblemsProblemsWhen the concentration of glycolic acid is 1.0 When the concentration of glycolic acid is 1.0 ×× 1010--4 M and the concentration 4 M and the concentration of ascorbic acid is 1.0 of ascorbic acid is 1.0 ×× 1010--5 M, the ratio of the signals was found to be5 M, the ratio of the signals was found to be

SSsamp,2samp,2//SSsamp,1samp,1 = 1.44= 1.44a.a. Using the ratio of the two signals, determine the value of the selectivity Using the ratio of the two signals, determine the value of the selectivity

ratioratioKKGA,AAGA,AA = = kkAAAA//kkGAGA

b.b. Is the method more selective toward glycolic acid or ascorbic acid?Is the method more selective toward glycolic acid or ascorbic acid?c.c. If the concentration of ascorbic acid is 1.0 If the concentration of ascorbic acid is 1.0 ×× 1010--5 M, what is the smallest 5 M, what is the smallest

concentration of glycolic acid that can be determined such that the error concentration of glycolic acid that can be determined such that the error introduced by failing to account for the signal from ascorbic acid is less introduced by failing to account for the signal from ascorbic acid is less


introduced by failing to account for the signal from ascorbic acid is less introduced by failing to account for the signal from ascorbic acid is less than 1%?than 1%?

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KIMIA KIMIA ANALISA ANALISA 33 SKS SKS - Website Personalpersonal.its.ac.id/files/material/2852-heru-che-KA03.pdf · Graphite furnace atomic absorption spectroscopy Pb in Water Pb