$lURBIfl LAffr B,$6emstrv

riJi'l :,ji{-irf ni.}. }

*"T*r kei$btien

rNrRcDufnor*
fuotsrfls are separated frmr sotr*tion by tlE prfi*ss of
BreeipE'atinn. FreciplHtion is b:arg&t
about by a glreutt number of cfremkal reagents such as: safts of t*eavy metab, alkabilJal
reagents, akohol, acetore, irrerganrc agOsl and satufilted solrltaon of ammonium sutfate.
Precipi?atbn of prmeirs r* dso Jnormal fumcilan ef tt?e body. An enampn !s the change of
fibrinogen to ftbrin in Hood clotting and th* precipitathn of rasein # curd during dE$hon of
rnilk.

Since arntno acds in proteins exist in tle Zwitterisr form, they will ionize as negative sFcies
above tlreir isoehctrrc point. The po6itive ions d l-eavy metd satE, strh ffi Ag", Hgr*, Cd2* ard
Pb'" {Qmbiffi vsith t}E prctesn anft:n b furn an hsiiunk rnet Fro*e,fl*e, T?e preclpitatiorl
wrth heavy rnetal salts may be iltLEtrated as:

I
+As +
:i i:-
n-l-.-coo ng. + HNoi
l*
NHr

At pH below their isoelectrk point, the prseins erist as pffiitive spefles. wlth alkabical
reagents such as tannr aciC, prcrk aciJ ard turEstic acid. B€ proteifts are precipitat€d from
solutiofl as follows:
H i
+-l-
R-**"1*-co0 +
I
1.1 :: pmrate R-*f-COOH
Nlit
l* ---+ 1+
t{1"t3 picrate

Pr*teim ffi
rery sefisiti$e to pH c&arrys. Adetifn* of afi e(Ees$,i*si1anie *cid srch aal Hf,lO;,
HCl, H;SO4 disrupts the salt bri@es which stabilee tfts teftrary strudure of profieim. \rVhen a
np&ule aquires E hrye poqitfrc drrge, tte bniabh gFErrpE Fres€nt rep*l wtr e{fuer at*
reduces the awilabilrty of tfE profiein mobsrE to interact with tre soh,ent mohcuE.

Ethanol coagulates proteins. The addftiofl of etfunol or acetone reduces tfR dehctric constant
or mnic strengtfi of the aqueos sofiltion. The water motscuhs whhh form a protectffe cffiting
arourd the prffiin moEcu& hydrqffi-bor$s *iS* $e akshol mcheuh* resultiflg ir1 an
increased ekctrostatic atractiofl betrareen pE*eln rnokcuhs The protein nrobcuies then
a*Bce,

H6ft&ql kerrcg;. CIepsrtrrc$t Ao@ 0f Wue ars In{mnatcn ftrhmte;A@

Aamboarqn Crty, Pfulrppmes,
I
* &uElI* lg&: Bffitlef{"lsitry

A fffit*r nrfr$h aff*.ct$ prsteifi$ at yaryi*S @ree* ic kat. Fl€atr*g sa aqir€0{S sa}q *{
*
Sratein *}ay earisE pr*eiflta'tlun. **clp*Xft,* tire ts *{.,e t* tfie diserqa*#atien *f tt}e
pr*{*+rr
m*lecute*" -rf* f'ty*i"gei b"rd;, i*i(-6qn*s an* nydr*pktrc m{de Jre desrryed hecau*e *f
irureaged n*ieeuhr-*b.attnn-and the pr+tem *nder€**$ intrarn*lee$tsr rE*rr*inqers*{1ir
t re*s*ring it i**dubls' hut rn*re readily *qestible" Sr'lch is cajld d*rat*res prtte'irr*

