Вы находитесь на странице: 1из 91

LaboratoryExperimentsinAnalyticalChemistry

CHEM3332
AnalyticalChemistryDivision
DepartmentofChemistry
UniversityofConnecticut
Storrs,CT062693060

ContributingAuthors:
JamesStuart,JaneKnox,PedroCidAgueroandAbhayVaze

TableofContents
SyllabusfortheLaboratoryPartofCHEM3332
Introduction
Safety
Attendance
Notebook
Reports&Paper
Grades
Materials
Assignments
Beforeyoustart
Policies&Procedures
ExperimentSchedule
AcademicHonesty
TheLaboratoryNotebook
1.UsingLaboratoryNotebook
a.Safekeeping
b.OrganizationandReadabilityoftheLabNotebook
c.QualityoftheRecordKeeping
d.GuidelinesforKeepingtheNotebook
2.ContentsoftheNotebook
a.TableofContents
b.Dates
c.TitleofExperiment
d.Experimental
e.Data
f.DataTables
ExampleofaTable

S16

g.Notes
h.Calculations
i.Results
3.GradingoftheLaboratoryNotebook
a.PhysicalAspects
b.Format
c.Neatness
d.Organization
e.Completeness
f.AppropriateRecordofData
4.HowtheLabNotebookwillbegraded?
WarningaboutAcademicMisconduct!
PreLabReport
PostLabReport
LaboratoryPaper
WarningaboutAcademicMisconduct
WritingthePaper
SubmittingReportsandLabPaper
LatenessPolicyandLabPreparedness
a.PrelabReport
b.PostlabReport
c.LabPaper
UsingAnalyticalBalances
GeneralRules
GA1AnalyticalTechniquesandMeasurements
UseoftheAnalyticalBalances
a.PrecisionofVariousBalances
b.UseoftheDigitalBalance
c.SourcesofDeterminateWeighingErrors

S16

CalibrationofVolumetricGlassware
a.BuoyancyCorrections
b.DirectCalibrationof25mLPipet
c.DirectCalibrationofaDigitalPipet
DeterminationoftheDensityofAntifreeze
Questions/TopicsfortheQuiz
Calculations&/orQuestionsforPostlabReport
Datasheets
GA2AcidBaseTitration
SolutionsandChemicals
Standardizationof~0.1MNaOHagainstthePrimaryStandardKHP
DeterminationofPercentKHPinanUnknownMixture
CalculationsforPrelabReport
Calculations&/orQuestionsforPostlabReport
GA4AnalysisofCalciumbyEDTA
SolutionsandChemicals
PreparationandStandardizationofa0.03MEDTASolution
PreparingCalciumStandard
DeterminationoftheIndicatorBlank
Standardizationof~0.03MEDTA
DeterminationoftheCalciuminanUnknownCalciumSample
WaterSample
Questions/TopicsfortheQuiz
Calculations&/orQuestionsforPostlabReport
GA5IodometricTitrationofAscorbicAcidinVitaminCTablets
SolutionsandChemicals
Glassware
PreparationoftheThiosulfateSolution
PreparationofthePotassiumIodateSolution

S16

StandardizationoftheThiosulfateSolutions
Equations
AnalysisofascorbicacidintheVitaminCTablets
Questions/TopicsfortheQuiz
Calculations&/orQuestionsforPostlabReport
GA7PotentiometricTitration
SolutionsandChemicals
Equipment
DeterminationoftheEquivalencePoint
StandardizationofthepHMeterwithTwoStandardBuffers
StandardizationofHClbyPotentiometricTitration
PotentiometricTitrationofPotassiumAcidPhthalate(KHP)
PotentiometricTitrationofaDiproticAcid
Questions/TopicsfortheQuiz
Calculations&/orQuestionsforPreLabReport
Calculations&/orQuestionsforPostlabReport
GA8 Ion Selective Electrode: Measurement of Fluoride Ion Activity in Mouthwash,
Toothpaste&DrinkingWater
SolutionsandChemicals
Equipments
Glassware
PreparationofStandardSolutions
AdjustingtheZerooftheMillivoltmeter
CautionsConcerningUseofElectrodesandMeters
DeterminationofPotentialsfortheCalibrationCurve
DeterminationofFluorideinToothpaste(optional)
DeterminationofFluorideintheMouthwash
DeterminationofFluorideinWaterSamples
DeterminationofFluorideinUnknown

S16

Questions/TopicsfortheQuiz
Calculations&/orQuestionsforPostlabReport
GA16AStudyofAtomicAbsorption
SolutionsandChemicalsRequired
Equipment
InstrumentWarmup
PreparationofSolutions
a.CalciumSolutions
b.UnknownSolutions
CleaningUp
Questions/TopicsfortheQuiz
Calculations&/orQuestionsforPostlabReport
GA20 High Speed Liquid Chromatographic Separation of Various Compounds Found in
Coffee,TeaandSoftdrinks
Instrumentation
Equipment
SolutionandChemicals
Glassware
GettingStarted
a.AdjustmentofSolventFlow
b.FillingtheSampleLoop
c.Injectionofthesample
d.IntroductionofSampleintotheColumn,andRecordingofthePeak
e.IdentificationofComponentsinMixtures
f.TheoreticalPlates&PeakSymmetry
g.TheSoftDrinkorCoffeeand/orteasample
InstrumentShutdown
RecordingAllExperimentalConditions
CalculationsandQuestionsfortheLabPaper

S16

GA21SpectrophotometricDeterminationofIron
SolutionsandChemicals
PreparationofStandardSolutions
PreparationofUnknownSolution
Spectrophotometermeasurements
TopicsfortheQuiz
Calculations&/orQuestionsforPostlabReport

S16

SyllabusfortheLaboratoryPartofCHEM3332
DepartmentofChemistry,UniversityofConnecticut
QuantitativeAnalyticalLaboratory
Chemistry3332

Instructor:DrAbhayVaze
Email:abhay.vaze@uconn.edu
Officehours(T414):Wednesday&Thursday9AM5PM,byappointment

Introduction
Chemistry 3332 introduces a number of experiments oriented toward developing experience in
performing accurate and precise chemical measurements by both, classical and instrumental
techniques. These experiments will give you skills in solutions preparation, understanding of
instrumentation,datainterpretation,resultsevaluationandpreparationofscientificreports.

Safety
In many of the experiments, you will be handling toxic or dangerous chemicals. Special
precautions will be needed, so followtheinstructionscarefully. SafetyGogglesmustbewornat
all times in the lab. You must wear Solid Shoes(nosandalsoropentoeshoes).Failuretodoso
will result in your dismissal from the lab period without make up options. It is your
responsibility to clean your glassware. For more information, please see: Page 30,Quantitative
ChemicalAnalysis,edition8thbyDanielHarris,W.H.FreemanandCompanyNY.

Attendance
Attendance is Mandatory and all students should be on time.Onlywelldocumentedabsencesto
thelaboratorywillbeconsidered,andmakeupswilldependonavailabilityofinstructors.

S16

Notebook
A bound notebook, with numbered pagesandcapableofproducingcarboncopies,isrequiredby
the first lab period in which experiments are performed. All data and observations must be
directly recorded in your notebook (except for GA1 experiment, see experiment handout for
details). All entries must be madeinink.Referto"TheLaboratoryNotebook"sectionforformat
and editorial style. The notebook must not be removed from the laboratory until the end of the
semester.

Reports&Paper
PreLab report: To assure advance preparation, you will be required to prepare a prelaboratory
report. Prelab report needs to be submitted electronically (PDF) to the lab instructor at least
TWOdayspriortothescheduledexperiment.SeesectiononPrelabReportfordetails.

Post Lab report: is required for each experiment. See section on Experiment Report for
details. Eachstudentwillsubmithis/herownindependentreport.Noplagiarismwillbetolerated.
Aftercompletionoftheexperiment,labreportisdueinthefollowingweek.
Electroniccopy(PDFdocument)oftheExperimentReportisdueon:

ForMonday/WednesdaySection,reportisdueonWednesdayby10am.

ForTuesday/ThursdaySection,reportisdueonThursdayby10am.

If the Experiment Report is late, there will be a cumulative 10 points deduction per day the lab
reportisoverdue.
Late reports due to a valid and welldocumented emergency will beconsideredonanindividual
casebasis.

Laboratory Paper: A full paper will be required for one assigned experiment. See section on
Laboratory Paper for details. Each student will submit his/her own independently written
paper. See Experiment Schedule to know the experiment assigned for the lab paper.
LaboratoryPaperisdueonLaboratoryPaperisdueonThursday,April282016,at10.00am.

S16


10

Grades
CHEM3332CourseGrade:Coursegradewillbebased:

52%onexaminations(4examinations,eachwillconstitute13%ofthecoursegrade)

35%ontheLaboratory
PreLabReport
PostLabReport
PostLabReport
LabPaper
LabNotebook

GA1,2,7
GA4,5,8,16,21
GA20

5%
15%
60%
15%
5%

Laboratory notebook will be assessed and graded, without priors notice, several times
duringthesemester.

13%onQuiz(8quizzesinasemester).
Quizzes will be based on Laboratory (see Experiment handouts for topics/questions) and Class
Material.

Materials
AlllaboratorymaterialisavailableonHuskyCT.

S16


11

Assignments
Before beginning any experiment, you must have read the assigned background material from
lab manual, as well as from the indicated chapters in the course textbook (Harris: 7th or 8th
Edition)

1. GA1AnalyticalTechniquesandMeasurements(Chapters14)
2. GA2AcidBaseTitration1(Chapters711)
3. GA7AcidBasePotentiometricTitration(Chapters711)
4. GA4AnalysisofCalciumByEDTAtitration(Ed7th:12&Ed8th:11)
5. GA5 Iodometric Titration of Ascorbic Acid in Vitamin C Tablets (Ed 7th: 15 & Ed 8th:
16)
6. GA8 Ion Selective Electrode: The Measurement of Fluoride Ion ActivityinMouthwash,
Toothpaste,DrinkingWater(Ed7th:14,15&Ed8th:13,14)
7. GA16AStudyofCabyAtomicAbsorptionSpectroscopy(Ed7th:21&Ed8th:20)
8. GA20 High Speed Liquid Chromatographic Separation of Various Compounds found in
soda,CoffeeorTea(Ed7th:23,25&Ed8th:22,24)
9. GA21SpectrophotometricDeterminationofIron(Ed7th:1820&Ed8th:17,18)

Beforeyoustart
Pleaseseefollowingsectionsofthelabmanual:
PreLabReport
SubmittingReportsandLabPaper
Safety
Attendance

S16


12

Policies&Procedures
http://policy.uconn.edu/
PolicyagainstDiscrimination,HarassmentandInappropriateRomanticRelationships
http://policy.uconn.edu/?p=2884
Studentswithdisabilities
Please contact me during office hours to discuss academic accommodations that maybeneeded
during the semester due to a documented disability. The Center for Students with Disabilities
(CSD) engages in an interactive process with each student and reviews requests for
accommodations on an individualized, casebycase basis. Depending on the nature and
functional limitations of a students documented disability, he/she may be eligible for academic
accommodations. CSD collaborates with students and their faculty to coordinate approved
accommodations and services for qualified students with disabilities. If you have a documented
disability for which you wish to request academic accommodations and have not contacted the
CSD, please do so as soon as possible. TheCSDislocatedinWilburCross,Room204andcan
be reached at (860) 4862020 or at csd@uconn.edu. Detailedinformationregardingtheprocess
torequestaccommodationsisavailableontheCSDwebsiteatwww.csd.uconn.edu.
http://www.csd.uconn.edu/

S16


13

ExperimentSchedule
Date

Section

T1

T2

T3

T4

T5

T6

T7

T8

19Jan

Tu/Th

Meeting

20Jan

M/W

Meeting

25Jan

M/W

GA1

GA1

GA1

GA1

GA1

GA1

GA1

GA1

26Jan

Tu/Th

GA1

GA1

GA1

GA1

GA1

GA1

GA1

GA1

27Jan

M/W

GA1

GA1

GA1

GA1

GA1

GA1

GA1

GA1

28Jan

Tu/Th

GA1

GA1

GA1

GA1

GA1

GA1

GA1

GA1

1Feb

M/W

GA2

GA2

GA2

GA2

GA2

GA2

GA2

GA2

2Feb

Tu/Th

GA2

GA2

GA2

GA2

GA2

GA2

GA2

GA2

3Feb

M/W

GA2

GA2

GA2

GA2

GA2

GA2

GA2

GA2

4Feb

Tu/Th

GA2

GA2

GA2

GA2

GA2

GA2

GA2

GA2

RelatedChaptersCh.2,3&Ch.4

Date

Section

T1

T2

T3

T4

T5

T6

T7

T8

8Feb

M/W

GA20

9Feb

Tu/Th

GA20

10Feb

M/W

GA20

11Feb

Tu/Th

GA20

15Feb

M?W

GA20

16Feb

Tu/Th

GA20

17Feb

M/W

GA20

18Feb

Tu/Th

GA20

RelatedChaptersCh.22&Ch.24
LaboratoryPaperisdueonThursday,April282016,at10.00am.

Date

Section

T1

T2

T3

T4

T5

T6

T7

T8

22Feb

M/W

GA7

GA7

GA7

GA7

GA7

GA7

GA7

GA7

23Feb

Tu/Th

GA7

GA7

GA7

GA7

GA7

GA7

GA7

GA7

24Feb

M/W

GA7

GA7

GA7

GA7

GA7

GA7

GA7

GA7

25Feb

Tu/Th

GA7

GA7

GA7

GA7

GA7

GA7

GA7

GA7

29Feb

M/W

GA4

GA4

GA4

GA4

1Mar

Tu/Th

GA4

GA4

GA4

GA4

2Mar

M/W

GA4

GA4

GA4

GA4

3Mart

Tu/Th

GA4

GA4

GA4

GA4

RelatedChaptersCh.811

Continuedonthenextpage

S16


14

Date

Section

T1

T2

T3

T4

T5

T6

T7

T8

7Mar

M/W

GA4

GA4

GA20

8Mart

Tu/Th

GA4

GA4

GA20

9Mar

M/W

GA4

GA4

GA20

10Mar

Tu/Th

GA4

GA4

GA20

21Mar

M/W

GA4

GA4

GA20

22Mar

Tu/Th

GA4

GA4

GA20

23Mar

M/W

GA4

GA4

GA20

24Mar

Tu/Th

GA4

GA4

GA20

RelatedChaptersCh.11Ch.22&Ch.24
LaboratoryPaperisdueonThursday,April282016,at10.00am.

Date

Section

T1

T2

T3

T4

T5

T6

T7

T8

28Mar

M/W

GA5

GA5

GA8

GA8

29Mar

Tu/Th

GA5

GA5

GA8

GA8

30Mar

M/W

GA5

GA5

GA8

GA8

31Mar

Tu/Th

GA5

GA5

GA8

GA8

4Apr

M/W

GA8

GA8

GA5

GA5

5Apr

Tu/Th

GA8

GA8

GA5

GA5

6Apr

M/W

GA8

GA8

GA5

GA5

7Apr

Tu/Th

GA8

GA8

GA5

GA5

RelatedChaptersCh.1315

Date

Section

T1

T2

T3

T4

T5

T6

T7

T8

11Apr

M/W

GA16

GA16

GA21

GA21

12Apr

Tu/Th

GA16

GA16

GA21

GA21

13Apr

M/W

GA16

GA16

GA21

GA21

14Apr

Tu/Th

GA16

GA16

GA21

GA21

18Apr

M/W

GA21

GA21

GA16

GA16

19Apr

Tu/Th

GA21

GA21

GA16

GA16

20Apr

M/W

GA21

GA21

GA16

GA16

21Apr

Tu/Th

GA21

GA21

GA16

GA16

25Apr

M/W

26Apr

Tu/Th

27Apr

M/W

Checkout

28Apr

Tu/Th

Checkout

RelatedChaptersCh.17,19,20
LaboratoryPaperisdueonThursday,April282016,at10.00am.

