Determining Concentration of Allure Red Food

Coloring dye in Gatorade

Valentin Uzunov (author) & Gavin Koenig (lab partner)
PSU ID#: 96!""#$$
%&' (($ )ab (6!
ABSTRACT
*he purpo+e o, thi+ e-peri.ent /a+ to 0eter.ine the 1on1entration o, 2llura 3e0 (a ,oo0 1oloring 04e)
pre+ent in ("oz Gatora0e u+ing +pe1trophoto.etri1 anal4+i+5 *he 1on1entration /a+ 0eter.ine0 ,ro. a
1alibration 1urve o, ab+orban1e v+ 1on1entration5 2 linear relation+hip i+ ob+erve0 bet/een ab+orban1e
an0 1on1entration in a11or0an1e /ith the BeerLambert Law. *he re+ult o, the anal4+i+ +ho/e0 the
1on1entration o, 2llura 3e0 in (" oz o, Gatora0e to be 65" - !"
7$
'8 or (!( 9g
INTRODUCTION
:oo0 1oloring 04e+ are o,ten a00e0 to beverage+ to in1rea+e vi+ual appeal8 an0 ta+te per1eption8
ho/ever their health e,,e1t+ have been ;ue+tione08 an0 their +a,et4 ,or hu.an 1on+u.ption i+ largel4
un<no/n5 2+ +u1h the ob=e1tive o, thi+ e-peri.ent /a+ to 0eter.ine the 1on1entration o, 2llura 3e08 a
parti1ular ,oo0 1oloring 04e ,oun0 in Gatora0e8 a 1o..er1ial +port+ 0rin< pro0u1t5 *he 1on1entration
o, the 04e /a+ 0eter.ine0 u+ing +pe1trophoto.etr4 to prepare a 1alibration 1urve bet/een ab+orban1e
v+ 1on1entration5 *he relation+hip bet/een ab+orban1e an0 1on1entration i+ be+t 0e+1ribe0 b4 the >eer?
)a.bert la/5 *he >eer?)a.bert la/8 +tate+ that there i+ a logarith.i1 0epen0en1e bet/een
tran+.i++ion an0 ab+orban1e @A;uation !B5
Beer-Lambert Law [Euation !"
/here T i+ tran+.ittan1e8 /hi1h i+ the ratio bet/een the inten+it4 o, the tran+.itte0 light8 I
1
an0 the
inten+it4 o, light +our1e8 I
0 5
It i+ i.portant to note that tran+.ittan1e 0i.ini+he+ e-ponentiall4 a+ it
travel+ through a .aterial8 a+ +ho/n b4 the e-pan+ion o, b4 >eer?)a.b la/ [Euation #"$
[Euation #"
Chen 0ealing /ith li;ui0+ in parti1ular8 tran+.i++ion i+ o,ten e-pre++e0 in ter.+ o, ab+orban1e ,or a
given /avelength5 :or thi+ e-peri.ent a /avelength o, $"n. /a+ u+e0 a+ thi+ /avelength
1orre+pon0+ to the highe+t ab+orban1e ,or 2llura 3e0 ba+e0 on it+ ab+orption +pe1tru. @(B5 :or the
BeerLambert Law to hol0 there are 6 ne1e++ar4 1on0ition+ @!B:
!$ *he ab+orber+ in +olution .u+t a1t in0epen0entl4 o, ea1h otherD
#$ *he ab+orbing .e0iu. .u+t be ho.ogeneou+ ( +olution+)
%$ *he ab+orbing .e0iu. .u+t not +1atter the ra0iation
&$ *he in1i0ent ra0iation .u+t 1on+i+t o, parallel ra4+8 ea1h traver+ing the +a.e length in the
ab+orbing .e0iu.5
'$ *he in1i0ent ra0iation +houl0 be .ono1hro.ati18 or have at lea+t a /i0th that i+ narro/er
than that o, the ab+orbing tran+itionD an0
($ *he in1i0ent ,lu- .u+t not in,luen1e the ato.+ or .ole1ule+D it +houl0 onl4 a1t a+ a non7
A =log
!"
(T)
(T ) =
I
!
I
"
=!"
l c
inva+ive probe o, the +pe1ie+ un0er +tu045 In parti1ular8 thi+ i.plie+ that the light +houl0 not
