Chapter I

II.
AUTOMATIC ANALYTICAL METHODS FOR ENVIRONMENTAL MONITORING AND CONTROL

Laboratory exercises
I. ANALYTICAL TECHNIQUES FOR
ENVIRONMENTAL MONITORING AND
CONTROL
Laboratory Exercises
1
II. AUTOMATIC ANALYTICAL METHODS FOR ENVIRONMENTAL MONITORING AND CONTROL
I.1. SPECTROPHOTOMETRIC DETERMINATION OF
AMMONIUM BY THE INDOPHENOL BLUE METHOD
This method is appi!a"e to the dete#mi$atio$ o% the ammo$i&m !o$te$t i$
'ate#s (e.). p#e!ipitatio$* 'ithi$ the #a$)e +.+, to -.+ m) NH
,
.L.
Principle
I$ a$ a/ai$e so&tio$ (pH 1+.,011.1* ammo$i&m io$s #ea!t 'ith h2po!ho#ite
to %o#m mo$o!ho#ami$e. I$ the p#ese$!e o% phe$o a$d a$ e3!ess o% h2po!ho#ite4 the
mo$o!ho#ami$e 'i %o#m a "&e !oo&#ed !ompo&$d4 i$dophe$o4 'he$
$it#op#&sside is &sed as !ata2st. The !o$!e$t#atio$ o% ammo$i&m is dete#mi$ed
spe!t#ophotomet#i!a2 at 56+ m$.
Reagent an! Intr"#entati$n
A the !hemi!as m&st "e o% #e!o)$i7ed a$a2ti!a )#ade. The 'ate# &sed %o#
di&tio$ a$d #i$si$) sho&d "e do&"e0distied o# de0io$i7ed a$d distied.
8he$o (C
5
H
1
OH*
Sodi&m $it#op#&sside (Na
-
Fe(NO* (CN*
1
9 -H
-
O*
Sodi&m h2d#o3ide (NaOH*
Sodi&m h2po!ho#ite so&tio$ (NaOC* 1M: ma/e a so&tio$ !o$tai$i$) app#o3.
6.1; a!ti<e !ho#i$e (61 ).L* i$ +.1 M NaOH
Ammo$i&m !ho#ide (NH
,
C*
Sodi&m thios&phate (Na
-
S
-
O
6
*
Rea)e$t A: disso<e 6.1 ) phe$o a$d +.+,+ ) sodi&m $it#op#&sside i$ 1++ mL
'ate#. Sto#e the so&tio$ #e%#i)e#ated i$ the da#/. I% the !oo&# o% the so&tio$
t&#$s )#ee$ish4 it m&st "e dis!a#ded4 a$d a %#esh so&tio$ p#epa#ed.
Rea)e$t =: disso<e 1.> ) sodi&m h2d#o3ide i$ some 'ate# i$ a 1++ mL
<o&met#i! %as/. Add ,.+ mL 1 M sodi&m h2po!ho#ite so&tio$4 a$d di&te 'ith
'ate# to the ma#/. Sto#e the so&tio$ #e%#i)e#ated i$ the da#/. I% the so&tio$ is
sto#ed %o# 'ee/s4 the !o$!e$t#atio$ sho&d "e !he!/ed "2 tit#atio$ 'ith a sodi&m
thios&phate so&tio$.
Sta$da#d ammo$i&m so&tio$ I4 1++ m) NH
,
?
.L:
Ammo$i&m !ho#ide m&st "e d#ied %o# o$e ho&# at 1++ @C.
Disso<e +.-A51 ) o% the d#ied sat i$ 'ate# i$ a 1+++ mL <o&met#i! %as/. Di&te
to the ma#/ 'ith 'ate#. The so&tio$ is sta"e %o# si3 mo$ths 'he$ sto#ed i$
a #e%#i)e#ato#.
Sta$da#d ammo$i&m so&tio$ II4 , m) NH
,
?
.L: "2 mea$s o% a pipette4 t#a$s%e#
-+.+ mL o% sta$da#d ammo$i&m so&tio$ I to a 1++ mL <o&met#i! %as/. Di&te
-
'ith 'ate# to the ma#/. This sta$da#d ammo$i&m so&tio$4 a$d the ammo$i&m
so&tio$s made %o# p#epa#i$) the !ai"#atio$ !&#<e4 m&st "e %#esh2 made.
Spe!t#ophotomete# 'ith opti!a !e o% 1+ mm.
Bate# "ath 'ith the#mostat4 1+ @C
Test t&"es: 6+ mL
Vo&met#i! %as/s: 1+4 1++ a$d 1+++ mL
8ipettes: 1.+4 -.+4 ,.+4 1.+4 1+.+4 -+.+4 -1.+4 1+.+ mL.
Mi!#opipette: -1+ CL
Cali%rati$n
8#epa#atio$ o% !ai"#atio$ !&#<e:
1. T#a$s%e# to 1++ mL <o&met#i! %as/ +.+4 1.+4 -.+4 1.+4 1+.+4 -1.+ a$d 1+.+ mL o%
sta$da#d ammo$i&m so&tio$ II. Di&te to the ma#/ 'ith 'ate#. The
!o$!e$t#atio$s o% these so&tio$s a#e +.++4 +.+,4 +.+>4 +.-4 +.,4 1.+ a$d -.+ m)
NH
,
?
.L. T#a$s%e# 1.+ mL o% ea!h o% these sta$da#d so&tio$s a$d 1.+ mL o% 'ate#
to a 6+ mL test t&"e.
-. Add to the test t&"e -1+ CL #ea)e$t A &si$) a mi!#o pipette4 a$d mi3 'e. Add
the$ -1+ CL #ea)e$t = &si$) a mi!#o0pipette a$d mi3 'e. Co<e# the ope$i$) o%
the t&"e 'ith some i$e#t mate#ia. 8a!e the t&"e i$ the 'ate# "ath at 1+ @C %o#
t'o ho&#s.
6. Coo the so&tio$ to #oom tempe#at&#e4 a$d t#a$s%e# it to a 1+ mm !e. Meas&#e
the a"so#"a$!e at 56+ $m.
,. 8#epa#e a !ai"#atio$ !&#<e "2 potti$) the a"so#"a$!e o% ea!h o% the sta$da#d
so&tio$s a)ai$st its !o$!e$t#atio$ o% ammo$i&m. 8#epa#e a $e' !ai"#atio$ !&#<e
%o# ea!h se#ies o% sampes.
1. I$ o#de# to !he!/ %o# ammo$i&m i$ the #ea)e$ts4 ta/e a photomet#i! #eadi$) o%
the "a$/ (+.++ m) NH
,
?
.L* a)ai$st 'ate#. The a"so#"a$!e sho&d $ot e3!eed
+.+-+.
Anal&tical Pr$ce!"re
T#a$s%e# 1.+ mL o% the sampe a$d 1.+ mL o% 'ate# to a 6+ mL test t&"e.
8#o!eed a!!o#di$) to the i$st#&!tio$s p#ese$ted at poi$ts - a$d 6 o% a"o<e. Co$<e#t the
spe!t#ophotomet#i! #eadi$)s o% the sampe to m) NH
,
?
.L "2 mea$s o% the !ai"#atio$
!&#<e. The !o$!e$t#atio$ ma2 "e e3p#essed as m) N.L "2 m&tip2i$) 'ith +.DD>.
Sampes !o$tai$i$) mo#e tha$ -.+ m) NH
,
?
.L m&st "e di&ted. Bith s&ita"e
eE&ipme$t the FI$dophe$o methodG !a$ "e made a&tomati!.
Inter'erence
I#o$ (III* ma2 i$te#%e#e i% the !o$!e$t#atio$ is mo#e tha$ - m).L. I% the pH0
<a&e o% the sampe is o'e# tha$ 64 the sampe sho&d "e $e&t#ai7ed.
6
I% the sampe is t&#"id4 "oth the sampe a$d the "a$/ sho&d "e %ite#ed th#o&)h a
'hite "a$d %ite#.
,
I.(. DETERMINATION OF CHLORIDE IN )ATERS
The method !a$ "e &sed %o# di#e!t dete#mi$atio$ o% the !ho#ide io$ !o$te$t i$
'ate# sampes 'ithi$ the #a$)e +.+1 to 1 m).L.
Principle
Cho#ide io$s 'i s&"stit&te the thio!2a$ate io$s i$ &$disso!iated me#!&#2
thio!2a$ate. The #eeased thio!2a$ate io$s #ea!t 'ith %e##i! io$s %o#mi$) a da#/ #ed
i#o$0thio!2a$ate !ompe3.
The a"so#"a$!e is meas&#ed at ,5+ $m.
- C
0
? H)(SCN*
-
H H)C
-
? -SCN
0
SCN
0
? Fe
6?
H Fe(SCN*
-?
D&#i$) the a$a2sis4 &se o$2 !hemi!as o% #e!o)$i7ed a$a2ti!a )#ade a$d
o$2 do&"e0distied o# deio$i7ed a$d distied 'ate#.
8e#!ho#i! a!id (HCO
,
* D-; I
Me#!&#2 (II* thio!2a$ate (H)(SCN*
-
*I
I#o$ (III* $it#ate $o$ah2d#ate (Fe(NO
6
*
6
9 AH
-
O*I
Sodi&m !ho#ide (NaC*I
Etha$o (C
-
H
1
OH*I
8e#!ho#i! a!id4 1:1. Mi3 1 <o&me D-; pe#!ho#i! a!id 'ith 1 <o&me o% 'ate#I
Me#!&#2 (II* thio!2a$ate so&tio$4 sat&#ated. Sha/e 1 ) H)(SCN*
-
'ith 1+++ mL
etha$o. Fite# the so&tio$ a%te# -, ho&#s. The so&tio$ ma2 "e sto#ed i$ a )ass
"otte at #oom tempe#at&#eI
I#o$ (III* $it#ate so&tio$4 5;. Disso<e 5 ) Fe(NO
6
*
6
9 AH
-
O i$ 1++ mL 1:1
pe#!ho#i! a!id. Fite# the so&tio$ a%te# -, ho&#sI
Sta$da#d !ho#ide so&tio$ I4 1+++ m).L. Disso<e ,1-.1 m) NaC d#ied at 1,+0
-++ @C4 i$ 'ate# a$d %i it &p to -1+ mL 'ith 'ate#I
Sta$da#d !ho#ide so&tio$ II4 1+ m).L. Di&te 1+.+ mL sta$da#d !ho#ide so&tio$
I to 1++ mL 'ith 'ate#.
Preparati$n $' t*e Cali%rati$n C"r+e
1. T#a$s%e# -.14 1.+4 D.14 1+4 114 -+ a$d -1 mL o% sta$da#d !ho#ide so&tio$ II to 1+
mL <o&met#i! %as/s4 a$d %i &p to the ma#/ 'ith 'ate#. These so&tio$s !o$tai$
+.14 1.+4 1.14 -.+4 6.+4 ,.+ a$d 1.+ m) C
0
.L.
1
-. T#a$s%e# -1 mL o% the !ai"#atio$ so&tio$s to 1++ mL E#e$me2e# %as/s. To ea!h
%as/ add 'ith pipettes 1 mL me#!&#2 (II* thio!2a$ate so&tio$ a$d - mL i#o$ (III*
$it#ate so&tio$. Mi3 'e "et'ee$ a$d a%te# the additio$s. A%te# -+ mi$&tes4
meas&#e the a"so#"a$!e i$ 1+ mm !es at ,5+ $m. As #e%e#e$!e4 &se -1 mL 'ate#
mi3ed 'e 'ith 1 mL o% #ea)e$t (-* a$d - mL o% #ea)e$t (6*.
8ot the #eadi$)s a)ai$st the !o$!e$t#atio$s a$d d#a' the !ai"#atio$ !&#<e.
T#a$s%e# -1 mL o% the p#e!ipitatio$ sampe to a 1++ mL E#e$me2e# %as/.
8#o!eed as a"o<e. Read the !ho#ide !o$te$t o% the sampe %#om the !ai"#atio$ !&#<e.
Inter'erence
=#omide a$d iodide 'i )i<e the same a"so#"a$!e as the eE&i<ae$t amo&$t
o% !ho#ide.
Re'erence
1. I'asa/i4 I.4 Uts&mi4 S.4 a$d O7a'a4 T. (1A1-* Ne' !oo#imet#i! dete#mi$atio$ o%
!ho#ide &si$) me#!&#i! thio!2a$ate a$d %e##i! io$. Bull. Chem. Soc. Japan, 25,
--5.
-. Ja4 M.4 Fishe#4 D.4 a$d Game#4 K. (1A15* 8hotomet#i! dete#mi$atio$ o%
!ho#ides i$ 'ate#. Anal. Chem., 28, 15510155>.

5
I.,. DETERMINATION OF THE SO
(
CONCENTRATION IN AIR
Intr$!"cti$n
A se#io&s e!oo)i!a p#o"em is the atmosphe#i! po&tio$ "2 s&ph&# dio3ide
'hi!h o#i)i$s a#e d&e to the !om"&stio$ o% s&ph&# !ompo&$d !om"&stio$ i$ e$)i$es
o# "oie#s. The so' %i)ht a)ai$st this po&tio$ sta#ted #e!e$t2I i$ %a!t e)a #&es a#e
<e#2 #est#i!ti<e 'ith the !o$!e$t#atio$ o% s&ph&# i$ %&es.
The p#ese$t method %o# s&ph&# dio3ide dete#mi$atio$ i$ the ai# is "ased o$
the a"so#ptio$ a$d o3idatio$ o% s&ph&# dio3ide i$ the sampe p#od&!i$) s&ph&#i!
a!id. T#appi$) is pe#%o#med "2 mea$s o% the ai# "&""i$) th#o&)h a$ h2d#o)e$
pe#o3ide disso&tio$ at %i3ed pH d&#i$) a !e#tai$ pe#iod (-, h*.
To dete#mi$e the s&ph&#i! a!id !o$!e$t#atio$ %o#med this is p&t i$ !o$ta!t
'ith a 'e0/$o'$ amo&$t o% "a#i&m a$d tho#i$e sat. This #ea!tio$ p#od&!es "a#i&m
s&phate p#e!ipitati$) a$d the #esid&a "a#i&m io$s 'i %o#m a !ompe3 !oo&#ed "2
#ea!tio$ 'ith the tho#i$e that !a$ "e meas&#ed "2 a"so#ptio$ at 1-+ $m.
The di%%e#e$!e "et'ee$ the amo&$ts i$itia a$d %i$a o% "a#i&m io$s 'hi!h
!o##espo$ds 'ith the s&phate !o$!e$t#atio$ i$ the so&tio$ )i<es the amo&$t o%
s&ph&# dio3ide o3idi7ed i$ the sampe.
Rea!tio$ i$to the "&""i$) <esse:
SO
-
? H
-
O
-
H
-
SO
,
Fo#matio$ o% !oo&#:
=a
-?
? tho#i$e (=a 0 tho#i$e*
#ed !oo&#
(=a0tho#i$e* ? H
-
SO
,
=aSO
,
? -H
?
? tho#i$e ? (=a
#esid&a
0tho#i$e*
#ed
Target
To dete#mi$e the dai2 a<e#a)e !o$!e$t#atio$ o% SO
-
i$ sampes o%
atmosphe#i! ai#.
Ha$de !o##e!t2 a ma$&a !o$t#o statio$ %o# the ai# E&ait2.
To "e t#ai$ed o$ the !o##e!t &se o% mate#ias a$d p#o!esses 'ith possi"e
!hemi!a #is/.
D
Glassware Reagents
Sampi$) set
(ma$&a !o$t#o statio$*
Spe!t#ophotomete#
Gass !e4 1+ mm i)ht path
Bate# E&ait2 Mii0K
H
-
O
-

HCO
,
D-;
H
-
SO
,
A>;
=a(CO
,
*
-
a$h2d#e
1 mi!#opipet4 -1+ L
1+ <o&met#i! %as/s 1++ mL
- pipettes 1 mL Tho#i$e
- pipettes 1+ mL Dio3a$e
Manual station of control
A ma$&a statio$ !o$sists o% the %oo'i$) eeme$ts:
0 Ai# e$t#a$!e: !o$i!a %&$$e %o# sampi$) o% 601 !m4 made o% po2<i$2 !ho#ide
a$d pa!ed <e#ti!a2 'ith the mo&th do'$'a#ds. It sho&d "e pa!ed %a# a'a2 %#om
'as a$d othe# dist&#"i$) eeme$ts.
0 Meas&#e# the sampe <o&me a$d !o$t#o o% the %o'0#ate.
0 Aspi#ati$) p&mp: the %o'0#ate sho&d "e adL&sted to - m
6
da2.
0 S2stem o% %it#atio$: a 1 !m
-
a#ea0t#ap %o# pa#ti!es %o#med "2 a %ite#i$) de<i!e
#etai$i$) pa#ti!es o<e# the #a$)e 101 Cm 'ith a$ e%%i!ie$!2 as !ose as possi"e to
1++;. This %ite# 'i "e apt %o# a %o'0#ate o% - m
6
da2.
0 =&""i$) de<i!e 'ith !apa!it2 %o# a <o&me o% 1++ mL.
0 Co$$e!to#s: po2<i$2 !ho#ide t&"i$) 'ith a$ i$te#$a diamete# o% > !m. A<oid
!&#<es a$d i% $ot possi"e the st#i!t2 $e!essa#2 m&st ha<e a #adi&s hi)he# tha$ 1
!m.
A so&tio$s p#epa#ed 'ith 'ate# E&ait2 o% mii0K.
Caution: The %oo'i$) #ea)e$ts m&st "e !a#e%&2 ha$ded: pe#!ho#i! a!id4
"a#i&m pe#!ho#ate4 dio3a$e a$d tho#i$e.
1* -D06+ ; H
-
O
-
('.'* .
-* +.1 mo L
01
pe#!ho#i! a!idI ta/e ,.6 mL o% D-; pe#!ho#i! a!id a$d e<e to 1++
mL.
6* 6* +.+1 mo L
01
pe#!ho#i! a!id: -1mL o% +.1 mo L
01
pe#!ho#i! a!id sho&d "e
e<eed to -1+ mL.
,* T#appi$) so&tio$. Di&te 1+ mL o% disso&tio$ 1 o% -D06+ ; H
-
O
-
&p to 1+++
mL 'ith 'ate# E&ait2 mii0K. AdL&st pH o<e# the #a$)e ,.+ M ,.1 (pH0mete#*
"2 d#oppi$) the so&tio$ +.1 mo L
01
o% pe#!ho#i! a!id. This so&tio$ !a$ "e
!o$se#<ed d&#i$) a mo$th i$ the #e%#i)e#ato#.
1* =a(CO
,
*
-
so&tio$. So<e +.1-1 ) o% a$h2d#o&s "a#i&m pe#!ho#ate i$ a sma
<o&me o% so&tio$ 6 (+.+1 mo L
01
pe#!ho#i! a!id*. This so&tio$ is t#a$s%e##ed to
<o&met#i! %as/ o% -1+ mL a$d e<e 'ith the same a!id.
5* So&tio$ o% =a(CO
,
*
-
.dio3a$e. Di&ted 1 mL o% the so&tio$ 1 (=a(CO
,
*
-
* 'ith
,+ mL o% 'ate# mii0K a$d t#a$s%e# i$to a <o&met#i! %as/ o% 1+++ mLI the$
made &p 'ith dio3a$e.
>
D* Sta$da#d so&tio$ o% s&phate +.1++ mo L
01
. Di&te -> mL o% A>; o% H
-
SO
,
&p to
1+++ mL a$d tit#ate 'ith NaOH so&tio$ (the h2d#o3ide has "ee$ tit#ated <s the
a!id potassi&m phthaate*. 1 mL o% this so&tio$ s&phate !o##espo$ds 'ith 6- m)
o% SO
-
.
>* Sta$da#d so&tio$ o% s&phate +.1-1 mo L
01
. -1 mL %#om so&tio$ D (+.1 mo L
01
*
a#e ta/e$ a$d di&ted 'ith mii0K 'ate# i$to a <o&met#i! %as/ 1+++ mL. 1 mL o%
this so&tio$ !o##espo$ds 'ith >++ ) o% SO
-
.
A* Sta$da#d s&phate so&tio$ +.++1-1 mo L
01
. Di&te 1+ mL o% the sto!/ so&tio$
(+.1-1 mo L
01
* i$ a 1++ mL <o&met#i! %as/ 'ith 'ate# (1 mL o% this so&tio$
!o##espo$ds to >+ ) o% SO
-
*.
1+* Tho#i$e so&tio$ -.1 ) L
01
. Disso<e +.1-1 ) o% tho#i$e i$ -+ mL o% so&tio$ A
(s&phate sta$da#d +.++1-1 mo L
01
* a$d p&t i$to a <o&met#i! %as/ o% 1+ mL a$d
e<e it 'ith mii0K E&ait2 'ate#.
Note: This so&tio$ sho&d "e %#esh2 p#epa#ed dai2.
1* Sampi$) a$d sampe p#e0t#eatme$t. 8a!e 1++ mL o% the t#appi$) so&tio$
(so&tio$ ,* i$to the "&""i$) %as/ a$d sta#t &p the sampi$) eE&ipme$t4 a$ ai#
<o&me e3a!t2 meas&#ed is %o#!ed th#o&)h the "&""i$) de<i!e4 so that the
s&ph&# dio3ide !o$!e$t#atio$ #emai$s 'ithi$ the !ai"#atio$ i$ea# #a$)e. I% it is
$e!essa#2 pe#%o#m the #eE&i#ed di&tio$. B#ite do'$ the !o$ditio$s o% the
sampi$)
-* Cai"#atio$ )#aph.
-.1* S&phate sta$da#d so&tio$s. A se#ies o% s&phate sta$da#d so&tio$s is
p#epa#ed "2 di&ti$) 1.+4 -.+4 6.+4 ,.+4 1.+4 5.+4 D.+4 >.+4 A.+ a$d 1+.+ mL
o% the +.++1-1 mo L
01
sto!/ so&tio$ (so&tio$ A* i$ <o&met#i! %as/s o%
1++ mL a$d made &p 'ith 'ate#. The #es&ti$) !o$!e$t#atio$s a#e4
#espe!ti<e2: +.>4 1.54 -.,4 6.-4 ,.+4 ,.>4 1.54 5.,4 D.- a$d >.+ ) mL
01
.
-.-* =a$/ sta$da#d so&tio$. 8#epa#e a so&tio$ "2 mi3i$) ,.+ mL o% t#appi$)
so&tio$ (so&tio$ ,* 'ith 1+ mL o% "a#i&m pe#!ho#ate.dio3a$e so&tio$
(so&tio$ 5* a$d +.-1 mL %o# tho#i$e so&tio$ (so&tio$ 1+*. This "a$/
so&tio$ p#ese$ts the ma3im&m a"so#"a$!e. The tho#i$e so&tio$ aiE&ot
sho&d "e added immediate2 "e%o#e #e!o#di$) the a"so#"a$!e. This
so&tio$ #emai$s &$!ha$)ed %o# 6+ mi$&tes p#ote!ted a)ai$st #oom i)ht.
6* De<eopme$t o% the !oo#imet#i! so&tio$ 'ith the aid o% the sta$da#d s&phate
so&tio$s. =2 &si$) ,.+ mL o% ea!h sta$da#d s&phate so&tio$s (poi$t -.- o%
a"o<e* a$d "2 app2i$) the p#o!ed&#e #epo#ted %o# the "a$/ assa2 (poi$t -.- o%
a"o<e*. The #es&ti$) a"so#"a$!e sho&d "e #e!o#ded "e%o#e te$ mi$&tes.
,* De<eopme$t o% the !oo#imet#i! #ea!tio$ i$ ai# sampes. AdL&st the <o&me i$ the
"&""i$) %ash to 1++ mL. Ta/e a$ aiE&ot o% ,.+ mL to "e mi3ed 'ith 1+ mL o%
A
"a#i&m pe#!ho#ate.dio3a$e (so&tio$ 5* a$d +.-1 mL o% tho#i$e so&tio$ (so&tio$
1+* as #epo#ted at the poi$t -.- o% a"o<e.
1* Meas&#eme$t o% the a"so#"a$!e. AdL&st the 'a<ee$)th at 1-+ $m a$d test the
appa#at&s 'ith a "a$/ so&tio$ adL&sti$) the a"so#"a$!e at +.>++I pa!e the
sta$da#d so&tio$s a$d #e!o#d the !o##espo$di$) a"so#"a$!e <a&es. Fi$a24 #ead
the sampe so&tio$ "e%o#e te$ mi$&tes.
5* Ca!&atio$ o% the SO
-
!o$!e$t#atio$
.
Ca!&ate the st#ai)ht !ai"#atio$ i$e "2
dotti$) a"so#"a$!e <s the di%%e#e$t !o$!e$t#atio$s o% SO
-
. D&e to the de<eoped
#ea!tio$4 the "a$/ dispa2s the ma3im&m a"so#"a$!e a$d the sope is $e)ati<e.
D* Fi$a2 a$d to !a!&ate the SO
-
!o$!e$t#atio$ i$ the "&""i$) de<i!e as most
a!!&#ate as possi"e4 the #a$)e (%#om the !ai"#atio$ )#aph* &sed sho&d "e + a$d
5 ) mL
01
. I% the !o$!e$t#atio$ is o<e# the #epo#ted #a$)e4 ma/e the
!o##espo$di$) di&tio$.

Re"lt
1* Dispa2 the !o$ditio$s o% the sampi$) p#o!ess:
0 Meas&#eme$t o% the )as !o&$te# "e%o#e to sta#t the sampi$) (m
6
*
0 Data a$d ho&# o% the sampi$) (sta#t*
0 Meas&#eme$t o% the )as !o&$te# at the sampi$) e$d (m
6
*
0 Data a$d ho&# o% the sampi$) (e$d*
-* Dispa2 the a"so#"a$!e <a&es a$d depi!t these o"tai$ed a"so#"a$!e <a&es vs the
s&ph&# dio3ide !o$!e$t#atio$ a$d adL&st "2 #e)#essio$ the o"tai$ed !&#<e.
Ca!&ate de ai# !o$!e$t#atio$ o% the po&ta$t i$ ) m
06
.
Relate! -"eti$n
0 N&sti%2 the $e)ati<e <a&e o% the sope o% the st#ai)ht i$e #ep#ese$ted i$ this a"
e3e#!ise.
0 Bhi!h te!h$iE&e sho&d "e s&ita"e to o"tai$ i$%o#matio$ i$ #ea time o$ the
!o$!e$t#atio$ o% SO
-
i$ the ai#O
0 Mai$ me!ha$isms o% o3idatio$ that !a$ &$de#)o the SO
-
i$ the atmosphe#e.
0 Des!#i"e the e%%e!ts o% the SO
-
o$ the di%%e#e$t #e!epto#s.
0 E3pai$ the p#o"ems de#i<ed %#om po&tio$ d&e to Fa!id #ai$P4 its o#i)i$s4 e%%e!ts
a$d possi"e so&tio$s.
0 Comme$ts o$ the pa#ti!e F!o$!e$t#atio$G i$ s&spe$sio$ a$d the !o$!e$t#atio$ o%
SO
-
4 a!!o#di$) to the E&#opea$ e)a #&es.
1+
I... DETERMINATION OF SULPHATE IN )ATERS
This method is appi!a"e to the dete#mi$atio$ o% s&phate i$ 'ate#s 'ithi$ the
#a$)e +.+1 m) S.L to , m) S.L. Sampes !o$tai$i$) hi)he# !o$!e$t#atio$s m&st "e
di&ted p#io# to the a$a2sis.
Principle
=a(CO
,
*
-
is added i$ e3!ess to p#e!ipitate the s&phate as "a#i&m s&phate i$
a$ o#)a$i! so<e$t. The o#)a$i! so<e$t 'i mi$imi7e the so&"iit2 p#od&!t o% "a#i&m
s&phate.
The e3!ess !o$!e$t#atio$ o% "a#i&m (II* io$s i$ the so&tio$ is dete#mi$ed
spe!t#ophotomet#i!a2 at 1-+ $m th#o&)h the #ea!tio$ 'ith tho#i$ (the sodi&m sat o%
,0(o#tho0a#se$ophe$20a7o*060h2d#o320-4D0$aphtae$e0dis&pho$i! a!id*.
Se<e#a o#)a$i! so<e$ts ma2 "e &sed. The most %a<o&#a"e !ai"#atio$ !&#<e
is o"tai$ed 'ith dio3a$e.
A !hemi!as4 e3!ept tho#i$4 m&st "e o% #e!o)$i7ed a$a2ti!a )#ade. The
'ate# &sed %o# di&tio$ a$d #i$si$) m&st "e do&"e distied o# deio$i7ed.
S&ph&#i! a!id (H
-
SO
,
* +.+1 M
8e#!ho#i! a!id4 (HCO
,
* D- ;
=a#i&m pe#!ho#ate (=a(CO
,
*
-
*4 a$h2d#o&s
Dio3a$e o# isop#opa$o
Tho#i$ (disodi&m sat*
Catio$ e3!ha$)e #esi$4 st#o$)2 a!idi! (e.). Do'e3 1+ B 3 >4 1+01++ mesh*.
+.1 M pe#!ho#i! a!id (HCO
,
*.
+.+1 M pe#!ho#i! a!id (HCO
,
*.
=a#i&m pe#!ho#ate sto!/ so&tio$ -1+.+ m) a$h2d#o&s "a#i&m pe#!ho#ate4
(=a(CO
,
*
-
*

4 is disso<ed i$ +.1 M HCO
,
to a <o&me o% 1++ mL i$ a <o&met#i!
%as/.
=a#i&m pe#!ho#ate #ea)e$t so&tio$ 1+.+ mL o% sto!/ so&tio$ is di&ted to
1+++ mL 'ith dio3a$e o# isop#opa$o.
Tho#i$ #ea)e$t so&tio$ 1-1.+ m) o% the disodi&m sat is disso<ed i$ 1 mL
+.+1 M HCO
,
a$d di&ted to 1+ mL i$ a <o&met#i! %as/. A %#esh so&tio$ sho&d
"e p#epa#ed ea!h da2.
S&phate sta$da#d so&tio$ 61.-1 mL +.+1 M H
-
SO
,
is di&ted to 1+++ mL i$ a
<o&met#i! %as/. The !o$!e$t#atio$ is eE&a to 1+ m) S.L.
Spe!t#ophotomete# %o# meas&#i$) a"so#"a$!e at 1-+ $m.
11
Opti!a )ass spe!t#ophotomete# !esI -+ mm
Mi!#o pipette: -1+ CL
=&" pipettes: 14 -4 64 ,4 14 54 D4 >4 A a$d 1+ mL
=&#ette: 1+ mL
Io$ e3!ha$)e !o&m$s: 11 !m e$)th4 1 !m diamete#
Test t&"es: 6+ mL
Vo&met#i! %as/s: 1+4 1++ a$d 1+++ mL
A )ass'a#e sho&d "e o% "o#osii!ate a$d sho&d "e tho#o&)h2 #i$sed i$
distied 'ate# "e%o#e &se.
Cali%rati$n C"r+e
8#epa#e a se#ies o% sta$da#d so&tio$s !o$tai$i$) +4 +.14 1.+4 1.1 ..... , m) S.L
"2 di&ti$) +4 14 -4 64 ..... > mL o% the s&phate sta$da#d so&tio$ to 1++ mL 'ith 'ate#
i$ <o&met#i! %as/s. T#a$s%e# , mL o% ea!h o% these sta$da#d so&tio$s to a test t&"e.
Add 1+ mL "a#i&m pe#!ho#ate #ea)e$t so&tio$ a$d -1+ CL tho#i$ so&tio$. Use a
mi!#opipette %o# the tho#i$ so&tio$. Mi3 tho#o&)h2 (do $ot &se #&""e# stoppe#sQ*
T#a$s%e# the so&tio$s to opti!a !es. The spe!t#ophotomete# 'a<ee$)th is
set at 1-+ $m4 a$d +; t#a$smissio$ is adL&sted a!!o#di$) to the p#o!ed&#e i$ the
ma$&a o% the photomete#. The$ )ai$ a$d.o# sit 'idth is adL&sted to )i<e a #eadi$) o%
+.>+ a"so#"a$!e &$its 'ith the "a$/ (+ m) S.L* i$ the sampe !ompa#tme$t. Meas&#e
the a"so#"a$!e o% the so&tio$s 'ithi$ 1+ mi$&tes a%te# additio$ o% the tho#i$ so&tio$.
This is espe!ia2 impo#ta$t %o# o' !o$!e$t#atio$s o% s&phate a$d %o# the "a$/
"e!a&se the "a#i&m0tho#i$ !ompo&$d ma2 p#e!ipitate %#om the so&tio$.
A !ai"#atio$ )#aph is !o$st#&!ted %#om the a"so#"a$!e #eadi$)s o"tai$ed %#om
the sta$da#d so&tio$s. The !ai"#atio$ !&#<e is $ot i$ea# "eo' +.1 m) S.L. This is
s&pp#essed "2 addi$) s&phate i$ a E&a$tit2 !o##espo$di$) to +.1 m) S.L to a
sampes a$d "a$/s. The dete!tio$ imit is the$ +.+1 m) S.L.
Catio$s a#e #emo<ed "2 t#eati$) the sampe 'ith a st#o$)2 a!idi! !atio$
e3!ha$)e #esi$.
T#a$s%e# , mL o% the p#e0t#eated sampe to a test t&"e a$d p#o!eed a!!o#di$)
to a"o<e. Dete#mi$e the s&ph&# !o$!e$t#atio$ o% the sampe %#om the a"so#"a$!e
#eadi$) "2 mea$s o% the !ai"#atio$ !&#<e.
Bith s&ita"e eE&ipme$t4 the "a#i&m pe#!ho#ate0tho#i$ method !a$ "e made
a&tomati!.
Inter'erence
8hosphate 'i i$te#%e#e 'ith this method.
Re'erence
1-
1. 8e#sso$4 G.A. (1A55* A&tomati! !oo#imet#i! dete#mi$atio$ o% o' !o$!e$t#atio$s
o% s&phate %o# meas&#i$) s&ph&# dio3ide i$ am"ie$t ai#. Air Water Pollut., 10, >,10
>1-.
16
I./. DETERMINATION OF ATMOSPHERIC LEAD 0II1
Intr$!"cti$n
The ead is a #ee<a$t atmosphe#i! po&ti$) a)e$t. Its mai$ a$th#opo)e$i!
so&#!e 'as4 &$ti 2ea# -++14 the emissio$ o% )ases %#om the )asoi$e &sed "2 e$)i$e
<ehi!es 'hi!h &sed ead tet#aeth2 as a$ a$ti0deto$atio$ additi<e. I$ additio$4 it is
$e!essa#2 to !o$side# impo#ta$t ead so&#!es i$ some pa!es4 i/e "atte#2
ma$&%a!t&#e#s4 pai$ti$)s4 i$se!ti!ides4 meta smeti$) a$d meta #e!o<e#2. The
atmosphe#i! ead is deposited o$ the s&#%a!e o% the ea<es o% <e)eta"es o!ated i$ the
p#o3imities o% a hi)h'a2 o# o$ d&st pa#ti!es at the ed)e o% the hi)h'a2s. Its
asso!iated pe#siste$!e to those d&st pa#ti!es 'i "e o% se<e#a 2ea#s.
The atmosphe#i! ead !o$!e$t#atio$ o!!&#s at sma !o$!e$t#atio$s4 $o#ma2
asso!iated to pa#ti!es4 'hi!h mea$s i% 'e %o#!ed a hi)h ai# <o&me to pass th#o&)h a
%ite#I the deposited pa#ti!es !o$tai$ ead e$o&)h to "e dete!ted. I$ these !o$ditio$s
the iE&id0iE&id e3t#a!tio$ p#o!ed&#e is s&ita"e to isoate a$d !o$!e$t#ate the
#e!o<e#ed a$a2te.
As empi#i!a appi!atio$ o% this p#o!ed&#e4 the ead deposited o$ the s&#%a!e
o% the ea<es o% a pa$t o# t#ee o# i$ the pape# o% %ite# disso<es i$ $it#i! a!id. To
eimi$ate the i$te#%e#e$!es o% othe# metas the !ompe3i$) a)e$t CN
0
is added.
Dithi7o$e (o# diphe$2thio!a#"a7o$e* #ea!ts 'ith ma$2 metai! io$s to %o#m
!heates 'ith i$te$se !oo#atio$4 'hi!h !a$ "e e3t#a!ted "2 o#)a$i! so<e$ts. Bhe$ a
so&tio$ o% dithi7o$e i$ di!ho#ometha$e is p&t i$ !o$ta!t 'ith a$ aE&eo&s so&tio$ o%
a meta at the app#op#iated pH4 the meta0dithi7o$ate is %o#med a$d hosted i$to the
o#)a$i! a2e#. The #ea)e$t (dithi7o$e* e3!ess is passed i$to the aE&eo&s phase "2
a/ai$i7i$).
The ma3im&m o% a"so#ptio$ o% the ead0dithi7o$e !ompe3 i$to the o#)a$i!
phase is at 1-+ $m.
Target
To dete#mi$e the !o$!e$t#atio$ o% the ead asso!iated to pa#ti!es.
To i&st#ate the ead po&tio$ o$ di%%e#e$t <e)eta o#)a$isms d&e to e$)i$e
)asoi$e.
The &se o% a$ easi2 a<aia"e methodoo)2 "ased o$ the iE&id0iE&id
e3t#a!tio$ !om"i$ed 'ith the <isi"e photomet#2.
A$ empi#i!a test o% the $o$0aE&eo&s so<e$ts a$d thei# most &s&a
appi!atio$s.
1,
A) Reagents
The )ass'a#e sho&d "e p#e<io&s2 'ashed 'ith +.+1 moes L
01
$it#i! a!id
to eimi$ate a$2 possi"e #esid&e o% ead.
Glassware and apparatus Reagents
1++ mL po2eth2e$e %as/s (-* a$d
ate3 )o<es
Sta$da#d so&tio$ 1+++ m).L o% 8"(II*
1+ mL <o&met#i! !2i$de# (1* Dithi7o$e
1++ mL "ea/e# (1* HNO
6
Sampi$) set Di!ho#ometha$e
Gass %i""e# %ite# -1 ; NH
6
(aE*
8et#i pates. Na
-
SO
6
A$a2ti!a "aa$!e RCN
S!isso#s Na
-
SO
,