S16


15

AcademicHonesty
Statement on Academic Honesty(PartscopiedfromPage1516oftheUniversityofConnecticut
pamphlet entitled: Student Conduct Code revised May 10, 1996 & available from the Vice
PresidentforStudentAffairs,352MansfieldRoad,U121)

All students have the right to pursue their academic careers inanatmospherebasedonhonesty
and trust. Acts of academic misconduct destroy that atmosphere, violate that trust, and are
therefore subject to penalty... A fundamental concept of all educational institutions is academic
honesty... Misrepresentation of someone elses work as ones own is a most serious offense in
any academicsetting...Noacademicmisconduct,includinganyformsofcheatingandplagiarism,
can be condoned. Academic misconduct includes but is not limited to providing or receiving
assistance in any manner not authorized by the instructor in the creation of work to be
submitted for academic evaluation including papers,projects,examinations,etc.,presentingas
ones own the ideas or words of another for academic evaluation without proper
acknowledgment, doing unauthorized academic work for which another person will receive
credit or be evaluated and presenting the same or substantially the same papers or projects in
two or more courses without the explicit permission of the instructors involved.... Also, one is
not allowed to cooperate or be an accessory to anothers academic misconduct. Thus a student
who writes a paper or does an assignment foranotherstudentisanaccompliceandmustbeheld
accountable justasseverelyastheother.Itisperhapslessobvious,butitisequallylogical,thata
student who knowingly permits another to copy from his or her own paper, examination, or
projectshouldbeheldasaccountableasthestudentwhosubmitsthecopiedmaterial.

Part of the Facultys responsibility and usual penalty (refer to the full text found in the
StudentConductCode)
...When an instructor believes there is sufficient evidence to demonstrate a clear case of
academic misconduct, the instructor shall notify the student in writing, and also orally if
possible, that unlessthestudentrequestsahearingtocontesttheinstructorsbelief,theinstructor

S16


16

shall impose the appropriate academic consequences warranted by the circumstances..... In


reference to the imposed consequences, theinstructorsofthiscoursemayconsiderthefollowing
opinions of (1) awarding a Failure on the actual academic work in which the academic
misconduct was found (2) awarding an immediate Failure for the entire course, which might
include the recommendation to the Dean of Students for the immediate dismissal of the student
fromtheUniversity,ifthecircumstancesdictate.

S16


17

TheLaboratoryNotebook
The purpose of the laboratory notebook is to keep a permanent record of your laboratory work,
which can be read and understood at a future time by both you and other scientists.
Experimental work should be able to be reproduced by someone else directly from the
descriptionthatyouprovideinyourlaboratorynotebook.

1.UsingLaboratoryNotebook
There are three major concerns about the notebook: (a) the safekeeping of the labnotebook,(b)
theorganizationandreadabilityofthelabnotebook,and(c)thequalityoftherecordkeeping.
a.Safekeeping
In laboratory jobs, the laboratory notebook belongs to the organization for whom the work is
being done. There will be very specific guidelines about exactly how and where the notebook
must be kept, (in any case itstaysattheworksite!) Generally,ifyouaredoingresearchallday,
you will not havetimeortheinclinationtoworkonyourlabnotebookoutsideofthelabtomake
it look nice. You need to get the important observations into the lab notebook as you are doing
the work. To help you get into this habit, we require you to leave your lab notebook in the
laboratory. You will be making a carbon copy of everything that goes into the book, and that
copymaybetakenwithyouasyouleavethelab.
b.OrganizationandReadabilityoftheLabNotebook
The main concern is the permanent nature of the record of your laboratory activity. A bound
notebook (as opposed to spiral) minimizes the chance of losing pages. Good quality, black ink
minimizes the chance for erasures, obscuring of written material and provides for a clearer
photocopyintheeventthatitisnecessary.
c.QualityoftheRecordKeeping
The importance of being able to find your past experimental work in one place cannot be
overemphasized. No one likes to redo, or rethink past work that had been successfully

S16


18

completed! Without complete information, work cannot be reproduced and verified. Without
wellorganizedinformation,thesignificanceofexperimentalresultsmightnotberealized.

d.GuidelinesforKeepingtheNotebook
With the above points in mind, the following guidelines for keeping laboratory notebook have
beendeveloped.
1. The notebook must contain bound, numbered pages. A carboncopyoftheprimarypage
shouldbemadeontotheduplicatelynumberedpage.
2. Entries must be made in the notebook with good quality ballpoint pen, preferably black.
Avoid writingoverdata.Ifanentryismadeinerror,crossoutthaterrorwithasingleline
andaddasimplelegibleexplanation(forexample,scalemisread,etc.)
3. Allentriesmustbemadeinduplicateusingthecarbonpaper.
4. TheLabNotebookmustbekeptinthelabatalltimes.
5. All data taken in the laboratory must be directly written into the laboratory
notebook.
6. Noscrapsofpaperwillbetolerated!
7. At the end of each lab period or any time thereafter, the carboncopy pages of
materialmaybetakenhome.
8. After submitting experiment report (or lab paper) electronically, please stop by the
laboratoryduringlaboratoryhourstosubmitstapledcarboncopypages.
9. The Lab Notebook shouldbeleftonthedesignatedshelfasyouleavetheLaboratory. Its
presence will be checked after each laboratory period. Notebook is evaluated at least
twice a semester (see the part on grading of the laboratory notebook at the end of this
section).

S16


19

2.ContentsoftheNotebook
a.TableofContents
This is the only part of the notebook, which does not have to be duplicated. Leave two ormore
pages at the beginning of the book and be sure to keep this table uptodate. Youmayuseshort
title(e.g.,GA1)intheTOC,alsoincludedateandpage#.
b.Dates
Always date the work in a prominent position where it can easily be seen so it is clear where a
newday'sworkbegins.

c.TitleofExperiment
The title identifies the experiment in a few words. It should precede anything related to that
experiment that goes into the book. One should never open thebookandfindabunchofdataor
experimentaldescriptionwithoutatitle.
d.Experimental
Any changes to the procedure given in the manual, raw data such as amounts weighed out,
solutionvolumesmeasured,unknown#,Purityfactorsofprimarystandards,etc.
e.Data
The data should be entered into a table directly as it is taken! Nothing should go onto scrapsof
paper!
f.DataTables
Tables should be constructed with a straight edge, should have a title, and should have clearly
labeled columns and rows. Note that data is read directly from the measuring device. Numbers
calculated from a measurement is a result, but as noted below may be subsequently added to a
table.

S16


20

Referring to the following table as an example, a table may contain both raw data (weights
obtained from a balance) as well as calculated results (calculated weightsofsampleobtainedby
differencesandby%sulfatecalculatedusinganequation).
ExampleofaTable

g(Vial+Sample)
g(VialSample)
g(Sample)
Avg.g(Crucible+Precipitate)
Avg.g(EmptyCrucible)
g(Precipitate)
%Sulfateinsample

Sample1

Sample2

Sample3

g.Notes
Any notes necessary to clarify, emphasize or interpret the data should be entered in the
Laboratory Notebook as the experiment progresses. Be especially careful to document any
changefromthedescribedprocedure.
h.Calculations
Ifyoudocalculationsduringthelab,dothemintheLabNotebook.
i.Results
IfyouhavethecalculationsinyourLabNotebook,theresultsshouldfollowthecalculations.

3.GradingoftheLaboratoryNotebook
The Lab Notebook grade is separate from the lab report and lab project grades and is an
evaluation of your record keeping. Each time a lab report is graded, an evaluation will also be
made of the material on the carbon copies of the lab book. In addition, the lab notebooks
themselves will be collected and evaluated as a whole at least twice a semester this collection
willnotnecessarilybeannounced!Thisgradeisbasedonthefollowingcriteria:

S16


21

a.PhysicalAspects
Is the notebook bound with numbered duplicate pages? Is the writing recorded in permanent
ink?Isthereanyunexplainedcrossingoutofdata?
b.Format
Are all the required sections included such as table of contents, dates, title, any changes to
experimentalprocedurefromthelabmanual,etc?
c.Neatness
Does the book have a neat appearance? Is there a lot of writing or crossing out of data,
scribbling, etc.? This is a working notebook so you are writing as you are doing therefore, we
donotexpectittobeflawless,butneithershoulditbeatotalmess!
d.Organization
Isthematerialwellorganized?Canwefindwhatwearelookingforeasily?
e.Completeness
Isalltheinformationthere,whichisnecessaryforonetoevaluatethequalityofyourwork?
f.AppropriateRecordofData
Arethetablesclearandwelldesigned?Dothenumbersincludeunits?

4.HowtheLabNotebookwillbegraded?
Grade
10
9
8
7
6
5
4

Quality
Justaboutperfecthardtofindmuchwrong
Verygoodbookwithafewminorproblems
Goodbookbuthasdefiniteareasthatneedimprovement
OKbutmanythingsneedtobestraightenedout
Mediocrecanfindmostinformationbutwithalotofwork
Poorjustaboutacceptable
Unacceptable

S16


22

WarningaboutAcademicMisconduct!
Pleasepayparticularattentiontothefactthatalllaboratoryreportsandlaboratorypaperthatyou
submittobegradedfallunderthegeneralcategoryofstudent'soriginalwork.Anyacase
thatitisobservedthattherehasbeendeliberatecopyingfromanotherstudentsreportandboth
reportsarehanded,bothwillbeconsideredasactofplagiarismwhichwouldbedealtunderthe
ruleofAcademicMisconductasspecifiedintheStudentHandbook.

S16


23

PreLabReport
Read the experiment carefully before coming to the lab. The experimental procedure is in
reasonably explicit detail. When you read this part beforehand,trytoimaginewhatitisyouwill
bedoing.Also,attempttofigureoutthewhysoftheexperimentalsetupandprocedure.

1. Theprelabreportreferstofollowingaspects(expectedlength11.5pages).
Objective(s)(fewsentences)
Indicatekeyequations.
Experimental requirements: Indicate the instrumentation and/or special apparatus
neededfortheexperiment.
Include a general outline of the experimental procedure, by indicatingthemainsteps
tofollow.Noneedtogivedetailsastheyarealreadyinthelabmanual.
Precalculations: If solution preparations are needed, make sure you have all the
calculationsdone(orfiguredout)toinsureanefficientprocedure.
Identify critical steps in the experiment procedure that can be determinant in its
successfulresults.
Safety:Listthehazardsandthepersonalprotectiveequipmentneededtoavoidthem.

There should be one, single document for the prelab report. This means that equations and
precalculations need to be typed in using equation editor. Document Margins (Left 1, Right
2.5,TopandBottom1).LineSpacing1.5.

Prelab report needs to be submitted electronically (PDF document) to thelabinstructoratleast


TWOdayspriortothescheduledexperiment.

PleaseNote
PrelabReportwillnotbegradedifwritteninessayformat.

S16


24

PostLabReport
Post lab Report MUST be submitted electronically via email as a PDF. Thereshouldbeone,
single document containing answers, tabulated data, plots, sample calculations, etc. No
handwrittenassignmentswillbeaccepted. Samplecalculationsaremustandneedtobetypedin
Microsoft Word equation editor (or formula editor in Openoffice word processor or
equation editor in Google Docs). Plots from spreadsheet software (Excel or Google
Sheets) should be pasted into the word processor document (Word or Google Docs, etc)
before creating a single PDF document for submission. Document Margins (Left 1, Right
2.5,TopandBottom1).LineSpacing1.5.

No formal lab report: Just answer questions from the experiment handout and show
calculations.

PleaseNote
PostLabReportwillnotbegradedifwritteninessayformat.

S16


25

LaboratoryPaper
WarningaboutAcademicMisconduct
Please pay particular attention to the fact that all laboratory reports and laboratory papers that
you submit to be graded fall under the general category of student's original work. Any a
case that it is observed that there has been deliberate copying from another students reportand
both reports are handed, both willbeconsideredasactofplagiarismwhichwouldbedealtunder
theruleofAcademicMisconductasspecifiedintheStudentHandbook.

Assignments for this course (Prelab Report, Post lab Report and Lab Paper) MUST be
submitted electronically via email as a PDF. There should be one, single document
containing answers, tabulated data, plots,samplecalculations,etc. Nohandwrittenassignments
will be accepted. Sample calculations are must and need to be typed in Microsoft Word
equation editor (or formula editor in Openoffice word processor or equation editor in Google
Docs). Plots from spreadsheet software (Excel or Google Sheets) should be pasted into
the word processor document (Word or Google Docs, etc) before creating a single PDF
document for submission. Document Margins (Left 1, Right2.5,TopandBottom1). Line
Spacing1.5.

WritingthePaper
General guidelines are given below. See experiment assigned for the laboratory paper for
specificsoncontentofsectionsTheory,Experimental,Results,andDiscussion.
Summary(aparagraphorso)
(The summary of a paper serves several purposes. One is to give the reader a briefintroduction
of why the workwasdone.Alsoitshouldgiveaverybriefoverview(executivesummary)ofthe
major findings of the work. In the case of the teaching lab, the summary should also help the
marker to determine the writer's understanding of the main thrust of the lab experiment while
againsummarizingtheresultsthatwereobtained.)

S16


26

Theory
Experimental
Results
Discussion
Conclusions
A conclusion should briefly summarize the key findings reported in the Discussion section and
suggestions for the future work. Basically the Conclusion is a statement of the information
obtained what was foundout.Acommonerroristoputtoomuchinformationintheconclusion,
which really belongs in the discussion. If your conclusion is more than a paragraph, it should
reallybeinthediscussion.
References
Appendices
Appendix1shouldcontainallcarboncopiesofthedatapagesfromtheLabNotebook.
Appendix 2 should contain the original or goodquality, photocopies of the recorder or
computer outputs. These should be taped or pasted onto 8 x 11.5 inch paper. Never include a
longpartialrollofrecorderorintegratoroutputwiththereport!

Appendices 1 & 2 (carbon copies, recorder outputs, etc.) can be scanned and included in the
word document as images.

Use appropriate options on the desktop scanner or (on the

smartphone app) to limit the image size (& so PDF document file size). Keep PDF document
filesize2Mb.

S16


27

SubmittingReportsandLabPaper
Assignments for this course (Prelab Report, Post lab Report and Lab Paper) MUST be
submitted electronically via email as a PDF. There should be one, single document
containing answers, tabulated data, plots,samplecalculations,etc. Nohandwrittenassignments
will be accepted. Sample calculations are must and need to be typed in Microsoft Word
equation editor (or formula editor in Openoffice word processor or equation editor in Google
Docs). Plots from spreadsheet software (Excel or Google Sheets) should be pasted into
the word processor document (Word or Google Docs, etc) before creating a single PDF
document for submission. Document Margins (Left 1, Right2.5,TopandBottom1). Line
Spacing1.5.
YouneedtosubmitasinglePDFdocument.
Filename syntax should include experiment name, section#, team# andyourlastnamein
the filename. Please see example shown below, note a space intentionally included
inbetweensyntaxterms.
Filenamesyntaxisveryimportantforsuccessfulsubmissionofprelabreport,postlab
reportandoflabpaper.
PleaseNote
You will not be notified about incorrect syntax, and submitted report/paper PDF file with
incorrectsyntaxwillnotbesavedinthesystemandsoitwillnotbegraded.

PleaseemailPrelabReporttototheLabInstructor(abhay.vaze@uconn.edu)
PleasenamePDFdocumentas:
Forexample:GA1PreS1T1Sandberg
PleaseemailPostLabReporttotheLabInstructor(abhay.vaze@uconn.edu)
Forexample:GA1PostS1T1Sandberg
PleaseemailLabPapertotheLabInstructor(abhay.vaze@uconn.edu)
PleasenamePDFdocumentas:
Forexample:GA20PaperS1T1Sandberg
Pleasedontaddanyotherword,suchasFinalorLabReporttothefilenamesyntax.