1au+e opti1al +aturation or opti1al pu.ping8 +in1e +u1h e,,e1t+ /ill 0eplete the lo/er level
an0 po++ibl4 give ri+e to +ti.ulate0 e.i++ion5
:or thi+ e-peri.ent8 the .o+t +uitable ,or. o, the >eer?)a.bert la/ 1an be e-pre++e0 a+:
A
)
* + l c [Euation %"
Chere A
)
i+ the ab+orban1e ob+erve0 8 + i+ the e-1itation 1oe,,i1ient8 /hi1h repre+ent+ the a.ount o,
light ab+orbe0 0uring tran+.i++ion through the .aterial ( E i+ 1hara1teri+ti1 o, the ab+orber8 an0 varie+
/ith /avelength )5 l i+ path length8 /hi1h repre+ent+ the 0i+tan1e light ha+ to travel through the .aterial
(length o, 1uvette)8 an0 c i+ the 1on1entration o, the +olution5 :or a given +ub+tan1e at a parti1ular
/avelength E an0 l are 1on+tant+ an0 A;uation ! 1an be +i.pli,ie0 to:
A
)
* , c [Euation &"
/here < F E l8 an0 repre+ent+ the +lope o, the 1alibration 1urve5 U+ing thi+ relation+hip the
1on1entration o, a 0ilute0 +a.ple Gatora0e /a+ 0eter.ine08 an0 the pro1ee0ing re+ult+ u+e0 to
appro-i.ate the a1tual 1on1entration+ o, 2llure re0 in a (" oz bottle5
*he 1alibration 1urve /a+ 0e,ine0 ,ro. ab+orban1e an0 1on1entration+ o, G G" .) +tan0ar0
+olution prepare0 ,ro. a "5"""! ' +to1< +olution o, 2llura 3e08 an0 one blan< +a.ple 1ontaining onl4
/ater5 :or ea1h +olution the ab+orban1e /a+ .ea+ure0 u+ing a %olori.eter 1alibrate0 u+ing the blan<
+olution to 1orre1t ,or the ab+orption o, /ater5
DATA
*2>)A ! ? Stan0ar0 Solution 2b+orban1e 3ea0ing+ an0 %on1entration+
Sa.ple
2llura 3e0
("5"""! ')
(.))
Cater
(.))
%on1entration (
.ol H ))
2b+orban1e *ran+.ittan1e (I)
>lan< " G" "5" "5""" !""
Stan0ar0 ! # $ 65" - !"
76
"5"#$ 9!59(
Stan0ar0 ( G G !5" - !"
7G
"5"66 6G59
Stan0ar0 # $ # !5 - !"
7G
"5"9( 6"59#
Stan0ar0 9 ! !56 - !"
7G
"5!!" $$5GG
Stan0ar0 G !" " (5" - !"
7G
"5!( $G5(G
Gatora0e
Jun<no/nJ
Dilute !" " 95 - !"
76
"5"6" 6$5"9
Un0ilute0 (" oz 65" - !"
7$
Table 1- Standard solutions prepared from a .1 ! stoc" solution
RESU-TS
A;uation o, *ren0line +olving ,or 2b+orban1e
[Euation '"
3earrange A;uation to +olve ,or %on1entration
[Euation ("
Dilution A;uation
[Euation .a"
rearran#ed to sol$e for un"nown !
[Euation ./"
!$ Calculating concentration of in m-0 for Standard u1ing [Euation 'a"
Figure 1 - %rap& s&ows t&e linear relations&ip between absorbance and
concentration ' mol ( L ). T&e line of best fit is defined f'*) + ,1-,..1,* / .10112
wit& a coefficient of determination '34) + ...15
0.000000 0.000005 0.000010 0.000015 0.000020 0.000025
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
Calibration curve for Allura Red analysis
Concentration (mol / !
A
b
s
o
r
b
a
n
c
e
f (*) =6!6659!- + "5""!$!G
f (*) =
* "5""!$!G
6!6659!
!
!
6
!
6
(
= !
(
!
!
6
!
= !
(
6
(
"5"""!! 6
!
="5"G L 6 *!"
6