> metai! s&ppo#ts
> metai! !i#!es
>4 -1+ mL )ass %&$$es
1 pipettes 1 mL
-4 1 mL pipettes
Fite# pape#
> !o$i!a %as/s
test t&"es (>*
> t&"es 'ith !o#/ o% spi#a o% 1+ mL
Spe!t#ophotomete# (VISI=LE*
B) Solutions to be prepared
1* 1 m) L
01
8"(II* so&tio$. The sta$da#d so&tio$ is p#epa#ed "2 addi$) -.1 mL %#om
the sto!/ so&tio$ !o$tai$i$) 1+++ m) L
01
a$d e<ei$) to 1 L 'ith +.1
mo L
01
HNO
6
. The sto!/ so&tio$4 1+++ m) L
01
8"(II*4 is p#epa#ed "2
so<i$) 1.1A>+ ) 8"(NO
6
*
-
(#ea)e$t a$a2sis* 'ith +.1 mo L
01
HNO
6
a$d
the$ e<ei$) &p to 1+++ mL 'ith the same so<e$t.
-* A/ai$e !ompe3i$) so&tio$. Fo# thei# p#epa#atio$ D1+ mL o% -1; NH
6
(aE* a#e
mi3ed 'ith 1.+ ) o% RCN a$d 1.1 ) o% Na
-
SO
6
a$d is e<eed to 1 L 'ith p&#e 'ate#.
6* So&tio$ o% dithi7o$e (diphe$2thio!a#"a7o$e* -1 m).L. +.+-1 ) o% dithi7o$e i$ 1
L o% di!ho#ometha$e.
Caution: - a$d 6 so&tio$s m&st "e p#epa#ed i$to the sa%et2 %&$$e.
1.a* Sampi$) a$d sampe p#e0t#eatme$t o% the ead o$ pa$t ea<es (sampe
A*. 8i!/ !a#e%&2 > medi&m ea<es $ea# a hi)h'a2 o# 7o$e 'ith de$se t#a%%i!4 'hi!h
do $ot sho' <isi"e ea#th pa#ti!es $o# othe# <isi"e po&ti$) a)e$ts. 8&t the ea<es
11
i$to a po2eth2e$e "otte p#e<io&s2 'ashed 'ith $it#i! a!id ($ot to 'ash $o# to #&"
the ea<es* a$d !o$se#<e them i$ a d#2 pa!e &$ti the mome$t o% the a$a2sis.
Add i$to the sampe #e!ipie$t -+ mL o% +.1 mo L
01
HNO
6
a$d sha/e
<i)o#o&s2 d&#i$) a !o&pe o% mi$&tes. T#a$s%e# the !o$te$t i$to a 1++ mL )ass
"ea/e# a$d 'ash the sampi$) "otte (th#ee times* 'ith , mL o% +.1 mo L
01
HNO
6
a$d
add the 'ashi$) $it#i! to the "ea/e# !o$tai$i$) the so<ed ead.
Note Do $ot th#o' a'a2 the ea<es.
1."* Sampi$) a$d sampe t#eatme$t %o# a$a2sis o% atmosphe#i! ead t#apped
o$ the s&#%a!e o% a pa#ti!e %ite# (sampe =*. Bith the aid o% the sampi$) de<i!e %o#!e
s&%%i!ie$t ai# <o&me (a"o&t - m
6
* th#o&)h the %ite# to o"tai$ the ead amo&$t
deposited o$ the %ite# e$o&)h to "e i$to de !ai"#atio$ #a$)e. O$!e %i$ished the
sampe !oe!tio$4 the %ite# is !a#e%&2 t#a$sated (&se !amps* i$to a 8et#i pate. The$
it is t#a$s%e##ed to the a"o#ato#24 i$t#od&!ed i$to a "otte a$d adds -+ mL o% +.1 mo
L
01
o% HNO
6
. 8#o!eed as #epo#ted i$ the p#e<io&s se!tio$.
-* Meas&#eme$t o% the a#ea o% ea<es. I$ o#de# to dete#mi$e the tota s&#%a!e
o% the ea<es o$ 'hi!h the ead is deposited4 #i$se the ea<es 'ith distied 'ate# a$d
d#ied "2 p#essi$) "et'ee$ %ite# pape#. D#a' its !o$to&# o$ a meas&#eme$t pape#4 !&t
the pape# a$d 'ei)h. A$ sE&a#e pie!e o% pape#4 1+ !m o% side4 is aso 'ei)hed to
!a!&ate the a#ea i$ !m
-
o% the tota s&#%a!e o% the sampe o% ea<es.
6* E3t#a!tio$ a$d dete#mi$atio$ o% ead. Bith ea!h o$e o% the sampe
so&tio$s A a$d = ope#ate as %oo's: mi3 i$to the e3t#a!tio$ %&$$e the !o$te$t o% the
"ea/e#4 1 mL o% the a/ai$e !ompe3i$) so&tio$ a$d 1 mL o% the dithi7o$e so&tio$.
The mi3t&#e is sha/e$ <i)o#o&s2 d&#i$) 1 mi$&tes. 8a!e the %&$$e E&iet d&#i$) 1
mi$&tes to ao' the phases (aE&eo&s a$d o#)a$i!* sepa#atio$. The %&$$e is ope$ed
a$d the o#)a$i! phase is passed i$to a d#2 test t&"e. Add a$ e$d o% spat&a o%
a$h2d#o&s Na
-
SO
,
i$to the test t&"e4 it is sha/e$ a$d the o#)a$i! phase is %ite#ed
th#o&)h a pape# %ite#4 )athe#i$) the %it#ate o$ a d#2 test t&"e 'hi!h is immediate2
!osed to a<oid the e<apo#atio$ o% pa#t o% the so<e$t.
Caution: test t&"es a$d the photomete# !e m&st "e !ompete2 d#2: &se hot
ai# i% #eE&i#ed. AdL&st 'a<ee$)th at 1-+ $m a$d p&t 7e#o a"so#"a$!e 'ith
di!ho#ometha$e.
,* Cai"#atio$ )#aph. Ne3t ta"e depi!ts the #ea)e$ts (i$ this o#de#* to "e
mi3ed %o# p#epa#i$) the sta$da#d so&tio$s set.
F&$$e: 1 - 6 , 1 5
mL o% sta$da#d so&tio$4 1 m) L
01
+ +.1 1.+ 1.1 -.+ -.1
mL o% +.1 mo L
01
HNO
6
-+ -+ -+ -+ -+ -+
mL o% a/ai$e !ompe3i$) so&tio$ 1 1 1 1 1 1
mL o% -1 m) L
01
dithi7o$e 1 1 1 1 1 1
S8"T (m) L
01
* o#)a$i! a2e# + +.1 1.+ 1.1 -.+ -.1
15
As #epo#ted %o# the sampe so&tio$s4 the mi3t&#e is sha/e$ <i)o#o&s2 d&#i$)
1 mi$&tesI a%te# 1 mi$&tes ("oth a2e#s a#e sepa#ated* a$d the %&$$e is ope$ed to
dei<e# the o#)a$i! phase i$to a d#2 test t&"e. Add a$ e$d o% spat&a o% a$h2d#o&s
Na
-
SO
,
. A%te# %ite#i$) the o#)a$i! phase th#o&)h a pape# %ite# the #es&ti$) o#)a$i!
so&tio$ is /ept i$ a test t&"e ti)ht2 !osed to a<oid so<e$t e<apo#atio$.
Caution: test t&"es a$d the photomete# !e m&st "e !ompete2 d#2: &se hot
ai# i% #eE&i#ed. AdL&st 'a<ee$)th at 1-+ $m a$d p&t 7e#o a"so#"a$!e 'ith
di!ho#ometha$e.
A<oid i$ a$2 mome$t the !o$ta!t o% the !2a$ide so&tio$ 'ith a!idi! so&tio$sI
e<o&tio$ o% !ia$h2d#i! a!id is e3t#eme2 da$)e#o&s (smei$) to "ite amo$ds*.
Do $ot th#o' a'a2 the #esid&a so&tio$sI it sho&d "e pa!ed i$to the
!o##espo$di$) deposits.
Relate! -"eti$n
8#opose othe# te!h$iE&es o% a$a2sis to dete#mi$e ead i$ the atmosphe#e.
I$di!ate ad<a$ta)es a$d disad<a$ta)es.
List othe# hea<2 metas that !a$ "e dete#mi$ed Loi$t2 'ith the p#oposed
te!h$iE&es.
Ma/e a ist o% the di%%e#e$t e3pe#ime$ta <a#ia"es asso!iated to the a$a2sis4
i$di!ati$) possi"e so&#!es o% e##o#.
1D
I.2. SEPARATION BY E3TRACTION AND
SPECTROPHOTOMETRIC DETERMINATION OF MERCURY
FROM )ASTE )ATERS
Principle
The method p#oposed %o# the sepa#atio$ a$d dete#mi$atio$ o% me#!&#2 is
"ased o$ the see!ti<e p#e!ipitatio$ o% me#!&#2 'ith the #ea)e$t Cadio$ A (,0
$it#ophe$2 dia7oami$oa7o"e$7e$e* a$d the e3t#a!tio$ o% the %o#med !ompe3 'ith
to&e$e. =2 meas&#i$) the a"so#"a$!e o% the e3t#a!t i$ to&e$e a$d &si$) a !ai"#atio$
!&#<e the E&a$tit2 o% me#!&#2 %#om the a$a27ed sampe !a$ "e dete#mi$ed. The
%o#matio$ o% the me#!&#2 !ompe3 ta/es pa!e at a pH o% the aE&eo&s phase o% 1.1 M
11. The me#!&#2 (II* %o#m 'ith the Cadio$ a !ompe3 'ith the %oo'i$) st#&!t&#e:
N N N
N
N NO
-
N N N
N
N NO
-
H)
Bith the )i<e$ 'o#/i$) !o$ditio$s the #ea!tio$ is hi)h2 see!ti<e. No
i$te#%e#e$!es appea# %#om the io$s o% C&(II*4 A&(III*4 =e(II*4 S$(II*4 Cd(II*4 T (I4 III*4
UO
-
(II*4 8"(II*4 M$(II*4 Fe(II4III*4 =i(III*4 C#(III*4 Co(II*4 Ni(II*4 Rh(III*4 8d(II*4
8t(IV*4 8O
,
60
4 MoO
,
-0
a$d othe#s. The io$ A)(I* i$te#%e#es %o#mi$) a #ed !ompe3 'ith
the #ea)e$t. The io$ C
0
does $ot i$te#%e#e &p to a #atio C
0
.H)(II* o% 1+. The io$s o%
=#
0
4 I
0
4 SCN
0
4 S
-0
a$d EDTA i$te#%e#e %o#mi$) sta"e !om"i$atio$s 'ith the me#!&#2
(II*. Fo# the domai$ +.1M, ) H)(II*.mL is o"se#<ed a i$ea# depe$de$!e o% the
a"so#"a$!e %&$!tio$ o% the me#!&#2 !o$!e$t#atio$. The <a&e o% the moa# a"so#pti<it2
is o% ,++++ L mo
01
!m
01
. =e!a&se the %o#matio$ o% the me#!&#20!adio$ !ompe3 ta/es
pa!e i$ a $e&t#a e$<i#o$me$t4 to the #ea!tio$ media is added a so&tio$ o% sodi&m a$d
potassi&m ta#t#ate4 'hi!h th#o&)h the !ompe3es %o#med 'i p#e<e$t the p#e!ipitatio$
o% some io$s.
The #ea)e$t has the a"so#ptio$ ma3im&m at ,+- $m4 a$d the me#!&#2
!ompe3 at ,-D $m. The a"so#"a$!e meas&#eme$ts4 %o# the dete#mi$atio$ o% me#!&#2
'e#e do$e at ,A+ $m4 a 'a<ee$)th at 'hi!h the a"so#"a$!e o% the #ea)e$t is sma.
1>
0 Sta$da#d so&tio$ o% me#!&#2 !ho#ide 'ith the !o$!e$t#atio$ o% 1+ ) H)(II*.mL
i$ distied 'ate#I
0 So&tio$ o% Cadio$ A (,0$it#ophe$2 dia7oami$oa7o"e$7e$e* +.+11 ; i$ metha$oI
0 So&tio$ o% sodi&m a!etate +.- MI
0 So&tio$ o% sodi&m a$d potassi&m ta#t#ate -+ ; i$ distied 'ate#I
0 C2i$de#s o# test t&"es 'ith the <o&me o% 1+ M 1+ mL p#o<ided 'ith p&)4
mi!#opipette o% 1+++ ULI pipettes o% 14 - a$d 1 mL4 <o&met#i! !ai"#ated %as/ o%
1++ mLI
0 The a"so#"a$!e dete#mi$atio$s a#e made 'ith a spe!t#omete# %o# moe!&a#
a"so#ptio$ i$ UV0VIS.
Cali%rati$n C"r+e
F#om the so&tio$ o% 1+ ) H)(II*.mL a#e ta/e$ 'ith a mi!#opipette +.1I +.-I
+.,I a$d +.5 mL 'hi!h a#e "ei$) i$t#od&!ed i$ !osed !2i$de#s. Bith a pipette a#e
the$ i$t#od&!ed 1 mL o% distied 'ate#4 'ashi$) the 'as o% the !2i$de# to !a##2 the
'hoe E&a$tit2 o% me#!&#2 i$ the aE&eo&s so&tio$. A%te#'a#ds a#e i$t#od&!ed: 1 mL
sodi&m a!etate so&tio$ +.- M a$d 1 mL #ea)e$t so&tio$ +.+11 ;I the mi3t&#e is
sti##ed a$d e%t to sette %o# 1+ mi$ %o# the %o#matio$ o% the !ompe3.
A%te# this a#e i$t#od&!ed 1 mL to&e$e a$d the mi3t&#e is sti##ed st#o$)2 %o# 1
mi$. It is impo#ta$t that a%te# the %i#st se!o$ds4 o% sti##i$) it is stopped a$d the p&) is
#emo<ed to #eease the to&e$e <apo#s4 a%te# 'hi!h the sti##i$) is !o$ti$&ed. A%te# the
phase sepa#atio$ (a"o&t 1+ M 11 mi$* a pa#t o% the to&e$e e3t#a!t is #emo<ed 'ith a
d#2 pipette a$d it is i$t#od&!ed di#e!t2 i$ the spe!t#ophotomete# !e. The a"so#"a$!e
dete#mi$atio$s a#e do$e a)ai$st a #e%e#e$!e sampe p#epa#ed i$ the same ma$$e# as
the sta$da#ds (distied 'ate#4 sodi&m a!etate so&tio$4 #ea)e$t a#e i$t#od&!ed a$d the
to&e$e e3t#a!tio$ is e3e!&ted*. I$ the #e%e#e$!e sampe $o me#!&#2 is i$t#od&!ed.
Ces 'ith the opti!a path o% 1 !m a$d a 'a<ee$)th o% ,A+ $m 'i "e &sed.
The a"so#"a$!e <a&es meas&#ed a#e #ep#ese$ted )#aphi!a2 %&$!tio$ o% the me#!&#2
E&a$tities ta/e$ %o# a$a2sis. I$ this ma$$e# is o"tai$ed a !ai"#atio$ !&#<e 'hi!h4 i%
the p#o!ed&#e 'as %oo'ed p#ope#24 'i "e a st#ai)ht i$e passi$) th#o&)h the o#i)i$.
Merc"r& Anal&i
The a$a27ed sampe ma2 !o$tai$ i$ additio$ to the me#!&#2 io$s (II* a#)e
E&a$tities o% io$s o% C&(II*I Co(II*I Ni(II*I Fe(II4 III*I J$(II*I Ca(II*I M)(II*4 et!. F#om
this sampe is #e!ei<ed %o# a$a2sis a !e#tai$ E&a$tit2 i$ a <o&met#i! %as/ o% 1++ mL.
The <o&met#i! %as/ is %ied to the ma#/ 'ith distied 'ate# a$d the mi3t&#e is
homo)e$i7ed. F#om this sampe a#e the$ ta/e$ sampes (t'o o# th#ee* that a#e
i$t#od&!ed i$ the !2i$de#s %o# the e3t#a!tio$. The sampes ta/e$ %o# a$a2sis m&st "e
"et'ee$ +.1 M 1 mL. I$ ea!h !2i$de# a#e the$ i$t#od&!ed 1 mL o% sodi&m a$d
potassi&m ta#t#ate so&tio$4 'hi!h 'i %o#m !ompe3es 'ith some io$s4 'hi!h mi)ht
1A
p#e!ipitate %#om the #ea!tio$ media. S&"seE&e$t2 the 'o#/i$) p#o!ed&#e %o# the
!ai"#atio$ !&#<e 'i "e &sed.
Interpretati$n $' t*e E4peri#ental Data
The a"so#"a$!e <a&es meas&#ed %o# the sampes ta/e$ %o# a$a2sis ao'4
'ith the aid o% the !ai"#atio$ !&#<e4 the dete#mi$atio$ o% the E&a$tit2 o% me#!&#2 %#om
these sampes. R$o'i$) the E&a$tit2 o% me#!&#2 %#om ea!h sampe a$d aso its
pe#!e$ta)e %#om the tota <o&me o% the sampe ta/e$ %o# a$a2sis4 mea$i$) 1++ mL4
the E&a$tit2 o% me#!&#2 #e!ei<ed %o# a$a2sis !a$ "e dete#mi$ed. The #es&t is )i<e$ i$
m) o% me#!&#2.
Observations: The to&e$e is i)hte# tha$ 'ate#4 a$d a%te# sti##i$)4 it is
sepa#ated i$ the &ppe# pa#t. The a"so#"a$!e meas&#eme$ts a#e a'a2s do$e a)ai$st a
#e%e#e$!e sampe that does $ot !o$tai$ me#!&#2.
-+
I.5. DETERMINATION OF LEAD BY ATOMIC ABSORPTION
SPECTROMETRY )ITH ELECTROTHERMAL
ATOMI6ATION 0ET7AAS1
Maria 7 Critina BRATU
Intr$!"cti$n
The metai! ead a$d its !ompo&$ds a#e to3i!I the2 !a$ e$te# the o#)a$ism "2
i$)estio$4 i$hai$)4 o# a"so#ptio$ th#o&)h the s/i$ (the a!&te ead poiso$i$) is #a#e d&e
to the poo# a"so#ptio$*. The !o$ti$&o&s assimiatio$ o% sma E&a$tities o% ead is "2
%a# the most da$)e#o&s. I$itia24 the ead is 'ea/2 "o$ded to the e#2th#o!2tes a$d
o$2 a sma pa#t is eimi$ated th#o&)h the &#i$e4 most o% it "ei$) a!!&m&ated i$ the
"o$es.
F#om a the o#)a$i! !ompo&$ds o% ead4 the t#ia/2 !ompo&$ds ha<e the
st#o$)est $e&#oto3i! e%%e!t o$ mammas. A%te# assimiatio$4 the tet#aa/2 !ompo&$ds
a#e E&i!/2 meta"oi7ed to the t#ia/2 !ompo&$ds.
I$ the e$<i#o$me$t4 the mai$ so&#!e o% o#)a$i! ead !ompo&$ds is thei# &se as
a$tio3ida$t a)e$ts %o# )asoi$e. As a #es&t o% the a's4 i$ most !o&$t#ies this po&tio$
so&#!e is de!#easi$). O$!e e$te#ed i$ the atmosphe#e4 the tet#aa/2 !ompo&$ds o%
ead a#e de!omposed photo!ata2ti!a24 a p#o!ess that !a$ "e <e#2 E&i!/ i$ the )i<e$
!o$ditio$s. The de!ompositio$ p#od&!ts a#e ead !ompo&$ds that a#e t#i0 di0 a$d
mo$oa/2ated. The ead is a$ om$ip#ese$t po&ti$) eeme$t i$ a a#eas o% the
"iosphe#e.
Ra#e24 the $at&#a 'ate#s !o$tai$ a !o$!e$t#atio$ a#)e# tha$ 1 C).L 8"4 "&t
hi)he# !o$!e$t#atio$s o% ead 'e#e #epo#ted. A$othe# so&#!e o% ead i$ the tap 'ate#
a#e the ead pipes th#o&)h 'hi!h it !i#!&ates.
Fo# the dete#mi$atio$ o% ead !a$ "e appied a !oo#imet#i! method $ame2 the
dithi7o$e method4 atomi! a"so#ptio$ spe!t#omet#2 'ith %ame a$d #espe!ti<e2
ee!t#othe#ma atomi7atio$ ()#aphite %&#$a!e*4 a$d the atomi! emissio$ spe!t#omet#2
'ith i$d&!ti<e2 !o&ped pasma. The atomi! a"so#ptio$ spe!t#omet#2 'ith %ame
p#ese$ts a #eati<e2 $ot <e#2 hi)h dete!tio$ imit4 #eE&i#i$) the app2i$) o% a$
e3t#a!tio$ p#o!ed&#e %o# the a$a2sis o% the d#i$/i$) 'ate# i$ 'hi!h o'
!o$!e$t#atio$s o% ead a#e %o&$d. The atomi! a"so#ptio$ spe!t#omet#2 'ith
ee!t#othe#ma atomi7atio$ is m&!h mo#e se$siti<e %o# o' !o$!e$t#atio$s o% ead a$d
does $ot #eE&i#e a$ e3t#a!tio$ step. The atomi! emissio$ spe!t#omet#2 'ith
i$d&!ti<e2 !o&ped pasma has a simia# se$siti<it2 to the atomi! a"so#ptio$
spe!t#omet#2 'ith %ame.
-1
I$ this 'o#/ is p#ese$ted a p#o!ed&#e %o# the dete#mi$atio$ o% ead %#om the ai#
o% a 'o#/pa!e e3posed to this !hemi!a po&ta$t4 "2 the method o% atomi! a"so#ptio$
spe!t#omet#2 'ith ee!t#othe#ma atomi7atio$.
Reagent an! Apparat"
0 Nit#i! a!id 51 ; S&p#ap&# (Al!rich"4 ammo$i&m dih2d#o)e$ phosphate
(#erc$*4 distied 'ate# (!o$d&!ti<it2 , CS.!m*.
0 Nit#i! a!id +.- ;. It is p#epa#ed "2 di&ti$) 6.+D5 mL HNO
6
S&p#ap&# 'ith
1+++ mL distied 'ate#.
0 Mat#i3 modi%ie# %o# the )#aphite %&#$a!e M a#e disso<ed +., ) o% ammo$i&m
dih2d#o)e$ phosphate i$ 1+ mL o% distied 'ate#.
0 Comme#!ia2 a<aia"e ead sta$da#d 'ith the !o$!e$t#atio$ o% 1+++ m).L
%Al!rich".
0 Lead sta$da#d 'ith the !o$!e$t#atio$ o% 1+ m).L: i$ a <o&met#i! %as/ o% -1
mL o% Te%o$ a#e di&ted -1+ CL o% ead sta$da#d 'ith the !o$!e$t#atio$ o% 1+++
m).L 'ith -1 mL o% S&p#ap&# HNO
6
+.- ;.
0 Lead sta$da#d 'ith the !o$!e$t#atio$ o% 1++ C).L: i$ a <o&met#i! %as/ o% -1
mL o% te%o$ a#e di&ted -1+ CL o% ead sta$da#d 'ith the !o$!e$t#atio$ o% 1+ m).L
'ith -1 mL o% s&pe#0p&#e HNO
6
+.- ;.
0 Lead sta$da#d 'ith the !o$!e$t#atio$ o% 1+ C).L: i$ a <o&met#i! %as/ o% -1
mL te%o$ a#e di&ted -1+ CL o% ead sta$da#d 'ith the !o$!e$t#atio$ o% 1++ C).L
'ith -1 mL o% S&p#ap&# HNO
6
+.- ;.
0 The ead sta$da#ds 'ith the !o$!e$t#atio$s o% - a$d , C).L a#e o"tai$ed "2 the
a&tomated di&tio$ made "2 the a&tosampe# o% the atomi! a"so#ptio$
spe!t#ophotomete# F8e#/i$ Eme#G 4 mode FA$a2st D++G o% the ead sta$da#d 'ith
the !o$!e$t#atio$ o% 1+ C).L 'ith $it#i! a!id +.- ;.
A the ead dete#mi$atio$s 'e#e do$e &si$) a$ atomi! a"so#ptio$
spe!t#ophotomete# F8e#/i$ Eme#G4 mode FA$a2st D++G4 eE&ipped 'ith :
0 ra!iation source M !o$sisti$) i$ a i)ht so&#!e emitti$) the i$e spe!t#a
!ha#a!te#isti! %o# the eeme$t "ei$) a$a27ed (8"*: a amp 'ith a hoo' !athode
(pa!ed o$ a$ a&tomated t#a$spo#t s2stem 'ith ei)ht positio$s %o# amps*.
0 correction source &or the 'ac$(roun!: a de&te#i&m amp.
0 monochromator0 &sed %o# the isoatio$ o% a$ a"so#ptio$ i$e %#om the
!ha#a!te#isti! i$e spe!t#a %o# the a$a27ed eeme$t.
0 soli! !etector asso!iated 'ith a$ electronic ampli&)in( s)stem a$d measurin(
e*uipment.
0 atomi+ation source #ep#ese$ted "2 the (raphite &urnace 'ith o$)it&di$a
heati$).
The <ota)e is appied ao$) the )#aphite t&"e4 pa#ae 'ith the #adiatio$
"eam. The s&pp2 so&#!e4 the ee!t#o$i! s2stems4 a$d the p$e&mati! !o$t#o o% the
i$e#t )as %o# the )#aphite %&#$a!e a#e i$!o#po#ated i$ the spe!t#omete#.
--
The )#aphite t&"e is mai$tai$ed i$ positio$ i$ the %&#$a!e "2 t'o )#aphite
!o$ta!ts that e$s&#e the ee!t#i!a !o$ta!t %o# the heati$) o% the t&"e. The )#aphite t&"e
is ma$&%a!t&#ed %#om p2#o2ti! )#aphite a$d p#ese$ts a$ i$te)#ated pat%o#m LV<o<
that dete#mi$es a the#ma eE&ii"#i&m "et'ee$ the sampe a$d the atmosphe#e i$side
the t&"e d&#i$) the atomi7atio$ step.
To p#e<e$t the o3idatio$ o% the )#aphite t&"e at hi)h tempe#at&#es a$d to
p&#)e %#om the t&"e the <apo#s a$d %&mes4 is &sed a$ i$e#t )as (a#)o$* "oth i$side the
t&"e a$d a#o&$d its o&te# she. The !ooi$) o% the )#aphite t&"e is do$e "2 !o&pi$) it
to a !od 'ate# so&#!e4 a$ optima tempe#at&#e a$d %o' #ate o% the 'ate# !o&d
a!hie<e the !ooi$) o% the %&#$a!e i$ -+ se!o$ds.
0 the autosampler M !o$tai$s a the me!ha$i!a a$d ee!t#i!a !ompo$e$ts #eE&i#ed %o#
the i$ta/e o% a !o##e!t <o&me o% so&tio$ %#om a see!ted !o$tai$e#4 its i$Le!tio$ i$ the
)#aphite t&"e a$d the 'ashi$) o% the i$Le!tio$ $eede.
The &se o% the a&tosampe# %o# the i$Le!tio$ o% the so&tio$s i$ the )#aphite
t&"e eads to a !o$side#a"e imp#o<eme$t o% the a$a2ti!a #es&ts !ompa#ed to the
ma$&a pipette meas&#i$).
The meas&#ed <o&me is "et'ee$ 1 M AA CL.
The a&tosampe# aso !o$tai$s a t#a2 'ith >> positio$s4 i$ 'hi!h a#e pa!ed
the po2p#op2e$e <ias &sed %o# the #e%e#e$!e so&tio$s4 the mat#i3 modi%ie#s
so&tio$s4 the sampe so&tio$s a$d the di&e$t so&tio$s. The positio$ o% ea!h <ia is
$&m"e#ed o$ the t#a2.
, computer M 'ith spe!i%i! so%t'a#e FAA Bi$La"G4 that !o$t#os the
spe!t#ophotomete#.
)$r8ing Pr$ce!"re
The ai# !omp#esso# 'hi!h !o$t#os the p$e&mati! ope#atio$ o% the
spe!t#omete# is sta#ted. The a#)o$ !2i$de# is ope$ed a$d the$ the )#aphite %&#$a!e
!ooi$) !i#!&it is ope$ed. The atomi! a"so#ptio$ spe!t#ophotomete# a$d the !omp&te#
a#e !o$$e!ted to the $et'o#/. The spe!i%i! so%t'a#e %o# this i$st#&me$t is oaded4 FAA
Bi$ La"G. A%te# a$ a&tomated eE&ipme$t <e#i%i!atio$ test4 %#om the method i"#a#24 is
see!ted the ead dete#mi$atio$ method "ased o$ the &se o% the )#aphite %&#$a!e 'ith a
mat#i3 modi%ie# (ammo$i&m dih2d#o)e$ phosphate*. The hoo' !athode amp a$d the
de&te#i&m amp (%o# the "a!/)#o&$d !o##e!tio$* a#e s'it!hed o$ M a$d et to heat &p
%o# ha% a$ ho&#.
I$ the method edito# a#e esta"ished: the E&a$tit2 o% sampe.sta$da#d i$Le!ted
(-1 CL*4 the E&a$tit2 o% mat#i3 modi%ie# added (1CL* a$d its positio$ o$ the
a&tosampe# t#a24 positio$ o% the di&e$t (HNO
6
+.- ;*4 $&m"e# o% #epi!ates4 t2pe o%
!ai"#atio$ !&#<e (i$ea#.$o$i$ea#*4 the !o$!e$t#atio$s o% the sta$da#ds &sed %o# the
!ai"#atio$ a$d the positio$ o% the most !o$!e$t#ated sta$da#d %#om 'hi!h "2
a&tomated di&tio$ the othe# sta$da#ds a#e p#epa#ed a$d the p#o)#am o% the )#aphite
%&#$a!e.
-6
T*e Te#perat"re Pr$gra# $' t*e 9rap*ite F"rnace
-1 CL o% "a$/.sta$da#d.sampe a#e i$t#od&!ed i$ the atomi7atio$ &$it a$d the
tempe#at&#e is i$!#eased )#ad&a2 to #emo<e the so<e$t a$d i$te#%e#e$ts as 'e as
possi"e "e%o#e the atomi7atio$. I$ the ta"e "eo' is p#ese$ted the tempe#at&#e
p#o)#am o% the )#aphite %&#$a!e optimised %o# the dete#mi$atio$ o% ead.
The optimised tempe#at&#e p#o)#am %o# ead dete#mi$atio$
Step
:
Te#perat"re
0;C1
Ra#p
0ti#e71
H$l!
0ti#e71
Internal
ga 'l$<
0#L=#in1
T&pe $' ga
0n$r#=pl.1
Rea!ing
1 1++ 1 -+ -1+ N No
- 1,+ 11 11 -1+ N No
6 D++ 1+ -+ -1+ N No
, 1>++ + 1 + N Yes
1 -5++ 1 6 -1+ N No
5 -1+ N No