S16


28

LatenessPolicyandLabPreparedness
a.PrelabReport
Prelab Report is mandatory and needs to besubmittedelectronically(PDFdocument)tothelab
instructoratleastTWOdayspriortothescheduledexperiment.
b.PostlabReport
After the experiment is completed, report is due in the following week. Electronic copy (PDF
document)oftheLabReportisdueon:
10. ForMonday/WednesdaySection,reportisdueonWednesdayat10am.
11. ForTuesday/ThursdaySection,reportisdueonThursdayat10am.
If the lab report is late, there will be a cumulative 10 points deduction per day the lab reportis
overdue.
Along with the lab report you need to submit carbon copies of your notebook pages. Those
copiesneedtobeturnedinonthedaythelabreportisdue.
c.LabPaper
Electronic copy (PDF document) of the Lab Paper is due on the day and time specified in the
syllabus.LabPaperwillnotbeacceptedifnotreceivedbythetimeitwasdue.

S16


29

UsingAnalyticalBalances
Analytical balances, especially thefourorfiveplace,digitalorsubstitutionbalances,arecapable
of measuring the weight of an object to 0.0001 or even 0.00001 g, i.e. (0.1 mg or 0.01 mg).
These balances if properly calibrated and maintained are possibly the most accurate and
precise measuring instrument found in any laboratory. Whether those balances might bein
analytical, physical, advanced inorganic and organic and whether they might be in academic or
insuchindustriallaboratoriesas:biotechnology,forensic,pharmaceuticalorpolymerlaboratory.
Hence it is imperative that our shared analytical balances be kept in excellent operating
condition. I should be noted that each balanceintheAdvancedChemistryCoursesoftheDept.
of Chemistry of UConn is checked daily by a member of the teaching staff for its proper
operation andgeneralcleanliness. Alsoeachyear,anoutsidebalanceservicingcompanyishired
tocomeintoproperlyserviceandifnecessarilyperformmajorrepaironeachbalance
It is absolutely imperative that each student needs to first hear Introductory Balance
Talk and do the introductory about five minute weighing experiment conducted by a
qualified laboratory instructor or teaching assistant. Theneachstudentneedstoadhereto
the rules that are given below. Repeatedfailuretodoso,afterhavingreceivedaclearwarning
from their proper teaching staff member, will cause that student tobeassignedtodoweightings
on the older substitution balance, (which takes about2.0min.)toaccomplishthesameweighing
measurement that otherwise could be done on the newer digital balances (within about 30
seconds).
(Note these substitution balances are just as accurate but perhaps not as reproducible as the
newer digital balances). And as is evident take at least four time longer to achieve the same
weighing.

GeneralRules
1. No one may use a balance withoutfirsthearinganintroductorybalancetalkgivenbya
laboratoryinstructororteachingassistant.

S16


30

2. Use only the balance to which you are assigned. This mustbedoneeveniftheassigned
balance is in use by another assignee at this particular time. (Rememberitusuallytakes
within 30 seconds. to accomplish one weighing or only a matter of a few minutes to
complete a series of weighings). Posted near the balance will bealistofthosestudents
thathavebeenassignedtousethatparticularbalanceforthecurrentsemester.
3. If there seems tobeaproblemwiththebalance,reportitimmediately,totheinstructoror
teachingassistant.Donotuseanotherbalance,unlessinstructedtodoso.
4. It is imperative that you keep the balance and the immediate area around the
balance clean! Clean up all chemical spills and excess chemical after each use of the
balance.DonotleaveoldKimwipesorcrumpledpapersonthebalancetable.
When the balances are not in use, the balance should bealwaysturnedoffandinthecaseofthe
substitutionbalancesthebeamarrested,allweightsonzeroandthedoorsclosed.

Please
Use electronic balance carefully as it is a calibrated & expensive instrument. To prevent
corrosion, be careful nottospillChemicalsorWater.Formoreinformation,pleasesee:Page
31, Quantitative Chemical Analysis, edition 8th by Daniel Harris, W. H. Freeman and
CompanyNY.

S16


31

GA1AnalyticalTechniquesandMeasurements
This first laboratory exercise has two primary goals. The first goal is to give you a chance to
practice the classical techniques of weighing andofusingvolumetricglassware. Nomatterhow
sophisticated an analytical method is, one must still begin by weighing a standard material and
usually dissolving it in an appropriate solvent in volumetric glassware. For much of analytical
chemistry, the accuracy of the entire analysis depends on the accuracytowhichthestandard(or
standards) are compared. The second goal is to introduce and utilize certain concepts of
statistical treatment of data. Carefully obtained results are useless unless their values and their
limitations are known. The following statistical concepts are introduced in this laboratory
exercise: accuracy and precision (absolute and relative) estimation of error (deviation)
propagation of error significant figures Gaussian (normal) distribution average (mean)
medianstandarddeviationandconfidenceinterval.
Therearethreepartstothisexperiment
1.

UseoftheAnalyticalBalance

2.

CalibrationofVolumetricGlassware

3.

DeterminationoftheDensityofAntifreeze

Fillin sheets, labeled F1 to F4 will be provided in the lab (also included at the end of this
experimental writeup). When you have completed the data collection, these sheets and any
supplementary calculation sheets may be taken home, attached together and handed in for
marking when you have completed each section. Starting with the second experiment, all
laboratory data is to be recorded in the laboratory notebook at the time that the data is
collected. Losesheetsofnumberswillnotbeallowed!Thelaboratorynotebookstaysinthe
laboratory at all times (this will be checked), but the carbon copies of the data from the
notebookmaybetakenfromthelaboratoryandusedinpreparingthelaboratoryreport.

S16


32

UseoftheAnalyticalBalances
There are three types of analytical balances available for use in this laboratory: the digital
(electronic) balance, a top loading balance and a simple triple beam balance. In certain
instances, the less precise (and quicker) top loading or triple beam balances should be used.
However, since every analysis involves at least one, accurate weighing step, it is essential that
youareabletousetheanalyticalbalancesaccuratelyandreproducibly

This experiment will give you practice in using all three of the balances. Each balance has an
inherent accuracy (the number of significant figures to which youfeelconfidentinreportingthe
weight measured by each balance) and a precision (or replicate value that you obtain if you
continued to weigh the same object several times). In mostcases,itisassumedthatyourezero
the balance between each subsequent weighing. Evenwiththemostexpertuse,thesameobject
will not necessarily weigh the same amount each time that it is weighed. Thebestvalueforthe
weight of the object is generally consideredtobetheaveragevalue. Precision(deviation)isthe
measure of the variation among replicate measurements.

The inherent precision (or

reproducibility) of a balance can be expressed by either an absolute deviation or a standard


deviation.

As more andmoretrialsaremade,theprecisionimprovesandthemeanvaluegenerallybecomes
a better and better estimate of the true value. However, good precision alone does not insure
good accuracy! Systematicerrorscanoccurwhichcausetheexperimentalvaluetodeviatefrom
the true value. There are many kinds of systematic or determinate errors, but they all can, in
principle, be identified and then be either eliminated or corrected for. For example, one source
of a determinate error is inaccurate weights in the balance itself. In principle, this could be
eliminatedbymakingsurethattheabsoluteweightsandvolumesofthesolutionmeasuredareall
accurate or traceable to acceptable standards. In this experiment, experimental causes, such as
touching the object to be weighed with ones hands, which may be detrimental when using the
mostpreciseanalyticalbalances,willbeinvestigated.

S16


33

a.PrecisionofVariousBalances
There are three types of balances in the analytical lab: the digital balance (DB), the top loading
(TLB), and the triple beam balance (TBB). A weighing bottle and its cap, both are in your
locker top shelf are needed forthispartofexperiment. Makesurethatyoupickuptheseobjects
only with crucible tongs or tweezers so none of yourfingersgreaseormoistureispickedupon
the object between weighings. Since, you will be doing experiment with your lab partner, just
one weighing bottle & a cap between two of you is required, so each one will have exact same
data.
Weigh your object oneachofthethreedifferenttypesofbalance,consecutively. Recordexactly
the weight (to the proper number of significant figures) you obtained by reading the balances
graduated scale. Make sure you remove the object and rezero the balance between each
successiveweighing.
Reweightheobjectatotalofthreetimesoneachbalance. Makesureyouremovetheobjectand
rezerothebalancebetweeneachsuccessiveweighing.
b.UseoftheDigitalBalance
In this section, you will first weigh a weighing bottle and then its lid using only the digital
balance. Then you will weigh both together on the pan at the same time. The sum of the first
two weights should be the same within the precision of the balance. If you are still not able to
reduce the experimental differences between the two weighings, after a secondchecking,please
consultthelaboratoryinstructorand/orteachingassistant.
1. Handling it with crucible tongs, determine the weight of a clean, dry weighing bottle to
thenearest0.1mg(0.0001g)
2. Determinetheweightofthelidtothenearest0.1mg
3. Determinethetotalweightofthebottleplusthelidtothenearest0.1mg.
4. Find the sum of the results of #1 and #2 and compare with #3. The two should agree to
within0.5mg.

S16


34

c.SourcesofDeterminateWeighingErrors
1. Roll the weighing bottle in your hand, handle it, and finger it. Then reweigh it and
compareitwiththepreviousresult.
2. Wipe the weighing bottle with a Kimwipe. Weigh it and compare with the previous
results.
3. Breathe on the weighing bottle weigh it again and compare this resultwiththeprevious
results.
4. Initial the weighing bottle with a #2 pencil on the ground glass patch (do not use ink it
cannotberemoved).Reweighandcomparewithpreviousresults.
Place the weighing bottle in a drying oven for about three minutes, just enough to warm it.
Remove the bottle from the oven with tongs and reweigh it immediately while it is still warm.
Follow the change in the apparent weight for several minutes. Record the weight every 30
seconds.

Please

Use electronic balance carefully as it is a calibrated & expensive instrument. To prevent


corrosion, be careful nottospillChemicalsorWater.Formoreinformation,pleasesee:Page
31, Quantitative Chemical Analysis, edition 8th by Daniel Harris, W. H. Freeman and
CompanyNY.

CalibrationofVolumetricGlassware
Since the exact volume of a piece of volumetric glassware is not necessarily identical with that
printed on it, glassware must be calibrated when it is to be used in extremely accurate
quantitative work. Glassware calibration is most often performed by weighing the amount of
water contained by or delivered from the glassware. Knowing the density of water at the
appropriatetemperature,onemaycalculateitsactualvolume.
1.

Calibrationofa25mLPipet

2.

CalibrationofadigitalPipet

Inthisexperiment,apipet(nominalvolume25ml)andadigitalpipetwillbecalibrated.

S16


35

a.BuoyancyCorrections
Obtaining the true weight of water delivered from volumetric glassware requires weighing the
sample, and then mathematically correcting for the fact that the balance can give a true weight
only for materials having the same density as the weights. This differenceindensitycausesthe
apparent weight of the water to be less than the actual weight since a certain amount of air has
been displacedbytheitembeingweighed. Formostsolids,nocorrectionisrequiredbecausethe
density of the solid is sufficiently similar to the density of the weights. But, for liquids and
gases,howeverabuoyancycorrectionmustbemade,whenhighaccuracyisdesired.
The equation belowisanexpressionforthecorrectedweightoftheobjectinavacuum,basedon
its measured, apparent, weight in air. Derivation of this equation may be found in various
analytical texts. The weight of water in vacuum, or thecorrectedweight,shouldbeusedforthe
calculationsofglasswarevolume.

WV=WeightofWaterinVacuum

Wv =

W o {(Dw Do )Da }
Dw Do

WO=WeightofObject(Water)inAir

+ W o

D0=DensityofObject(Water)
Da=DensityofAir
DW=DensityofWeight(8.00g/mL)

DO, the density of water, is temperature dependent. Values at various temperatures may be
found in Table 27 for more details refer to the text, D. C. Harris, Quantitative Chemical
Analysis,7thedition,Chapter2,page327thEdition,page42,page42.
Temperature(0C)

DensityofAir(g/mL)

20

0.001184

21

0.001180

22

0.001176

23

0.001172

24

0.001168

25

0.001164

Da,thedensityofairatroom
temperaturemaybefoundintheadjacent
tabletakenfromtheHandbookofChemistry
andPhysics,52ndedition,197172,The
ChemicalRubberCo.,Cleveland,OH.

Toobtainthedensityofairatother
temperatures,calculatethatvalueassuminga
simplelinearprogressionofthedatafoundin
theabovetable.

S16


36

Precaution
Before beginning the calibration of glassware, be sure that the glassware is scrupulously
clean. Clean glassware shows nowaterdropletswhenfilledwithwaterandthenemptied.To
clean, first try washing the glassware with a very dilute solution 13 per cent of detergent in
water. Follow by rinsing each piece of glassware with about five successive rinses of tap
water then with three rinses ofthelaboratorydeionizedwater.Ifthisdoesnotworkandthere
still remains droplets (called hang) on the inside of the glassware,askthe instructor and/or
teaching assistant to cleanthe glassware with the available, lowenvironmental impacting
detergent.

b.DirectCalibrationof25mLPipet
Proper pipetting will be demonstrated in one of the laboratory briefings. It is highly
recommended that you perform several practice runs including checking the weight delivered
fromsubsequent25mlpipetdeliveries.Practicepipettingwiththe25mlpipetasfollows:

Precaution
Do not take deionized water from the tap, since it will change temperature astheexperiment
proceeds. From one of the largejugssittingattheendofseveralofthebenches,filla400mL
beaker with about 300 mL of deionized water that has already been stored at room
temperature.Measureandrecordthetemperatureofthewaterthroughouttheexperiment.

Please
At other times do not use this temperatureequilibrated water as it is used as a backup
deionized water supply when the house water supply runs dry. These large jugs that contain
thetemperatureequilibratedwaterareverydifficulttorefill.

1. Fill the pipet past the calibration mark with deionized waterusingapipetbulb. Remove
the bulb, and by manipulating the index finger, not the thumb,adjustthebottomlevelof
themeniscusuntilitcoincidesexactlywiththecalibrationmark.
2. While carefully maintaining the pipet in a vertical position,touchthetipofthepipettoa
glasssurfacetoremoveanyextradropletofwateradheringtotheoutsideofthetip.
To emptythepipet,removetheindexfingerandletthewaterflowintoaglasscontainer. Letthe
pipet drain for a minimum of 10 seconds after thefreeflowofwaterfromthepipethasstopped.
Infactitisusuallysuggestedtoallowfora1secondwaitforeachmLofvolumedelivered.Then

S16


37

touch the tipofthepipettothewallofthecontainertoremoveanyhangingwaterdropsfromthe


tip. However, do not force out by either blowing or shaking any water thatremainsinthe
tip. The remaining water in the tip has been taken into accountinthedesignofthepipetbythe
manufacturer. The total volume delivered by these TD (to deliver) pipets then becomes
reproducible. But of course, it may not be completely accurate! When pipetting technique has
beensatisfactorilymastered,thecalibrationcanbestarted.

Weigh (using the digital analytical balance) a clean 125 mL Erlenmeyer flask equipped with
the proper rubber stopper to the nearest milligram (0.001 g). Note:accuraciesof0.1mgarenot
needed for the glassware calibrations. Always handle the flask with tongs or a strip of paper.
The flask need not be dry on the inside, only on the outside, but the inside neck in which the
rubberstopperininsertedmustbekeptdry.(Why?)

Carefully, transfer an aliquot (the contents) of the 25 mL pipet into the flask holding the pipet
vertically with its tip well inside the flask to avoid water splashing onto the neck, or out. It is
important that the neck of the flask remain dry. As in the practice runs,allowthepipettodrain
thoroughly. Then touch the tip of the pipet to the side of the flask. Withdraw the pipette
(pipettingshouldbeperformedattheworktableandnotintheBalanceRoom).Weightheflask.
Repeat by weighing additional aliquots of water deliveredintotheErlenmeyerflasksubsequent
aliquots may be added to the volume of water already in the flask until the capacity of the
balanceisexceeded.
Perform at least four trials or enough to obtain a precision in terms of relative average
deviationof1ppt(partperthousand)or0.1%.
c.DirectCalibrationofaDigitalPipet
This section is designed to give you practice on using a digital pipet. Please see the Instructor
about which pipet to use. Obtain the proper tip and practice the volume delivery, making sure
that the liquid drops do not leak from the tip during transfer. If this occurs it usually indicates
that the tip is not tightly secured or that the inner seal of the pipet is not lubricated or needs
replacing.