!"""mL
! L
=#mL
# $Calculating concentration of dilute Gatorade 1olution u1ing [Euation ("
rea0ing ,ro. graph i+ appro-i.atel4 95" - !"
76
!
u+ing BeerLambert Law [Euation %" F 95$ - !"
76
'
%$ Calculating concentration of Allura Red in #2 oz of Gatorade u1ing [Euation ./"
8/here ("oz F "5G9 )
&$ Calculating mole1 of Allure Red in #2 oz of Gatorade
8/here ("oz F "5G9 )
'$ Calculating num/er of molecule1 of Allure Red in #2 oz of Gatorade
($ Calculating ma11 of Allure Red in #2 oz of Gatorade

.olar .a++ o, 2llure 3e0 ( %
!6
&
!
K
(
L
6
S
(
(7

) F G"5 g H .ol
DISCUSSION
*he purpo+e o, thi+ e-peri.ent /a+
to u+e +pe1trophoto.etr4 to 0eter.ine the
1on1entration o, 2llura 3e08 a ,oo0
1oloring 1o.poun0 o,ten u+e0 in
1o..er1ial +port+ beverage+ +u1h a+
Gatora0e5 :ro. the re+ult+ o, the
e-peri.ent it /a+ 0eter.ine0 that the the
1on1entration o, 2llure 3e0 in a (" oz
bottle o, Gatora0e i+ appro-i.atel4 65" -
!"
7$
'8 /hi1h i+ e;uivalent to (!( 9g o,
2llura 3e08 or (56 - !"
!$
.ole1ule+ o, the
1o.poun05 *he lo/ 1on1entration+
0eter.ine0 +ee. rea+onable8 in light o,
the potential health ri+<+ a++o1iate0 /ith
the 1on+u.ption o, +u1h 1o.poun0+ a+
0etaile0 in a report b4 the %enter ,or
S1ien1e in the Publi1 Intere+t @B5
2+ /ith an4 e-peri.ent potential +our1e o, error ari+e /hen a++u.ption are .a0e5 In thi+ e-peri.ent
an a++u.ption i+ .a0e that the ab+orban1e o, an a;ueou+ +olution o, 2llura 3e0 (a+ u+e0 ,or +tan0ar0+
in preparing the 1alibration 1urve) a0e;uatel4 repre+ent+ the ab+orban1e o, a 0ilute +olution o,
Gatora0e5 *hi+ o, 1our+e i+ not entirel4 a11urate a+ even a 0ilute +olution o, Gatora0e 1ontain+ +everal
f ("5"6) =
"5"6 "5""!$!G
6!6659!
= 95 * !"
6
!
95 * !"
6
! "5"G L
"5G9 L
=65" * !"
$
!
"5G9 L
65" * !"
$
mol
! L
=5$ * !"
$
mol
65"(( !"
(#
! mol

5$ * !"
$
mol
! L
= (56 !"
!$
molecules
5$ * !"
$
mol
G"5 #
! mol

!"
6
7#
! #
=(!( 7#
8i#ure 1 - S&ows Structural formula and ball and stic"
model of disodium ,-&9dro*9-1-'':-met&o*9-1-met&9l-
5-sulfop&en9l)a;o)-:-nap&t&alenesulfonate2 also "now
as Allura 3ed2 and 3ed 52 wit& a molecular formula of
<a:=1->15<:?-S:
other ingre0ient+ along /ith 2llure 3e0 an0 /ater5 >4 not 1alibrating the in+tru.ent to ta<e into
a11ount all po++ible ab+orber+8 a higher ab+orban1e rea0ing+ i+ .o+t li<el4 .ea+ure0 an0 thu+
1al1ulation+ /ill 4iel0 a po++ibl4 higher 1on1entration5 In or0er in1rea+e the a11ura14 o, the
.ea+ure.ent the %olori.eter /oul0 nee0 to 1alibrate0 to +ubtra1t the ab+orban1e ,ro. all other
potential ab+orber+ in the Gatora0e +olution e-1lu0ing the 2llure 3e05
2nother po++ible +our1e o, error to 1on+i0er i+ that volu.e in the e-peri.ent i+ treate0 a+ being
a00itive8 /hen the .i+1ibilit4 o, the t/o +olution i+ un<no/n an0 the re+ulting 0ilution+ .a4be not be
ver4 pre1i+e5 *hi+ i.pa1t+ the +ub+e;uent a11ura14 o, the 1on1entration+ 0eter.ine05 &o/ever given
the +.all 1on1entration+ u+e0 ,or the +tan0ar0 +olution+8 it i+ po++ible the error in1urre0 i+ largel4
in+igni,i1ant8 thu+ it+ not e-pe1t to have ha0 an4 +igni,i1ant i.pa1t on the re+ult+5 :inall4 there i+
al/a4+ the 1on+i0eration ,or hu.an error an0 ina11ura14 an0 pre1i+ion in te1hni;ue /hi1h 1an o,ten be
eli.inate0 /ith .ore trial+5
CITATION
@!B "eer#ambert la$. (201%& 'ovember 22!. (n Wikipedia, The Free Encyclopedia. Retrieved 21)08& 'ovember 24&
201%& from *tt+)//en.$i,i+edia.or-/$/inde..+*+/title0"eer12218013%ambert4la$5oldid058282%045
@(B ChemLab - Chemistry 6 - Week 4 - The Spectra of Conjugated Dyes and Inestigation of !eer"s La#.
(2011, September 20). Retrieved November 11, 2013, from
http://www.dartmouth.edu/~chemab/chem!/d"e#/fu$te%t/chemi#tr".htm
@#B $fficia% Site for &epsiCo !eerage Information ' Co%ors. (2013, &ctober 2'). Retrieved November 2(,
2013, from http://www.pep#icobevera)efact#.com/coor#.php
@B *e+ter for Scie+ce i+ the ,ubic -+tere#t (2010). (ood Dyes) * +ainbo# of +isks (.). Retrieved from
http://c#pi+et.or)/+ew/pdf/food/d"e#/rai+bow/of/ri#0#.pdf
8i#ure 1 - S&ows Structural formula and ball and stic" model of disodium ,-&9dro*9-1-'':-met&o*9-1-
met&9l-5-sulfop&en9l)a;o)-:-nap&t&alenesulfonate2 also "now as Allura 3ed2 and 3ed 52 wit& a
molecular formula of <a
:
=
1-
>
15
<
:
?
-
S
: