AS245: Bachelor in Science Applied Applied Chemistry Title: Analysis Title: Analysis of Riboflavin Riboflavin in Energy Drink Course: Spectrochemical Methods of Analysis (CHM!"# Name: Mohamad $or Amir%l A&har b' amis Matrix no:
)"*+,+-.++ (AS)+ .S#
Date of experiment: *th April )"* Date of submission: )/th May )"* Group members: *' $%rnailah bt' $oora&lan )' $or Amirah bt' Ahmad A&%an .' $oramira bt' Saad It is my responsibility as a student of iTM to ad!ere to trut!fulness and a"oid dis!onesty# fraud or de$eit of any type in $onne$tion %it! %rite up and $ondu$t of t!is experiment& Si'nature:(((((((((( Date:
TIT)*:
Determination of amo%nt riboflavin in energy drinks samples'
A+ST,ACT: Sample %sed in this e0periment is the energy drinks (livita# that 1as dil%ted into smaller concentration 1ith dil%tion factor of )"2 so that the intensity 1ill be in the range range of standa standard rd sol%ti sol%tions ons'' 3l%ore 3l%oresce scence nce Emissi Emission on Spectr Spectrosc oscopy opy 1as %sed %sed to analysed the analyte of riboflavin in the sample to kno1 the concentration of the riboflavin in the sample' 4he sample has been triplicate to ens%re the acc%racy and getting more acc%rate data' 4he average concentration of riboflavin in the triplicate sample is /'-)ppm (act%al concentration# 1ith relative standard deviation of *"',, 1ith average amo%nt of riboflavin of "'+!,.mg'
INT,-DCTI-N: Riboflavin 1ith molec%lar form%la of C*-H)"$+5, is also kno1n as vitamin B ) is a part of vitamin B gro%p' 6t 1as formerly kno1n as vitamin 7 1ith yello1ish orange solid s%bs s%bsta tanc nce e that that poor poorly ly sol% sol%bl ble e in 1ate 1ater' r' 4he 4he pers person on 1ho 1ho took took this this vita vitami min n as s%pplement can be vis%ally observed from the colo%r the %rine that 1ill be yello1 in colo%r' Riboflavin can be obtained from many so%rces s%ch as milk2 liver2 kidneys2 yeast2 m%shrooms and almonds' 3or this e0periment2 the riboflavin is get from the energy drinks' RDA recommends riboflavin intake for ad%lt men and 1omen 1ith *'.mg8day and *'*mg8day respectively' 6nfants and children are recommended to take riboflavin in lighter amo%nt of "'.9"'+mg8day for infants and "',9"'/mg8day for children' 4oo m%ch riboflavin 1ill ca%se stomatitis incl%ding painf%l red tong%e 1ith sore throat2 chapped and fiss%red lips (cheilosis# and inflammation of the corners of the mo%th (ang%lar stomatitis#'
-+.*CTI/*S: *' 4o determin determine e the concentr concentration ation of riboflav riboflavin in in energy energy drinks' drinks'
*01*,IM*NTA): A' :reparation of reagents; *' :repar :reparati ation on of < aceti acetic c acid acid sol%tio sol%tion; n; a' )m= )m= of concent concentrat rated ed acetic acetic acid sol%tio sol%tion n 1as pipett pipetted ed into a ""m= ""m= vol%metric flask and 1as dil%ted %ntil the calibration mark by %sing de9 ionised 1ater' B' :reparation of standards sol%tions; *' :reparatio :reparation n of *""ppm *""ppm riboflavin riboflavin stock sol%tion; sol%tion; a' *"mg of riboflav riboflavin in 1as dissolv dissolved ed by %sing %sing < acetic acetic acid sol%tion sol%tion'' b' 4he sol%ti sol%tion on then 1as transfe transferre rred d into into a *""m= *""m= vol%me vol%metric tric flask flask and 1as marked %p by %sing < acetic acid sol%tion' )' :rep :repar arat atio ion n of *"pp *"ppm m ribo ribofla flavi vin n sol%t sol%tio ion n from from *""p *""ppm pm ribof riboflav lavin in stoc stock k sol%tion; C*>* ? C)>) *"">* ? *"("# >* ? m= a' m= m= of *""p *""ppm pm ribofl riboflav avin in stoc stock k sol% sol%ti tion on 1as pipe pipett tted ed into into a "m= "m= vol%metric flask and 1as marked %p by %sing < acetic acid sol%tion' .' :repar :reparati ation on of standa standard rd sol%t sol%tion ions; s; Riboflavin 1orking sol%tion ? *"ppm Standard
Con$entration ppm3
/olume of ppm ribofla"in %or6in'
Blank * ) . +
" "'") "'"+ "'", "'"! "'*"
solution pipetted m)3 " "'* "') "'. "'+ "'
a' 4he 4he vol% vol%me me of *"pp *"ppm m ribo ribofl flav avin in 1ork 1orkin ing g sol%t sol%tio ion n 1as 1as pipe pipett tted ed into into each standard of "m= vol%metric flasks based on the table above' b' 4he < acetic acetic acid acid sol%tion sol%tion 1as 1as %sed %sed to mark %p the sol%tion sol%tion %ntil %ntil the calibration mark of "m=' Sample calculation of standard solution 1: C 1V 1 = C 2 V2 2 V 2 10V 1 = 0.02(50) V 1 = 0.1mL
C' :reparation of sample; a' "')m "')m= = of samp sample le 1as 1as pipe pipett tted ed into into a "m= "m= vol% vol%me metr tric ic flask flask and and 1as 1as marked %p %ntil the calibration mark by %sing < acetic acid sol%tion' Calculation of dilution factor for sample: 50
DF=
0.2
= 250
,*S)T: a' E0cita E0citation tion data data at +*',) +*',)nm nm (@ (@ma0#
Blank ("'""ppm# Standard "'")p "'")ppm Standard "'"+ppm Standard "'",ppm Standard "'"!ppm Standard "'*"ppm Sample A Sample B Sample C
Intensity *!'*. **"'.) *,.')) ).!'" .*"'+.-"'!/ *!'+" *-!', *"'!