S16


38

Into the weighed Erlenmeyer flask, carefully deliver the full contents of the digital pipet. On
FillIn sheet, make sure that you record the name of the manufacturer of the digital pipet,
whether it has fixed or variable volume delivery, and the full volume delivery that you used.
Obtaintheweightofthewaterdelivered.
Repeat this procedure at least four times in order to obtain a meaningful mean value and
appropriateprecision.

DeterminationoftheDensityofAntifreeze
This part ofthelaboratoryexerciseutilizestheanalyticalbalanceandthecalibrated,25mLpipet
todetermineaccuratelythedensityofcommercialantifreeze.
Apart from calibrated 25 mL pipet, you alsoneeda250mLbeakeranda125mLflaskallthree
needtobedry,freeofanytracesofwater.
Measureoutintoa250mlbeakerabout100mlofantifreeze.
First, carefully rinse your 25 ml pipet at least three times with small volumes of the
antifreeze sample. Please collectallrinsingandexcessantifreezeinthesamebeakerthat
contains antifreeze sample. Carefully deliver a representative sample of this antifreeze
usingyourcalibrated25mlpipetintoapreweighedErlenmeyerflask.
Repeat this procedure another three or more times in order to obtain a mean and a clear
measurement of the precision of your weight deliveries. Subsequent volumes of the
antifreeze may be added to the original volume found in the Erlenmeyer flask. The
precisionshouldbewithin1ppt(partperthousand)or0.1%.

Questions/TopicsfortheQuiz
1. Defineprecisionandaccuracy
2. Give formula for (a) Average Deviation (b) Relative Average Deviation (c) Standard
deviation(d)RelativeStandardDeviation.

Calculations&/orQuestionsforPostlabReport
a.UseoftheAnalyticalBalance

S16


39

1. Whichbalanceismostprecise?Whichbalanceismostaccurate?Justifyyouranswer!

b.WeighingErrors
1. What precautions should one take to insure the best precision and accuracy when
weighing an object? Whichoftheaboveproceduresdonotaffectprecisionandaccuracy
(ifany)?Whichhavethegreatesteffectonprecisionandaccuracy?
2. Compare the sum of the weights of a weighing bottle and lid with the experimental
weight of these two objects together. What individual errors involved in the weighing
process can you expect? Specifically, how is the weight affected? Clearly explainyour
answersinthereportthatyouhandinwiththisdatasheets.

c.DirectCalibrationof25mLPipette
1. For each weight of water, find theweightinvacuum(correctedweight)bycorrectingthe
weightinairforthebuoyancyofair.Showsamplecalculations.
2. Using the density of water at the experimental temperature and the corrected weight of
thewater,findthevolumeofthepipet.
3. Findthemeanandmedian(centermost)valuesforthepipetvolume.
4. Find the standard deviation of the pipet volume. To how many significant figures the
average volumedeliveredbyyour25mLpipetshouldbereported? (Hint:RealRulefor
Significant Figures Chapter Experimental Error in Quantitative Chemical Analysis
byDanielHarris).

Remember

Always use Real Rule of Significant of Figures when reporting calculated mean and
standarddeviation

d.DeterminationoftheDensityofAntifreeze
Calculate standard deviation of the density of antifreeze byusingtheconceptofpropagationof
errors.Showsamplecalculations.

S16


40

Datasheets
F1
Name
Section

Date

Part1UseoftheAnalyticalBalance

BalanceType

DigitalBalance

TopLoading
Balance

Manufacturer

Mettler

Denver/Mettler

OHaus

Weighing

Mass(g)

Mass(g)

Mass(g)

TripleBeamBalance

Trial1

Trial2

Trial3

MeanMass

AverageDeviation

StandardDeviation

RelativeAverage
Deviation(RAD)

RelativeStandard
Deviation(RSD)

AbsolutePrecision

RelativePrecision

S16


41

F2
MassofWeighingBottleobtainedusingDigitalBalance
Part1BWeighingErrors

Mass(g)

Massofweighingbottlealone

MassofLid

Sumof1&2(MassofWeighingBottlealone+MassofLid)

Massofbottleandlid

Differencebetween3&4(shouldagreetowithin0.5mg)

Part1CSourcesofWeighingErrors
1

Massofweighingbottleafterhandling

Massofweighingbottleafterwiping

Massofweighingbottleafterbreathingonit

Massofweighingbottleafterpencilmark

Massofhotweighingbottleeverythirtyseconds

S16


42

F3
Part2CalibrationofVolumetricGlassware
25mLPipet
TemperatureofWater=

DensityofWater=

Weightofwaterandcorrectedweightofwaterdeliveredfrom25mLpipet

InitialWeight(g)

FinalWeight(g)

ActualWeight(g)

CorrectedWeight(g)

Calculatedvolumeforthe25mLpipet

CorrectedVolume(mL)

Meanvalue=

MedianValue=

Standarddeviation=

RelativeStandardDeviation=

DigitalPipet
DigitalPipetManufacturer=

FixedorVariable=

DigitalPipetCapacity=

DensityofWater=

WeightofwaterandcorrectedweightofwaterdeliveredfromtheDigitalPipet

ActualWeight(g)

CorrectedWeight(g)

CalculatedvolumefortheDigitalPipet

CorrectedVolume(mL)

Meanvalue=

MedianValue=

Standarddeviation=

RelativeStandardDeviation=

S16


43

F4
Part3DensityofAntifreeze

WeightofAntifreezedeliveredfrom25mLpipet
InitialWeight(g)

FinalWeight(g)

ActualWeight(g)

Meanvalue=

MedianValue=

Standarddeviation=

RSD=

MeanVolume&StandardDeviationof25mLpipetfromprevioussection(F3)=

AntifreezeDensity=

StandardDeviation*=

*Usepropagationoferrorforcalculating
Standarddeviation

S16


44

NotesonCalibratingVolumetricFlaskandBurette(excludedfromGA1)
Your 250 volumetric flask should be washed, then rinsed twice with small amounts of acetone
and left upside down, hanging from a tripod, to dry for about 30 minutes, so it will be ready
whenneeded.
A nominal 250 ml volumetric flask can be calibrated byanindirectmethod,whichisfasterthan
the direct method, and sufficient for our purposes. This is done by delivering ten aliquots of
water from the(same)25mLpipetintothe250mLvolumetricflask. Thelocationofthebottom
part of the waters meniscus correspondingtothetotaladditionofthe1025mlpipetadditions
is then marked. If the flask is subsequentlyfilledexactlytothecalibrationmark,thenone25ml
pipet full of solution will contain exactly

1
10

of the original amount of solute in the solution.

Thisisausefulcalibrationmethodevenifthepipetisnotdirectlycalibrated.
Burets may be calibrated at specific volume intervals. For example, the 50 mL buretsmightbe
calibrated every 10 ml by comparing the nominal volume of water delivered (read from the
buret) with the actual volume water delivered. The latter is obtained by weighing the water and
determining the volume from the density. You will not need to calibrate your burets because,
except in the most exacting work, it is not worth the time it takes to do the calibration.
Furthermore it has been generally noted that beginning analytical workers tend to report larger
errorsinthevolumesdeliveredbyburetsthanareactuallypresent.

S16


45

GA2AcidBaseTitration
GA2 AcidBase Titration is the determination of the percentage of potassium acid phthalate
(KHP) in an impure solid mixture (the "unknown"). The weakly acidic KHP, which is water
soluble, can be titrated withasolutionofNaOHwhoseconcentrationisprecisely,andhopefully,
accuratelyknown.

SolutionsandChemicals
~ 0.1M NaOH, phenolphthalein indicator, primary standard KHP, unknown KHP (Potassium
HydrogenPhthalate)

Standardizationof~0.1MNaOHagainstthePrimaryStandardKHP
The primary standard KHP will be used to standardize the sodium hydroxide solution. The
amount of primary standard KHP used for each titration is dictated by the requirement to keep
the relative error, resulting from the reading of the buret, as low as possible. Since the total
precision of the volume reading of the 50 mL buret is 0.04 mL, a volume of 3545 mL
deliveredwillresultinarelativeerrorofabout1ppt(partperthousand,i.e.,0.04/40.00X1000).
Assuming an end point is tooccuruponthedeliveryfrom35to45mLof0.1MNaOH,calculate
the range in the sample weight of primary standard KHP (FW 204.23) that should be weighed
out.

AmountofKHPthatwillrequire40mLof~0.1MofNaOHtoreachendpoint
0.04L ~ 0.1M N aOH = _________moles204.23KHP F W = _________g KHP
Primary Standard KHP is in the desiccator placed near digital weighing balance. To open a
desiccator, slide lid sideway and do not try to pull it straight up as it is greased to make an
1

airtightseal .

Page 41, Quantitative Chemical Analysis,edition8thbyDanielHarris,W.H.FreemanandCompany

NY.

S16


46

1. Weighoutfoursamplesofpure,dry,primarystandardKHPofknownpurityfactorintofour,
numbered 250 mL Erlenmeyer flasks. Onceweighedout,thesemaybeleft,undissolvedbut
stoppered, in your lab benchuntilreadyforuse. Besuretolabeltheflaskssothatyouknow
whichsampleisineachflask.
2. To the KHP in the flask, addabout50mLofdeionizedwaterand2dropsofphenolphthalein
indicator. It will be necessary to warm the solution gently to about 500C and swirl to
dissolvetheKHP.
3. Before filling completely the 50 mL buret, first rinse it three times with ~5 mL portions of
0.1M NaOH solution. Carefully read and record the initial buret reading. Note: all buret
readings should be read to the nearest 0.01 mL. Takecaretoreadtheburetproperly. Refer
to the pictorial illustration located at the end of GA2 showing how to read the water
meniscus of the 50 mL buret. Note that significant additional error in the buret reading
occursiftheburethastoberefilledasecondtime.Whyisthat?

4. Titrate with the NaOH solution until JUST BEFOREthefirstpermanentappearanceoflight


pink color is seen throughout the solution. STOP! At this point, continue the titration by
adding NaOHsolutiondropwiseorbysplittingdropsuntilthefirstpermanentappearanceof
apinkcolorpersiststhroughoutthesolutionforatleast30seconds.
5. Repeat by carefully titrating the other three samples. Check that the relative average
deviation of these four trials does not exceed 2 ppt. If the precisionistoopoor,higherthan
about three ppt., titrate two additional samples until the required precision is obtained.
EmploytheQtestinanefforttorejectthemostoutlyingvalue.
6. What if the deionized water used to dissolve the primary standard KHP were contaminated
with an interfering ion? All results are suspect unless a "blank" (control) titration is done.
To measure the blank carry out a separate titration following exactly the same procedure as
used for the sample except do NOT add the KHP to the Erlenmeyer flask which is used as
the titration vessel. Record the volume of NaOH required to reach the phenolphthalein
endpoint. Check to see that this value is less than 0.10 mL (or less thantwodrops). Ifnot,
pleaseinformtheinstructor.

S16


47

DeterminationofPercentKHPinanUnknownMixture
You need to weigh out amount of KHP thatwillrequire25mLofstandardizedNaOH. Amount
ofunknownKHPsamplerequiredforeachtrial:
0.025L ~ 0.1M N aOH = _________moles204.23KHP F W = _________g KHP

AssumingKHPpurityof~40%,theamountofKHPtobeweighedoutforeachtitration:

KHP unknown __________g


P urityF actor

=_________g

Obtain an unknownKHPsamplefromtheinstructor. Theinstructorwillhavecodeditwithyour


name.YouwillreceiveonlyoneunknownKHPsample.
1. Weigh out four samples of KHPunknown into four 250 mL Erlenmeyer Flasks. Follow the
procedure given in the previous part to titrate KHP solution against the standardized NaOH
using2dropsofphenolphthaleinasindicator.
2. The average relative deviation for the per cent KHP in the unknown sample should not
exceed 2 ppt, which correspondstoarelativestandarddeviationofabout3.6pptfora50per
centKHPsample.

Precaution
After using alkali solutions, glassware needs to be thoroughly washed. Please thoroughly
washburet,pipetandfunnelwithtapwaterandthenrinsewithDIwaterafteruse.

CalculationsforPrelabReport
1. Standardizationof~0.1MNaOHagainstthePrimaryStandardKHP
Amount of KHP that will require 40 mL of ~0.1M of NaOH toreachend
point
0.04L ~ 0.1M N aOH = _________moles204.23KHP F W = _________g KHP

S16


48

2. DeterminationofPercentKHPinanUnknownMixture
You need to weigh out amount of KHP that will require 25 mL of standardized NaOH.
AmountofunknownKHPsamplerequiredforeachtrial:
0.025L ~ 0.1M N aOH = _________moles204.23KHP F W = _________g KHP
AssumingKHPpurityof~40%,theamountofKHPtobeweighedoutforeachtitration:

KHP unknown __________g


P urityF actor

=_________g

Calculations&/orQuestionsforPostlabReport
1. SpecifyUnknown#.Reportwillnotbegradedifunknown#isnotspecified.
2. Calculate molarity of the NaOH solution. Calculatetherelativeaveragedeviation(RAD)in
ppt and standard deviation (SD) for NaOH molarity. No need to show sample calculations
for RAD, SD. Present experimental data in a table format: data for each trial titrate
volume,titrantvolume,andcalculatedmolarityforthattrial.
3. UseoneofthetrialstoshowsamplecalculationsforNaOHmolarity
4. PerformttesttocompareestimatedMNaOHtothegivenvalue.
5. Titration of Unknown KHP: Present experimentaldatainatableformat:dataforeachtrial
weight of unknown KHP sample, titrant volume, and for that trial. Calculate %KHP and
uncertaintyinitsestimationbyPropagationofErrorusingprovidedhandout.

S16


49

SeeD.C.Harris:QuantitativeChemicalAnalysis,8thedition,Chapter2,page35.

S16


50

GA4AnalysisofCalciumbyEDTA
A ~ 0.03M solution of EDTA is prepared and then standardized in order to analyzethecalcium
in an unknown sample and to determine total hardness of water. First, the ~0.03M EDTA
solution is standardized using a primary standard, CaCO3. This is done using a substitution
reactioninwhichasmall,butreproducibleamountofMgEDTAsolutionisadded. Thechemical
reactions employed in this experiment will beexplainedinalaboratorybriefing. Inthistitration
with EDTA, first the calcium ion and then the displaced Mg ion are titrated in a
complexforming titration which uses Eriochrome Black T to indicate the end point. The
percentage of calcium in the unknown sample is then similarly determined using the
standardized EDTA solution. As an illustration of the practicality of this analysis, the widely
used EDTA titration for the total hardness (both free calcium andmagnesiumion)indrinking
water, using the Eriochrome Black T indicator end point, will be conducted. For more details
refer to the text, D. C.Harris:QuantitativeChemicalAnalysis,7thedition,Chapter12,page228
8thEdition,page236.

SolutionsandChemicals
(a) Solid, reagent grade Na2EDTA (b) Solid,primarystandardCaCO3,previouslydriedat1100C
(c) Unknown calcium sample, previously dried at 1100C (d) 5M NaOH,1MNaOH,1MHCl(e)
Methyl red indicator (f) pH 10 ammonia buffer (g) Mg2+EDTA complex (h) EriochromeBlack
T indicator, Keep in the refrigerator or on ice (i) Toluene used to preserve the aqueous calcium
solution(j)Special10mLburet

PreparationandStandardizationofa0.03MEDTASolution
Into a 250 mL Erlenmeyer flask, accuratelyweighbydifferencetothenearest0.0001g,5.6gof
the reagent Na2EDTA.

Add about 80 mL of deionized water to the 250 mL flask and gently

warm the solution on the hot plate (not with a Bunsen burner) to a temperature no higher than
600C. It is necessary to heat the solution to dissolve the Na2EDTA, but the Na2EDTA will
permanently breaks down over 800C. Transfer the solution withseveraldeionizedwaterrinsing

S16


51

through a regular funnel to your500mLvolumetricflask. Quicklycoolthissolutionbyrunning


cool tap water around the side of the 500 mL volumetric flask. When the solution has reached
near room temperature, carefully dilute the solution with more deionized water to the flasks
calibration mark. Since EDTA can extract ions from the glass container, the EDTA solution
needs to be transferred to a 500 mL plastic bottle onthesamedaythatitismade. Rememberto
labelplasticbottlecontainingEDTAwithyourname,sectionanddate.

