[Analytical Techniques (605) Practical File]

EXPERIMENT # 04 15
th
April,2008
OBJECT:
Analysis of glycerin
a) TO CHECK THE SPECIFIC GRAVITY OF GIVEN SAMPLE
GLYCERINE
APPARATUS
Pyknometer
Weighing balance
FORMULA TO BE USED:
Specific ga!i"#$
water of wt
sample of wt
OBSERVATIOS:
FOR SAMPLE A
Wt of empty Pyknometer = 24.21 gm
Wt of Pyknometer+ water = 49.24gm
Wt of Pyknometer +glycerin = 55.02gm
Therefore,
Wt of water = 25.0 gm
Wt of glycerin = 0.!1 gm
FOR SAMPLE B
Wt of empty Pyknometer = 24.21 gm
Wt of Pyknometer+ water = 49.24 gm
Wt of Pyknometer +glycerin = 55.0" gm
Therefore,
Wt of water = 25.0 gm
Wt of glycerin = 0.!5 gm
Alishba Sana M.Sc (Final) Seat # H0401008
[Analytical Techniques (605) Practical File]
CALCULATIOS
#pecific gra$ity of #ample %
0 . 25
!1 . 0
=

=1.231
#pecific gra$ity of #ample &
0 . 25
!5 . 0
=

=1.22
RESULT:
#pecific gra$ity of #ample % i' fo(n) to *e=1.21
#pecific gra$ity of #ample % i' fo(n) to *e=1.22
%) TO CHECK THE RESI&'E ON IGNITION OF GIVEN SAMPLE
APPARATUS:
+hina )i'h
,e'icator
&(rner
Weighing *alance
OBSERVATIOS:
Alishba Sana M.Sc (Final) Seat # H0401008
[Analytical Techniques (605) Practical File]
RESULT:
-e'i)(e on ignition for 'ample % i' fo(n) to *e = 0.9.
-e'i)(e on ignition for 'ample & i' fo(n) to *e = 0.594.
OBJECT:
c) TO CHECK FATTY ACI& AN& ESTERS IN GIVEN SAMPLES
APPARATUS:
&(rette
+onical fla'k
&eaker
Pipette
C!EMICALS:
0.5 / /a01
0.5 / 1+2
OBSERVATIOS:
INITIAL REA&ING()*) FINAL REA&ING()*) &IFFERENCE()*)
FOR BLA" READI#
0.0 0." 0."
0." 0.12 0."
FOR SAMPLE A
Alishba Sana M.Sc (Final) Seat # H0401008
FOR SAMPLE A FOR SAMPLE +
Wt of empty china )i'h ".30 gm ".31 gm
Wt of china )i'h+glycerin "!.39 gm "!.3" gm
Wt of glycerin 5.09 gm 5.05 gm
Wt of china
)i'h+glycerin4after ignition
".32 gm ".34 gm
Wt of glycerin lo'' 0.02 gm 0.0 gm
%Residue=
100 x
sample of wt
loss wt
0.02 5100
5.09
0.9.
0.0 5100
5.05
0.594.
[Analytical Techniques (605) Practical File]
0.12 0.19 0.3
0.19 0.!9 0.3
FOR SAMPLE B
0.!9 1.9 0.5
1.9 1.!9 0.5
Alishba Sana M.Sc (Final) Seat # H0401008
[Analytical Techniques (605) Practical File]
+ee,La)%e" La-
&eer62am*ert 2aw, more commonly known a' &eer7' 2aw, 'tate' that the optical
a*'or*ance of a chromophore in a tran'parent 'ol$ent $arie' linearly with *oth the 'ample
cell pathlength an) the chromophore concentration. &eer7' 2aw i' the 'imple 'ol(tion to
the more general )e'cription of 8a5well7' far6fiel) e9(ation' )e'cri*ing the interaction of
light with matter. :n practice, &eer7' 2aw i' acc(rate eno(gh for a range of chromophore',
'ol$ent' an) concentration', an) i' a wi)ely ('e) relation'hip in 9(antitati$e
'pectro'copy.
%*'or*ance i' mea'(re) in a 'pectrophotometer *y pa''ing a collimate) *eam of light at
wa$elength ; thro(gh a plane parallel 'la* of material that i' normal to the *eam. <or
li9(i)', the 'ample i' hel) in an optically flat, tran'parent container calle) a c($ette.
%*'or*ance 4%;= i' calc(late) from the ratio of light energy pa''ing thro(gh the 'ample
4:0= to the energy that i' inci)ent on the 'ample 4:=>
%; = 6log 4:?:0=
&eer7' 2aw follow'>
%; = @;*c
@; = molar a*'orpti$ity or e5tinction coefficient of the chromophore at
wa$elength ; 4the optical )en'ity of a 16cm thick 'ample of a 1 8 'ol(tion=.
@; i' a property of the material an) the 'ol$ent.
* = 'ample pathlength in centimeter'
c = concentration of the compo(n) in the 'ample, in molarity 4mol 2
61
=
:n an a*'or*ance e5periment, light i' atten(ate) not only *y the chromophore, *(t al'o *y
reflection' from the interface *etween air an) the 'ample, the 'ample an) the c($ette, an)
a*'or*ance *y the 'ol$ent. The'e factor' can *e 9(antifie) 'eparately, *(t are often
remo$e) *y )efining :0 a' the light pa''ing thro(gh a 'ample A*lankA or A*a'elineA or
reference 'ample 4for e5ample, a c($ette fille) with 'ol$ent *(t Bero concentration of the
chromophore i' ('e) a' the *lank=.
Alishba Sana M.Sc (Final) Seat # H0401008
[Analytical Techniques (605) Practical File]
8any factor' can affect the $ali)ity of &eer7' 2aw. :t i' ('(al to check for the linearity of
&eer7' 2aw for a chromophore *y mea'(ring the a*'or*ance of a 'erie' of 'tan)ar)'.
Thi' Acali*rationA can al'o remo$e error' in the e5periment, the e9(ipment, an) the
*atch of reagent' 4'(ch a' c($ette' of (nknown pathlength
+a.ic Pi/cip*e. 0f Spec"0p10"0)e"#
% 'pectrophotometer con'i't' of two in'tr(ment', namely
% spectrometer for pro)(cing light of any 'electe) color 4wa$elength=.
% photometer for mea'(ring the inten'ity of light. The in'tr(ment' are arrange)
'o that li9(i) in a c($ette can *e place) *etween the 'pectrometer *eam an) the
photometer. The amo(nt of light pa''ing thro(gh the t(*e i' mea'(re) *y the
photometer. The photometer )eli$er' a $oltage 'ignal to a )i'play )e$ice,
normally a gal$anometer. The 'ignal change' a' the amo(nt of light a*'or*e) *y
the li9(i) change'.
D$t$r%i&'ti(& () C(&*$&tr'ti(&
:f )e$elopment of color i' linke) to the concentration of a '(*'tance in 'ol(tion then that
concentration can *e mea'(re) *y )etermining the e5tent of a*'orption of light at the
appropriate wa$elength. <or e5ample hemoglo*in appear' re) *eca('e the hemoglo*in
a*'or*' *l(e an) green light ray' m(ch more effecti$ely than re). The )egree of
a*'or*ance of *l(e or green light i' proportional to the concentration of hemoglo*in.
SPECTROP!OTOMETER
Alishba Sana M.Sc (Final) Seat # H0401008
[Analytical Techniques (605) Practical File]
#pectrophotometry in$ol$e' the ('e of a 'pectrophotometer. % 'pectrophotometer i' a
photometer 4a )e$ice for mea'(ring light inten'ity= that can mea'(re inten'ity a' a
f(nction of the color, or more 'pecifically, the wa$elength of light. There are many kin)'
of 'pectrophotometer'. %mong the mo't important )i'tinction' ('e) to cla''ify them are
the wa$elength' they work with, the mea'(rement techni9(e' they ('e, how they ac9(ire
a 'pectr(m, an) the 'o(rce' of inten'ity $ariation they are )e'igne) to mea'(re. 0ther
important feat(re' of 'pectrophotometer' incl()e the 'pectral *an)wi)th an) linear range.
The 'pectrophotometer mea'(re' 9(antitati$ely the fraction of light that pa''e' thro(gh a
gi$en 'ol(tion. :n a 'pectrophotometer, a light from the lamp i' g(i)e) thro(gh a
monochromator, which pick' light of one partic(lar wa$elength o(t of the contin(o('
'pectr(m. Thi' light pa''e' thro(gh the 'ample that i' *eing mea'(re). %fter the 'ample,
the inten'ity of the remaining light i' mea'(re) with a photo)io)e or other light 'en'or,
an) the tran'mittance for thi' wa$elength i' then calc(late).
Se23e/ce 0f e!e/".
:n 'hort, the 'e9(ence of e$ent' in a 'pectrophotometer i' a' follow'>
1. The light 'o(rce 'hine' thro(gh the 'ample.
2. The 'ample a*'or*' light.
. The )etector )etect' how m(ch light the 'ample ha' a*'or*e).
4. The )etector then con$ert' how m(ch light the 'ample a*'or*e) into a n(m*er.
5. The n(m*er' are either plotte) 'traight away, or are tran'mitte) to a comp(ter to
*e f(rther manip(late) 4e.g. c(r$e 'moothing, *a'eline correction=
Alishba Sana M.Sc (Final) Seat # H0401008
[Analytical Techniques (605) Practical File]

Spectrpht!etric cells

+ell' 4or c($ette'= are '(pplie) with $arying light path' an) )ifferent 9(alitie' of
gla''. 8o't 'pectrophotometric work i' con)(cte) with 'tan)ar) rectang(lar cell' of 1
cm path length. The'e are '9(are in cro''6'ection an) a*o(t 4 time' a' high a' they are
wi)e. They generally ha$e a pathlength tolerance of 60.01 mm. :n a))ition, rectang(lar
cell' of 0.1, 0.5 an) 4 cm are commercially a$aila*le. <or more )il(te 'ol(tion',
cylin)rical cell' of 5 an) 10 cm are a$aila*le. The'e ha$e two filler neck' into which fit
fl(oropolymer 'topper'. The a*ility to 'eal the'e cell' i' an attracti$e feat(re for $olatile
or haBar)o(' 'ample'. The cylin)rical cell' may *e '(pplie) with fl(oropolymer co$er',
*(t the'e )o not make a $ery goo) 'eal. -ectang(lar cell' with e5tra thick 'i)e wall' are
al'o a$aila*le for the analy'i' of 'mall $ol(me' of li9(i). :n a))ition, $ario(' type' of
flow thro(gh an) Cackete) cell' are a$aila*le for kinetic 't()ie', contin(o(' monitoring,
an) temperat(re 'en'iti$e work.
+ell' for a*'orption 'pectrophotometry ha$e oppo'ing optically poli'he)
win)ow' with fro'te) gla'' 'i)e wall'. &e '(re to orient the cell' 'o that the optic
win)ow' are perpen)ic(lar to the light path. <l(ore'cence cell' are ma)e with all 4or
a)Cacent= optically poli'he) win)ow'.
+ell' 'ho(l) *e cleane) like other la*oratory gla''ware. :f )etergent' pro$e
ineffecti$e, they may *e 'oake) in chromic aci) cleaning 'ol(tion. /ote that the
win)ow' an) 'i)e wall' are f('e) forming an aci)6proof 'eal. :n a))ition, 9(artB cell'
may al'o *e cleane) *y immer'ing in concentrate) nitric aci) followe) *y (ltra'onic
agitation or 10615 min(te' of *oiling in water.
Alishba Sana M.Sc (Final) Seat # H0401008