b' Emissi Emission on data data at ."' ."'*n *nm m (@ma0#
Blank ("'""ppm# Standard "'")ppm Standard "'"+ppm Standard "'",ppm Standard "'"!ppm Standard "'*"ppm Sample A Sample B Sample C
Intensity *',) *"!'+) *,*' ).,'*+ ."'++ .,-'!/ *,'. *-,',* *+!'!)
Graph of emission intensity versus ver sus concentration 400 350
f(x) = 3467.14x + 25.82 R² = 1
300 250
Intensity
200 150 100 50 0 0
0.02
0.04
0.06
0.08
0.1
0.12
Concentration (ppm)
Sample calculation of concentration of sample: sample intensit! = 15".#5 ! = #$"%.1& ' 25.2 15".#5 = #$"%.1& ' 25.2 & = 0.0#%"ppm
Intensity *,'. *-,',* *+!'!)
Sample A Sample B Sample C
Con$entration ppm3 "'".-, "'"+. "'".
0.0376 + 0.0435 + 0.0355
Average concentration concentration of sample ?
3
? "'".!/ppm
See appendi0 for graph that sho1 ho1 the concentration of sample is obtained'
Stan Standa dard rd dei deiat atio ion n =
√
1
N
∑ ( x − mean) N − 1
2
i
i
=
√
1 3
−1
2
2
2
[( 0.0376 −0.0389 ) +( 0.0435− 0.0389 ) + ( 0.0355−0.0389 ) ]
= $.1$%* + 10 ,#
-elatie standard de deation //-SD
=
standard standard deviation + 100/ mean
−3 4.1479 × 10
=
0.0389
+ 100/
= 10.""
Act%al concentration of sample sample (average#
? "'".!/ppm )" (dil%tion factor# factor# ? /'-)ppm
mg Average amo%nt amo%nt of riboflavin in sample ? /'-) L "'"=
? "'+!,.mg
DISCSSI-N: 4he process of fl%orescence begin 1ith the absorption of > radiation by molec%le of analyte2 and the molec%le 1ill be e0cited to the higher electronic state from the gro%nd state2 contin%e 1ith vibrational rela0ation and internal conversion bet1een same m%ltiplicity and finally falls do1n back to the gro%nd state 1hile emitting the
fl%ore fl%oresc scenc ence' e' 3or e0cit e0citati ation on ma0im% ma0im%m m 1avele 1avelengt ngth h determ determina inatio tion2 n2 the emiss emission ion 1avelength 1as fi0ed 1hile the e0citation 1avelength 1as varied in a range' 3rom the e0periment2 the ma0im%m 1avelength for e0citation lies on +*',)nm' hen perfor performin ming g the emissi emission on analys analysis2 is2 the e0cit e0citati ation on 1avele 1aveleng ngth th 1as 1as fi0ed fi0ed and the emission 1avelength 1as varied in range' 4he ma0im%m 1avelength of emission is ."'*nm that is at longer 1avelength compared to e0citation 1avelength' As all kno1ing that energy is inversely proportional to the 1avelength2 it is proven that emiss emission ion of fl%ore fl%oresce scence nce occ%r occ%r at lo1er lo1er energy energy as compar compared ed to absorp absorptio tion n of radiation by referring to the ma0im%m 1avelength of e0citation and emission' 4he ma0im%m 1avelength is also as the %alitative analysis for specific analyte' Each analyte has its o1n specific ma0im%m 1avelength to e0cite and emit the radiation in order to determine them' 3rom the graph2 it is sho1n a good standard sol%tion preparation d%e to nearer of the val%e of R) to *' By %sing the e%ation of the straight line2 the concentration of each each sample sample can be determ determine ined' d' 4he 4he relati relative ve standa standard rd deviat deviation ion perce percenta ntage ge sho1ing the val%e of *"',, that indicates lo1 precision bet1een the triplicate sample intensity val%es' 4he good precision 1ill bring the
C-NC)SI-N: 4he act%al average concentration of riboflavin in the livita energy drink sample is /'-)ppm' 4he average amo%nt of riboflavin in the triplicate sample is e%al to "'+!,.mg' ,*7*,*NC*S: *' Riboflavin Riboflavin22 Retrieved April April ),2 from http;88en'm' http;88en'm'1iki 1ikipedia pedia'org81 'org81iki8Rib iki8Riboflav oflavin in )' 3' F' Holl Holler er22 D' A' Skoo Skoog2 g2 S' R' Cro% Cro%ch ch ()"" ()""-# -#22 :rin :rinci cipl ples es of 6nst 6nstr% r%me ment ntal al Analysis ,th ed'#2 nited States2 Brooks8Cole Cengage =earning'