PreparingCalciumStandard
Weigh about 2.2 g of primary standard CaCO3 into a dry weighing bottle. In your laboratory
notebook, record the purity factor of the pure CaCO3 from the bottles label. Into a 250 mL
Erlenmeyer flask, accurately weigh by difference 1.0 to 1.1 g of the dry CaCO3 known to the
nearest0.0001g.

To dissolve the sample, add about 20 mL of deionized water followed by dropwise addition of
concentrated HCl (found in several of the hoods) until the effervescence (loss of CO2) stops.
SwirlandgentlyboilthesolutionusingahotplatetoexpeltheCO2untilthesolutionsvolumeis
about of its original volume. Do not heat to dryness. If a solid precipitate appears, add
enough deionized water to redissolve it.

It should redissolve! Ifitdoesnot,addthreemore

drops of concentrated HCl andcontinueheating. Iftheprecipitatepersists,notifythelaboratory


instructor. Rinse down thesidesoftheflaskwithdeionizedwater. Addenoughdeionizedwater
sothetotalvolumeisabout50mL.

Precaution
If you have insufficient time (about 1 hour is needed) to titrate this calcium solution during
the present laboratory period, stopper the Erlenmeyer flask with a clean rubber stopper,
leaving the calcium solution in the acidic state until the next laboratory period. The reason
for this is that a (harmlesstohumans) water mold tends to grow in nonacidic calcium
solutions inourbuildingsdeionizedwatersupply. Ifthishappens,thecalciumsolutionmust
bediscardedandanewonemade.

S16


52

When ready to begin the titrations of the standard calcium solution with the EDTA solution,
carefully neutralize the strongly acidic solution as follows so that later additions of the pH 10
buffer will be effective. First, add two drops of methyl red indicator to the calcium solution.
Then, slowly add 5M NaOH solution (provided in a dropping bottle) until the red color of the
indicator just turns yellow. Bringthecolorbacktoredusingafewdropsof1MHCl. Thenvery
carefully add 1M NaOH until the solution just turns yellow again. Do notaddalargeexcessof
NaOH or a white precipitate of Ca(OH)2 will form which is difficult to redissolve with acid.
With deionized water, carefully transfer the entire calcium solution through a funnel to your
calibrated 250 mL volumetric flask. Add a drop of toluene to inhibit the growth of the water
mold. Carefully dilute the solution to the calibration mark. Shake well, usually 10 times,
endoverend.

DeterminationoftheIndicatorBlank
An appreciable indicator blank exists for the titration of calcium with EDTA in the presence of
free Mg ion. Usually from 0.30 to 0.60 mL of the EDTAisneededtojustchangetheindicator.
Todeterminethisindicatorblankproceedasfollows:
Take 50 mL of deionized water, measured with a graduated cylinder, add specified number of
drops of Eriochrome Black T indicator (the number of dropsofindicatorneededdependsonthe
batch of indicator used), add 5.0 mL of the pH 10 buffer, then using provided digital pipet,add
2.0 mL of the MgEDTA solution. Titrate with the EDTA titrant from a color change of red
through purple to anendpointofapureroyalbluewithnoreddishtinge.Repeatatleasttwo
times. Agreement in the mL of EDTA titrant should be within 0.03 mL. The average value of
thisblankmustbesubtractedfromthevolumeofEDTAdeliveredfromsubsequenttitrations.

Standardizationof~0.03MEDTA
Carefully pipette exactly one 25 mL aliquot of the standard calcium solution into a 250 mL
Erlenmeyer flask. Add specified number of drops of EriochromeBlackTindicator,add5.0mL
of the pH 10 buffer, then using provided digital pipet, add 2.0 mL of the MgEDTA solution.

S16


53

For precise results, it is very important to be consistent in delivering the same number of
indicator drops as was used fortheIndicatorBlank! Titratethecalciumsolutionwiththe0.03M
EDTA solution to the samecolorchangefromredthroughpurpletoanendpointofpureroyal
blue with no reddish tinge. Since the change in the indicators color is slow, the EDTA should
be added slowly and the solution swirled well. Repeat the determination at least three more
times. Calculate the molarity of the EDTA solution. Results should be within 3.0 parts per
thousand. Ifnot,youshouldprobablyperformanothertwocalibrationruns,perhapsuseaQtest
torejectthemostoutlyingresult(s)andcheckyournewprecision.Thisisnotaneasyendpoint!

Dontforgetto
Pleasethoroughlywashburet,pipetandfunnelwithtapwaterandthenrinsewithDIwater
afteruse.

DeterminationoftheCalciuminanUnknownCalciumSample
Before this part is started, the standardization of EDTA with standard calcium solution must be
successfully completed sinceitisnecessarytodiscardthestandardcalciumstandardandtoreuse
the calibrated 250 mL volumetric flask. Discard the standard calcium solution and carefully
rinseyour250mLvolumetricflask,firstwithtapwaterandthenwithdeionizedwater.
Unknown calcium sample is predried at 1100C for an hour. Into a 250 mL Erlenmeyer flask,
accurately weigh 1.0 to 1.2 g, known to the nearest 0.0001 g, of the unknown calcium sample.
Dissolve this calcium sampleexactlyasthestandardCaCO3wassolubilizedinthepreviouspart.
Continue following the procedure, performing all steps through the final dilutionofthesolution
to 250 mL but now with your unknown calcium sample. Titratetheunknowncalciumsolution
following the same procedure as for the standard calcium solution. Do atleastfourtrials. The
indicator blank need not be repeated as long as the same batch of indicator is used, but do not
forget to subtract the volume of the indicator! All the equationsneededforthecalculationwill
be given in a laboratory briefing. The results should be reproducible to within 3.0 parts per
thousand.

S16


54

WaterSample
The laboratorywateristhesameasmostdormitoriestapwater.AllofUniversityswatersupply
is taken from below the Fenton River. There will be certain other real drinking water samples
also available. Be sure to record in your laboratorybookthesourceofyourtapwater.Askyour
instructor for a 10 mL buret for EDTA. Using a 100 mL pipet,transfera100mLaliquotofthe
tap water into a clean 250 mL Erlenmeyer flask. Acidify the water sample with two drops of
concentrated HCl and gently boil the solution to remove the CO2. Rapidly cool the sample by
running tap water around the outside of the Erlenmeyer flask. Add two drops of methyl red
indicator. Neutralize the solution with 1M NaOH until the yellow color just appears. Add 5.0
mLofthepH10bufferandtheusualnumberofdropsoftheEriochromeBlackTindicator. But
do not add the 2.0 mL of MgEDTA. This analysis of total hardness involves titrating boththe
free magnesium as well as calcium in drinking water. Using the 10 mL buret, titrate the water
sample with the standard 0.03M EDTA solution to the same pure blue end point. If the color
change is very sluggish, it could be because of the absence of free Mg ion in the water. Inthat
case, add 2.0 mL of the MgEDTA complex. Repeat at least twice more. If the MgEDTA
solution is used, then the previously determined indicator blank needs to be subtracted fromthe
totalvolumeofEDTAdelivered.Theresultsshouldbereproducibletowithin3.0ppt.

Dontforgetto
Pleasethoroughlywashburet,pipetandfunnelwithtapwaterandthenrinsewithDIwater
afteruse.Carefullywash&rinsethe10mLburetafterusebecauseEDTAsolution,when
dried,tendstoclogthefinertipofthese10mLburets.

Questions/TopicsfortheQuiz
1. AcidbasepropertiesofEDTA.
2. Defineformationconstantandconditionalformationconstant
3. Explain the role of (a) auxiliary complexing agent, triethanolamine (b) pH 10 buffer (c)
MgEDTA used inthistitration.(Formoredetailsrefertothetext,D.C.Harris:Quantitative
ChemicalAnalysis,7thedition,Chapter12,page2288thEdition,page236.)

S16


55

Calculations&/orQuestionsforPostlabReport
For the Report, just answer questions in the order they are asked, report will not be graded if
writtenintheessayformat.
1. SpecifyUnknown#.Reportwillnotbegradedifunknown#isnotspecified.
2. Calculate the average molarityoftheEDTAsolution.CalculatetheStandardDeviation(SD)
and Relative Standard Deviation (RSD) for EDTA molarity. No need to show sample
calculations for RSD, SD. Present experimental data in a table format: data for each trial
titrate volume, titrant volume, and calculated molarity for that trial. Also, includeRSDand
SD.
3. UseoneofthetrialstoshowsamplecalculationsforEDTAmolarity
4. Calculate the percentage of calcium the unknown sample. CalculatetheStandardDeviation
and Relative Standard Deviation for the percentage of calcium in the unknown sample. No
need to show sample calculations for RSD,SD. Presentexperimentaldatainatableformat:
data for each trial titrate volume, titrant volume, and calculated %Ca for that trial. Also
includeRSDandSD
5. Useoneofthetrialstoshowsamplecalculationsfor%Ca
6. CalculatetheCaCO3ppminthewater.
7. Present experimental data in a table format: data for each trial titrate volume, titrant
volume, and calculated CaCO3 ppm for that trial. Use one of the trials to show sample
calculationsforppmofCaCO3.

S16


56

GA5IodometricTitrationofAscorbicAcidinVitaminCTablets
The objective of this laboratory experiment is to determine the ascorbicacidcontentofVitamin
C tablets using an iodometric titration. Ascorbicacidisamildreducingagentthatreactsrapidly
and quantitatively with iodine to reduce it to iodide ion. In this experiment, a known excess of
iodine isformedbythereactionofanaccuratelyweighedamountofiodateioninthepresenceof
excess iodide ion. Once the ascorbic acid has quantitatively reacted with the iodine, the
remaining iodine is back titrated withthiosulfate. Thetitrant,thiosulfate,isstandardizedagainst
iodine using the same iodate and iodide ion reaction.

To indicate the end point, the

disappearance ofabluestarchcolorisused.Thebluestarchcoloriscausedbytheformationofa
colored complex between iodine and the amylose molecule found in the starch. For more
details refer to the text, D. C. Harris, Quantitative Chemical Analysis, 6th edition, Chapter 16,
pages355,3613657thEdition,page334,3403438thEdition,page347,351357

SolutionsandChemicals
Sodium thiosulfate (Na2S2O3), Sodium carbonate, Potassium iodide (KI) and potassium iodate
(KIO3), all in solid form. Starch indicator (containing HgI2 as the preservative), 0.50M and
0.30Msulfuricacid

Glassware
(a) 50 mL volumetric pipette for Potassium Iodate solution willbeprovided. (b)Usethe10mL
graduated cylinder for starch and 100 mL graduated cylinder for acid fromyourowndrawer(c)
From your own drawer, get 250 mL Volumetric flask for Sodium Thiosulfate and 500 mL
Volumetric flask for Potassium Iodate solutions(d)VitaminCtabletseachcontainingeither250
or 500 mg of vitamin C. PleaseconfirmVitaminCcontentofthetabletbeforeuse(e)Mortar&
Pestle

S16


57

PreparationoftheThiosulfateSolution
Thiosulfate solution (~ 0.100 M) is prepared using freshly boiled water. Keep two 500 mL
beakers containing 200 mL deionized water in each on the hot plate it takes about 20 min for
water to boil. Bringwatertoroomtemperaturebyfirstholdingbeakersundertapwaterandthen
in the ice bath. Once, water is at room temperature, then weigh out about 6.20 g of solid
Na2S2O3 and about 0.025 g of Na2CO3. Add both reagents to the 250 mL volumetric flask and
dissolve by adding small portion of freshly boiled water once dissolved, make it up to the
volume. Store this solution in your amber bottle (rinse the bottle out ahead of time with the
freshlyboileddeionizedwater).Keepthebottletightlycapped.

PreparationofthePotassiumIodateSolution
Prepare a 0.013 _ _M Potassium Iodate solution by carefully weighing out 1.39 _ _ g of
predried, primary standard KIO3. Carefully dissolve the weighed amount and quantitatively
dilute in your 500 mL volumetric flask. This solution can be kept in your 500 mL volumetric
flask.

StandardizationoftheThiosulfateSolutions
Carefully pipet 50.00 mL of the KIO3 solutions into a 250mL Erlenmeyer flask. Add
approximately 2.0 gofsolidKIand10mLof0.50Msulfuricacid. Swirlandimmediatelytitrate
withthethiosulfateuntilthesolutionhaslostalmostallofitscolor(hasastrawyellowcolor).

Equations
StandardizationoftheNa2S2O3
I O3 + 8I + 6H + 3I 2 I + 3H 2 O
I 2 I + 2S 2 O32 3I + S 4 O62
Precaution
Ifthesolutionisclearandnobluecolorisseenafteradditionofthestarch,thetrialmustbe
discardedandredone.

S16


58

At this point add 2 mL of the starch indicator and carefully completethetitrationuntilonedrop


of titrant removes the blue color. Repeat this titration with at least two additional 50.00 mL
volume of KIO3 solution. Titrations should be performed carefully but rapidly to minimize air
oxidationoftheiodideion.

Calculate the Normalityofthethiosulfatesolution. Therelativeaveragedeviationshouldbeless


than3partsperthousand.TheformulaweightofNa2S2O3is158.11gandofKIO3is214.00g.

AnalysisofascorbicacidintheVitaminCTablets
Use pestleandmortartocrushvitaminCtabletintoafinepowder. Weighoutabout0.20__gof
fine powder using the analytical balance. Transferfinepowderintoa250mLErlenmeyerflask,
add 60 mL of 0.3Msulfuricacidandswirltomix,somesolidbindingmaterialmaynotdissolve.
Add about 2.0 g of KI, swirl to dissolve then add 50.00 mL of thestandardKIO3solutionusing
pipet provided. Swirl to mix and then carefully titrate with the standardized thiosulfateasdone
in previous part. Remember to add the 2 mL of starch only when thesolutionislightyellowin
color. Perform the analysis three times. Calculate % Vitamin C for each trial. A suggested
relative average deviation is less than 3 parts per thousand should be possible. The formula
weightofascorbicacidis176.12g.

Dontforgetto
Pleasethoroughlywashburet,pipetandfunnelwithtapwaterandthenrinsewithDIwater
afteruse.

Questions/TopicsfortheQuiz
1. DifferencebetweenIodimetryandIodometry,specifytitrantusedineachtechnique.
2. ReactionsrelatedtostandardizationwithI2Isolution.
3. Explain (a) why the pHofsolutionsiscriticalforthosechemicalequations(b)Thiosulfateis
stored in capped bottle (c) It isimportanttoboilwater(d)Whystarchisaddedattheend(e)
HowexcessKIhelps(f)WhyNa2CO3wasaddedtothiosulfatesolution.

S16


59

Calculations&/orQuestionsforPostlabReport
For the Report, just answer questions in the order they are asked, report will not be graded if
writtenintheessayformat.
1. Calculate Normality of thiosulfate calculate Relative Average Deviation (RAD), Standard
Deviation (SD). No need to show sample calculations for RAD, SD. Present experimental
data in a table format: data for each trial titrate volume, titrant volume, and calculated
normality for that trial. Also, include RAD and SD. Use one of the trials to show sample
calculationforThiosulfateNormality.
2. Calculate % Vitamin C in tablet calculate RAD (ppt), SD. Present experimental data in a
table format: data for each trial titrate volume, titrant volume, andcalculated%VitaminC
for that trial. Also, include RAD and SD. Use one of the trials to showsamplecalculation
for%VitaminC.
3. Compareexperimentallydetermined%VitaminCinthetablettothegivenvalue.
4. If you were working for the manufacturer of the vitamin tablets would you be comfortable
withyourresults?Yes,no?Why?

