20152016FALLSEMESTER

MARMARAUNIVERSITY
DEPARTMENTOFBIOENGINEERING

CHEM213
BIOCHEMISTRYI
EXPERIMENT1
PREPARATIONOFBUFFERSOLUTIONS
SelenENAL150813035
HandeBEKLEN150813025
AbuzerESEN150812035
ZeynepEcemBASKI150813034

DATEOFEXPERIMENT:
06.10.2015
DATEOFSUBMISSIONOFTHEREPORT:
13.10.2015
SUBMITTEDTO:
ASSOC.PROF.BERNASARIYARAKBULUT
RESEARCHASSISTANTS:
FATMAGZEMAVCI,CEYDAKULA
ONURSERNOLUZEYNEPYILMAZ

INTRODUCTION
pH
ThepHhasthreedefinition.Therearesimple,theoreticalandexperimental.Simpledefinition
isthatpHisalogarithmicmeasureofhydrogenionconcentration.Thiswasdefinedby
Danishbiochemist
SrenPeterLauritzSrensenin1909.CompactOxfordEnglishDictionary
thepmeanspowerinGermanthiswordispotenz,sopHisanabbreviationforpower
ofhydrogen

pH=log[
H+

]
+
+
Thetheoreticaldefinitionismoreaccurately.pHisdefinedaspH=loga
where
a
H
H isthe
hydrogenionactivity.

TheexperimentaldefinitionisthatIUPAChasapprovedapHscalebasedonastandard
bufferofknownpHusingelectrochemicalmeasurements.[1]
Buffer
Abuffersolutionismixtureofaweakacidanditsconjugatebaseoraweakbaseandits
conjugateacid.IthelpstoresiststochangesinpHwhensmallamountsofacidorbaseadded
toit.Inadditiontopreapareabuffersolutionitmustcontainlargeconcentrationofacidto
reactitsconjugatebaseandalsomustcontainsimilaramountofbasetoreactitsconjugate
acidandtheacidandbasecomponentsmustbeneutralizeeachother.Therearetwotypesof
buffersolution.Firstoneisacidicbuffersolutionandsecondonealkalinebuffersolution.
AcidicbuffersolutionhasapHlessthan7.Itcontainsaweakacidanditssaltconjugatebase.
AlkalinebuffersolutionhasapHgreaterthan7.Itcontainsaweakbaseandoneofits
conjugateacid.[2]Buffersolutionsareusedseveralareas.Theseare

infermentation
intextileindustry
inchemicalanalysis
calibrationofpHmeters
keepingpHatanearlyconstant

OntheotherhandtomeasurepHthereisaequationthatcalledasHendersonHasselbach.
TheHendersonHasselbachhelpstomeasurepHofabuffersolution.Thebasicequationas
shown:
HA(weakacid)

(saltconjugatebase)+
H+

Ka=[
H+

][
A

]/[HA]whentakenpbothsideandleavealonethepH,theequationtoget:
pH=pKa+log[
A

]/[HA]
HoweverthereisaequilibriumthatcalledasLeChateliersprinciple.Whensomestrongacid
isaddedtoanequilibriummixtureoftheweakanditsconjugatebase,theequilibriumis
shiftedtotheleftinaccordancewiththisprinciple.

This experiment was performed tounderstandcalculatingthepHofbuffersolutionsand how

adding different solutions effect the pH changes.Also,observingthedilutionprocesswasone
ofthemainpurposes.
MATERIALS

Erlenmeyerflask
pHteststrips
Washingbottle
Volumetricpipet
Graduatedcylinder

0.10M50mLNaOH(sodiumhydroxide)
0.10M50mLHCl(hydrochloricacid)

0.50M200mL
C
H
O

2
4
2
2(aceticacid)

0.50M200mL
KCH
COO
(potasiumacetate)
3
0.50M200mL
NH
Cl

(ammoniumchloride

4
0.50M200mL

NH
OH(ammoniumhydroxide)
4
Enoughdeionizedwater

METHOD
PreparationandpHmeasurementofbuffersolutions

1)Firstofall,classwassplittedintotwogroups.Aceticacidandpotasiumacetatewere
inspectedbyfirstgroup.Ammoniumchlorideandammoniumhydroxidewereviewedby
secondgroup.

