pH MEASUREMENT AND BUFFER PREPARATION Jarvis Kelly C. Tan, Christian Roel M. Tribujenia, Joebert Edison B. Velasco, Camille V.

Viray, Daniel Carlo P. Won !rou" # $E Medical Technolo y Biochemistry %aboratory

ABSTRACT
The aim of this experiment is to able to prepare different buffer solutions, to determine the pH of the buffers and samples colorimetrically by using different liquid indicators and electrometrically using the pH meter and also to calculate the buffer capacity of the prepared buffer solutions. In this experiment, secondary sodium phosphate di-hydrate (Na H !"# $ H "% and phosphoric acid (H&!"#% 'as used in order to create the buffer solution. In electrometric method, the pH meter 'as used to read the pH of the prepared buffer solution( 'hile in colorimetric method, the pH of the prepared buffer solution 'as measured by using the acid-base indicators gi)ing the results of blue for Thymol blue, blue )iolet for bromophenol blue, light blue for bromocresol green, purple for bromocresol purple, pin* for phenol red, none for methyl red, orange for methyl orange and colorless for phenolphthalein. +nd due to the the lac* of time, determination of buffer capacity 'as not possible.

INTRODUCTION ,)ery biological process is greatly affected by pH. In food processing, pH measurement is )ery important because it is applied to a)oid health problems to the consumers and to produce products 'ith consistent 'ell defined properties-./. In blood cells, the pH of the blood should be maintained at 0.#. If the pH drops belo' 1.2 or rises abo)e 0.2, death may occur-1/. The term pH is a 'ay of expressing lo' concentrations of ions.-3/ In the pH measurement, 4p4 stands for the 5erman 'ord poten6, meaning po'er, and the 4H4 for hydrogen.-./ It is used to determine the acidity or basicity of a solution. If the pH of a solution is 0 then the solution is neutral( any pH lo'er than 0 is considered acidic 'hile any pH higher than 0 is considered as basic. The pH (hydrogen ion potential% of a solution is defined in (7ig. 3% 'here -H8/ is the hydrogen ion concentration.-#/ pH 9 - log3:-H8/ 7igure 3. pH ,quation In determining the pH of a substance is 'ith the use of pH meter. It consists of a special type of glass electrode connected to an electronic meter that measures and displays the pH reading. + buffer solution is a mixture of a 'ea* acid or base and its con;ugate base or acid that can 'ithstand sudden changes in pH 'hene)er amounts of acid or base is added. It also neutrali6es excess hydrogen ion. -3/ 7or buffers, the HendersonHasselbalch equation in (7ig. % relates the "& o'
an acid and the "( o' the solution. Based on the solution, the e)uation can be used to calculate the amount o' acid and its conju ate base.*+,

Fi !re ". Hen#ers$n%Hassel&al'( E)!a*i$n

E+PERIMENTA, A. C$-p$!n#s *es*e#

Distilled -ater, concentrated (.P/0 123.45 -6-7, concentrated (Cl 1.8..5 -6-7, lacial acetic acid 1##.85 -6-7, 9a($P/0.($/, 9a$(P/0.8($/, 9a/( "ellets and the acid:base indicators 1thymol blue, bromo"henol blue, bromocresol reen, bromocresol "ur"le, "henol red. Methyl red, methyl oran e and "henol"hthalein7 -ere the com"ounds used in the e;"eriment.

B. PROCEDURE .. PREPARATION OF REA/ENTS

< =olume o' 344m% o' a)ueous solution -as "re"ared by usin a bea>er then it is trans'erred to a container to be labelled "ro"erly.

". BUFFERS

Each rou" -as assi ned to "re"are $34m% 4.+4M o' bu''er solution de"endin on their rou" number.

2.a. Prepara*i$n $0 '$l$!r s*an#ar#s !sin *(e &!00er s$l!*i$ns

Ta&le .. Prepara*i$n $0 B!00er S$l!*i$n

!rou"
+ $ . 0 3,B,8 2 #

Bu''er ?olutions
Phos"hate@ "& A $.+$ Phos"hate@ "& A $.+$ <cetate@ "& A 0.84 Phos"hate@ "& A 8.$+ Phos"hate@ "& A 8.$+ Phos"hate@ "& A 8.$+ Phos"hate@ "& A +$..$

Desired "(
$.44 ..44 3.44 8.44 8.34 2.4 +$.44

Pre"are B test tubes. Then label each test tube -ith the "( o' bu''er and acid: base indicator to be added. Then, "i"ette 3 millilitre o' a bu''er a bu''er o' a certain "( into the test tube and add $ dro"s o' an acid:base indicator. Then, sha>e the mi;ture and "ro"erly obser=e and note the chan e in its colour. <'ter-ards, Pi"ette 3 millilitre o' a bu''er o' a di''erent "( into another test tube and add $ dro"s o' the same acid:base indicator. Per'orm this "rocedure on all the "re"ared bu''ers. Then, "re"are another set o' B test tubes. Re"eat the second ste" but di''erent acid: base indicator should be used li>e, thymol blue bromo"henol blue, bromocresol reen, bromocresol "ur"le, "henol red, methyl red, methyl oran e, and "henol"hthalein.

