SCHOOL OF PHARMACEUTICAL SCIENCE
FAR 313/4: Pharmaceutical Analysis
Experiment 4 Ultraviolet and Vii!le Spe"trop#otometr$ % Aa$ o& T#eop#$lline in Aminop#$lline Ta!let Ta!let 'ro(p ) *+ Name
Matri" No.
TAN TAN LI MA2 TAN SIU SIEN TAN 2I 2EN TA2 SUE CH2EN
Le"t(rer
**430+ **4304 **4303 **4300
, -R. LAI CHOON SHEEN
-ate o& S(!miion , /0t# NOVEM)ER /1*4 1.0 Aims
1
*. To t(d$ t#e (tiliation o& (ltraviolet and vii!le pe"trop#otometr$ in 5(antitative anal$i. /. To "al"(late t#e amo(nt o& t#eop#$lline in one aminop#$lline ta!let. 2.0 Introduction
Ultraviolet and vii!le pe"tro"op$ re&er to a!orption pe"tro"op$ in t#e (ltraviolet6vii!le pe"tral re7ion 89illiam Re("#: /1*+;. It i a met#od (ed to mea(re t#e a!orption o& (!tan"e radiation in t#e
to t#eir 7ro(nd ener7$ tate. Ultraviolet pe"tro"op$ i a ver$ e&&i"ient tool &or 5(alitative a nono
2
a expreed !$ t#e &orm(la, T = I / I o <#ere t#e lo7arit#m &or t#e re"ipro"al o& T i "alled t#e a!or!an"e: A: <#i"# i expreed a, A = log (1/T). T#e prin"ipal !e#ind pe"trop#otometer i t#e a!orption la<: t#e )eer6Lam!ert la<. It 5(antitativel$ #o< #o< t#e amo(nt o& atten(ation depend on t#e "on"entration o& t#e a!or!in7 mole"(le and t#e pat# len7t# over <#i"# a!orption o""(r. A""ordin7 to )eer6Lam!ert la<: a!or!an"e A i linearl$ related to t#e "on"entration o& t#e a!or!in7 pe"ie 8c; and t#e pat# len7t# 8l ; o& t#e a!or!in7 medi(m: at "ertain nono
3.0 Procedure A) a!elen"th #ontrol
*. T#e
*. An amo(nt o& potai(m di"#romate [email protected] m7 % 0+.1 m7; <#i"# #ad !een dried at *=11C ; at t#e
a!elen"th &nm)
A &1 'ercent( 1 cm)
a*imum tolerance
3
/+3 /3@ +*+ +31
*/4.3 *44.1 4=.0 *10.0
*//.D % */0./ *4/.4 % *43.@ [email protected] % 31.+ *14.D % *1=./
#) +heo'hylline Assay in Amino'hylline +a%let I. ,'ectrum and #ali%ration #ur!e o- +heo'hylline
*. T#eop#$lline to"> ol(tion ol(tion . 4. T#e
II. Assay o- the +a%let
*. Ten &or *1 min(te. +. -itilled . 3. T#e a!or!an"e o& t#i ol(tion
4
T#e
,'eci-ied a!elen"th &nm)
%ser!ed a!elen"th &nm)
i--erence &nm)
/4*./
6
6
/=@./
/=@.*0
61.14
+0*.3
+01.=4
61.00
3+0.+
3+0.+0
61.10
Permitted toleran"e * nm &or (ltraviolet re7ion 8/11 6 411nm; + nm &or vii!le re7ion 8B 411 nm;
$. A%sor%ance #ontrol
T#e a!or!an"e and molar a!orptivit$ o& potai(m di"#romate ol(tion in (lp#(ri" a"id
5
Table 2: Absorbance control by using potassium dichromate
a!elen"th( nm
A 8*:*"m;
A%sor%ance &A)
,'eci-i
%ser!e
,'eci-ic
*'erimenta
a*imum +olerance
c
d
/+3.1
/+4.31
1.@+@=3
*/4.3
*//.D3
*//.D to */0./
/[email protected]
/30./@
1.=3==*
*44.1
*4+.*1
*4/.4 to *43.@
+*+.1
+*/.3@
1./=@@0
4=.0
[email protected]
[email protected] to 31.+