The %i<e steps i$ the tempe#at&#e p#o)#am o% the )#aphite %&#$a!e ha<e the
%oo'i$) p&#pose:
1.-. 0 d#2i$) M ta/es pa!e a%te# the i$Le!tio$ o% the sampe i$ the %&#$a!e.
The sampe m&st "e d#ied at a !o##espo$di$) o' tempe#at&#e to a<oid its
de!ompositio$. Fo# the aE&eo&s so&tio$s4 the d#2i$) is appied at
tempe#at&#es o% 1++ M1,+ WC. The &se o% a tempe#at&#e #amp e$s&#es the
i$!#ease o% the tempe#at&#e i$ a 'e dete#mi$ed i$te#<a. I$ the !ase o% the
)#aphite t&"es 'ith i$te)#ated pat%o#m (the !ase o% the p#ese$t 'o#/*4 the
time !o##espo$di$) to the tempe#at&#e #amp is sho#te#4 a o$)e# time is
appied %o# the atomi7atio$ o% the sampe %#om the t&"e 'as. A%te# the
tempe#at&#e #amp4 the %&#$a!e is mai$tai$ed at the see!ted d#2i$)
tempe#at&#e4 &$ti %& d#2i$). =e!a&se o$2 a %e' mi!#oite#s o% sampe a#e
&sed4 the time %o# mai$tai$i$) at !o$sta$t tempe#at&#e is &$de# 1 mi$&te.
D&#i$) the d#2i$) p#o!ess4 the i$te#$a )as %o' is mai$tai$ed at its
ma3im&m <a&e (-1+06++ mL.mi$* %o# the p&#)i$) %#om the t&"e o% the
<apo#i7ed so<e$t.
6. p2#oisis M is &sed %o# the <oatii7atio$ o% the o#)a$i! a$d i$o#)a$i!
!ompo&$ds o% the mat#i3 see!ti<e2 %#om the sampe4 ea<i$) the a$a27ed eeme$t i$
a ess !ompe3 mat#i3 %o# a$a2sis. D&#i$) this step4 the tempe#at&#e is i$!#eased as
m&!h as possi"e to <oatii7e the mat#i3 !ompo$e$ts4 "&t &p to the a$a2te
de!ompositio$ tempe#at&#e. The i$te#$a )as %o' is mai$tai$ed at -1+06++ mL.mi$
%o# the #emo<a o% the <oatii7ed mat#i3 !ompo$e$ts.
,. atomi7atio$ M is &sed %o# the p#od&!tio$ o% the atom pop&atio$
!o##espo$di$) to the eeme$t to "e a$a27ed4 that 'i ao' the meas&#i$) o% the
atomi! a"so#ptio$. I$ this step4 the tempe#at&#e is i$!#eased &p to the poi$t o%
-,
disso!iatio$ o% the <oatii7ed moe!&a# spe!ies. The atomi7atio$ tempe#at&#e is a
!ha#a!te#isti! o% the a$a27ed eeme$t. Fo# the atomi7atio$ is desi#ed the %astest
possi"e tempe#at&#e i$!#ease. That is 'h2 the time !o##espo$di$) to the tempe#at&#e
#amp 'i "e set to the mi$im&m <a&es that 'i e$s&#e the hi)hest tempe#at&#e
i$!#ease #ate. At the "e)i$$i$) o% this step4 is a!ti<ated the F#eadi$)G ope#atio$ o% the
spe!t#omete# "2 meas&#i$) the a"so#"a$!e o% the i)ht #adiatio$.
1. !ea$i$) M a%te# the atomi7atio$4 the )#aphite %&#$a!e !a$ "e heated to
hi)he# tempe#at&#es4 so that a$2 sampe #esid&e #emai$i$) i$ the %&#$a!e 'i "e
"&#$ed.
9enerating $' t*e Cali%rati$n C"r+e
FJe#oG poi$t o% the i$st#&me$t is #ead "2 meas&#i$) the a"so#"a$!e
!o##espo$di$) to the !ai"#atio$ "a$/ (HNO
6
+.- ;*. The <a&e o% the
a"so#"a$!e !o##espo$di$) to the !ai"#atio$ "a$/ 'i "e a&tomati!a2
s&"t#a!ted %#om the a"so#"a$!es !o##espo$di$) to the sta$da#ds a$d the
sampes.
=oth %o# #eadi$) o% !ai"#atio$ "a$/ a$d sampes is #eE&i#ed the additio$ to
the a&tomated p#epa#atio$ o% sampes o% 1 CL o% ammo$i&m dih2d#o)e$ phosphate
(pa!ed i$ the positio$ >5 o% the a&tosampe#* as a mat#i3 modi%ie#.
Fo# the )e$e#ati$) o% the !ai"#atio$ !&#<e a#e &sed ead sta$da#ds 'ith the
!o$!e$t#atio$s o% -4 ,4 1+ C).L. The ead sta$da#ds 'ith the !o$!e$t#atio$s o% - a$d
#espe!ti<e2 , C).L a#e o"tai$ed "2 the a&tomated di&tio$ made "2 the a&tosampe#
o% the spe!t#ophotomete# o% 1 CL a$d #espe!ti<e2 1+ CL o% ead sta$da#d 'ith the
!o$!e$t#atio$ o% 1+ C).L (pa!ed i$ the positio$ >D o% the a&tosampe#* 'ith -+ CL
a$d #espe!ti<e2 11 CL o% $it#i! a!id +.- ; (the positio$ o% the di&e$t i$ the
a&tosampe# M >>*.
The a"so#"a$!es !o##espo$di$) to the th#ee ead sta$da#ds a#e #ead4 'ith t'o
#epi!ates ea!h at ->6.6 $m. The !ai"#atio$ !&#<e is )e$e#ated a&tomati!a2 "2 the
so%t'a#e o% the i$st#&me$t4 i$ a $o$i$ea# t2pe. The <a&es !o##espo$di$) to the
!o##eatio$ !oe%%i!ie$t a$d the sope !o##espo$di$) to the !ai"#atio$ !&#<e a#e
!a!&ated a&tomati!a2 a$d dispa2ed o$ the s!#ee$ o% the !omp&te# !o$$e!ted to the
a$a2ti!a i$st#&me$t.
Deter#inati$n $' t*e Lea! C$ncentrati$n 'r$# t*e En+ir$n#ent $' a
)$r8place
The ead !o$!e$t#atio$ %#om a$ e$<i#o$me$t e3posed to this po&ta$t is
dete#mi$ed: i$side the i$ot2pe 'o#/shop %#om t2po)#aph2.
I$ o#de# to dete#mi$e the ead !o$!e$t#atio$ at a #espi#ato#2 e<e4 the ead
is !oe!ted %#om th#ee ai# sampes4 at th#ee di%%e#e$t mome$ts4 &si$) %o#
this4 a sampi$) p&mp %o# )aseo&s sampes FAmete/G. The th#ee ai#
sampes a#e !oe!ted 'ith a %o' #ate o% a"o&t 1 .mi$4 %o# te$ mi$&tes.
The sampi$) %o' #ate is meas&#ed 'ith a %o' #ate !ai"#ato# 'ith soap
-1
"&""e FGii"#ato# -G4 %o# ea!h sampe #e!o#di$) the e3a!t <a&e o% the
meas&#ed %o' #ate4 to dete#mi$e the <o&me o% the !oe!ted ai#:
Vo&me
ai#
(V
ai#
* H Fo' (D*X time(t* (1*
Fo# the !oe!tio$ o% the ai# sampes is &sed the sampi$) <esses4 i$ 'hi!h
a#e i$t#od&!ed 1+ mL o% $it#i! a!id +.-;4 a!id i$ 'hi!h is "&""ed a
<o&me V o% ai#4 #etai$i$) i$ the a"so#"e$t so&tio$ the ead.
Ea!h o% the th#ee a"so#"e$t so&tio$s ($it#i! a!id +.- ;* i$ 'hi!h the ead 'as
#etai$ed is E&a$titati<e t#a$s%e##ed i$ a <o&met#i! %as/ o% 1+ mL4 'hi!h is %ied to
the ma#/ 'ith $it#i! a!id +.- ;. These so&tio$s a#e i$t#od&!ed i$ th#ee <ias o% the
a&tosampe# o% the spe!t#omete# a$d t'o #epi!ates %o# ea!h o$e a#e a$a27ed. -1 CL
o% so&tio$ 'i "e a$a27ed &si$) the atomi! a"so#ptio$ spe!t#ophotomete# FA$a2st
D++G4 a$d the a"so#"a$!es %o# ea!h #epi!ate a#e i$te#poated o$ the !ai"#atio$ !&#<e4
th&s dete#mi$i$) the !o##espo$di$) !o$!e$t#atio$s.
To !a!&ate the ead !o$!e$t#atio$ e3p#essed as m).m
6
o% ai# is appied the
%o#m&a:
1+++ X * (
X
* . (
6
- .
c .
m m( c
air
!il
P'
=
(-*
'he#e V
di.
H 1+ mL.I
Repa!i$) the eE&atio$ (1* i$ the #eatio$ship (-*4 is o"tai$ed:
1+++ X t X D
! X V
* m . m) ( !
di 6
8"
= (6*
'he#e D H %o' #ate (L.mi$*I t H time (mi$*.
Th&s4 %o# the dete#mi$atio$ o% the ead !o$!e$t#atio$ %#om the !oe!ted ai#
sampes a#e !o$side#ed the ead !o$!e$t#atio$ o"tai$ed o$ the !ai"#atio$ !&#<e (!0
e3p#essed i$ C).L*4 the <o&me to 'hi!h the sampe a"so#"e$t so&tio$ 'as di&ted
(V
di.
H 1+ mL*4 the sampi$) %o' #ate o% the ai# (D0 e3p#essed i$ L.mi$ * a$d the
sampi$) time o% the ai# i$ the a"so#"e$t so&tio$ (t0 e3p#essed i$ mi$*.