S16


60

GA7PotentiometricTitration
The potentiometric titration is a useful means of characterizing an acid. ThepHofasolutionis
measured as a function of the amount of titrant added. The change in pH is small until theend
point where there is a sharp change. Thestrengthofanacidorbasedeterminesthesharpnessof
the change. The end point is found by graphing the points and determining the location of the
sudden change inpH. Itisnotnecessarytoaddthetitrantdropwiseaswasrequiredtoobtainthe
exact equivalence point using a visual indicator. The important thing is that a good graph with
increments such as 2.00, 1.00, 0.50, 0.20 and 0.10 (2 drops) mL of titrant be added and the
resulting pH readings obtained. (Refer to Harris: 6th Edition, Chapter 713 7th Edition,Chapter
7118thEdition,Chapter810)

SolutionsandChemicals
(a) Standardized 0.1M HCl (b) Standardized 0.1M NaOH from GA2 (c) Buffer solution, pH
7.00andpH4.00tostandardizethepHmeter(d)SolidKHP(e)Unknowndiproticacid

Equipment
(a) pHmeter(b)Magneticstirrermotor

DeterminationoftheEquivalencePoint
The hydrogen ion activity of a solution is conveniently measured using a glass indicator
electrode. The reference is usually a calomel or silversilver chloride electrode.

In this

laboratory, you will be using a combination electrode that incorporates both electrodes
surrounded byaprotectiveplasticsleeve. ThepHmeter,ahighinputimpedancemillivoltmeter,
iscalibratedbysettingthepHtothatofaknownstandardbuffer.

Inthisexperiment,threetitrationcurveswillbeobtained:

S16


61

Strongacid(HCl)withastrongbase(NaOH)
Weakmonoproticacid(KHP)withastrongbase(NaOH)
Anunknowndiproticacid(H2A)withastrongbase(NaOH)

A titration curve has a characteristic sigmoid curve. The partofthecurvethathasthemaximum


change marks the equivalence point of the titration. In the case of an acid titration, the slopeis
increasing before the equivalence point and decreasing after the equivalence point (just the
oppositeistrueforthetitrationofabase). Notethattheslopechangesfastestjustbeforeandjust
after the equivalence point. Attheequivalencepoint,itdoesnotchangeatall,andwesaythere
isaninflectionpoint.Therateofchangeoftheslopeiszero.
pH 2 pH 1
V2 V1

= pH

The equivalence point can be found in three ways. One method is to look at the sigmoidcurve
and estimate wherethecentralpartoftheriseis. Thesecondmethodistomakeafirstderivative
plot. The first derivative, pH
is the slope of the curve, and can be obtained simply from
V
Equation1.
pH

pH

( mL )2 ( mL )1
V 2 V 1

pH
V 2

Each first derivative point is plotted against V' where V' is the average of the two volumes, V1
and V2. The endpoint occurs at the volume, V', where

pH
V

has the maximumvalue. Thethird


2

way istomakeasecondderivativeplot. Thesecondderivative, VpH2 istherateofchangeofthe


slope and can be obtained from Equation 2. Each second derivative point is plotted against V"
2

where V" is theaverageof V 1 and V 2 . Theendpointoccursatthevolume,V",where VpH2 is


zero. See last pages of GA7 for an example of first and second derivative data and derivatives
2

plots .

StandardizationofthepHMeterwithTwoStandardBuffers
Just before beginning the first titration, standardize the pH meter using the buffers provided.
See the instruction sheet provided with the pH meter for its standardization. Remember to

Forthelabreport,useExcelforFirstandSecondDerivativeCalculations&Plots:Harris,8thEd,page216

S16


62

restandardize the pH meter with the buffer beforeeachtitration. Youshouldnoteandrecordin


your lab notebook the reproducibility of the pH meter readings. If there is any questions about
the performance of the electrodes or meter during the titration, please consult either the
laboratoryinstructororteachingassistant.

StandardizationofHClbyPotentiometricTitration
Fill the buret with standardized 0.1____M NaOH.

Using second burette, carefully deliver

25.00 mL of the~0.1MHClintoa250mLbeaker. Addabout25mLofdeionizedwater. Place


a magnetic stirrer bar in the beaker. Place the beaker on the magnetic stirrer. Position the
electrode system in the solution making sure the bottom sensing portion of the combination
electrode is covered with solution. It might be necessary to add alittledeionizedwater. Outof
interest, add a few drops of phenolphthalein indicatortothetitrationsolution. Recordthetitrant
volume & pH at which the indicator changes color. This is to be added to the graph in the lab
report.

With the electrodes inserted and the stirrer bar turning, read the initial pH when 0.00 mL of
NaOH has been added. During the titration, keep the pH meter ON and maintain a constant
stirring rate. Carefully titrate with standardized NaOH solution startingwithincrementsof2.00
mL and gradually reducing the increments to2drops(about0.10mL)atatimeinthevicinityof
the equivalence point where the pH is changing rapidly. It will simplify graphingifincrements
are kept constant over the appropriate ranges. For example,add2.0mLincrementsatfirst. As
the slope of the titration curve increases, decrease to 0.5 then 0.20 and finally to 0.10 mL near
theequivalencepoint.

Addition of NaOH is controlled by monitoring two factors (a)changeinpHby0.2pHunitsand


(b) volume added should not exceed 2 mL at any interval. Stop when eitherpHchangesby0.2
pHunitor2mLNaOHisaddedwhichevercomesfirst.

At the first sign that the pH is changing faster, decrease the NaOHincrement.Recordallvalues
of volume NaOH solution added and pH. Continue beyond the equivalence point with first

S16


63

smaller and then larger increments as the change decreases. Stop when the pH has been at or
near a pH of 11 for several, consecutivemilliliterssothatyouhaveatleast57datapointswhen
pH is about 11. Because the pH meter is precise to only 0.02 pH units, there is no needtoboil
solutions priortotheequivalencepointduringpotentiometrictitrations. TurnoffthepHmeterat
theendofthetitrationremoveandrinsetheelectrodes.

PotentiometricTitrationofPotassiumAcidPhthalate(KHP)
It will be necessary to know the exact weight, to 0.1 mg, the weight of KHP used. Weigh 2.5
mmol of KHP to 0.1 mg, by difference into a 250 mL beaker. Add 50.0 mL of deionized
water.

Warm slightly to dissolve KHP. Again add 50.0 mL more of deionized water and

quickly cool the solution running tap water around the outside of the beaker until it is close to
room temperature. Add phenolphthalein. Titrate as done before, recording pH and volume
values.Recordthetitrantvolume&pHatwhichtheindicatorchangescolor.

PotentiometricTitrationofaDiproticAcid
In this experiment, one of the five diprotic acids listed in the table will be investigated by your
team. See table below for diproticacidswiththeirformulaweightandpKavalues. Eachofthese
acids exhibits a different shaped titration curve. From the shape of the curve that is obtained,
one candeterminetheformulaweightandtheequilibriumconstant(s)oftheacid. Asyourgroup
performs the titration, have one of the partners prepare a rough plot. After the titration is over,
plottitrationcurveusingExceltoconfirmthattwoequivalencepointsareclearlyvisible.
Weigh by difference the specified weight of thediproticacidintoa250mLbeaker. Add50mL
of deionized water to dissolve. Add phenolphthalein indicator. Titrate as before, recording pH
and volume values. Record the titrant volume & pH at which the indicator changes color. Itis
suggested that you first perform a rapid, crude titration in order to determine where the
equivalence point(s) is/are. Add the titrant in2.00mLincrementsatatimeatfirstrecordingthe
pH after each addition. As the pH rises more rapidly, add smaller increments. After the first
equivalence point isobserved,useonceagainlargerincrementsuntilavolumebeyondtwicethat
required for the first equivalence point has been added. Continue adding the NaOH for an

S16


64

additional 10 or 15 mL. It is necessary touseall50mLNaOHinordertogetapropertitration


curve.
Dontforgetto
Pleasethoroughlywashburet,pipetandfunnelwithtapwaterandthenrinsewithDIwater
afteruse.

PossibleDiproticAcid
Acid

FormulaWeight

pKa1

pKa2

GlutamicAcid

147.13

4.07

9.47

MalonicAcid

104.06

2.83

5.69

MaleicAcid

116.07

1.92

6.27

SuccinicAcid

118.09

4.16

5.61

TartaricAcid

150.09

2.98

4.34

Questions/TopicsfortheQuiz
DiscussErrorsinpHmeasurement(Seepage319ofHarrisEd.8,Ch.14)

Calculations&/orQuestionsforPreLabReport
Study Excel worksheet shown on page 216 (of Harris, Ed8,Ch.10). YouneedtouseExcelfor
plotting first derivative andsecondderivativetitrationcurves. Seeexampleonpage217. Verify
thederivativeincellD7ofFigure106.

Calculations&/orQuestionsforPostlabReport
1. SpecifyUnknown#.Reportwillnotbegradedifunknown#isnotspecified.
2. Titration Curves (dont include pH vs. NaOH volume data) for HCl, KHP anddibasicacids
with:
Agoodtitle
Labeledaxes
Asmoothlydrawncurve

S16


65

Indicationofthevisualindicatortransition
PositionofpKawhereapplicableandvolume(s)atequivalencepoint(s)
Firstderivativeplotsforallacids
SecondderivativeplotforKHPonly(forjustonetrial)

(Using Excel for First and Second Derivative Calculations & Plots: Harris, 8th Ed, page
216).

3. Includethetitrationcurvefortheunknowndiproticaciddonebyyourgroup.
4. Fromthedataandthetitrationcurve,calculatetheconcentrationofHCl
5. CalculatethepKaofKHPfromthedataandthetitrationcurve.
6. CalculatetheformulaweightofKHP.
7. Calculate the absolute and relative error for theFWandpKaoftheKHP(acceptedvaluefor
FWis204.23andforpKais5.41).
8. DeterminetwopKasofyourdiproticacidfromthedataandthetitrationcurve.

Calculatetheformulaweightofyourdiproticacid,seefollowingequations:
MonoproticAcidFW:
mgof Acid
M N aOH mLN aOH

= _________ gmacid

mol

DiproticAcidFW:
Acid
2 M mgofmL
N aOH

N aOH

= _________ gmacid

mol

9. IdentifyyourdiproticacidusingcalculatedFWandpKa1andpKa2.
10. Calculate the absolute and relative error for FWandpKa1andpKa2fortheidentifieddiprotic
acidbycomparinggivenvaluesintheTable.

S16


66

Example:TitrationCurves

S16


67

Example:TitrationCurveData

S16


68

GA8 Ion Selective Electrode: Measurement of Fluoride Ion Activity


inMouthwash,Toothpaste&DrinkingWater

In this experiment, a fluoride, solidstate type, ion selective electrode is used to measure the
potential that arises when itisimmersedinaqueoussolutionshavingdifferentfluorideactivities.
Using a simple millivolt/pH meter, first the potentials of a series of aqueous standards having
known fluoride activities (concentrations as each solution has the same ionic strength) are
measured with the fluoride ion selective electrode, versus a standard calomel (reference)
electrode (SCE). A plot of these potentials vs. the log of the concentrations of fluoride ion,
usually over the 100 to 1.00 ppm fluoride concentration range, gives a straightlinewhoseslope
approximates a 0.0592 volt change per decade. Below 1.0 ppm the data points arefoundnotto
be as linearbutarestillneededinordertomeasurethefluorideionconcentrationtypicallyfound
in most fluoridated drinking waters. Diluted samples of fluoridated mouthwash, toothpaste, an
unknown fluoride solution received from the instructor, and tap water are measured using the
fluoride ion selective electrode. The fluorideconcentrationsofthosesolutionsarereadfromthe
standard curve. In the case of the mouthwash and toothpaste,appropriatedilutionequationsare
neededtoobtaintheactualpercentfluorideintheweighedsample.

SolutionsandChemicals
NaF, NaCl, NaOH, and 1,2diaminecyclohexaneN,N,N',N'tetracetic acid monohydrate
(CDTA), 400ppmFluoridesolutions(asNaF)storedinplasticbottles. TheTotalIonicStrength
Adjustment Buffer (TISAB) solution. Unknown fluoride solution. Fluoridated toothpaste and
mouthwash.

Equipments
Millivolt/pH Meter (Digital with three digits) or expanded scale analog millivolt/pH meter
Fluoride ion selective electrode, (Orion Part No. 9409 or equivalent) Saturated Calomel

S16


69

(reference) Electrode (SCE)

Magnetic stirrer with stirring bar Stirring bar retriever

Thermometer Hotplatetohastenthedissolvingofthetoothpaste Analyticalbalance,readable


to0.1mg.

Glassware
Beakers:15 30 mL, 1400 mL beaker, 1250 mL beaker Volumetric flasks: 8 100 mL, 1500
mL Volumetric Pipets: 1 50 mL, 125 mL, 3 10 mL Graduated pipet: 1 10 mL and 1
pipetbulb.

PreparationofStandardSolutions
Prepare following fluoride (F) standard solutions by first diluting the 400 ppm to a 100 F
solution usingdeionizedwaterin100mLvolumetricflask. Then,prepare40ppmFstandardby
dilutingthe100ppmFstandardusingdeionizedwaterina100mLvolumetricflask.

Usethe40ppmFstandardanddiluteasfollowsinother100mLvolumetricflasks:
Use40ppmFstandard

FstandardPrepared

50.00mLdiluteto100mL

20

25.00mLdiluteto100mL

10

10.00mLdiluteto100mL

5.00mLdiluteto100mL

Prepare the following F standard solutions by diluting the indicated volumes of 10 ppm F
standardprepared,
Use10ppmFstandard

FstandardPrepared

10.00mLdiluteto100mL

5.00mLdiluteto100mL

0.500

2.50mLdiluteto100mL

0.250

S16


70

Into the nine,30mLlabeledbeakers,pipet10.0mLof0.250,0.500,1.00,2.00, 4.00,10.0,20.0,


40.0, and 100.0 ppm F solution, respectively. Then,intoeachoftheseninebeakers,carefully
pipet,exactly10.00mLofTISAB.

AdjustingtheZerooftheMillivoltmeter
Have the Instructor or the Teaching Assistant show you how to zero the Millivoltmeter to 0.0
mV. At the same time, using the thermometer provided, measure the temperature of the 100.0
ppm F solution. Have the instructorexplaintoyouhowtoadjustthetemperaturecompensation
dial(circuit)ofthemillivoltmetersothatitcorrectsforthedifferenceintemperatureintheslope
(theNernsttermfactor).

CautionsConcerningUseofElectrodesandMeters
The fluoride electrode is used with a saturated calomel electrode. The former is stored in a
fluoride solutionandthelatterinaKClsolution. Besuretoreturnthemtothecorrectsolutionat
theendoftheexperiment.
Remove each electrode from its storage flask and rinse with deionized water. BLOT each
electrode with a Kimwipe. Do not rub the surface of the fluoride electrode since this sort of
treatment could damage it (the fluoride electrode costs about $250 the SCE referenceelectrode
costs about $50). Place each electrode in theelectrodeholderandimmersetheelectrodesystem
in the first solution to be measured. After the measurement, switch meter to stand by mode
and then remove electrodes from solution, rinse both with deionized water, wipe and put into
nextsolutiontobemeasured.

Solution is stirred using magnetic stirrer during F measurement. Take care not to allow the
stirring bar tohittheelectrodessincethismaycausedamage. Astirringbarretrieverisprovided
to withdraw the stirring bar from the solution after measurement. Rinse the stirring bar with
deionized water. After removal, dry with a Kimwipe, and drop it into the nextsolutiontobe
measured.

S16


71

DeterminationofPotentialsfortheCalibrationCurve
Measure the potential of all the standard solutions starting with the highest fluoride
concentration, 100 ppm, and working downtothelowest,0.250ppm. (Recordthesign:positive
or negative) of the resulting millivolt readings as measured against the reference electrode.
Unlike a pH electrode,thefluorideelectrodedoesnotreachequilibriumimmediately,anditmay
be necessary to wait a minute or so before the potential stops drifting. The drift will be more
pronouncedastheconcentrationdecreases.
While still on this laboratory experiment, before discarding the known solutions or dismantling
the measuring equipment, make a rough plot of mV (yaxis) vs. Log [F ppm] (xaxis) using
Excel. Make sure the electrode is behaving linearly. If there are some obvious data points
deviating from the expected linear relationship you might want to recheck one or more of the
potentialreadings.

