Journal of Scientific Research Vol. XXXX No.

1, June, 2010

ISSN 0555-7674

COMPARATIVE STUDY FOR THE DETERMINATION OF METALS IN MILK SAMPLES USING FLAME-AAS AND EDTA COMPLEXOMETRIC TITRATION
!i" Hu##!in$ A%%!r! N!&ir$ U%'r S(!)i*u'$ Mu(!%%!d S!+%!n Institute of $hemistr!, 4niversit! of the 5un6a%, 7ahore-545.0 Abstract: 8 comparative stu ! "as carrie out for the etermination of calcium an magnesium in 5 ifferent %ran s of milk 10alee%, Milk 5ak, 9lpers, :air! ;ueen an Nurpur3# 8fter r! ashing an treatment "ith concentrate nitric aci , calcium an magnesium "ere etermine %! 2lame 8tomic 8%sorption Spectrometr! 1288S3 an complexometric titration "ith <:(8# <richrome =lack-( an patton an ree er in icator "ere use for calcium an magnesium etermination %! complexometric titration metho # (he values o%taine for $a an Mg "ere foun to %e "ithin >09 limts Keywords: calcium? magnesium? complexometric titration? flame atomic a%sorption spectrometr!# Introduction Milk is a complex material consisting of several components, "hich have a significant role even though present in lo" concentrations# Increase a"areness of the influence of iet on human health has prompte pro ucers to pro uce foo of higher &ualit! "hich shoul %e rich in nutrients an vitamins# 'oo &ualit! measurements are essential to control, maintain pro ucts an process &ualit!, in manufacturing, tra e an research# (his is important, since ra" materials for foo pro uction are %ecoming poor in ) essential minerals# Milk an air! pro ucts have %een recogni*e "orl "i e as a goo source of "holesome nutrition for the human %o !# Milk contains a large variet! of essential nutrients for the evelopment , an maintenance of a salutar! life# It is an outstan ing source of calcium an phosphorus, an can suppl! mo erate amounts of magnesium, *inc, iron an /,4 copper# (hese essential metals are of interest to etermine the a e&uate ail! intake %! an organism# 0o"ever, ue to an increase in environmental pollution, it is necessar! to etermine an monitor the
1-For Proof and Correspondence: Zaib Hussain

level of toxic metals in milk, such as ca mium an lea , as the! ma! have a verse effects on human health# $alcium contri%utes to structural functions in %ones an teeth along"ith regulating man! %iological functions# More recentl!, focus on calcium has centere on its role in preventing 5 osteoporosis# Magnesium is the fourth most a%un ant cation in the human %o ! after so ium, potassium an calcium# Most of the magnesium in the %o ! is 6 eposite in %ones# (he un erstan ing of milk magnesium composition is important for nutritional management 7 uring earl! life# $opper has great %iological importance in milk "here%! higher copper contents in milk contri%ute to"ar s acceleration of lipi oxi ation# +ecentl! the action of copper in several -,. important iseases has %een reporte # (he function of nickel is not completel! un erstoo as an essential element in the human %o ! %ut its importance in the human %o ! nee s to %e further researche # (his "ork com%ines anal!sis using atomic a%sorption spectroscop! for the etermination of metals 1$a, Mg, 2e, Ni, $u an $ 3 in 5 ifferent milk %ran s

Zaib Hussain

10alee%, Milk 5ak, 9lpers, :air! ;ueen, an Nurpur3 an a complexometric titration "ith <:(8 for etermination of $a an Mg# 8 comparison "as ma e to measure the levels of $a an Mg in milk an stan ar eviation# @alues "ere calculate for these metals 1$a, Mg, 2e, Ni, $u an $ 3 an compare to >09 )0 limits# M!t'ri!+# !nd M't(od# St!nd!rd So+ution#-2or anal!sis of the samples %! 88S, stan ar solutions of $a, Mg, 2e, Ni, $u an $ of anal!tical gra e 1Merck3 "ere use an the appropriate ilutions "ere ma e accor ingl!# Pr',!r!tion o) S!%,+' So+ution#-5 ifferent milk %ran s of 0alee%, Milk 5ak, 9lpers, :air! ;ueen an Nurpur "ere purchase from local markets of 7ahore to prepare the sample solutions# Dr/in. o) S!%,+'-8)00 ml covere porcelein cruci%le "as use in "hich, 5 ml of milk sample "as evaporate to r!ness in a r!ing oven o 1=in er3 at )00 $# (his allo"e for po" ere milk to %e o%taine "hich is easil! preserve # (he shelf life of po" ere milk is longer than li&ui milk an so there is no nee to refrigerate it ue to its lo" moisture content# A#(in. o) S!%,+'-(he rie po" ere milk "as then place in a porcelein cruci%le an transferre to a graphite furnace 1=oxt!pe +esistance 2urnace Mo el SC-5),3# (he temperature "as increase o slo"l! at a maximum rate of 50 $Dhr to o 450 $ an heating "as continue until the ash turne "hite# (he sample "as coole overnight an then groun using a pestle an mortar into finel! ivi e po" ere form#