The com"onents o' each -ea> acid and conju ate -ere identi'ied. By usin the (enderson:(asselbalch e)uation amounts o' bu''er com"onents -as calculated. The "re"ared solution -as trans'erred to a container and -as "ro"erly labelled.

2.A De*er-ina*i$n Sa-ples

$0

*(e

pH

$0

1. E,ECROMETRIC OF pH

DETERMINATION

Determine the "( o' the 'ollo-in sam"les usin distilled -ater and the assi n sam"le -hich is 'ruit juice. The, use 3 millilitre o' the sam"le and add $ dro"s o' an acid:base indicator. <'ter-ards, sha>e the mi;ture "ro"erly and obser=e and ta>e note o' its colour. Then re"eat the "rocedure but this time use a di''erent indicator.

Calibrate the "( meter at "( 0,8, and +4. Then measure the "( o' a $4 millilitre "ortion o' the "re"ared bu''er solution, distilled -ater and the assi ned sam"le. Then adjust the "( =alue o' the "re"ared bu''er solution to its desired "( by monitorin it on the "( meter and adjust it by addin +.4M o' (Cl or 9a/(.

3. DETERMINATION CAPACIT4

OF

BUFFER

2. CO,ORIMETRIC OF pH

DETERMINATION

Calibrate the "( meter at "( 0,8, and +4. Then, "re"are +44 millilitre o' each 4.4+M (Cl and 4.+4M 9a/(. <'ter-ards, by addin 4.+4m (Cl in + millilitre "ortions 'rom a burette to $4 millilitre "ortions 'rom a burette to $4 millilitre o' the bu''er until its "( chan es as monitored -ith the "( meter the bu''er

ca"acity can be determined. Then, ta>e note o' the =olume added. Then, re"eat usin 4.+4 M 9a/(. Then, re"eat this "rocedure on all bu''ers.

RESU,TS AND DISCUSSION .. E,ECTROMETRIC DETERMINATION OF pH

Ta&le .. Res!l*s #e*er-ina*i$n $0 pH $0 ele'*r$-e*ri'

1. DETERMININ/ $0 BUFFER CAPACIT4

?ince there is a shorta e o' time, doin this "rocedure is not "ossible.

pH Dis*ille# 8a*er Assi ne# sa-ple 9s$#a: 3... $.B8

5H67 0.B2 ; +4:B

$.+0 ; +4:.

C$-p!*a*i$ns; Distilled -aterC *(D, A antilo *:3..., *(D, A 0.B2 ; +4:B ?am"le1soda7C *(D, A antilo *:$.B8, *(D, A $.+0 ; +4:. En table one, by usin the "( meter, the "( o' distilled -ater is 3... -hich mean that it is a little acidic -hile the 'ruit juice has a "( o' $.B8 -hich is acidic. The hydro en "resent in Distilled -ater is 0.B2 ; +4:B -hile the hydro en in sam"le1soda7 is $.+0 ; +4:. .

". CO,ORIMETRIC OF pH

DETERMINATION

Ta&le ". Res!l* $0 '$l$ri-e*ri' De*er-ina*i$n $0 pH

Acid-base indicator Thymol blue ?romphenol blue ?romocresol green ?romocresol purple !henol red Aethyl orange !henolphthalein 2.0 <ed @ello' "range @ello' "range <ed =olorless 3.0 !ale yello' >ight 5reen !ale yello' yello' @ello' <ed orange =olorles s 5.0 !ale yello' ?lue )iolet ?lue ?lue )iolet @ello' "range =olorless 7.0 !ale yello' ?lue )iolet ?lue ?lue )iolet @ello' "range =olorless 7.5 =olorless ?lue )iolet ?lue ?lue )iolet "range @ello' =olorless 8.0 >ight yello' ?lue )iolet >ight blue !urple !in* "range =olorless 12.0 ?lue ?lue )iolet >ight blue !urple !in* "range <ed )iolet Distilled water !ale yello' ?lue )iolet ?lue @ello'green @ello' "range =olorless

Re0eren'es
From boo>C *+, Crisostomo, <.C., Daya, M.%., de !uia, R.M., Farro-, F.%., !abona, M.!., %iu, M.<.D., PeGa, !.T., PeGa, %.%., ?antia o, %.<., ?antia o, M.R., ?arile, <.?., Torres, P.C., Var as, <.!., H Israel, M.C. 1$4+47. Laboratory Manual in General Biochemistry. 2.# ED?<, ?outh Trian le JueKon City, Phili""inesC C H E Publishin Enc. *$, Farell, ?ha-n /., Cam"bell Mary &. 1$4+$7.Biochemistry 8th Edition.<sia.Broo>s6Cole CE9!<!E %earnin . From internetC *.,httpBCC'''.foodmanufacturing.comCarticlesC ::0

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