-5
I.>. DETERMINATION OF MERCURY BY COLD ?APOUR
ATOMIC ABSORPTION SPECTROMETRY
Maria 7 Critina BRATU
Intr$!"cti$n
The to3i! metas a#e $ot "iode)#ada"e4 ha<i$) the te$de$!2 to a!!&m&ate i$
the <ita o#)a$s o% h&ma$s 'he#e the2 a!t %o# a o$) pe#iod o% time. The po&tio$ o%
the e$<i#o$me$t 'ith to3i! metas is d&e to the i$d&st#ia e%&e$ts4 as 'e as to the
dis!ha#)e o% 'aste'ate# %#om di%%e#e$t so&#!es.
The me#!&#2 is a t#a!e eeme$t4 'hi!h i$ sma E&a$tities !a$ p#od&!e se<e#e
to3i! e%%e!ts. I$ the !ase o% h&ma$s that a#e $ot e3posed to me#!&#2 th#o&)h thei# Lo"s4
the most p#o"a"e so&#!e %o# this eeme$t is the diet. The #epo#ted E&a$tit2 o% me#!&#2
i$ %oodst&%% is #eati<e2 o'4 app#o3imate2 +.+- C).)4 "&t a a#)e <a#ia"iit2 e3ists
d&e to the t2pe o% p#od&!t4 its )eo)#aphi!a o#i)i$4 a$d the i$d&st#ia a$d a)#i!&t&#a
te!h$iE&es %#om that a#ea. The me#!&#2 a!!&m&ates th#o&)ho&t the %ood !hai$4
espe!ia2 i$ the aE&ati! e$<i#o$me$t. The o#)a$i! a$d i$o#)a$i! !ompo&$ds o%
me#!&#2 !a$ "e p#ese$t i$ the $at&#a 'ate# a$d !a$ "e !o$!e$t#ated i$ di%%e#e$t
o#)a$isms4 i/e %ish. That is 'h2 the %ish !a$ ha<e a hi)he# !o$te$t o% me#!&#2 as
!ompa#ed 'ith othe# %oodst&%%4 "&t it is di%%i!&t to #epo#t a mea$ !o$!e$t#atio$ o% this
eeme$t4 "e!a&se it depe$ds o$ spe!ies4 a)e4 si7e a$d 'ate# E&ait2 i$ the #eside$!e
a#ea.
F#om a the methods &sed %o# the dete#mi$atio$ o% me#!&#24 the !od <apo&#
atomi! a"so#ptio$ spe!t#omet#2 (CVAAS* has "e!ome p#o"a"2 the most &sed
te!h$iE&e o% a$a2sis "ei$) !ha#a!te#ised "2 a spe!ia se$siti<it2 a$d see!ti<it2.
I$ this 'o#/ is des!#i"ed a method o% a$a2sis "ased o$ the !od <apo&#
atomi! a"so#ptio$ spe!t#omet#2 (CVAAS* &sed %o# the dete#mi$atio$ o% me#!&#2 %#om
a 'o#/i$) e$<i#o$me$t e3posed to this !hemi!a po&ta$t4 a$d %#om a %ish sampe.
E4peri#ental Pr$ce!"re
Reagents and apparatus
0 Ti$ !ho#ide