Pr',!r!tion o) So+ution-(o po" ere "hite ash, 50ml of eionise "ater an 5 ml of concentrate nitric aci "as a e # (he solution "as gentl! stirre an heate to issolve the ash contents completel! after "hich the solution "as filtere to remove the un issolve particles# (he solution "as then transferre into a )00 ml volumetric flask an the volume "as ma e upto the mark using eionise "ater# R'!.'nt# !nd So+ution#-8ll reagents use "ere of anal!tical gra e 1Merck3# :eionise "ater "as use for the preparation of all solutions# (riethanol amine an eth!l alcohol 18nal!tical 'ra e3 "ere use for the preparation of organic reagent <riochrome =lack-(, as an in icator, an <th!lene iamine tetreacetic aci 1<:(83, "as use as a complexing agent# 8n alkaline %uffer of ammonium chlori e-ammonia 1p0 )03 "as use to maintain p0 an concentrate 0N9 / "as use for the igestion of organic matter# 5atton an ree er in icator "as use in the complexometric titration of calcium along "ith A90 for p0 a 6ustment# Pr',!r!tion o) 0 1 0 2 M EDTA So+ution-0#0) M <:(8 solution "as prepare %! i s s o l v i n g / # 7 7 , 4 g < : (8 1Na,0,B#,0,93 in 500 ml eionise "ater "ith constant stirring for )5 min in a )7 volumetric flask# >hen the soli cr!stals of <:(8 "ere completel! issolve , the volume "as ma e upto the mark# Pr',!r!tion o) 3u))'rSo+ution-8n alkaline %uffer of ammonium chlori e-ammonia 1p0 )03 "as prepare %! issolving )7#5g of ammonium chlori e 1N04$l3 in 50 ml eionise "ater in a ,50 ml volumetric flask# (o this, )4, ml of concentrate li&ui N0/ "as a e an the volume "as ma e )0

Comparati e Stu!" for the #etermination of $etals in $il% Samples

upto the mark to form a %uffer of p0 )0# Pr',!r!tion o) Erioc(ro%' 3+!c4 T Indic!tor-2or preparation of this in icator, 0#,g of <:( 1/-h! rox!-4-F 1)-h! rox!-,naphth!l3 a*o-7-nitro-)-naphthalenesulphonic aci , so ium salt3 "as issolve in )5 ml of triethanolamine "ith the a ition of 5 ml of a%solute ethanol to o%tain fresh in icator# (he solution "as filtere an store %et"een o 0 an 5 $# In#tru%'nt!tion-(he a%sor%ance of the solution "as measure "ith atomic a%sorption spectrophotometer 10itachi-5olarise

Eee man 8t omic 8%sorption Spectrophotometer Mo el E-5003# <lectric =alance 1Scout 5ro 9084S3 "as also use for "eighing purposes# G'n'r!+ Proc'dur'-S!%,+' An!+/#i# "/ Ato%ic A"#or,tion )or C!$ M.$ F'$ Cu$ Ni !nd Cd 8 0itachi-5olari*e Eeeman 8tomic 8%sorption Spectrophotometer, E-5000 "as use for the etermination of $a, Mg, 2e, $u, Ni an $ in ifferent %ran s of milk (he samples "ere run in triplicate# (a%le ) an , sho"s the "orking parameters for each element anal!se #

(a%le )G >orking parameters H stan ar con itions for 0itachi-5olarise 1Eeeman 8tomic 8%sorption Spectrophotometer 1Mo el E-50003#

Metal $a Mg 2e $u Ni $

>avelength 1nm3 4,,#,-5#/ ,4-#/ /,4#,/, ,,-#-

Slit "i th 1nm3 )#/ )#/ )#/ )#/ 0#, )#/

(ime constant 1s3 )#0 )#0 )#0 )#0 )#0 )#0

7amp current 1m83 .#0 .#0 .#0 .#0 ), .#0

(a%le ,G 5arameters of 88S anal!sis

Metal $a Mg 2e $u Ni $

8tomi*er Stan Stan Stan Stan Stan Stan ar ar ar ar ar ar

2lame t!pe 8ir- $,0 , 8ir- $,0 , 8ir- $,0 , 8ir- $,0 , 8ir- $,0 , 8ir- $,0 ,

2uel flo" 1)Dmin3 ,#, ,#, ,#, ,#, ,#, ,#,

9xi ant 1k5a3 )60 )60 )60 )60 )60 )60

=urner height 1mm3 7#5 7#5 7#5 7#5 7#5 7#5

))