I PROCEOT'RE

*. klts *f ,kieaw !{etats

*
&) Fla€ t ml ef *eufral ta& albumin sot*lon in wh *f 3 test tub€s' Add tl1€
fdl*!6littg r-eagents totlx te,st tuhe$ drsp hy drsp'
I tS ttrbe 1* 19c kad S$ acetate
tes tubs ? * I% reere.tt? tII) #tbrrde
test tube 3 - 1% cadmium nltratre
*qt
Shak* $* t# tube after sfu drop ie addd. cor.rnt the aqmber of *$'p5 tq
pr*iurJa Br.cle-tare. n-"* ae* in e*cess sf Sts r€*qent, t*o* wlether tli* arri*unt sf
pr*ipitat* ls iftr-r,ea*ed or dis$trd bY ac excess reagent"
r
B, AlkakrrdalReagents

prepare 4 test tube$ conhaming 2 mL etrh of 1% albumq soluttsn Add tf€ fotlowing
reubenrts b the mrresp+rd+ng kst tuhe usillg a dmF at 6 time and sh.aktn€ after every
addrtbn'
fe* rube t - saturded picftc acd slutio{l
* test tube 2 - 1C% tafink ecid sfiluliln
I test tube 3 - f$Eo trl*lhnonmtic Ec.ill solt thrt

{I
I
C. Concentrated inorPnic Aods

:preprc 3 *5t trJbs rCI*taifi*rq I rn[ Mr cf 1% filtered aft$*]in slut+on. #d t]*e '

f$k1"qdflS 1eapntt, drryurise, shalff€ careeJlv afrer sh adctiiion'

td tube I - mrEentrated hydrochlort acid

test tube 2 * corxentrated sulfuric acid
test tuh 3 - carenuatd nitric aerd

Note $rh€fher a precipiEte is fuflned. Tl*ry, .dd an s([ss of tfe reagent. Observe the
*fffi af tte €xre$sruageiltonf*e preci$itrte'

IcffiAg nepa*rm*,
tltahrEi C*egdTffitsrid inf$T*atfln Tachq@, A&*et $c ralr'*aafiS Ll*sffsrlY,
&nssarq* Ctq, Fftil*Pn€s'
*

il $i;t{-i11 1.. 1tg, iJ}i:#:!*irli1{lh

.4,Sqltt$p, S*oq+r

I {_}. H*}irg,s lssJ

Fb** , r:1- *f *.$9.i *lsxrnr* s*{ut*fi i{! * {*s. {*h*. LlsiE tfe t*$t ti*k and *eki
I talPfirt\' 4 ntt cocrentrdted nrtrr aud" DO r'll:f SrtaXf F*ote the whfe prec'prtafe .it
t ttx, tun(Eon flf lne t"!+o tirl,,;S

]P'tt the !{'fis,twry of thrS reafitnn hf rppeatrng th€ test On drfferent dilutorls Of i.tE'
fi^51'h altlum+n *'"4tf! diitilid rir*t€r. Fi€(; *eOiir*& r*B* at rtihi{h th&,s*}r€m* g*ex **{y
a falntly perc€pfibk rea{ron.

G Areohol

Hme t nrl *f if*e f*lbt+irrg ts solutk}+rs iri s€Sarat* te*t trh*s: l*& a-i**r:fn* i*&
qelatrr], and lYu E€BtorE *OO S mL of 95% abohol to each test trrbe antl r;rrx
thar*rqfrly. #berv* an$ fcrnratisn sf prsciFfiate. Test fE sotubllrly *f tk prmr$*taf* rn
water.

A*mat ths tesf ** try* athErnin *a{uti}n rsirg ?0% a*$i0f inste*d *f gS% ak*}r*}.
Compare the resutts.

H. Heat

put a pinch of porvoered

egg albumrn tn each of 2 clry test tubes lab€led l.to.1 and No.l.
To test tube Nc.1 - add ? mL drsnlkd water thrn piace bCIth test tubes rn a boirrng
rlater bath for 10 minutes, wth constant shaking. Rernove the test fubes, cool til rcarn
t€fnperat{trs afld thefl atrl ? rnt distilled $ater to test tuhe lrts"}" Filter the $Ehfrens aFd
test both fiitrates wi$r Biuret reageitt.