%/lu$a"4 h2d#o!ho#i! a!id %Ba$er"4 potassi&m pe#ma$)a$ate

%Spole$"4
s&%&#i! a!id %/lu$a"4 h2d#o!ho#i! h2d#o32ami$e %0eactivul"I distied 'ate#
(!o$d&!ti<it2 , CS.!m*.
0 Nit#i! a!id 51; p&#iss. p.a. 0 H) Y +4++++++1 ; %/lu$a"4 s&%&#i! a!id A>; p&#iss.
p.a. M H) Y +4++++++1 ; %/lu$a".
0 A!id mi3t&#e: $it#i! a!id p&#iss. p.a 1.1 ; a$d s&%&#i! a!id p&#iss. p.a 1.1 ;.
0 A"so#"e$t so&tio$ %o# the me#!&#2 <apo&#s: -., 3 1+
0-
; RM$O
,
i$ H
-
SO
,
, ;.
-D
0 S$C
-
1 ; i$ HC 1+ ; .
0 8otassi&m pe#ma$)a$ate so&tio$ 5;.
0 H2d#o32ami$e h2d#o!ho#ide so&tio$ -+ ;.
0 Sta$da#d so&tio$ o% me#!&#2 o% !o$!e$t#atio$ 1+++ m).L (Al!rich*.
0 Sta$da#d so&tio$ o% me#!&#2 o% !o$!e$t#atio$ 1+ m).L 0 p#epa#ed "2 di&tio$ o%
1++ CL o% sta$da#d so&tio$ o% me#!&#2 o% !o$!e$t#atio$ 1+++ m).L 'ith 1+ mL
o% a"so#"e$t so&tio$ %o# me#!&#2 <apo&#s made i$ a <o&met#i! %as/ o% 1+ mL.
0 Sta$da#d o% me#!&#2 o% !o$!e$t#atio$ 1++ C).L 0 p#epa#ed "2 di&tio$ o% 1++ CL
de sta$da#d o% me#!&#2 o% !o$!e$t#atio$ 1+ m).L 'ith 1+ mL o% a"so#"e$t
so&tio$ %o# me#!&#2 <apo&#s made i$ a <o&met#i! %as/ o% 1+ mL.
0 Sta$da#ds o% me#!&#2 o% !o$!e$t#atio$s o% - C).L4 1 C).L4 1+ C).L &sed %o#
the !ai"#atio$ o% the s2stem AAS0MHS 1+ M p#epa#ed "2 addi$) i$ 1+ mL
o% a"so#"e$t so&tio$ %o# me#!&#2 <apo&#s o%: -++ CL4 1++ CL #espe!ti<e2
1+++ CL o% me#!&#2 sta$da#d o% the !o$!e$t#atio$ 1++ C).L.
A the me#!&#2 dete#mi$atio$s "2 atomi! a"so#ptio$ a#e made &si$) a$ atomi!
a"so#ptio$ spe!t#ophotomete# F8e#/i$ Eme#G mode FA$a2st D++G !o&ped 'ith a
ma$&a h2d#ide )e$e#atio$ s2stem M mode FMHS01+G.
)$r8ing Pr$ce!"re '$r t*e Deter#inati$n $' t*e Merc"r& C$ncentrati$n
'r$# a )$r8ing En+ir$n#ent
The me#!&#2 !o$!e$t#atio$ %#om a$ e$<i#o$me$t e3posed to the po&ta$t is
dete#mi$ed: the e$!os&#e o% a met#oo)2 a"o#ato#2. Amo$) the a!ti<ities !a##ied o&t
i$ this a"o#ato#2 is the #epai# o% the#momete#s.
I$ o#de# to dete#mi$e the !o$!e$t#atio$ o% me#!&#2 i$ ai#4 th#ee ai# sampes
a#e !oe!ted4 at th#ee di%%e#e$t mome$ts4 &si$) %o# this a )as sampi$)
p&mp FAmete/G. The th#ee ai# sampes a#e !oe!ted 'ith a %o' #ate o%
a"o&t 1 L.mi$4 %o# te$ mi$&tes. The sampi$) %o' #ate is meas&#ed 'ith a
%o' #ate !ai"#ato# 'ith soap "&""e FGii"#ato# -G4 %o# ea!h sampe
#e!o#di$) the e3a!t <a&e o% the meas&#ed %o' #ate4 to dete#mi$e the
<o&me o% the !oe!ted ai#:
Vo&me
ai#
(V
ai#
* H Fo' (D*X time(t* (1*
Fo# the !oe!tio$ o% the ai# sampes is &sed the sampi$) <esse 'ith %#it o%
1++ mL <o&me4 i$ 'hi!h a#e i$t#od&!ed 1+ mL o% a"so#"e$t so&tio$ %o# me#!&#2
<apo&#s4 so&tio$ i$ 'hi!h is "&""ed a <o&me V o% ai#4 #etai$i$) i$ the a"so#"e$t
so&tio$ the me#!&#2.
=e%o#e the pe#%o#mi$) o% a set o% meas&#eme$ts4 is #eE&i#ed the !ai"#atio$ o%
the AAS0MHS01+ s2stem. Fo# the !ai"#atio$4 the a"so#"a$!es o% the me#!&#2
sta$da#ds 'ith the !o$!e$t#atio$s -4 14 1+ C).L a#e meas&#ed at the 'a<ee$)th o%
-16.D $m .
->
The me#!&#2 sta$da#ds o% !o$!e$t#atio$s -4 14 1+ C).L a#e p#epa#ed #i)ht
"e%o#e the meas&#eme$ts4 "2 i$t#od&!i$) i$ the #ea!tio$ <esse o% the h2d#ide
)e$e#ato# MHS M1+4 i$ 1+ mL o% a"so#"e$t so&tio$ %o# me#!&#2 <apo&#s4 o% -++ CL4
1++ CL a$d #espe!ti<e2 1+++ CL o% me#!&#2 sta$da#d 'ith the !o$!e$t#atio$ 1++
C).L.
Operati$n $' t*e MHS71@ S&te# C$"ple! in Serie <it* t*e At$#ic
A%$rpti$n Spectr$p*$t$#eterA
I$ the %oo'i$) %i)&#e is p#ese$ted the s!heme o% the s2stem MHS01+.
The s!heme o% the s2stem MHS M1+.
Bhe$ the s2stem MHS01+ is i$ sta$d"24 a$ a#)o$ %o' !i#!&ates !o$ti$&o&s2
%#om the i$p&t poi$t o% the <a<e (8* to'a#ds its o&tp&t poi$t4 (A* a$d %#om the#e
th#o&)h the %o' #est#i!to# F
6
(that dete#mi$es a $omi$a <a&e o% the %o' o% 51+
mL.mi$*.
Bhe$ the #ea!tio$ <esse !o$tai$i$) the me#!&#2 sta$da#d is !o$$e!ted to the
s2stem (!o$!e$t#atio$s -4 14 a$d #espe!ti<e2 1+ C).L*4 a %o' o% i$e#t )as !i#!&ates
th#o&)h the t&"e (e* to'a#ds the E&a#t7 !e4 #emo<i$) the ai# %#om the s2stem. A
!o$ti$&o&s %o' o% i$e#t )as passes th#o&)h the #est#i!to# F
1
('ith a $omi$a <a&e o%
-A
the %o' o% -1 mL.mi$* a$d the i$e ("* to'a#ds the imme#sio$ t&"e. The )as %#om the
imme#sio$ t&"e a$d the !o$i!a shape o% the #ea!tio$ <esse e$s&#e a )ood
homo)e$i7atio$ o% the so&tio$. The thi#d %o' o% i$e#t )as !i#!&ates th#o&)h the
#est#i!to# F
-
('ith a $omi$a <a&e o% the %o' o% ,++ mL.mi$* a$d the i$e (a* to'a#ds
the #ea!tio$ <esse.
The #emo<a o% ai# %#om the s2stem 'ith the aid o% the a#)o$ %o' is do$e %o#
1+ se!o$ds. The$4 to pe#%o#m the a$a2sis4 the pisto$ o% the de<i!e is /ept p#essed4 a$d
the %o' o% i$e#t )as that !i#!&ates th#o&)h F
6
is !&t o%%. A p#ess&#e is appied "et'ee$
the o&tp&t (=* o% the <a<e4 o$ the i$e (!* to'a#ds the #ed&!i$) a)e$t ta$/. Th&s the
#ed&!i$) a)e$t is p&shed th#o&)h the t&"e (d* to'a#ds the imme#sio$ t&"e4 th#o&)h
'hi!h is added to the sta$da#d.sampe so&tio$. As #ed&!i$) a)e$t is &sed a so&tio$ o%
S$C
-
1 ; p#epa#ed i$ HC 1+ ; 'hi!h is added to the so&tio$ to "e a$a27ed %o# -5
se!o$ds. The io$s o% H)
-?
%#om the so&tio$ to "e a$a27ed a#e #ed&!ed "2 the S$C
-
to
eeme$ta state me#!&#24 the #es&ted me#!&#2 <apo&#s "ei$) t#a$spo#ted "2 the a#)o$
to the 'i$do'ess E&a#t7 !e (pa!ed o$ a metai! s&ppo#t mo&$ted a"o<e the "&#$e#
o% the atomi! a"so#ptio$ spe!t#ophotomete# FA$a2st D++G*4 'he#e the2 a"so#" the
#adiatio$ %#om the me#!&#2 hoo' !athode amp o% the spe!t#ophotomete#. The
spe!t#ophotomete# FA$a2st D++G is !o$t#oed "2 !omp&te#4 a$d a%te# the additio$ o%
the #ed&!i$) a)e$t to the so&tio$ to "e a$a27ed4 is sta#ted the meas&#i$) o% the
a"so#"a$!e %o# 5+ se!o$ds. O$ the dispa2 o% the !omp&te# the a"so#"a$!e
!o##espo$di$) to the me#!&#2 <apo&#s is #e!o#ded as pea/ 'ith the hei)ht p#opo#tio$a
to the !o$!e$t#atio$ o% the eeme$t %#om the a$a27ed sampe.
A%te#'a#ds the a"so#"a$!es !o##espo$di$) to the ma3im&m o% the pea/s %o#
the me#!&#2 sta$da#ds 'ith the !o$!e$t#atio$s -4 14 a$d #espe!ti<e2 1+ C).L a#e #ead4
'ith t'o #epi!ates %o# ea!h sta$da#d4 a$d the$ the !ai"#atio$ !&#<e is )e$e#ated.
The me#!&#2 !o$!e$t#atio$ is dete#mi$ed %#om ea!h sampe o% a"so#"e$t
so&tio$ i$ 'hi!h the me#!&#2 'as #etai$ed.
Fo# this p&#pose4 %#om the 1+ mL o% a"so#"e$t so&tio$4 1+ mL o% so&tio$ a#e
a$a27ed 'ith t'o #epi!ates4 "2 &si$) the ma$&a h2d#ide )e$e#atio$ s2stem FMHS0
1+G !o&ped 'ith the atomi! a"so#ptio$ spe!t#ophotomete# FA$a2st D++G4 a$d the
a"so#"a$!es !o##espo$di$) to ea!h #epi!ate a#e i$te#poated o$ the !ai"#atio$ !&#<e4
th&s dete#mi$i$) the !o##espo$di$) !o$!e$t#atio$s.
To !a!&ate the me#!&#2 !o$!e$t#atio$ e3p#essed i$ m).m
6
o% ai# is appied
the %o#m&a:
1+++ X * (
X
* . (
. . 6
- .
c .
m m( c
air
sol a's
1(
=
(-*
'he#e V
.so. a"s.
H 1+ mL.
Repa!i$) the eE&atio$ (1* i$ the #eatio$ship (-*4 is o"tai$ed:
6+
1+++ X X
X
* . (
. . 6
t 2
c .
m m( c
sol a's
1(
= (6*
'he#e D H %o' #ate (L.mi$*I t H time (mi$*
Th&s4 to dete#mi$e the !o$!e$t#atio$ o% me#!&#2 %#om the !oe!ted ai#
sampes is ta/e$ i$to !o$side#atio$ the !o$!e$t#atio$ o% me#!&#2 o"tai$ed %#om the
!ai"#atio$ !&#<e (!0 e3p#essed i$ C).L*4 the <o&me o% the a"so#"e$t so&tio$ (V
a"s.so
H 1+ mL*4 the ai# sampi$) %o' #ate (D 0 e3p#essed i$ L.mi$ * a$d the ai# sampi$)
time i$ the a"so#"e$t so&tio$ (t0 e3p#essed i$ mi$*.
)$r8ing Pr$ce!"re '$r t*e Deter#inati$n $' t*e Merc"r& C$ncentrati$n
'r$# a Fi* Sa#ple
A sampe o% !od %iet a!E&i#ed %#om a s&pe#ma#/et is a$a27ed. Fo# this4 a
#ep#ese$tati<e E&a$tit2 o% sampe is !hopped &p4 'ith the aid o% a pasti! /$i%e. F#om
this 'e homo)e$i7ed sampe4 a#e 'ei)hed a"o&t +.D ) o% %ish (the e3a!t E&a$tit2 is
#e!o#ded*. T'o sampes a#e pe#%o#mi$) i$ pa#ae4 a$d to a thi#d sampe is added a
/$o'$ E&a$tit2 o% me#!&#2 sta$da#d. The !ai"#atio$ !&#<e is )e$e#ated &si$) "a$/
so&tio$ a$d me#!&#2 sta$da#ds p#o!essed "2 the same mi$e#ai7atio$ p#o!ed&#e as the
%ish sampes
Procedure for the Mineraliation of the !ish Sa"ples# the Blan$ Solution
and the Mercur% Standard &'(( )g*+ ,g)
I$ t'o E#e$me2e# %as/s 'ith p&) a#e i$t#od&!ed app#o3imate2 +.D ) o% %ish4
1 mL !o$!e$t#ated H
-
SO
,
. The <esses a#e !osed a$d pa!ed i$ the 'ate# "ath set at
D+ WC4 %o# o$e ho&#. The$4 the %as/s a#e !ooed o$ a$ i!e "ath4 addi$) i$ this positio$
1+ mL o% RM$O
,
5 ;. The %as/s a#e the$ i$t#od&!ed i$ the 'ate# "ath set at 11 WC4
%o# t'o ho&#s. The$4 the %as/s a#e et to !oo at #oom tempe#at&#e a$d a#e added 11
mL o% h2d#o32ami$e h2d#o!ho#ide -+ ;. F#om the #es&ted so&tio$s is dete#mi$ed
the me#!&#2 !o$!e$t#atio$ 'ith the aid o% the AAS0MHS 01+ s2stem4 a$a27i$) t'o
#epi!ates o% 1+ mL %o# ea!h so&tio$.
I$ the !ase o% the "a$/ so&tio$ &sed %o# the )e$e#ati$) o% the !ai"#atio$
!&#<e a$d %o# the p#epa#i$) o% the !ai"#atio$ sta$da#ds 'ith the !o$!e$t#atio$s -4 14
a$d #espe!ti<e2 1+ C).L a#e i$t#od&!ed i$ the E#e$me2e# %as/s 'ith p&) 1 mL o%
!o$!e$t#ated H
-
SO
,
4 the #est o% the p#o!ed&#e "ei$) ide$ti!a as %o# the mi$e#ai7atio$
o% the %ish sampes.
The !ai"#atio$ !&#<e is )e$e#ated &si$) the "a$/ so&tio$ a$d the sta$da#ds
that a#e p#o!essed "e%o#e the a$a2sis i$ the same ma$$e# as the sampes. The me#!&#2
sta$da#d o% !o$!e$t#atio$ 1+ m).L is p#epa#ed "2 di&tio$ o% 1++ CL !omme#!ia2
a<aia"e me#!&#2 sta$da#d 'ith the !o$!e$t#atio$ o% 1+++ m).L 'ith 1+ mL o% a
mi3t&#e o% $it#i! a!id 'itho&t H) 1.1 ; a$d s&%&#i! a!id 'itho&t H) 1.1 ; i$ a
<o&met#i! %as/ o% 1+ mL (i$ the %as/ a#e added %i<e d#ops o% RM$O
,
5; %o# the
61
sta"ii7atio$ o% the me#!&#2*. The me#!&#2 sta$da#d 'ith the !o$!e$t#atio$ o% 1++
C).L is o"tai$ed "2 i$t#od&!i$) i$ a$ E#e$me2e# %as/ 'ith p&)4 o% D++ CL me#!&#2
sta$da#d 'ith the !o$!e$t#atio$ o% 1+ m).L a$d 1 mL !o$!e$t#ated H
-
SO
,
4 %oo'i$)
the same mi$e#ai7atio$ p#o!ed&#e as %o# the %ish sampes. Bith a the #ea)e$ts added
i$ the mi$e#ai7atio$ p#o!ess4 the D++ CL o% me#!&#2 sta$da#d 'ith the !o$!e$t#atio$
o% 1+ m).L 'i "e di&ted 'ith D+ mL o% #ea)e$t mi3t&#e4 o"tai$i$) the me#!&#2
sta$da#d 'ith the !o$!e$t#atio$ o% 1++ C).L.
Fo# )e$e#ati$) the !ai"#atio$ !&#<e4 -++ CL4 1++ CL4 a$d #espe!ti<e2 1+++
CL o% me#!&#2 sta$da#d 'ith the !o$!e$t#atio$ o% 1++ C).L mi$e#ai7ed a#e i$t#od&!ed
i$ 1+ mL o% mi$e#ai7ed "a$/ so&tio$ p&t i$ the #ea!tio$ <esse o% the s2stem MHS0
1+4 i$ o#de# to o"tai$ the !ai"#atio$ sta$da#ds 'ith the !o$!e$t#atio$s -4 14 1+ C).L
H). A%te# the !o$$e!tio$ o% the #ea!tio$ <esse 'ith ea!h i$di<id&a sta$da#d4 to the
s2stem MHS M1+4 a#)o$ is "&""ed i$ the #ea!tio$ <esse %o# 1+ se!o$ds to #emo<e the
ai# %#om the s2stem. A%te#'a#ds the #ed&!i$) a)e$t is added i$ the #ea!tio$ <esse
(so&tio$ o% S$C
-
1 ; p#epa#ed i$ HC 1+ ;*4 %o# -5 se!o$ds. U$de# the a!tio$ o% the
#ed&!i$) a)e$t4 the H)
-?
io$s a#e #ed&!ed to eeme$ta H)4 the #es&ted me#!&#2
<apo&#s "ei$) t#a$spo#ted "2 the a#)o$ to the 'i$do'ess E&a#t7 !e 'he#e the2
a"so#" the #adiatio$ %#om the me#!&#2 hoo' !athode amp o% the spe!t#ophotomete#
FA$a2st D++G. The a"so#"a$!e !o##espo$di$) to ea!h sta$da#d is #ead 'ith t'o
#epi!ates. To #ead the F7e#o me#!&#2G poi$t o% the i$st#&me$t i$ o#de# to #emo<e the
i$%&e$!e o% the H) !o$tai$ed i$ the #ea)e$ts4 "e%o#e the #eadi$) o% the a"so#"a$!es
!o##espo$di$) to the me#!&#2 sta$da#ds 'ith the !o$!e$t#atio$s o% -4 14 1+ C).L H)4
i$ the #ea!tio$ <esse o% the de<i!e a#e added 1+ mL o% mi$e#ai7ed "a$/ so&tio$4
meas&#i$)4 a%te# the additio$ o% the #ed&!i$) a)e$t4 its a"so#"a$!e. S&"seE&e$t2 this
<a&e is a&tomati!a2 s&"t#a!ted %#om the a"so#"a$!es !o##espo$di$) to the me#!&#2
sta$da#ds a$d the sampes.
The a"so#"a$!es !o##espo$di$) to the me#!&#2 %#om the %ish sampes a#e
i$te#poated o$ the !ai"#atio$ !&#<e a$d the me#!&#2 !o$!e$t#atio$ %#om 1+ mL o%
a$a27ed so&tio$ is dete#mi$ed.
Calc"lati$n $' t*e Merc"r& C$ncentrati$n 'r$# t*e Fi* Sa#ple
To !a!&ate the me#!&#2 !o$!e$t#atio$ %#om the %ish sampes4 e3p#essed i$
m)./) sampe4 the %oo'i$) %o#m&a is appied:
1+++ X * (
X * (
* . (
.
( m
c m- .
$( m( c
sample
&inalsol
1(
=
(,*
'he#e V
%i$a so.
H D+ mL.
Th&s4 %o# the dete#mi$atio$ o% the me#!&#2 !o$!e$t#atio$ %#om the %ish
sampes is !o$side#ed the !o$!e$t#atio$ o% me#!&#2 o"tai$ed %#om the !ai"#atio$
6-
!&#<e (!0 e3p#essed i$ C).L*4 the <o&me o% the %i$a so&tio$ (a%te# the additio$ o% a
the #ea)e$ts "e%o#e the #eadi$) o% the sampes* a$d the E&a$tit2 o% %ish sampe
a$a27ed (e3p#essed i$ )#ams*.
Appl&ing t*e Met*$! $' Stan!ar! A!!iti$n '$r t*e ?eri'icati$n $'
Inter'erence
I$ a$ E#e$me2e# %as/ 'ith p&)4 a#e i$t#od&!ed app#o3imate2 +.D ) o% %ish4
61 CL me#!&#2 sta$da#d 'ith the !o$!e$t#atio$ o% 1+ m).L a$d 1 mL o% !o$!e$t#ated
H
-
SO
,
. The same p#o!ed&#e %o# the mi$e#ai7atio$ as des!#i"ed a"o<e is &sed. T'o
#epi!ates a#e the$ a$a2sed o% 1+ mL %#om the so&tio$ #es&ted a%te# the
mi$e#ai7atio$ o% the sampe.
The #e!o<e#2 2ied is !a!&ated (;*4 'ith the %o#m&a:
1++ X
!a!. Co$!.
det. .
(;* Re!o<e#2
Conc
= (1*
'he#e: Co$!. det. H !o$!e$t#atio$ dete#mi$ed4 o"tai$ed a%te# app2i$) eE&atio$ (,*I
Co$!. !a!. H !o$!e$t#atio$ !o##espo$di$) to the %ish sampe ?1 C).L (added me#!&#2
sta$da#d*.