Zaib Hussain

C!+ciu% !nd M!.n'#iu% D ' t ' r % i n ! t i o ni n M i + 4 " / Co%,+'5o%'tric Titr!tion 2or calcium etermination, ,0ml of sample "as taken in a conical flask an ,-/ pellets of A90 "ere a e # 8fter shaking the solution ) g of 5atton an ree er in icator 1calcon /-car%ox!lic aci 3 "as a e an the sample "as titrate against 0#0)M <:(8 solution until a color change from "ine re to %lue appeare # 2or magnesium, ,0ml of sample "as taken in a conical flask an ,-/ml of %uffer solution "as a e follo"e %! 45 rops of <riochrome =lack ( in icator# (he sample "as then titrate against 0#0)M <:(8 solution until the color change from "ine re to %lue# RESULTS C!+ciu% (he concentration of calcium in ifferent %ran s is ,#50, ,#0,, ,#,/, ,#-0 an ,#67 as etermine %! atomic a%sorption spectroscop!# @alues %! <:(8 titration are 4#-, 4, 5, 4 an 4# (he values of stan ar eviation %! atomic a%sorption an <:(8titration are 0#,- an 0#4/# M!.n'#iu% (he concentration of magnesium in ifferent %ran s of milk is )#64, )#,., )#5,, )#)) an )#)5 as etermine %! (a%le /G 8nal!sis of metals %! 88S anal!sis
E+'%'nt# $a Mg 2e $u Ni $ H!+''" 627 ,#50 )#64 0#56 0#,) 0#00#0Mi+4,!4 687 ,#0, )#,. 0#5. 0#)0#00#07

atomic a%sorption# @alues %! titration are /#/., /, 4#4, /#. respectivel!# (he stan ar values %! atomic a%sorption an titration are 0#,0 an 0#6,#

<:(8 an ,#50 eviation <:(8

Iron (he concentration of iron in ifferent %ran s of milk is 0#56, 0#5., 0#6,, 0#5an 0#56 as etermine %! atomic a%sorption# (he stan ar eviation is 0#0,# Co,,'r (he concentration of copper in ifferent %ran s of milk is 0#,), 0#)-, 0#)4, 0#)/ an 0#)/ as etermine %! atomic a%sorption# (he stan ar eviation is 0#0/# Nic4'+ (he concentration of nickel in ifferent %ran s of milk is 0#0-, 0#0-, 0#0-, 0#0an 0#0- as etermine %! atomic a%sorption# (he stan ar eviation is 0#,-# C!d%iu% (he concentration of ca mium in ifferent %ran s of milk is 0#0-, 0#07, 0#07, 0#07 an 0#07 as etermine %! atomic a%sorption# (he stan ar eviation is 0#,6#

Mi+4 3r!nd# O+,'r# 697 D!ir/ :u''n 6;7 ,#,/ ,#-0 )#5, )#)) 0#6, 0#50#)4 0#)/ 0#00#00#07 0#07

Nur,ur 6<7 ,#6) )#)5 0#56 0#)/ 0#00#07

),

Comparati e Stu!" for the #etermination of $etals in $il% Samples

(a%le 4G 8nal!sis of metals %! complexometric metho

E+'%'nt# $a Mg H!+''" 627 4#/#/. Mi+4,!4 687 4 /#/ Mi+4 3r!nd# O+,'r# 697 D!ir/ :u''n 6;7 5 4 4#4 /#. Nur,ur 6<7 4 ,#5

Di#cu##ion (he metho use for the igestion of organic matter is r! ashing or igestion# (here are t"o t!pes of igestions, "et igestion an r! igestion# :r! igestion is preferre over "et igestion %ecause "et igestion is time consuming an re&uires an excess of strong aci "hich interferes "ith the anal!*ers# 4se of nitric aci is preferre over the other aci s %ecause nitric aci is an accepta%le matrix for %oth flame an electrothermal anal!sis# Sulphuric aci , perchloric aci an h! rochloric aci interfere in the anal!sis of some metals an all provi e a poor matrix# 4sing 88S it "as foun that $a is highest in sample 4 an lo"est in sample ,# 8nal!sing %! complexometric titration sho"e higher concentration of $a in sample /# (he values for magnesium are )#64, )#,., )#5,, )#)) an )#)5 as etermine %! atomic a%sorption spectrometr!# (he higher value for magnesium is foun in sample num%er ) for atomic a%sorption an for complexometr! the higher value is foun for sample num%er /# (he values of calcium an magnesium %! titration are 4#-, 4, 5, 4, 4 an /#/., /, 4#4, /#,4 an ,#50 in the samples ), ,, /, 4 an 5 respectivel!# =oth the values as etermine %! atomic a%sorption spectroscop! an complexometric titration are "ithin the >09 limits# (he values o%taine %! titration are almost ou%le the values %! atomic a%sorption spectrometr! %ecause titration is an in irect metho an is not as precise as atomic a%sorption spectrometr!#