DeterminationofFluorideinToothpaste(optional)
Record the brand name in your laboratory notebook. As rapidly as possible, since volatile
materials are lost from the toothpaste sample, weigh about 0.2 g of toothpaste into the 250 mL
beakerprovided. Addabout50mLofdeionizedwaterandgentlyboilthismixtureontheheating
plate provided for about five minutes. Bring it to room temperature. Transfer to a 50 mL
volumetric flask and dilute to the mark using deionized water. Pipet 10.0 mL of the toothpaste
solution and 10.0 mL of TISAB into a 30 mL beaker. Insert the electrodes into the sample and
turnonstirring,whenthepotentialstopsdrifting,readit.

DeterminationofFluorideintheMouthwash
Record the brand name in your laboratory notebook. Weigh an empty 30 mL beaker of the
analytical balance. Carefully deliver into this weighed beaker exactly20drops(about1.00mL)
of the Mouthwash using the Pasteur pipet. Rapidly reweigh the beaker to obtain an accurate

S16


72

weight of the Mouthwash delivered. Add exactly 9.00 mL of deionized water and 10.00mLof
TISAB.Determinethepotentialasdiscussedintheprevioussection.

DeterminationofFluorideinWaterSamples
Pipet 10.00 mL of tap water to be tested and 10.00 mL of TISAB into a 30 mL beaker.
Determinethepotentialasdiscussedpreviously.

DeterminationofFluorideinUnknown
Obtain a sample of unknown fluoride from your instructor. Besuretorecordthenumberofthis
unknown. Into a 30 mL beaker pipet 10.00 mL of unknownand10.00mLofTISAB. Measure
thepotentialasdiscussedpreviously.

Questions/TopicsfortheQuiz
1. (a) Indicator Electrode types (b) Selectivity coefficient and equation describing response of
ISE in presence of interfering ions (c) the design of Fluoride ISE (d) role of TISAB and
significance of pH control. Refer to Harris: 7th edition, page 301 & 311314 8th Edition,
page311&323326

Calculations&/orQuestionsforPostlabReport
For the Report, just answer questions in the order they are asked, report will not be graded if
writtenintheessayformat.

1. Construct a calibration curve by plotting mV (Yaxis) vs. Log [F ppm] (Xaxis) of the
standardsolutions.Plotshouldincludeequationoftrendlineandregressioncoefficient.
2. Using those data points that clearly fall in the linear portion of the plot, usually in the 2 to
100 ppm range, determine the slope of thecurveinmV/decade. Calculatethedifferencesin
mVmeasured,forexamplefromthe100and10.0ppmfluoridestandardsorthedifferencein
mV measured for the 40 and 4.0 ppm fluoride standards. Does the electrode exhibit

S16


73

Nernstian or near Nernstian behavior? Manufacturer reported value is a difference of 54 to


58mVperdecade.
3. From the calibration plot, read determine concentrations in ppm of Fforallofthesolutions
measured: toothpaste, mouthwash, water sample, and unknown. Express your answer to 3
significant figures. Three significant figures are justified since there will be overa100mV
difference in mV readings between the 100 and 0.25 ppm fluoride standard. Sinceboththe
standard solutions and the solutions measured were diluted by half with TISAB, the
concentrationsofthe10.00mLsamplescanbereaddirectlyfromthecalibrationcurve.
4. Calculate the concentration of fluoride, expressed as % by weight of fluoride in the
toothpaste. Depending upon brands, usually a 0.07 to 0.20 % fluoride value is usually
obtained. Remember in your lab report to specify the brand name used. Realizing the
concentration measurement of the toothpaste solution was done on 10.0 mL of solution and
onefifth of the total solution was used, then the weight and percentage, of fluoride in the
toothpaste is calculated as follows (usually a value of 0.07 to 0.20 % fluoride expressedas
%NaFisobtaineddependinguponbrandoftoothpaste):
g
______ppmor mL
of F luoride 50.0mL
1mg = 0.______mg
10.0mL 1000g
0.______mg
%F luoride = ______mgT
100 = 0.______%
oothpaste

5. Calculate the concentration fluoride in the mouth wash sample. Express this as % fluoride
by weight. Depending upon brands of fluoridated mouthwash, a value of 0.02 to 0.05% is
usually obtained. Rememberinyourlabreportspecifythebrandnameused(usuallyavalue
of 0.02 to 0.05 % fluoride expressed as % NaF is obtained depending upon the brand of
mouthwash).
6

g
100
______ppmor mL
of F luoridesolution 10g g 10.0mL

1.0mL 0.______g

weightof 20drops

= 0.______%

6. Report the value, in ppm of F, for the amountoffluorideinyourunknownmeasuredandin


the drinking water tested. Remember to include fluoride unknown #. Do the potential
values measured for the amount of fluoride in the drinking water sample fall in the linear

S16


74

portion of the calibration plot? If not, would you expect those values tobeasaccurateasif
theywereinthelinearportionofthatplot?

S16


75

GA16AStudyofAtomicAbsorption
This lab is designed to be an introduction to the method of atomic absorption. In atomic
absorption, the absorption of light by atoms is utilized as the basis of an analytical technique.
Since atoms have no vibrational or rotational energy levels, absorption is the result only of
electronic transitions and the atomic spectra consist of discrete lines corresponding to these
transitions.
The light source for atomic absorption is a lamp whose cathode is made of the investigated
element. The lamp emits light of wavelengths characteristic of this element. In the path of the
light source, solutions to be tested are aspirated into a flame, which serves to atomize the
materials in the solution. Iftheelementinquestionispresentinthesolution,itwillabsorbsome
of the light being emitted from the lamp. After passing through a monochromatortoisolatethe
wavelength of interest, the transmitted light is detected. If the element is not present in the
solution, all of the light will pass through the flame and be detected. In the right concentration
range, Beer's law will be followed and the concentration of an unknown can easily be
determined. For a more detailed information, refer to Harris6thEdition,Chapter21,7thEdition,
Chapter21,8thEdition,andChapter20.

SolutionsandChemicalsRequired
(a) 1000 ppm calcium (standard) (b) 1000 ppm aluminum (c) 50,000 ppm lanthanum (d) 1.2M
HCl (e) Calcium containing water sample (You might want to bring one in!) (f) Unknown
calciumsample.

Equipment
(a) Atomic Absorption Spectrophotometer (Perkin Elmer 3100) (b) Burner Heads, airacetylene
andnitrousoxideacetylene(c)GA16GlasswareKit

S16


76

InstrumentWarmup
Check with the instructor or the teaching assistant to besurethattheinstrumenthasbeenturned
on and is warming up properly when you are about to finish preparing standards and start
preparing unknowns. Instructor will brief you on principles and operation of the instrumental
techniquewhenstartinganalysis.

PreparationofSolutions
Check out the glassware kit for GA16. Rinse all of the glassware with new deionized water
before starting. Be sure to shake all solutions well failure to obtain a homogeneous solutionis
often responsible for impreciseresults! Prepareallthesolutionstogetherandthenmeasurethem
all at the same time to insure that the unknown samples are measured under exactly the same
conditions as the standards. There are four sets of solutions to be prepared: calcium calcium
and lanthanum calcium and aluminum and calcium, lanthanum, and aluminum. Perform
analysesusingairacetyleneflame.

a.CalciumSolutions
Youhavetopreparefourseriesofstandardscontaining015PPMofCalcium
(a)Ca(b)Ca+La(c)Ca+Al(d)Ca+La+Al.
Add2mLof1.2MHCltoeachstandardflaskbeforeaddingCaandotherreagents.
Using 10 mL graduated pipet, add requiredvolumesof75PPMCalciumstandardstocksolution
into50mLvolumetricflask.
CaSTD[ppm]

12

15

CaStocksolution[mL]

10

CastandardscontainingAl:Add5mLof250ppmAlsolution
CastandardscontainingLa:Add5mLof50,000ppmLaCl3

S16


77

b.UnknownSolutions
Prepare unknown calcium solutions using your unknown calcium samples previously obtained.
This must be done so that your calcium solution falls within the linearportionofthecalibration
curve. Dissolve 50._ mg of solid Ca unknown sample in 5 mL 1.2M HCl (addmoreifneeded)
anddilutetothemarkwithDIwaterin200mLvolumetricflask(SolutionA). Pipetteout10mL
of the solution in a 100 mL volumetric flask, add2mLof1.2MHClanddilutetothemarkwith
DIwater(SolutionB).UsesolutionBforanalysis.
Also, locate a drinking water sample left near the GA16 setup or you may bring one in from
home. Pipette 10.00 mL of water into a 25 mL volumetric flask, add 1.00 mL of1.2MHCland
5.00mLof10,000ppmK.Diluteto25.00mLwithdeionizedwater.

CleaningUp
Whenyouarefinishedwiththeexperiment,emptyandrinseallglasswarethoroughly.

Questions/TopicsfortheQuiz
1. (a) nebulization (b) atomization (c) flame (d) design, operation & significance of hollow
cathodelamp.
2. AdvantagesofAtomicSpectroscopyoverMolecularSpectroscopy
3. ChemicalInterference.

Calculations&/orQuestionsforPostlabReport
For the Report, just answer questions in the order they are asked, report will not be graded if
writtenintheessayformat.
4. SpecifyUnknown#.Reportwillnotbegradedifunknown#isnotspecified.
5. Include Absorbance vs. Concentration data for Ca series. Compare plots of absorbance vs.
Calcium concentration forthefollowingcombinations:(a)Ca(b)Ca+La(c)Ca+Al(d)Ca
+La+Al.

S16


78

6. Report concentration of Ca2+ in the unknown as Ca% basedonunknownamount. Calculate


uncertainty in estimation of Ca% (Harris: 6th Edition, page 87, 7th Edition,page7172,8th
Edition, page 8990). ExcelorGooglespreadsheetmustbeusedtoobtainuncertaintiesinx,
y, b and m. Harris has an example about using Excel (applies to GoogleSheetsalso)toget
these quantities. Copy paste Excel/Google Sheet data into the document showing LINEST
output, x,ynandkvalues. Samplecalculationsforsxneedtobeshown. GiveamountofCa
and uncertainty in Ca expressed as (wt/wt %) based on the unknown amount weighed out.
Thus, you need to convert X and sx (mg/L or ppm) into (mg)andthendividewithunknown
amount.Remembertoincorporatedilutionfactor.
7. Report ppm of Ca2+ in the water sample and uncertainty in the estimated value, comment if
needed. Why 10,000 ppm of K+ was added to the water sample? (Harris: 7th Edition, page
467,8thEdition,page493)
8. CompareresultsofGA4andGA16forCaunknownforprecision.

S16


79

GA20 High Speed Liquid Chromatographic Separation of Various


CompoundsFoundinCoffee,TeaandSoftdrinks

High performance liquid chromatography (HPLC) has the following advantages over open
column chromatography: speed, better resolution, and the availability of oneormoredetectors.
HPLC has the following advantages over gas chromatography (GC): ability to analyze for
thermolabile or nonvolatile compounds and, with the availability of the proper column, to
separate high molecular weight compounds. HPLC can also be easily scaledup from nanogram
(with fluorescence, electrochemical and mass spectroscopy detectors), to microgram (withfixed
or variable wavelength absorbance detectors), to the milligram range (with refractive index
detector). Inthisexperiment,aknownsampleofthefollowingfivecompoundswillbeseparated
by HPLC: saccharin (S), aspartame (A), sodium benzoate (B), caffeine (C), and sorbic acid
(SA).

From the chromatogram of the Coffee & Soda the presence or absence of these

compounds will be determined. (Remember to carefully read the label ofthesodabottle,asthe


manufacturers by law are required to list the major components.) This experiment may be
modified for the analysis of caffeine in brewed coffee or tea by decreasing the strength of the
mobile phase from 19 to 10 per cent byvolumeoftheacetonitrile. Knownstandardsofcreatine
and theophylline as well as the caffeine should also be used. Below are the structures of the
compoundsthatmaybecontainedintheknownstandardmixture.

Aspartame

SodiumBenzoate

Caffeine

Saccharine

As with all chromatographic methods, HPLC relies on the distribution of analytes between the
stationary phase and the mobile phase. A detector is used to determine when each component

S16


80

elutes from thecolumn. Thetimeittakesforelutionofapeakiscalledtheretentiontime,trand


is characteristic of a given component under a given set of experimental conditions. For
example, if the detector is an ultraviolet spectrophotometer, a plot of absorbance at a given
wavelength vs. time is generated. Such a plot is calledachromatogram. Aseachcomponentof
the sample passes through the detector, a peak is generated. Ideally, both peak area and peak
height are proportional to the component's concentration in the sample. Standards are run for
each component in order to determine the relationship between peak size or peak area and
concentration.

Saccharin (S), aspartame (A), sodium benzoate (B)andcaffeine(C)areallpolarcompoundsnot


easily separated using "normal phase" HPLC columns of silica or alumina absorbent with an
organic solvent. However, "reverse phase" HPLC columns, where the solid phase is a support
altered by a coating or bonding with a nonpolar phase and the mobile phase is a polar solvent,
such as acetonitrile/water, produce good separation of such polar compounds. In reverse phase
HPLC, the mobile phase often consists of two solvents: one solvent in whichthesampleisvery
soluble (acetonitrile) and the other, water, in which the sample is less soluble. Byadjustingthe
proper ratio of "good" to "poor" solvent by trialanderror or by such modern methods as
"mixture design optimization statistics", a solvent mixture is foundthatadequatelyseparatesthe
componentswithinareasonabletimeframe.
The optimum solventmixtureforseparatingS,A,BandConthereversephasecolumnusedin
this experiment was found to be19%acetonitrileand81%deionizedwatercontaining2%acetic
acid and 0.5% ammonium acetate. The acetate buffer system (pH 3.9)isaddedtoprotonatethe
benzoateionandtoimprovepeaksymmetry.

Instrumentation
The HPLC apparatus consists of a highpressure pumping system, which draws mobile phase
fromasolventreservoirandforcesitthroughthechromatographiccolumn. Theinjectorusedfor
introducing a fixed volume of the sample is located between the pumpandthecolumn. Sample
components, separated on the column, are transported by the mobile phase through a detector
where they produce a signal. Since allofthecompoundsanalyzedabsorbUVlightat254nm,a

S16


81

fixed wavelength, UV detector is used in this experiment. (Note it is simply required that the
analyteshavesomeabsorbancebutnotnecessarilymaximumabsorbanceat254nm)
The recorder/Integrator plots the detector signal vs. time, producing a peak when a
UVabsorbing component passes through the detector. The elapsed time between injection and
the peak maximum is called the retention time (tr), and this time is characteristic of the
component under the experimental conditions. The size of the peak (height or area) is
proportionaltotheamountofthecomponentinjected.
In general, the mobile phase used must be (degassed) in order to remove entrappedairbubbles
which are deleterious to the proper
operation of the pump and the detector.
Degassing is most easilyaccomplishedby
stirring the mobile phase overnight.
Alternately themobilephasemaybemore
rapidly degassedbyspargingwithhelium,
placing in an ultrasonic bath, or slowly
revolvingonarotaryevaporator.
The HPLC system can be damaged by allowing unfiltered samples to enter the system. Hence,
the mobile phase and all solutions to be injected must be filtered through a submicron HPLC
filter disk (preferably 0.22 m for a 3 or 5 m size particle column). The laboratory staff will
have previously prepared mobile phase, solutions of the known standards, and the
multicomponent unknown solution so this experiment can be completed in a 3hour laboratory
period. Usually there are stainlesssteel filters inthepumpandattheentranceofthecolumnsto
keep out any impurities remaining after filtration. This is very necessary because the liquid is
being forced through a very narrow opening: the inner diameter of the interconnectingtubingis
0.007 to 0.010 inches. A clogwillresultinrapidbuildupofpressureinthesystem. Thestudent
should note the normalpressureofthesystem. MostHPLCwillautomaticallyshutdownifan
upper pressure limit is reached. The pump and detectorwillnotfunctionproperlyifthereareair
bubbles in the system. Air bubblescanusuallybeidentifiedbyerraticchangesinpressureorby
short,sharp"wanderings"ontherecorderoutputasshowninthefigure.