2)Aceticacidandpotasiumacetatesolutionsweremixedinthestatedamountof1,3and4
fromtable1.Later,ammoniumchlorideandammoniumhydroxidewerepreparedinthe
statedamountof1,3and4.Also,water(20mL)wasprepared.Afteramixingprocess,
solutionswerelabeled.

3)GroupmembersmeasuredthepHofthesolutionsandrecordedthem.

Solutionno

Volumeofaceticacidor
ammoniumchloride(mL)

Volumeofpotasiumacetate
orammonimhydroxide(mL)

10

10

15

18

18

15

Table1:requiredvolumestopreparebuffersolutions

Examinationoftheeffectoftheadditionofastrongacid/base

1)According to table 1, exact amount of solutions were created and named as 1A,3A,4Aand
WA.

2)Foreachsolution,pHwasmeasuredandrecorded.

3)0.10M 10mL
HCl was added the solution of 1,3,4 and water.After the firstprocess,0.10M
10mL NaOH was added to 1A,3A,4A and WA.Both of thesolutionsweremixedproperly.At
theendoftheprocedure,pHwasmeasuredandrecorded.

Examinatonoftheeffectofdilution
1)Each group applied the procedure of dilution for only solution 1(acetic acid and potasium
acetateorammoniumhydroxideandammoniumchloride)fromtable1.

2)10mL of 0.50M acetic acid and10mLof0.50Mpotasiumacetatewereput theflaskbyfirst

group.Then, 80mL deionized water supplemented to the solution for preparing 1:5 diluted
solution.

3) After this procedure, 10mL of 1:5diluted solution was poured into the beaker and 90mL
deionized water was added above the solution.Thusly group had 1:50 diluted solution.At the
same time, second group applied the same process by using ammonium hydroxide and
ammoniumchloride.

4)Intheend,pHofthedilutedsolutionsweremeasuredandrecorded.

RESULTS

1)
PreparationandpHmeasurementofbuffersolutions

For0.50M200mlacedicacidand0.50M200mlpotassiumacetate:
a)Forsolution1:
0.5Mx0.01Lacedicacid=0.005moleacedicacid
0.5Mx0.01Lpotassiumacetate=0.005molepotassiumacetate
log([base]/[acid])=log(0.005/0.005)=log1=0
b)Forsolution3:
3

0.5Mx0.018Lacedicacid=9x10moleacedicacid
3
0.5Mx0.002Lpotassiumacetate=1x10
molepotassiumacetate
3
3
log([base]/[acid])=log(9x10
/1x10
)=log(9)=0.95
c)Forsolution4:
3
0.5Mx0.002Lacedicacid=1x10
moleacedicacid
3
0.5Mx0.018Lpotassiumacetate=9x10
molepotassiumacetate
3

log([base]/[acid])=log(1x10/9x10)=log(0.11)=0.96

Plot 1

2)
Examinationoftheeffectoftheadditionofastrongacid/base

a)
Forsolution1:
5

CH

3COOHK

A=1.75X10

CH
+H

3COOHCH

3COO

+
K
]X[H
])/[CH
A=([CH

3COO

3COOH]

[CH
3COOH]=[C

2H
3KO
2]

[H
]=(K
AX[CH

3COOH])/[C

2H
3KO
2]

[H
]=K

A=1.75X10

+
pH=log[H
]

pH=log(1.75x10)=4,76
Forsolution3:
pH=pKa+log[base/acid]
pH=4.76+0,95=5,71
Forsolution4:
pH=pKa+log[base/acid]
pH=4.760.95=3.81
b)
For10mlof0.1MHClwasadded
[CH3COOH]=6x103/0.02=0.3M
[CH3COO]=4x103/0.02=0.2M
PH=pKa+log[base/acid]=4.76+log(0.2/0.3)=4.58
For10mlof0.1MNaOHwasadded
[CH3COOH]=4x103/0.02=0.2M
[CH3COO]=6.x103/0.02=0.3M

