Katz $ UV Spectrophotometric Analysis of Caffeine and Benzoate in Soft Drink Abstract The caffeine and benzoic acid concentration

of 2 samples, containin 2!""m# and $!""m# of %o&ntain De', 'ere determined to be $(!)*m +# and ))!("m +# for the more concentrated and *!,*m +# and $-!$2m +# for the more dil&te! These concentrations 'ere determined &sin UV spectrophotomery! Si. caffeine and benzoic acid standards 'ere created to enerate relati/ely linear calibration c&r/es! The data from these c&r/es, as 'ell as the obser/ed absorbance for the %o&ntain De' samples, 'ere &tilized to calc&late the caffeine and benzoic acid concentrations! Results To determine the amo&nt of caffeine and benzoate in %o&ntain De', caffeine and benzoic acid standard sol&tions 'ere made! Si. standards 'ere made for each component, the concentrations 'ere determined &sin the form&la 0$V$102V2! 2or the most dil&te caffeine standard, $!""m# stock caffeine sol&tion 3"!$"-( . $") m +#4 'as added to a 5"!"" m# /ol&metric flask and dil&ted to the calibration mark! 0$V$102V2 3"!$"-(.$")m +#43$!"".$"6)#410235"!"".$"6)#4 021$!"-(m +# 324 3)4 3$4

The absorbance for each standard sol&tion 'as obser/ed at both 2)" and 2,5 nanometers! The concentration and absorbance for the caffeine standards are displayed in table $! 7 2)"nm 2,5nm Table 1: Absorbance obser/ed for si. caffeine standard sol&tions $!"-(m +# -!$*-m +# *!)(*m +# $2!55m +# $(!,-m +# "!")$ "!$$"!2(5 "!)*, "!-*2 "!"5) "!2"( "!-)$ "!(-5 "!*-$ 2"!82m +# "!($, $!"()

Katz 2 The concentrations of the benzoic acid standards 'ere also calc&lated &sin 0$V$102V2! 2or the most dil&te benzoic acid standard, $!""m# of a stock sol&tion 3"!$"$*.$")m +#4 'as placed in a 2""!""m# /ol&metric flask and dil&ted to the calibration mark! 0$V$102V2 3"!$"$*.$")m +#43$!"".$"6)#410232""!"".$"6)#4 021"!5"8 m +# 354 3(4 3-4

The concentration for each standard sol&tion and the obser/ed absorbance at 2)" and 2,5 nm are displayed in table 2! 7 2)"nm 2,5nm Table 2: Absorbance obser/ed for si. benzoic acid standard sol&tions "!5"8m +# 2!")(m +# -!",2m +# (!$"*m +# *!$--m +# $"!$*m +# "!"82 "!22* "!-$( "!($" "!,8, "!8,* "!")5 "!"-$ "!"(" "!"*$ "!"8" "!$$(

The data from table $ and 2 'as &sed to constr&ct - calibration c&r/es, 'hich are sho'n in fi &res $6-!

Figure 1: Calibration C&r/e for Caffeine at 2,5nm

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Figure 2: Calibration C&r/e for Caffeine at 2)"nm

Figure 3: Calibration C&r/e for Benzoic Acid at 2,5nm

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Figure 4: Calibration C&r/e for Benzoic Acid at 2)"nm

The e.tinction coefficients for caffeine and benzoic acid at 2)" and 2,5nm 'ere determined from the calibration c&r/es! Beca&se the c&r/e depicts the relationship bet'een absorbance and concentration, the slope of the linear re ression line is e9&i/alent to the e.tinction coefficient! The e.tinction coefficients for each 'a/elen th are recorded in table )! Table 3: :.tinction Coefficients for Caffeine and Benzoic Acid 7 Caffeine Benzoic Acid 6$ 6$ 2)"nm "!"28-cm m # "!"82$cm6$m 6$# 6$ 6$ 2,5nm "!"5"*cm m # "!""*-cm6$m 6$# This information 'as then &sed to determine the concentration of caffeine and benzoic acid in 2 samples of %o&ntain De'! The absorbance for each of these samples at 2)" and 2,5nm are displayed in table -! Table 4: Absorbance for 2 %o&ntain De' sample sol&tions 2m# mo&ntain $ m# mo&ntain de'+5"!""m# de'+5"!""m# 2)"nm $!(-( "!*(5 7

Katz 5 2,5nm "!5** "!)$)

These absorbances and the e.tinction coefficients 'ere s&bstit&ted into the follo'in form&las to determine the concentration of caffeine and benzoic acid in the mo&ntain de' samples!

3,4

3*4 The calc&lated concentrations are displayed in table 5! Table 5: Concentrations of Caffeine and Benzoic Acid in 2 %o&ntain De' sol&tions 2m# mo&ntain de'+5"!"" $m# mo&ntain de'+5"!"" m# m# Caffeine Concentration $(!)( m +# *!,* m +# Benzoic Acid Concentration ))!(" m +# $-!$2 m +# The limit of detection 3#;D4 and lo'er limit of 9&antitation 3##;<4 'ere determined by cond&ctin 5 analyses of the most dil&te standard caffeine sol&tion at the 'a/elen th ma.im&m! The absorbances obser/ed for the fi/e trials, as 'ell as the standard de/iation are sho'n in table (! Table 6: 5 analyses of $!"-(m +# caffeine standard sol&tion at 2,5 nm Trial Absorbance

Katz ( $ 2 ) 5 Standard de/iation "!"5) "!","!",( "!",8 "!"*2 "!"$$

The #;D 'as fo&nd to be "!(5 m +# by di/idin the standard de/iation and the slope of the line in fi &re $ 3"!"5"*cm6$m 6$#4= the 9&otient 'as m&ltiplied by )! The ##;< 'as fo&nd to be 2!$, m +# by di/idin the standard de/iation and the slope of the line in fi &re $= the 9&otient 'as m&ltiplied by $"! Therefore concentrations of caffeine less than 2!$,m +# and reater than "!(5m +# cannot be 9&antified, b&t can be reco nized! The method allo'ed for the eneration of calibration c&r/es 'ith relati/ely linear relationships! The correlation coefficients ran ed from "!8*-(*6"!8888, sho'in that a stron linear relationship e.ists bet'een the data points! The relati/ely lo' correlation coefficient of the benzoic acid calibration c&r/e at 2,5nm is likely d&e to the fact that benzoic acid only absorbs minimally at this 'a/elen th! The lo' si nal co&ld ha/e affected the readin s of the spectrophotometer! Conclusion The content of caffeine and benzoic acid in a sample containin 2 m# of mo&ntain de' 'as determined to be $(!)* m +# and ))!("m +#, respecti/ely! The caffeine and benzoic acid concentration in the sample containin $ m# of mo&ntain de' 'as *!,*m +# and $-!$2m +#, respecti/ely! These concentrations 'ere determined &sin UV spectrophotometric analysis! This method allo'ed for the eneration of relati/ely linear calibration c&r/es and a determination of the #;D and ##;<!