S16


82

Equipment
High Performance Liquid Chromatograph, equipped with an injector and a fixed, 254 nm UV
detector,Recorderorintegrator,2magnetstirringmotors.

SolutionandChemicals
Acetonitrile, ammonium acetate, glacial acetic acid, solid saccharin(S),Aspartame(A),Sodium
Benzoate (B), and Caffeine (C). The proper name for aspartame is LAspartylLPhenyalanine
Methyl Ester.

Filtered mobile phase for the HPLC and for rinsing the syringe and injector

Individual solutions of S, A, B and C of known concentrations. Solution of a mixture S, A, B


andCofunknownconcentrations.

Glassware
(a) 25 L syringe for introducing samples into the injector (b) 10 mL glass syringe, equipped
with LuerLok to filter all samples (c) HPLC, 0.22 m, pore size filters for both mobile phase
and all individual (d) samples (MillexGV, 25 mm diameter, and Millex GV4, 4 mm
diameter, Millipore Corp., New Bedford, MA or comparable) Glassware kit containing:(a)30
mL beaker: 1 (b) 100 mLbeaker:1(c)2mLVolumetricflasktoquantitativelydilutethesample
for the spiking experiment (e) 1 mL volumetric pipets for the spiking experiment: 5 (f) 1 pipet
bulb (g) 2 stirring bars, Teflon coated (h) 7 Small glass vials (7 mL) with Teflon lined tops
(i)25mLvolumetricflaskstoholdtheconcentratedanddilutedstandardsolutions:12.

GettingStarted
a.AdjustmentofSolventFlow
Do not start this step without consulting the laboratory instructor. The lines from the solvent
reservoir to the pump should be carefully examined for air bubbles. Air must be removed
completely before starting the pump. Checktheflowratespecifiedfortheinstrumentationused.
If the pressure goes above 3000 psi, stop increasing the flow and alert the instructor. If air

S16


83

bubbles are observed inthelineduringtheexperiment,orifthemobilephasereservoirrunslow,


notifyinstructor.
b.FillingtheSampleLoop
A 50l syringe is used to fill a 10 or 20 L loop
sample loop. Filling must be done with the injector
valve in the load position. To introduce the sample
into the column, the valve is turned to the Inject
position. Do this switching in one smooth motion.
Stopping at an intermediate injection position between "load" and "inject" causes a rapid risein
system pressure & damages system. Figure illustrates the flowpathsthatoccurinthissocalled
sixportvalveandloopinjector.
c.Injectionofthesample
i.

Cleanthe25Lsyringebyrinsingseveraltimeswiththemobilephase.

ii. Rinsethesyringeseveraltimeswiththefirststandardcomponentsolution.
iii. Fill the syringe with standard solution. Invert the syringe and remove any air bubbles by
tapping and/or plunging the syringe needle into the solution to coax the bubbles out.
Alternately, make sure that any air bubbles in the sample stay directly underneath the
plunger andarenotinjectedintotheinjector. Expelasmalladditionalamountofliquidfrom
the syringe tip to be sure there are no bubbles hidden in theneedle. Gentlytapthebarrelof
the syringe so that any remaining air bubbles remain towards the top under plunger. Make
sure that these airbubblesarenotpushedintotheinjectorandsubsequentlyinjectedontothe
column.
iv. With the injector valve in the "load" position, fill the sample loop by expelling the solution
fromthesyringe.Makesurethatnoairbubblesareinjectedintotheloop!
v. Repeat Steps iii and iv twice, to be certain that any previous sample and mobilephasehave
beenflushedfromthesampleloop.RemoveSyringefromtheinjectionport.

S16


84

d.IntroductionofSampleintotheColumn,andRecordingofthePeak
Introduce the sample by moving thevalvefromthe"load"tothe"Inject"positioninonemotion.
Simultaneously, press the Start button on the integrator. Leave the valve in the "Inject"
position for at least one minute. Then return the valve to the "load" position. Once, sample is
completely eluted, press Stop and integrator will print results. Repeat sample injection to
check reproducibility of results. Be aware that one of the common errors in this laboratory
experimentistoinjectthenextsamplebeforetheentireprevioussamplehaseluted.

e.IdentificationofComponentsinMixtures
Analyze standard solutions to determine the retention times, analyte response (peak height) and
elutionorderofS,A,BandC.
Inject multicomponent unknown mixture to identify and quantify if S, A, B and C are present.
This will serve to check that the column is properly separating the 3 or 5component mixture.
Repeat the injection of this component mixture until reproducible peak heights are obtained.
Tworeproducibletrialsareneeded.

f.TheoreticalPlates&PeakSymmetry
For one trial only, increase thechartspeedbyafactorof4andinjectthissamemulticomponent
mixture. This chromatogram will be used fortheoreticalplateandasymmetrycalculationstobe
discussedlater.Allotherchromatogramsshouldberunattheslowerchartspeed.

g.TheSoftDrinkorCoffeeand/orteasample
Noteallsamplesmustbecooledorwarmedtoambienttemperatures!
SoftDrink:
Place about 50 mL of the sample in a 100 mL beaker and slowly stir magnetically until the
evolution of air bubbles (effervescence) ceases (1530 min). Filterthedegassedsamplethrough
a proper micron HPLC filter disc. (See instructor for technique.) Note all unknown samples,

S16


85

standards and even the mobile phase must be carefully filtered in HPLC to remove dust and
insolublematerial!
Inject the soft drink sample at least twice. There will be many peaks. Make sure the entire
samplehaselutedbeforeinjectingthenext.
Coffee:
Dissolve 30 mg of coffee in 10 mL DI water. Filter the sample throughapropermicronHPLC
filter disc. Inject the coffee solution at least twice. There will bemanypeaks,makesureentire
samplehaselutedbeforeinjectingnext.
Spikedcoffeesample:
Identify retention time for caffeine peak in the coffee sample. Itissuggestedthatyoudilutethe
same volume of the filtered sample withexactlythesamevolumeoftheknownstandards. Pipet
in 100 L of filtered coffee sample and 100 L caffeine standard in a small vial, mix well and
inject the spiked coffee sample. Note whether any additional peak appears or whether the
existing peaks either increase or decrease. (Explain your observations in the Result and
Discussion Section of your Laboratory Report). Repeat the injection of the spiked sample and
checkforreproducibility.Tworeproducibletrialsareneeded.

InstrumentShutdown
When you are sure that you have finished,injectthreeportionsofmobilephaseontothecolumn
in the normal way.

This should flush out all"junk"stillinthesystem. Thebaselineshouldbe

stable,andthereshouldbenoevidenceoffurtherpeakswhenyouarefinished. Informinstructor
whowillthenshutdownthemachine.

RecordingAllExperimentalConditions
Make sure you have recorded all concentrations ofknownsamples,andallparametersincluding
columnidentificationandmobilephasecomposition,chartspeed,attenuation,etc.

S16


86

CalculationsandQuestionsfortheLabPaper
AnswerfollowingquestionstowriteTheorysection:

1. Explainreversedphasechromatography
2. Discusstypesofchromatography(Harris:7thEdition,page5068thEdition,page542)
3. Discuss Isocratic and Gradient Elution. Criteria for selecting the separation mode (Harris:
7thEdition,page565,5668thEdition,page604,607)
4. Explain(a)ImportanceofmobilephasepH(b)design,featuresofUVdetector.
Experimental Section should include (a) changes made to the lab manual procedure (b)

amountsweighedouttopreparesolutions(c)unknown#ifany.
AnswerfollowingquestionstowriteResultssection:

5. SampleIdentificationandQuantitation:
a. Calculatetheaverageretentiontimeofeachcomponent.
b. Reporttheelutionorderofcomponentsinthemixture.
c. Calculatetheaverageconcentration(mg/ml)ofeachcomponentintheunknownmixture.
P eakHeightinU known
P eakHeightinStandard

mg

ConcentrationinU nknown( mL )
mg
ConcentrationinStandard( mL )

6. Identify any of thestandardcomponentswhichthesoftdrink,CoffeeorTeacontains. Using


the known spiking concentration, calculate the amount of at least one component present in
the soft drink, CoffeeorTea. (Alinearrelationshiphaspreviouslybeenestablishedbetween
peak heightandconcentrationforeachofthefourcompounds). Donotforget,however,that
in the process of spiking the soft drink soft drink, Coffee or Tea, the concentration was
decreased. Indicatewhetheranyadditionalpeakappearsorwhethertheexistingpeakseither
increaseordecrease.
Caf f eineP eakHeightinSpikedSample
Caf f eineP eakHeightinU nspikedSample

0.5(X Sample )+0.5(StandardConcentration)


(X Sample )

S16


87

AnswerfollowingquestionstowriteDiscussionsection:

7. Compare the sensitivity of this method for the determination of aspartame and caffeine.
Explainwhythesensitivitiesaredifferent.
8. PeakCharacteristics:

a. Findthepeakasymmetry,b/a,foreachcompound,asshowninFigure3. Reportwhether
eachpeakisaleading(b/a<1)ortailing(b/a>1)peak.
b. Calculate the number of theoretical plates, n (a measure of column efficiency) for each
2

peakinthemixture. n = 5.54( wtr ) .


1
2

tr =retentiontime&

w 1 =peakwidthathalfheight
2

Itisimportantthattheunitsusedinthisequationarethesameforboth tr and

w 1
2

Use ruler for measuring width and distance of unknown peaks from chromatogram
obtainedathigherchartspeed(4cm/min).

S16


88

GA21SpectrophotometricDeterminationofIron
Iron(II)formsastable,highlycoloredcomplexwith1,10phenanthroline.
Fe3+ does not form the complexandthusforthisanalysistobesuccessful,alloftheironmustbe

reduced from Fe3+ to Fe2+. In this experiment, the reduction is done with hydroxylamine
hydrochloride. Intensity of the color is independent of pH in the range 2 to 9 solution is
bufferedwithsodiumacetate.

SolutionsandChemicals
(a) Ferrous Ammonium Sulfate

hexahydrate (Mohrs Salt) (b) 10% hydroxyl amine

hydrochloride, (c) 0.1M Sodium Acetate (d) 1,10 phenanthroline monohydrate solution (2.5
mg/mL) (e) Unknown Iron samples (do not have to be dried). Special equipment: (a) 50 mL
volumetric flasks: 6 (b) 100 mL volumetric flasks: 2 (c) 1 mL volumetric pipet: 1 (d) 2 mL
volumetric pipet: 1 (e) 5 mL volumetric pipet: 2 (f) 10mLgraduatedpipet:1(g)10mLbeaker:
3(h)Spectrophotometer

PreparationofStandardSolutions
1. Weigh approximately 0.28 g of ferrous ammonium sulfate hexahydrate, [FeSO4
(NH4)2SO4]6H2O, to the nearest milligram on the analytical balance. As always, check the
purityfactor.
2. In a 250 mL volumetric flask, with deionized water, dissolve theferroussaltandthendilute
tovolume.Mixwell.

S16


89

3. Using your 25 mL pipet and a 100 mL volumetric flask, dilute 25 mL of the solution
preparedinstep2to100mLwithdeionizedwater.Mixwell.
4. To prepare the standard Fe solutions,pipetinsuccession1.00,2.00,3.00and4.00mLofthe
dilute Fe solution mixed in step 3 into separate 50 mL volumetric flasks. Into each of
these volumetric flasks, carefully pipet in the volumes ofhydroxylamineHClsolution,0.lM
sodium acetate solution, and 1, 10 phenanthroline solution shown in the following table.
Carefully dilute these solutions to volume (to the 50mLmark)withdeionizedwaterusinga
medicinedropper.Mixwell.Also,prepareablank,thesamewaybutcontainingnoiron.

FeSolution,mL

HydroxylamineHCl,mL

SodiumAcetate,mL

1,10Phenanthroline,mL

Complex1to4

Blank

PreparationofUnknownSolution
1. Weigh out 1.4 g to the nearest mg of unknown iron sample (does not have to be dried) and
dilutetovolumewithdeionizedwaterina250mLvolumetricflask.Mixwell.
2. Dilute 5.0 mL of the above solution to 100 mL with deionized water. Mix well. Thisisthe
stockunknownsolution.
3. Make a solution in the sixth 50mLvolumetricflask,asinstep4,using4mLofthestockFe
unknown solution and thesameamountsoftheothersolutionsaswasusedforthestandards.
Dilutetovolumewithdeionizedwater.Mixwell.

Spectrophotometermeasurements
1. Ask laboratory Instructor to help you with setting up the instrument. Spectrophotometer is
locatedinRoomT411.
2. Set the wavelengthtothevalueofthepeakmaximum515nmoftheFe1,10Phenanthroline
Complex.

S16


90

3. Rinse and fill each cuvette with deionized water to 70% capacityandplaceintheholdersin
thecellcompartmentofthespectrophotometer,closethelidandclicktheiconAutoZero.
4. Remove the front sample cuvette from the sample compartment. (The rear one is the
referenceandshouldremainthere.)
5. Fill the sample cuvette with blank solution to 70% capacity and then close the lid. Click
on the blank icon. Instrument reports 3 measurements of the same solution and then the
averagevalue.
6. Fill the cuvette with the less concentrated standard solution. Place this cuvette in the cell
compartment and measure the absorbance. Successively, measure the absorbance readings
oftheotherstandardsolutionsandtheunknownsolution.
7. Prepare a plot of Absorbance vs. mL Fe solution in Microsoft Excel or Google Sheets to
make sureitislinear. Dothiswhenyouarestillinthelaboratorywiththeinstrumentstillon
and solutions can be easily remade. To check linearity, it is not necessary to calculate
mg/mL Fe for the xaxis values. Simplyplottingabsorbancevs.mLFesolutionwilldo.(Do
youknowwhy?)
8. Are you satisfied with the linearity of the calibration obtained using the standards?
Generally a linear correlation coefficient (r) of 0.98 or better is necessary. If this is not
achieved, redo all or at least part of the calibration plot and again verify the linearity of the
standardsolutions.
9. If the absorbance of the unknown does not fall on the calibration curve make a more
concentratedordilutesolutionusingamoresuitablevolumeofstocksolution.

TopicsfortheQuiz
1. Spectrophotometry principles (Harris, Ed8 Ch. 172) & Instrumentation (Harris, Ed8 Ch.
19,page445446).
2. ImportantaspectsofthemethodusedfortheFedetermination.

S16


91

Calculations&/orQuestionsforPostlabReport
For the Report, just answer questions in the order they are asked, report will not be graded if
writtenintheessayformat.
1. SpecifyUnknown#.Reportwillnotbegradedifunknown#isnotspecified
2. Prepare a calibration plot of Absorbance vs. mg/mL of Fe (and not ferrous ammonium
sulfatehexahydrate).IncludeAbsorbanceandConcentrationdatainatable.
3. From the plot and the absorbance of the unknown solution, find the mg/mL of Fe in the
measuredunknownsolution.
4. Note the unknown Fe solution was first diluted to 250 mL, then that solution was diluted 5
mL to 100 mL. Finally 4.00 mL of the second solution was diluted to 50 mL. Find the
mg/mLoftheoriginalunknownsolution.
5. Knowing the volume of the original unknown solution, find the total mg Fe in the sample.
Calculate uncertainty (sx) in estimated amount of Fe. (Harris: 6th Edition, page 87, 7th
Edition, page 7172, 8th Edition, page 8990). Excel or Google spreadsheet mustbeusedto
obtain uncertainties in x, y, b and m. Harris has an example about using Excel (applies to
Google Sheets also) to get these quantities. Copy paste Excel/Google Sheets data into the
document showing LINEST output, x, y n and k values. Sample calculations for sx need to
beshown.
6. Give amount of Fe and uncertainty in Fe expressed as (wt/wt %) based on the unknown
amount weighed out. Thus, you need toconvertXandsx(mg/mL)into(mg)andthendivide
withunknownamount.Remembertoincorporatedilutionfactors.

S16

Вам также может понравиться