pH=pKa+log[base/acid]=4.76+log[0.3/0.2]=4.93
Examinationoftheeffectoftheadditionofastrongacid/base
Group 1 : (HCl added into 1A2A3A4Asolutionsforeachgroup,NaohaddedintoS1S2S3
S4solutionsforeachgroup)
G11A:10mlof0,5MCH3COOH10mlof0,5MCH3COOK+10mlof0,1MHCl
V230ml0,03L
[CH3COOH]=(0,5*0,01+0,1*0,01)/0,03=0,2M
[CH3COOK]=(0,5*0,010,1*0,01)/0,03=0,13M
pH=4,57
G12A:18mlof0,5MCH3COOH2mlof0,5MCH3COOK+10mlof0,1MHCl
[CH3COOH]=(0,5*0,018+0,1*0,01)/0,03=0,3M
[CH3COOK]=(0,5*0,0020,1*0,01)/0,03=0M
G1S1:10mlof0,5MCH3COOH,10mlof0,5MCH3COOK+10mlof0,1MNaOH
[NH4Cl]=(0,5*0,010,1*0,01)/0,03=0,13M
[NH4OH]=(0,5*0,01+0,1*0,01)/0,03=0,2M
pH=4,95
G1S2:18mlof0,5MCH3COOH2mlof0,5MCH3COOK+10mlof0,1MNaOH
[NH4Cl]=(0,5*0,0180,1*0,01)/0,03=0,26M
[NH4OH]=(0,5*0,002+0,1*0,01)/0,03=0,06M
pH=8,6

3)
Examinationoftheeffectofdilution

0.5 M acedic acid and 0.5M potassium acetate will be diluted.10ml of 0.5M acedic acid and
10ml of 0.5Mpotassiumacetatesolutiondilutedto100mlonebyone,diluterateis1:5.Later
the rate of dilution is 1:50 , 10 ml of each matter diluted in another beakers. The
concentrationofeachsolutionbecomes0.005M.Finally20mlbuffersolutionisobtained:
[CH
3COOH]=(0.05x0.01)/0.02=0.025

[C
2H
3KO
2]=0.025M

pH=4.76+log1=4.76
1/100dilutedsolutionsaremixed

[CH
3COOH]=(0.005x0.01)/0.02=0.0025

[C
2H
3KO
2]=0.0025M

pH=4.76

Solutionno

pH

HCl+pH

NaOH+pH

1:5

1:50

1/1A

5/4.5

4.5

3/3A

4/4

3.5

4/4A

6/6

5.5

6.5

W/WA

5/5

1.5

13

Table2:forpotasiumacetateandaceticacid

Solutionno

pH

HCl+pH

NaOH+pH

1:5

1:50

1/1A

10/10

9.5

10

10

3/3A

9.5/10

8.5

10

4/4A

11/11

10

11.5

W/WA

5/5

1.5

13

Table3:forammoniumhydroxideandammoniumchloride

DISCUSSION

In this experiment we observed the pH of buffer solutions.First of all we prepared buffer

solutions as asetic acid and potassium acetate or ammonium chloride and ammonium
hydroxide in different volumes and measured their pH of solutions. After this process we
added some strong acid and some strong base. After all these procedures groups observed
differences between ph of the solutions. This is because of the bubbles which stays in the
volumetric pipet also mistakes in the process of takenvolumes.Accordingtothisexperiment,
while buffer is prepared,if weak acid rate is more.pH valueis much closer to 0. If weak base
rate is more, pH value is much closer to 14. This means acidity and alkalinity is stronger.
When strong acid/strong base is added to buffer, pH values change very slightly.But when
strong acid/strong base is added to water, pH changes too big and it happens too quick.
Because waterhasnobuffercapacity.ItcannotresistpHchanges.Inbufferthereisweakacid
and its conjugate base or weak base and its conjugate acid.Whenacidorbaseisadded,buffer
neutralise acid or buffer. With this way, pH values keep almost constant.Dilution's effect on
bufferistoosmall.Afterdillution,pHvaluesarealmostsamewithundilutedbuffer'spH.

REFERENCES

[1]
http://antoine.frostburg.edu/chem/senese/101/acidbase/faq/whatispH.shtml#2
/received in
9.10.2015
[2]
http://chemwiki.ucdavis.edu/Physical_Chemistry/Equilibria/AcidBase_Equilibria/7._Buff
er_Solutions
/received in 9.10.2015