+31.1
+31./4
1.0+@@/
*10.1
*10./0
*14.D to *1=./
l
A%sor%ance( A = klc
<#ere A
a!or!an"e
k
rate "ontant
l c
pat# len7t# o& t#e ol(tion * "m "on"entration o& t#e potai(m di"#romate ol(tion in 7?L or
c
01.1*4 m7?L
#alculation
1.1011*4 7? *111 mL 1.11011*4
9#en
6
1.=3==*? 8* x 1.11011*4; 143.10 9#en
#. +heo'hylline Assay in Amino'hylline +a%let I. ,'ectrum and cali%ration cur!e o- theo'hylline
9ei7#t o& t#eop#$lline 1.1*/4 7 Con"entration o& t#e t#eop#$lline to"> ol(tion 81.1*/4 7?*11mL; x *11 1.1*/4 ol(tion 1 .1*/4 ol(tion M/ Con"entration o& t#eop#$lline ol(tion a&ter dil(ted V/ Final vol(me o& t#eop#$lline ol(tion a&ter dil(ted * mL o& to"> ol(tion ol(tion
7
M/ 4.D0 x *164 ol(tion ol(tion ol(tion
Table 3: The absorbance is measured at certain concentration of theophylline solution
Pre'aration
6olume o- +heo'hylline stoc7 solution( m8
9/! o- +heo'hylline
A%sor%ance( A
*
*.1
/.4= x *164
1.*4@/
/
/.1
4.D0 x *164
1./==+
+
+.1
@.44 x *164
1.4/+1
4
4.1
D.D/ x *164
1.33=/
3
3.1
*/.4 x *164
1.@*41
8
'rap# o& A!or!an"e ver( Con"entration o& T#eop#$lline 0.8 0.7
f(x) = 0.06x + 0 R² = 1
0.6 0.5
A!or!an"e: A
0.4 0.3 0.2 0.1 0
0
2
4
6
8
10
12
14
Con"entration o& T#eop#$lline 8 *164
)$ re&errin7 to )eer6Lam!ert e5(ation: A!or!an"e: A klc Sin"e l *"m: A kc: Sin"e t#e e5(ation o& t#e "(rve i $ mx G 1 A >" G 18/; )$ "omparin7 e5(ation 8*; 8/;: > "an !e m 8t#e 7radient o& t#e lope;. T#e linear e5(ation o!tained &rom t#e "ali!ration "(rve i $ 1.130Dx + 1.11/4: T#(: 7= 0.5 x104 8"1cm1
2. Assay o- the +a%let
T#e "om!ined no
8*;
9
)aed on t#e "ali!ration "(rve plotted: l Sin"e A klc: <#ere kl i 1.130D: A 1.+31+ " A?>l 1.+31+?1.130D 0.*0 x *164
Amo(nt o& t#eop#$lline in /31 mL ol(tion 0.*0 x*164 ? *11 mL x /31 mL *.3+D x *16+ 7
Amo(nt o& t#eop#$lline !e&ore dil(tion *.3+D x *16+ 7 ? 3 mL x /31 mL 1.1@@ 7
T#ere&ore: in 1.1@1 7 o& t#e po
)$ re&errin7 to t#e )riti# P#arma"e(ti"al Codex *D@+: t#e amo(nt or per"enta7e o& t#eop#$lline in aminop#$lline ta!let i a!o(t @= % =4. Ho
.0 iscussion A. a!elen"th control
)e&ore in7 a""ordin7l$. Hen"e: no
10
In t#i experiment: #olmi(m oxide t#at "o(ld !e een. 9e "o(ld not o!erve t#e preen"e o& an$ pea> &or t#e pe"i&ied
)eide: !e&ore tart a!or!an"e val(e o& potai(m di"#romate ol(tion i determined in "orrepond to repe"tive
C. Theophylline assay in aminophylline table 1. Spectrum and calibration curve of theophylline
11
Con"entration o& t#eop#$lline in aminop#$lline "an !e determined !$ a "ali!ration "(rve o& t#eop#$lline. From t#e 7rap# o& a!or!an"e ver( . T#i i !e"a(e t#i <#ile #i7#er "on"entration o& t#eop#$lline ol(tion prod("e too #i7# pea>. T#e maxim(m .
A&ter t#e a!or!an"e val(e o& all &ive ol(tion o& t#eop#$lline
eer!" #$%& $'"or'$nce i" ine$r *ro*ortion$ to the concentr$tion of the $'"or'ing "*ecie". he higher the concentr$tion of theo*hine& the gre$ter the $'"or'$nce o't$ined. -i&&erent val(e o& a!or!an"e o!tained &rom t#e 7rap# i 0.5 x104 8"1cm1
2. Assay of the tablet To o!tain t#e "on"entration o& t#eop#$lline in t#e prepared ol(tion: it
From o(r "al"(lation: it #o< t#at 1.1@@7 o& t#eop#$lline i preent in t#e 1.1@7 &ine po
12
i "ight higher fro, the r$nge& @= % =4 a "t$ted in the )riti# P#arma"e(ti"al Codex *D@+.
T#eop#$lline i a poiono( in7redient (ed to prevent and treat <#eein7 and ot#er !reat#in7 di&&i"(ltie "a(ed !$ l(n7 dieae ("# a at#ma. It indire"tl$ tim(late !ot# * and /re"eptor t#ro(7# releae o& endo7eno( "ate"#olamine. T#(: it i important to prevent t#eop#$lline toxi"it$ a it
hi" de-i$tion in re"t ,ight 'e de to "o,e error" th$t h$-e occrred dring the ex*eri,ent. here ,ight 'e "o,e in$ccr$cie" dring the dition *roce"" "ch $" not $dding the $ccr$te -o,e of "odi, hdroxide "otion c$"ing the theo*hine "otion 'eco,e" ,ore concentr$ted. here cod $"o 'e "o,e ,i"t$/e" dring the tr$tion of the theo*hine "otion "ch $" *re"ent of i,*ritie" $nd cont$,in$nt" %hich %i $ect the -$e of $'"or'$nce o't$ined. r'id or '''e" in the "otion %i $"o di,ini"hed the $ccr$c of re$ding". inger*rint "t$in" or "cr$tche" on the c-ette %$" ,$ $"o c$"e the de-i$tion in re"t $" the ,$ reect or $'"or' r$di$tion. ther th$n th$t& technic$ $nd *$r$$x error" $"o ,ight 'e exi"ted in thi" ex*eri,ent.
T#ere are ome pre"a(tion mea(re t#at ind o& mea(rement or e a !etter diol(tion. 4. La!elin7 m(t !e appropriatel$ done in order to avoid "on&(ion. 3. )e&ore ea"# mea(rement: t#e "(vet m(t !e "leaned t#oro(7#l$. T#e "(vet #o(ld !e properl$
relia!le and a!le to prod("e a""(rate re(lt.
13
2. T#e "on"entration o& t#eop#$lline in t#e ample o!tained &rom t#e "al"(lation !$ (in7 t#e "ali!ration
"(rve e5(ation i 5.15 * 104 9/!. 3. T#e amo(nt o& t#eop#$lline in one aminop#$lline ta!let o!tained &rom t#e "al"(lation !$ (in7 t#e "ali!ration "(rve e5(ation i 0.2;0" . 4. T#e per"enta7e o& t#eop#$lline in one aminop#$lline ta!let i 1109: <#i"# deviate &rom t#e ran7e tated in )riti# P#arma"opeia.
.0 Re-erences
*. 9illiam Re("#: /1*+. Vii!le and Ultraviolet Spe"tro"op$. C#emitr$.m(.ed(. Availa!le at, #ttp,??<<inelmer."om?...?446*+0=+DTCHValidatin7UVVii!le.pd& +. #ttp,??<<<.preervearti"le."om?/1*4***+/+/=D?"omplete6in&ormation6on6(ltraviolet6 pe"trop#otometr$.#tml 4. Prin"iple o& Spe"trop#otometr$: #ttp,??<<<.r(&.ri"e.ed(?Q!iola!?met#od?protein?pe"trop#otometer.#tml 3. Spe"trop#otometr$ /1*4. . ONLINE Availa!le at, #ttp,??<<