9ther than calcium an magnesium values for iron, copper, nickel an ca mium "ere also foun # @alues for iron in sample num%er ), ,, /, 4 an 5 are 0#56, 0#5., 0#6,, 0#5-, 0#56# @alues for copper in sample num%er ), ,, /, 4 an 5 are 0#,), 0#)-, 0#)4, 0#)/ an 0#)/ respectivel!# @alues for nickel in sample num%er ), ,, /, 4 an 5 "as foun to %e the same 10#0-3# @alues for ca mium in sample num%er ), ,, /, 4 an 5 are 0#0-, 0#07, 0#07, 0#07 an 0#07 respectivel!# Iron is present in highest concentration in sample num%er / an lo"est in samples ) an 5# 0igher concentration of copper has %een foun in sample num%er ) an lo"er in samples 4 an 5# (he value for nickel is the same in all the samples# 2or ca mium all the samples have the same value other than sample num%er ) "hich is higher than all# 8s these elements are present in ver! lo" concentrations so these elements cannot %e foun %! complexometric titration "ith <:(8# (he most a e&uate metho for etermining these metals is %! 8tomic 8%sorption Spectroscop!# (he values for the stan ar eviation for calcium an magnesium %! 8tomic 8%sorption Spectroscop! an complexometric titration %! <:(8 are 0#,-, 0#,0, 0#4/ an 0#6, respectivel!# (he values for stan ar eviation for iron, copper, ca mium an nickel are 0#0,, 0#0/, 0#,- an 0#,6 respectivel!# 8 graph is plotte %et"een the ifferent elements foun in milk i#e# calcium, magnesium, iron, copper, nickel an ca mium# $alcium is foun to %epresent in higher concentrations an ca mium is )/

Zaib Hussai

foun to %e present in lo"er concentrations# 0igher concentrations of calcium are esira%le as milk is a rich source of calcium# In the case of heav! metals, iron is present in higher concentrations an ca mium is present in lo"er concentrations# 8ll these concentrations are "ithin the >09 limits# (he values for all the minerals in icate that all the %ran s of milk are ver! "ell processe an are rinka%le# R')'r'nc'# )3 =irghila S, :o%rinas S, Stanciu ', Soceanu 8# :etermination of ma6or an minor elements in milk through I$5-9<S# <nviron# <ng# Manag# I# 7163G-05--0-, ,00-# ,3 2ennema 9, 2oo $hemistr!, /r e #, Marcel :ekker, Ne" Bork, )..6# 7ev! B, Eeharoa 8, 'rune%aum M, Nit*an M, Steinher* +# $opper eficienc! in infants foo fe co" milk# I# 5e iat# )06G7-67--, ).-5# 5ennigton I, >ilson :, Boung =, Iohnson +, @an erveen I# Mineral content of market samples of "hole milk# I# 8m# :iet# 8ssoc# -7G)0/6-)04,, ).-7# Mahan 7, Stump S, 8limentos, Nutricao an :ietuterapia, +oca,

Sao 5aulo, ,00,# 63 $ivitelli +, 8violi 7# $alcium, phosphate an magnesium a%sorption InG Iohnson 7+ e # 5h!siolog! of the 'astrointestinal tract, /r e # +aven 5ress, Ne" Bork, )..4# 2eele! +, <ittenmiller +, Iones I, =arnhart 0# $alcium, phosphorus an magnesium contents of human milk uring earl! lactation# I# 5e iatr# 'astroenterol# Nutr# ,G,6,-,76, ).-/# Aleva! 7, 2or%ush I# $opper meta%olism an epi emiolog! of coronar! heart isease# Nutr# +ep# Int#)4G,,)-,,/, ).76# E ! k a @# ' e o c h e m i c a l environment an the inci ence of cancer in Autna 0ora, S=# 'eol# @e # (echnol# 'eochem#))G)55)5., ).7)# Iorhem 7# :etermination of metals foo stuff %! atomic a%sorption spectrometr! after r! ashingG NMA7) inter-la%orator! stu ! of lea , ca mium, *inc, copper, iron, chromium an nickel# I# 898$ Inter# 76G7.--)/, )../#

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