Perfrrrm Bruret test on t rnl of 196 albumrn stuton. Compare the results.

1ffi} orPR.OPER DISffitAL: Sispose sf ssl*ti*ns i$ $rc* pro$er u,aste ho&les tas aeid
ti, bBsi( waste${ and srsa$i{ *r inorpa*ic wiste$}.

ilrriildlHi:rlr&-'eearrffidt -Arfiiild.rqa
u,.!*6ns-ry,
/an ftra nga Cfiy. PhrilBpn$
 i{{"}BBI6 LABr &isdHrr$srr?

QUESTIOits

11. What is &*attratnn?

12. Wfty is egg whrte used as an antidote fior bal or fllercury porsonrts?
i3. Explarn the use of tannh afid ard picre acll m tfE trBatmtrlt of burns.
11" {an ft€et trssrrhtiCIn test he umotto *tst k preseme of #hrmir* in ur*re? Hx&}n"
15. Whir.h h mure etrBdiw as a disinfmbnt - g5Ya akoluler 7$% *ffirel? Why?

x-

fie#rtrnnL
1retr5a@ffi F't$lippiits, ij&e 6i5dffi; anii-rito
a*mmqga C#"
 $nrup Hn.** S*t* Perfol*ted 1-."-
SI,}RBTfr LA8
Facilibatarrs trnifiais :-
Hembers;
1.
t,
?

*eTIYtTv t{o. ?

FEOIEIIiISI Fr*eip,itatisn

*ata
illamref
a ai---
!!
Te*t iTrs
?,* *. a ,r._,1 F:xiti-"-
I i-.I-i+ i*i"rc'* i i

Fiericmid i

t_.",
I tona
l*a

i
-r**".***.
'--T-nCqgents
ffameof fist iest
,1 --*_.--.-*- - .,i--Sdu$otlf
- : ,L-fddgg|*
'B. rnorg#ic rlo ritemo ' mrrc' HCI
;;iG -- Ahumin I

;iiffi
i i
I, +**."*'-*
i6rc'
,HNO:
;
i_ -"i ----- 1-----"-.

ffi s#ffi ars i!}ft{r?ffidfi Ttrhress- 6teneo d*;aae*flns ljr}tv€!5rtrr

7s666rga €ttV' Pfii&Pirltr'

 I'{URBIS LA*: Beckrn*trv
firfa$fiFcf
-i***,**

l n" *i*ll€r-; ---ib:S%" --'--r,:{o,* *''-"]'-- i

felr*nrin
j:il
i
: i I

js.l??tu
!^.-.----..-*"-.,.... -..-.!

i.,..,-. ii
::.*-'..*.-.--.-...--.."...=..- -i*.:"-*-
i; Il ---'-..-...-'".i
.

'
Albumin :.! ,

iilrl

l{am*cf T#
*iii6ei *' *_-rj-:.jr Preipitate kr Wattr
*. I
I
l

i ;
1l
i:i
tm esii:n Iiiil I-"**""----l-*
lii
: - .--*'-i iii: i*.* -- ----J.-
l%Feptone' i .
:

tlame of Test Te$t neag€nts &-eEi:lt r.,tri1.h *xpl;:r:;];.tili:

SysEm Used Etri-*+-r
Eir$Et T*rt I
I
{

Egg Atbursin
ard
EistilH
filkrcdJ , tr&bt€r

1"i# hiheT Fcsdered

I {lrea,tea, &rg Alhnnifi
j rnixed, ard
itnen BUIEd
I
Watry l

|\taBrra+ E{i* separttrsfit {odege 0f Stierce and InfsifrBfiel ?trhtshgfJ A&n€o {e gilr{%afiEE urliver*lty,
Zsillborriga C#Y, PndPP4{te!'