66
I.B. DETERMINATION OF pH IN PRECIPITATION.
POTENTIOMETRIC METHOD
Principle
The method is "ased o$ the dete#mi$atio$ o% the pote$tia di%%e#e$!e "et'ee$
a$ ee!t#ode pai# !o$sisti$) o% a )ass ee!t#ode se$siti<e to the di%%e#e$!e i$ the
h2d#o)e$ io$ a!ti<it2 i$ the sampe so&tio$ a$d the i$te#$a %ii$) so&tio$4 a$d a
#e%e#e$!e ee!t#ode4 'hi!h is s&pposed to ha<e a !o$sta$t pote$tia i$depe$de$t o% the
!ompositio$ o% the imme#si$) so&tio$. The meas&#ed pote$tia di%%e#e$!e is
!ompa#ed 'ith the pote$tia o"tai$ed 'he$ "oth ee!t#odes a#e imme#sed i$ a so&tio$
o# "&%%e# 'ith /$o'$ pH o# h2d#o)e$ io$ !o$!e$t#atio$. The pH is de%i$ed "2 the
%o#m&a:
pH
(sampe*
H pH
(#e%e#e$!e*
? S(E
(sampe
* M E
(#e%e#e$!e*
T F.RT1$1+
'he#e E a#e the ee!t#ode pote$tias4 R is the &$i<e#sa )as !o$sta$t4 T the a"so&te
tempe#at&#e a$d F is the Fa#ada2 !o$sta$t.
This is a$ ope#atio$a2 de%i$ed pH. =&%%e#s o% /$o'$ pH a#e spe!i%ied "2 the
Natio$a I$stit&te o% Sta$da#di7ed Te!h$oo)2 (NIST*. The p#ima#2 sta$da#d a$d the
most 'ide2 &sed "&%%e# %o# pH0mete# !ai"#atio$ is +.+1 M potassi&m h2d#o)e$
phthaate4 'hi!h has a pH o% ,.++ at -+@ C4 a$d a h2d#o)e$ io$ a!ti<it2 o% 1+
0,
M. This
atte# h2d#o)e$ io$ a!ti<it2 is "ased o$ theo#eti!a !a!&atio$s (the =ates0
G&))e$heim !o$<e$tio$*.
I$ p#e!ipitatio$ sampes4 the io$i! st#e$)th 'i t2pi!a2 "e i$ the #e)io$ 1+
06
to 1+
01
. The a!ti<it2 !oe%%i!ie$t %o# mo$o<ae$t !atio$s s&!h as the h2d#o)e$ io$ 'i
the#e%o#e "e i$ the #a$)e +.A10+.AA. This !o##espo$ds to Y+.+- pH0&$its di%%e#e$!e
"et'ee$ pH a$d Mo)SH
?
T. M&!h mo#e !#iti!a is the ass&mptio$ o% a !o$sta$t
#e%e#e$!e ee!t#ode pote$tia 'he$ )oi$) %#om a #eati<e2 !o$!e$t#ated potassi&m
h2d#o)e$ phthaate so&tio$ to e3t#eme2 di&te p#e!ipitatio$s sampes. The p#o"em
a#ises "e!a&se o% the i$he#e$t possi"iit2 o% "&idi$) &p a iE&id L&$!tio$ pote$tia
"et'ee$ the i$te#$a so&tio$ o% the #e%e#e$!e ee!t#ode4 a$d the sampe so&tio$. This
iE&id L&$!tio$ pote$tia ma2 "e a#)e# i% the io$i! st#e$)th di%%e#e$!e "et'ee$ the t'o
so&tio$s is a#)e. It is #ed&!ed "2 ma/i$) the "o&$da#2 "et'ee$ the !o$!e$t#ated
%ii$) so&tio$ a$d the sampe as sha#p as possi"e. Va#io&s desi)$s o% pH !es
meeti$) this !#ite#io$ ha<e "ee$ p#oposed. Tests o% !omme#!ia ee!t#odes a)ai$st
di&te a!id so&tio$s a$d o' io$i! st#e$)th "&%%e#s 'ith /$o'$ pH o# h2d#o)e$ io$
!o$!e$t#atio$s ha<e sho'$4 ho'e<e#4 that this p#o"em has a#)e2 "ee$ o<e#!ome
'ith mode#$ pH i$st#&me$tatio$ a$d ee!t#ode s2stems.
6,
Ho'e<e#4 it is st#o$)2 #e!omme$ded to !he!/ the ee!t#ode s2stem at #e)&a#
i$te#<as4 "2 meas&#i$) the Fappa#e$t pHG o% a so&tio$ 'ith o' io$i! st#e$)th 'ith
/$o'$ pH o# h2d#o)e$ io$ !o$!e$t#atio$. The pH #eadi$)s sho&d "e 'ithi$ +.+- o#
+.+1 pH0&$its o% the Ftheo#eti!aG #es&t. I% this is $ot the !ase4 o# i% the #eadi$) is
&$sta"e d&#i$) sti##i$) o% the so&tio$4 the #e%e#e$!e ee!t#ode sho&d "e #epa!ed.
Ne' )ass ee!t#odes sho&d "e tested a)ai$st at east t'o "&%%e#s to see that the
#espo$se is Ne#$stia$.
The #e%e#e$!e ee!t#ode sho&d p#e%e#a"2 "e sto#ed i$ di&te potassi&m
!ho#ide so&tio$ (+.1M*.
The Natio$a I$stit&te o% Sta$da#di7ed Te!h$oo)2 (NIST* so&tio$s 'ith
/$o'$ pH.
1. +.+1 M potassi&m h2d#o)e$ phthaate (C
5
H
,
(COOH*(COOR*4
pH H ,.++ at -+ @C pH H ,.+1 at -1 @C. Disso<e 1+.1- ) potassi&m h2d#o)e$
phthaate4 C
5
H
,
(COOH* (COOR*4 d#ied at 1-+ @C4 i$ 1+++ mL distied 'ate#.
-. +.+-1 M potassi&m dih2d#o)e$ phosphate (RH
-
8O
,
* a$d +.+-1 M disodi&m
h2d#o)e$ phosphate (Na
-
H8O
,
* pH H 5.>> at -+ @C pH H 5.>5 at -1 @C . Disso<e
6.6A ) potassi&m dih2d#o)e$ phosphate4 RH
-
8O
,
4 a$d 16 ) disodi&m h2d#o)e$
phosphate4 Na
-
H8O
,
4 d#ied at 1-+@ C4 i$ 1+++ mL distied 'ate#. I$stead o% the
a$h2d#o&s disodi&m h2d#o)e$ phosphate4 ,.,6 ) o% &$d#ied dih2d#ate4 Na
-
H8O
,
9
- H
-
O4 ma2 "e &sed.
Comme#!ia a<aia"e "&%%e# so&tio$s ma2 aso "e &sed4 "&t sho&d "e
!he!/ed a)ai$st the p#ima#2 sta$da#d "&%%e#s des!#i"ed a"o<e. The "&%%e#s sho&d "e
/ept i$ the da#/ i$ 'e !osed "ottes o% "o#osii!ate o# po2eth2e$e.
pH0mete# 'ith the possi"iit2 o% #eadi$) to the $ea#est +.+- pH0&$its o#
p#e%e#a"2 to the $ea#est +.+1 pH0&$it.
A )ass ee!t#ode a$d a #e%e#e$!e ee!t#ode m&st "e &sed 'ith the pH0mete#.
The #e%e#e$!e ee!t#ode sho&d "e s&ita"e %o# meas&#eme$t i$ o'0io$i! st#e$)th
so&tio$s a$d p#e%e#a"2 "e o% the !aome t2pe %ied 'ith sat&#ated potassi&m
!ho#ide. Othe# #e%e#e$!e ee!t#odes o# !om"i$atio$ ee!t#odes ma2 "e &sed4 "&t a
ee!t#odes sho&d "e !he!/ed %o# a!!epta"e pe#%o#ma$!e.
Ma)$eti! sti##e#4 'ith te%o$ !oated sti##i$) "a#.
=ea/e#s &sed %o# the test so&tio$ sho&d "e made o% "o#osii!ate )ass o#
po2eth2e$e.
Cali%rati$n C"r+e
Cai"#ate the pH0mete# a!!o#di$) to the i$st#&!tio$ ma$&a %o# the i$st#&me$t
&si$) o$e4 o# p#e%e#a"2 t'o4 "&%%e# so&tio$s. The tempe#at&#e o% the "&%%e# so&tio$s
m&st "e /$o'$. The !ai"#atio$ sho&d "e !he!/ed a%te# ea!h set o% sampes.
61
Meas&#e the pH0<a&e o% the sampe a!!o#di$) to the i$st#&!tio$ ma$&a %o#
the i$st#&me$t. The so&tio$ ma2 "e sti##ed4 "&t $ot <i)o#o&s2. The tempe#at&#e o% the
sampe so&tio$ m&st "e the same as the tempe#at&#e o% the "&%%e# so&tio$ &sed %o#
!ai"#atio$.
Ri$se the ee!t#odes tho#o&)h2 'ith distied 'ate# "et'ee$ ea!h
meas&#eme$t4 a$d 'ipe o%% the e3!ess 'ate# 'ith a so%t pape#.
Sto#e the ee!t#odes i$ +.1 M RC0so&tio$ o# a!!o#di$) to the ma$&%a!t&#e#s
#e!omme$datio$s. The #e%e#e$!e ee!t#ode sho&d $ot "e sto#ed i$ distied 'ate#Q
Per'$r#ance Tet $' t*e Electr$!e Pair
The "eha<io&# o% the #e%e#e$!e ee!t#ode is the mai$ so&#!e o% e##o#s i$ pH0
meas&#eme$ts4 espe!ia2 i$ o' io$i! st#e$)th so&tio$s. I$ o#de# to !he!/ the
pe#%o#ma$!e o% the #e%e#e$!e ee!t#ode4 !o$t#o meas&#eme$ts sho&d "e made o$
so&tio$s o% di&te a!ids o# di&te "&%%e#s to <e#i%2 that !o##e!t <a&es a#e o"tai$ed %o#
so&tio$s o% o'e# io$i! st#e$)ths. A so&tio$ 'hi!h sho&d )i<e a pH Z,.++ !o&d "e
&sed %o# the test. A 1+
0,
M HC10so&tio$ sho&d )i<e a pH o% 6.AA [ +.+1.
Ee!t#ode pai#s sho&d aso sho' mi$ima di%%e#e$!es "et'ee$ meas&#eme$ts
made i$ sti##ed a$d &$sti##ed o' io$i! st#e$)th so&tio$s.
Us&a2 the iE&id L&$!tio$ "et'ee$ the so&tio$ a$d the sat&#ated RC0so&tio$ i$ the
#e%e#e$!e ee!t#ode is %o#med i$ a po#o&s p&t o% !e#ami! %i"#e. So' sti##i$) #emo<es
the !o$!e$t#ated RC0so&tio$ 'hi!h so'2 #&$s o&t th#o&)h this !apia#2.
I% the sti##i$) is too <i)o#o&s4 the io$i! medi&m i$ the p&) itse% ma2 "e
di&ted. This 'i i$!#ease the iE&id L&$!tio$ pote$tia4 a$d sho&d "e a<oided. The
iE&id L&$!tio$ pote$tia ma2 aso i$!#ease i% the po#o&s p&) is !o))ed &p "2
imp&#ities.
Re'erence
1. =ates4 R.G. (1A51* Dete#mi$atio$ o% pH4 theo#2 a$d p#a!ti!e. Ne' Yo#/4 Bie2.
-. Li$$et4 N. (1AD+* pH meas&#eme$ts i$ theo#2 a$d p#a!ti!e. Cope$ha)e$4
Radiomete#.
6. Best!ott4 C.C. (1AD>* pH meas&#eme$t. Ne' Yo#/4 A!ad. 8#ess.
,. Da<iso$4 B. a$d Boo%4 C. (1A>1* 8e#%o#ma$!e tests %o# the meas&#eme$t o% pH
'ith )ass ee!t#odes i$ o' io$i! st#e$)th so&tio$s i$!&di$) $at&#a 'ate#s. Anal.
Chem., 53, -15D0-1D+.
65
I.1@. DETECTION OF PESTICIDE USIN9 PRUSSIAN BLUE7
SCREEN PRINTED BIOSENSORS
Intr$!"cti$n
This a"o#ato#2 e3e#!ise p#ese$ts a $e' method "ased o$ Pr"ian Bl"e
#$!i'ie! creen7printe! electr$!e %o# s!#ee$i$) o% o#)a$ophospho#o&s a$d
!a#"amate pesti!ides i$ 'ate#.
8esti!ides a#e amo$) the most impo#ta$t e$<i#o$me$ta po&ta$ts "e!a&se o%
thei# hi)h to3i!it2 a$d thei# si)$i%i!a$t p#ese$!e i$ the e$<i#o$me$t. The &se o%
pesti!ides i$ a)#i!&t&#e has p#o)#essi<e2 i$!#eased si$!e Bo#d Ba# II 'ith a
!o$!omita$t i$!#ease i$ 'o#d %ood p#od&!tio$. I$ this !o$te3t4 i$d&st#ia emissio$ o%
pesti!ides d&#i$) thei# p#od&!tio$4 a$d mo#e impo#ta$t2 the p#ese$!e o% #esid&es o%
these !hemi!as a$d thei# meta"oites i$ %ood4 'ate# a$d soi4 has "e!ome a p#o"em
%o# so!iet2 at a#)e S1T. Amo$) the ma$2 methods #epo#ted %o# pesti!ide dete!tio$4
!h#omato)#aphi! methods s&!h as Hi)h 8e#%o#ma$!e LiE&id Ch#omato)#aph2 (H8LC*
a$d Gas Ch#omato)#aph2 (GC* a#e &sed as #e%e#e$!e methods S-46T. Th&s e$72mati!
methods ha<e "ee$ adopted as a$ ate#$ati<e to !assi!a methods (GC4 H8LC* %o#
%aste# a$d simpe# dete!tio$ o% some e$<i#o$me$ta po&ta$ts S,4 1T.
The &se o% !hoi$este#ase e$72mes %o# i$hi"itio$0"ased dete#mi$atio$ o%
po&ta$ts has sho'$ )#eat p#omise %o# e$<i#o$me$ta s!#ee$i$) a$a2sis S50>T.
Choi$este#ase o!!&# i$ $e#<e tiss&e a$d #ed "ood !es a$d pa2 a$ impo#ta$t
ph2sioo)i!a #oe as it is #espo$si"e %o# the h2d#o2sis o% a!et2thio!oi$e4 a
$e&#ot#a$smitte# ope#ati$) i$ the !oi$e#)i! s2$apses. The e$72mati! h2d#o2sis o%
a!et2!hoi$e i$ the p#ese$!e o% a!et2!hoi$este#ase (AChE* p#o!eeds as %oo':

AChE
CH
6
COO(CH
-
*
-
N
?
(CH
6
*
6
C
0
? H
-
O CH
6
COOH ? HO(CH
-
*
-
N
?
(CH
6
*
6
C
0
(1*
AChE is i##e<e#si"2 i$hi"ited "2 o#)a$ophospho#o&s a$d !a#"amate
pesti!ides a$d a !ompa#iso$ o% the a!ti<it2 o% AChE "e%o#e a$d a%te# e3pos&#e to
e$<i#o$me$ta sampes !a$ p#o<ide a$ e<a&atio$ o% the po&tio$ e<e. The
meas&#eme$t o% e$72mati! a!ti<it2 !a$ "e a!!ompished "2 ee!t#o!hemi!a S54 D4 AT4
!hemi&mi$es!e$t S1+T o# spe!t#ophotomet#i! methods S114 1-T. I$ #e!e$t 2ea#s m&!h
atte$tio$ has "ee$ de<oted to the desi)$ o% i$te)#ate ee!t#o!hemi!a "iose$so#s %o# the
dete!tio$ o% !ompo&$ds i$hi"iti$) the a!ti<it2 o% !hoi$este#ases. The ampe#omet#i!
dete!tio$ o% pesti!ide is mo#e a!!&#ate a$d se$si"e the$ pote$tiomet#i! methods4
"ased o$ meas&#eme$t o% the a!eti! a!id !o$!e$t#atio$ i$!#ease. A "ie$7imati!
ampe#omet#i! s2stem has "ee$ de<eoped &si$) the AChE to)ethe# 'ith the !hoi$e
6D
o3idase (ChO*I the h2d#o27ed !hoi$e is o3idi7ed "2 ChO a$d the ee!t#oa!ti<e
!ompo&$d (H
-
O
-
* is E&a$ti%ied ampe#omet#i!a2.
ChO
HO(CH
-
*
-
N
?
(CH
6
*
6
C
0
? O
-
HOOC(CH
-
*
-
N
?
(CH
6
*
6
C
0
? H
-
O
-
(-*
The "iose$so# p#oposed i$ this 'o#/ is "ased o$ the modi%ied 8#&ssia$ =&e
(8=* t#a$sd&!e#s !o&ped 'ith AChE4 as e$72me4 a$d a!et2thio!hoi$e as s&"st#ate.
The thio!hoi$e p#od&!ed "2 the e$72mati! #ea!tio$ is meas&#ed &si$) s!#ee$0p#i$ted
ee!t#odes (S8Es* modi%ied 'ith 8=. The meas&#eme$t is !a##ied o&t i$ d#op4 at ? -++
mV <s A).A)C.
I$ a p#e<io&s 'o#/ S16T it 'as demo$st#ated %o# the %i#st time the
ee!t#o!ata2ti! e%%e!t o% 8= to'a#ds some thios.
E4peri#ental Part
Principle of the "ethod
The p#omisi$) ad<a$ta)es o% 8= as !ata2st a$d the s!#ee$ p#i$ti$)
te!h$oo)2 has "ee$ !om"i$ed to assem"e se$so#s 'ith imp#o<ed !ha#a!te#isti!s %o#
the ampe#omet#i! dete#mi$atio$ o% thio!hoi$e.
The "iose$so#s %o# pesti!ide dete#mi$atio$ a#e "ased o$ the %oo'i$)
#ea!tio$s:
AChE
CH
6
COS(CH
-
*
-
N
?
(CH
6
*
6
C
0
? H
-
O CH
6
COOH ? HS(CH
-
*
-
N
?
(CH
6
*
6
C
0

The e$72mati! #ea!tio$ p#od&!es thio!hoi$e4 'hi!h is o3idi7ed at 8= modi%ied
s!#ee$0p#i$ted ee!t#odes. 8=0modi%ied S8Es !o<e#ed 'ith a$ app#op#iate e$72mati!
mem"#a$e a#e &sed i$ a$ ee!t#o!hemi!a d#op s2stem.
Reagents
0 G&ta#adeh2de -1; aE&eo&s so&tio$ (<.<* (Si)ma*
0 Na%io$ %#om F&/a4
0 A!et2!hoi$este#ase (AChE* (EC 6.1.1.D4 %#om ee!t#i! ee4 -,, U.m)* (Si)ma*
0 8otassi&m ha3a!2a$o%e##ate (III* !ho#ide (Ca#o E#"a*
0 I#o$ (III* !ho#ide (F&/a*
0 8a#ao3o$ (dieth20p0$it#ophe$iphosphate* (Si)ma*
As 'o#/i$) "&%%e# so&tio$ a potassi&m phosphate "&%%e# sai$e (+.+1M
phosphate "&%%e# ? +.1M RC4 pH D.,* is &sed.
A !hemi!as %#om !omme#!ia so&#!e 'e#e o% a$a2ti!a )#ade.
Apparatus
Ampe#omet#i! meas&#eme$ts a#e !a##ied o&t &si$) a 8amSe$s
Ee!t#o!hemi!a Se$so# I$te#%a!e4 8am I$st#&me$ts =V4 Nethe#a$ds.
6>
4lectro!es preparation
S!#ee$0p#i$ted ee!t#odes &sed i$ this e3pe#ime$ta 'o#/ a#e home0made 'ith
a -,1 DER (Be2mo&th4 E$)a$d* s!#ee$0p#i$ti$) ma!hi$e. G#aphite0"ased i$/
(Ee!t#oda) ,-1* %o#m A!heso$ Itaia$a (Mia$4 Ita2* 'as &sed to p#i$t the 'o#/i$)
ee!t#ode. The s&"st#ate is a %e3i"e po2este# %im (A&tostat HT1* o"tai$ed %#om
A&tot2pe Itaia (Mia$4 Ita2*. The ee!t#odes a#e p#od&!ed i$ %ois o% -+. The diamete#
o% the 'o#/i$) ee!t#ode is +.6 !m #es&ti$) i$ a$ appa#e$t )eomet#i! a#ea o% +.+D
!m
-
4 the !o&$te# i$ )#aphite0"ased i$/ a$d the #e%e#e$!e i$ si<e# !ho#ide.
Prussian Blue !eposition
8= modi%i!atio$ o% S8Es 'as a!!ompished "2 pa!i$) a d#op (1+ L o% tota
<o&me* o% p#e!&#so# so&tio$s o$to the 'o#/i$) ee!t#ode a#ea. This is a mi3t&#e
o"tai$ed addi$) 1 L o% +.1 mo L
01
%e##i! !ho#ide i$ 1+ mmo L
01
HC to 1 L o% +.1
mo L
01
R
6
SFe (CN*
5
T i$ 1+ mmo L
01
HC. The d#op 'as !a#e%&2 pa!ed e3!&si<e2
o$ the 'o#/i$) ee!t#ode a#ea4 i$ o#de# to a<oid the %o#matio$ o% 8= o$ the #e%e#e$!e
a$d !o&$te# ee!t#odes that !o&d $ota"2 i$!#ease the i$te#$a #esista$!e o% the
s2stem. A%te# 1+ mi$&tes the ee!t#odes 'e#e #i$sed 'ith 6 mL o% 1+ mmo L
01
HC.
The ee!t#odes 'e#e the$ e%t A+ mi$&tes i$ the o<e$ at 1++@C to o"tai$ a mo#e sta"e
a$d a!ti<e a2e# o% 8=.
The 8= modi%ied ee!t#odes 'e#e sto#ed d#2 at #oom tempe#at&#e i$ da#/.
4n+)me immo'ili+ation
The AChE 'as immo"ii7ed o$to the 8=0modi%ied ee!t#ode s&#%a!e "2 the
!#oss0i$/i$) method. Fo# this , L mi3t&#e o% )&ta#adeh2de (1 ; i$ 'ate#*4 Na%io$
(1 ; i$ a!oho*4 =SA (6; i$ 'ate#* a$d +.+1 U AChE 'e#e e%t to d#2 (a"o&t 6+ mi$*
o$to the ee!t#ode a#ea. A%te# p#epa#atio$ the "iose$so#s a#e /ept i$ "&%%e# so&tio$ at
,@C.
Pr$ce!"re '$r Petici!e Mea"re#ent
The dete!tio$ o% pesti!ide is !a##ied o&t meas&#i$) the e$72mati! a!ti<it2
"e%o#e a$d a%te# e3pos&#e the e$72me to the sampe. The e$72mati! a!ti<it2
meas&#eme$t "e%o#e the e3pos&#e to the sampe is !a##ied o&t as %oo':
1* 1+ L o% "&%%e# a#e pa!ed o$ the ee!t#ode a$d the$ the ? -++ mV pote$tia
is appied a$d the !&##e$t #e!o#ded (a"o&t 1 $A*. A%te# meas&#eme$t the "&%%e#
so&tio$ is #emo<ed %#om the "iose$so# s&#%a!e.
-* 1+ L o% 'o#/i$) "&%%e# so&tio$ !o$tai$i$) 6 mM thio!hoi$e a#e added
o$ the ee!t#ode a$d the stead20state !&##e$t is #e!o#ded (a"o&t >+ $A*.
The !&##e$t d&e to the o3idatio$ o% thio!hoi$e is the di%%e#e$!e "et'ee$ the
#e!o#ded !&##e$t i$ the step - mi$&s the #e!o#ded !&##e$t i$ step 1. This <a&e is the
i$te$sit2 o% !&##e$t o% e$72me &$i$hi"ited (I
+
*.
6* The "iose$so# is 'ashed 'ith "&%%e# so&tio$ a$d i$!&"ated %o# 6+ mi$&tes
i$ 1 mL sampe4 &$de# sti##i$). D&#i$) the i$!&"atio$ step the AChE is i$hi"ited i% i$
the sampe pesti!ides a#e p#ese$t.
6A
,* A%te# that4 the "iose$so# is 'ashed 'ith "&%%e#4 a$d the -
$d
step is #epeated
a$d the #esid&a e$72mati! a!ti<it2 !&##e$ is meas&#ed.
The !&##e$t d&e to the o3idatio$ o% thio!hoi$e4 p#od&!ed a%te# the e$72me
i$hi"itio$4 is the di%%e#e$!e "et'ee$ the !&##e$t #e!o#ded i$ step , mi$&s the !&##e$t i$
the step 1 ( I
1
*.
8e#!e$ta)e i$hi"itio$ 'as !a!&ated a!!o#di$) to the %o#m&a:
I;H S(I
+
0I
1
*.I
+
T91++
Bi$en$r Reacti+ati$n
This /i$d o% "iose$so#s a#e disposa"e4 "&t it is possi"e to meas&#e the
pesti!ide mo#e times &si$) the same ee!t#ode. I$ %a!t it is possi"e to &se the same
"iose$so# #ea!ti<ati$) the e$72me 'ith p2#idi$e0-0ado3ime methiodide (-08AM*.
A%te# the meas&#eme$ts4 the "iose$so# is 'ashed 'ith distiated 'ate# a$d the
i$hi"ited e$72me is #e)e$e#ated %o# 6+ se! 'ith a d#op 1 mM -08AM p#epa#ed i$
"&%%e# so&tio$. The$4 the "iose$so# is 'ashed a$d a$othe# d#op o% "&%%e# so&tio$
!o$tai$i$) s&"st#ate is added to meas&#e the e$72mati! a!ti<it2 a)ai$. This step
pe#mits to &se o$e "iose$so# mo#e times.
Fo# meas&#i$) a$ &$/$o'$ !o$!e$t#atio$ o% pesti!ide p#ese$t i$ a 'ate# is
$e!essa#2 to pe#%o#m a the , steps a$d !a!&ate the pe#!e$ta)e i$hi"itio$4
#espe!ti<e2. The pesti!ide !o$!e$t#atio$ is !a!&ated &si$) the !ai"#atio$ !&#<e
'hi!h sho&d "e pe#%o#med "e%o#e the sampe a$a2sis.
)*at !$ &$" *a+e t$ !$C
1. Cai"#atio$ !&#<e. Fo# the !ai"#atio$ !&#<e is $e!essa#2 to meas&#e a$d !a!&ate
the i$hi"itio$ #es&ted %o# the $e3t !o$!e$t#atio$s o% pa#ao3o$: +4 1+4 -+4 6+4 ,+
a$d 1+ pp". The pa#ao3o$ so&tio$s 'i "e p#epa#ed i$ distied 'ate# a$d the
steps 10, p#ese$ted at the p#o!ed&#e 'i "e %oo'ed %o# ea!h !o$!e$t#atio$ poi$t.
The de)#ee o% i$hi"itio$ d&e to the amo&$t o% pa#ao3o$ 'i "e )#aphi!a2
#ep#ese$ted a$d the pa#amete#s o% the i$ea# eE&atio$ 'i "e !a!&ated.
-. Sampe a$a2ses a$d !a!&atio$ o% the pesti!ide !o$!e$t#atio$. T'o sampes o%
d#i$/i$) 'ate# spi/ed 'ith pa#ao3o$ 'i "e a$a27ed %o# pesti!ide !o$!e$t#atio$
dete#mi$atio$. The #es&ted e$72mati! i$hi"itio$ o"tai$ed %o# ea!h sampe 'i "e
i$t#od&!ed i$ the eE&atio$ p#e<io&s2 o"tai$ed a$d the pesti!ide !o$!e$t#atio$ 'i
"e !a!&ated.
,+
Re'erence
1. FAO4 A)#i!&t&#e to'a#ds -+1+4 i$: C A6.A, Do!&me$t o% -D
th
Sessio$ o% the FAO
Co$%e#e$!e4 Rome4 1AA6.
-. Stan!ar! metho!s &or e5amination o& 6ater an! 6aste6ater -+
th
ed.I Ame#i!a$
8&"i! Heath Asso!iatio$I Bashi$)to$41AA>I pp 5.>105.A+.
6. Lis/a I.I So"od$i/ N. J.Cromato(r.A 1BB24 3774 -610-1>.
,. 8#ei$i$)e# C.I Bo%"eis O.S. Biosens. Bioelectron. 1BB24 114 A>10AA+.
1. =a)i#o<a4 N.A.I She/ho<tso<a4 T. N.I <a$ H&2stee4 R. =. 8alanta (@@14
5. =e#$a"ei M.I Chia<a#i$i S.I C#emisi$i C.I 8aes!hi G. Biosens. Bioelectron. 1BB,4
84 -510-D1.
D. C#emisi$i4 C.I Di Sa#io4 S.I Mea4 N.I 8ioto$4 R.I 8aes!hi4 G. Anal. Chim. Acta
1BB/4 7114 -D60->+.
>. Ha#t A.L.I Coie# B.A.I Na$sse$ D. Biosens. Bioelectron. 1BB54 124 5,1051,
A. So\ S.I Me#/o!i A.I Ae)#et S. Crit. 0ev. in Anal. Chem. 3 (@@,4 77%2"4 A01-5.
1+. Da$et A.F.I =adea M.I A"o&0E$ei$ H.Y. Biopol)mers %Biospectr." (@@@4 534 6D0
,-.
11. 8o)a]$i/ L.I F#a$/o M. Biosens. Bioelectron. 1BBB4 1,4 15A01D>.
1-. Ema$4 G.L.I Co&#t$e2 R.D.I A$d#es VI Feathe#sto$e R.M. =io!hem. 8ha#ma!o.
1B214 34 >>0A1.
16. Ri!!i4 F.I A#d&i$i4 F.I Ami$e4 A.I Mos!o$e4 D.I 8aes!hi4 G. J. 4lectroanal.
Chem. (@@.4 1564 --A0-6D.
1,. Ri!!i4 F.I Ami$e4 A.I T&ta C.I Ci&!& A.4 L&!a#ei F.I 8aes!hi. Anal. Chim. Acta
(@@,4 9854 11101-+.
,1
I.11. DETERMINATION OF STRON9 AND )EAD ACIDS IN
PRECIPITATION. COULOMETRIC TITRATION METHOD
Intr$!"cti$n
This method is appi!a"e to dete#mi$atio$ o% st#o$) a!ids i$ p#e!ipitatio$
sampes 'ithi$ the !o$!e$t#atio$ #a$)e 1+
01
to 1+
06
M. The o'e# !o$!e$t#atio$ imit
is !ose to the !o$!e$t#atio$s at "a!/)#o&$d sites 'itho&t a/ai$e mi$e#a d&st.
Principle
I$ the !o&omet#i! tit#atio$ method (Li"e#ti et a.4 1AD-*4 the a!id is tit#ated at
!o$sta$t !&##e$t 'ith h2d#o32 io$s i"e#ated at a pati$&m ee!t#ode4 a si<e#0si<e#
"#omide ee!t#ode se#<i$) as the !o&$te# ee!t#ode. The o<e#a #ea!tio$ is:
=#
0
? A) ? H
-
O A)=# ? OH
0
? ^H
-
The EMF o% a )ass0!aome ee!t#ode pai# is #ead at i$te#<as a$d the #es&ts
a#e &sed to !o$st#&!t a G#a$Vs pot (G#a$4 1A1-I Rosotti a$d Rosotti4 1A51*4 'hi!h
)i<es the e$dpoi$t o% the tit#atio$ "2 e3t#apoatio$ o% the st#ai)ht pa#t o% the !&#<e.
The o$2 $e!essa#2 modi%i!atio$ is the additio$ o% a !o$sta$t4 /$o'$ amo&$t
o% a!id to the sampe "e%o#e the tit#atio$4 i$ o#de# to %a!iitate the tit#atio$ o% 'ea/2
a!idi! o# a/ai$e sampes 'itho&t i$te#%e#e$!e %#om !a#"o$ dio3ide.
Reagents
D&#i$) a$a2sis4 &se o$2 #ea)e$ts o% #e!o)$i7ed a$a2ti!a )#ade. The 'ate#
&sed %o# di&tio$ a$d #i$si$) m&st "e do&"e0distied o# de0io$i7ed a$d distied.
0 Nit#o)e$ )as (N
-
* AA.A;
0 8otassi&m "#omide (R=#*
0 S&ph&#i! a!id (H
-
SO
,
* +.+1M
0 =&%%e# so&tio$ pH H ,.++
0 So&tio$ I: 1 M R=# a$d -.1 9 1+
06
M H
-
SO
,
(t#a$s%e# 1-+.+ ) R=# a$d
e3a!t2 1+ mL o% +.+1M H
-
SO
,
to a 1+++ mL <o&met#i! %as/. Fi &p to the ma#/
'ith 'ate#.*.
-nstru"entation
0 E3pa$ded0s!ae pH0mete# (Radiomete# 8HM -5 o# a$ i$st#&me$t 'ith simia#
spe!i%i!atio$s*.
0 Co$sta$t !&##e$t so&#!e (-01+ mA adL&sta"e*
,-
0 A ,.1 V d#2 "atte#2 'ith a$ adL&sta"e se#ies #esista$!e a$d a mA mete# is
s&%%i!ie$t %o# meas&#eme$ts4 "&t F!o&omete#sG a#e a<aia"e !omme#!ia2 (e.).
Met#ohm*.
0 Tit#atio$ <esse4 1++ mL. This sho&d ha<e a s&ita"e id 'ith hoes to se#<e as
s&ppo#t %o# the ee!t#odes a$d the $it#o)e$ i$et4 a$d "e s&ppied 'ith a the#mostat
La!/et.
0 The#mostat (-1 @C [ 1 @C*
0 Se$si$) ee!t#odes. A$ o#di$a#2 )ass ee!t#ode (pH #a$)e +01+* a$d a !aome
#e%e#e$!e ee!t#ode4 o# a !om"i$ed ee!t#ode.
0 Bo#/i$) ee!t#odes. The pati$&m ee!t#ode (- 3 - !m
-
* is made o% "#i)ht pati$&m
(sheet o# $et*. The si<e# ee!t#ode is made %#om AA.A; p&#e si<e#4 1.+ mm
diamete# 'i#e4 a"o&t 6+ !m o$) a$d !oied to a s&ita"e dime$sio$.
0 8ipette: 1+ mL
0 Mi!#o pipettes: +.14 1.+ mL
0 Vo&met#i! %as/: 1+++ mL
T&#$ o$ a i$st#&me$ts4 a$d ao' heati$) %o# ^ ho&#. AdL&st the pH0mete# to
pH H ,.++4 &si$) the "&%%e# so&tio$. T#a$s%e# 1+ mL o% the sampe i$to the
the#mostated tit#atio$ <esse4 a$d add 1 mL o% so&tio$ I. Sta#t $it#o)e$ p&#)i$) a$d
adL&st %o' to )i<e !o$ti$&o&s a)itatio$ o% the so&tio$. The "&""es sho&d $ot dist&#"
the so&tio$ "et'ee$ the se$si$) a$d the 'o#/i$) ee!t#odes. Meas&#e the pH o% the
so&tio$. I% the pH o% the sampe is a"o<e 1.5 it ma2 "e $e!essa#2 to add mo#e tha$ 1
mL o% so&tio$ I.
Bait &$ti pH #eadi$) is !o$sta$t. S'it!h pH0mete# to #ead mii<ots (#a$)e
+0-,+ mV 'ith )ass ee!t#ode positi<e* a$d sta#t the ee!t#o2sis !&##e$t.
Read the )ass ee!t#ode pote$tia <s the !aome ee!t#ode e<e#2 -+ se!o$ds a$d
!o$ti$&e &$ti the pote$tia !ha$)es si)$ (at pH !a. >*. Stop the ee!t#o2sis.
8ot G#a$Vs %&$!tio$4 Y4 at -1@C (see the %oo'i$) ta"e* a)ai$st ee!t#o2sis
time (i$ se!o$ds*. The pot i$te#!epts the a"s!issa at the eE&i<ae$!e poi$t4 t
e
(F H
Fa#ada2s !o$sta$t4 R H the &$i<e#sa )as !o$sta$t4 T H a"so&te tempe#at&#e*.
,6
G#a$Vs %&$!tio$
E
#?
& E
#?
& E
#?
& E
#?
&
1 1.+, ,1 ,.A6 >1 -6., 1-1 111
- 1.+> ,- 1.16 >- -,., 1-- 111
6 1.1, ,6 1.66 >6 -1.6 1-6 1-+
, 1.1D ,, 1.11 >, -5.6 1-, 1-1
1 1.-- ,1 1.DD >1 -D., 1-1 16+
5 1.-5 ,5 1.A> >5 ->., 1-5 161
D 1.61 ,D 5.-- >D -A.5 1-D 1,+
> 1.65 ,> 5.,D >> 6+.D 1-> 1,5
A 1.,- ,A 5.D1 >A 6-.+ 1-A 11-
1+ 1.,> 1+ D.++ A+ 66.6 16+ 11>
11 1.1, 11 D.-> A1 6,.5 161 15,
1- 1.5+ 1- D.1D A- 65.+ 16- 1D1
16 1.55 16 D.>D A6 6D., 166 1DD
1, 1.D6 1, >.1A A, 6>.> 16, 1>1
11 1.>+ 11 >.11 A1 ,+., 161 1A-
15 1.A+ 15 >.>1 A5 ,-.+ 165 1AA
1D 1.A, 1D A.-+ AD ,6.5 16D -+D
1> -.+- 1> A.1D A> ,1.6 16> -15
1A -.1+ 1A A.A, AA ,D.- 16A --,
-+ -.1> 5+ 1+.6 1++ ,A.1 1,+ -66
-1 -.-5 51 1+.D 1+1 11.+ 1,1 -,-
-- -.65 5- 11.1 1+- 16.1 1,- -1-
-6 -.,1 56 11.5 1+6 11.- 1,6 -5-
-, -.1, 5, 1-.1 1+, 1D., 1,, -D-
,,
E
#?
& E
#?
& E
#?
& E
#?
&
-1 -.51 51 1-.1 1+1 1A.D 1,1 ->6
-5 -.D1 55 16.+ 1+5 51.A 1,5 -A,
-D -.>5 5D 16.1 1+D 5,., 1,D 6+5
-> -.AD 5> 1,.1 1+> 5D.+ 1,> 61>
-A 6.+A 5A 1,.5 1+A 5A.D 1,A 661
6+ 6.-1 D+ 11.- 11+ D-., 11+ 6,,
61 6.6, D1 11.> 111 D1.6 111 611
6- 6.,> D- 15.1 11- D>.6 11- 6D1
6, 6.51 D, 1D.1 116 >1.1 116 6>5
6, 6.D1 D, 1D.> 11, >,.D 11, ,+-
61 6.A+ D1 1>.1 111 >>.1 111 ,1>
65 ,.+5 D5 1A.6 115 A1.5 115 ,6,
6D ,.-6 DD -+.+ 11D A1.1 11D ,1-
6> ,.6A D> -+.> 11> A>.A 11> ,D+
6A ,.15 DA -1.D 11A 1+6 11A ,>A
,+ ,.D, >+ --.1 1-+ 1+5 15+ 1+D

E4prei$n $' Re"lt
The !o$!e$t#atio$ o% st#o$) a!id i$ the sampe is !a!&ated %#om the %o#m&a:
o#
'he#e
,1
i H ee!t#o2sis !&##e$t i$ ampe#es
t
e
H ee!t#o2sis time at eE&i<ae$!e poi$t (se!o$ds*
F H Fa#ada2s !o$sta$t (!o&om"s.mo*
V
o
H i$itia sampe <o&me (it#es*
N
H-SO,
H $o#mait2 o% added s&ph&#i! a!id
V
H-SO,
H <o&me o% added s&ph&#i! a!id (it#es*
Notes =o#osii!ate )ass !a$ "e &sed %o# sto#a)e o% sampes.
The )ass'a#e m&st "e t#eated 'ith hot di&te a!id a$d tho#o&)h2 soa/ed i$ distied
'ate# p#io# to &se. 1- ho&#s 'ith 1+; h2d#o!ho#i! a!id at A+@C %oo'ed "2 -, ho&#s
soa/i$) i$ distied 'ate# is !o$side#ed adeE&ate. Othe#'ise4 a/ai metas %#om the
)ass 'i di%%&se i$to the sampes.
Re'erence
1. G#a$4 G. (1A1-* Dete#mi$atio$ o% the eE&i<ae$!e poi$t i$ pote$tiomet#i! tit#atio$s.
8a#t II. Anal)st, 334 55105D1.
-. Li"e#ti4 A.4 8ossa$7i$i4 M. a$d Vi!edomi$i4 M. (1AD-* The dete#mi$atio$ o% the
$o$0<oatie a!idit2 o% #ai$ 'ate# "2 a !o&omet#i! p#o!ed&#e. Anal)st, :34 61-0615.
6. Rosotti4 F.N.C. a$d Rosotti4 H.N. (1A51* 8ote$tiomet#i! tit#atio$s &si$) G#a$Vs pots.
Chem. 4!uc., 924 6D106D>.
,5
,D
I.1(. DETERMINATION OF BEN6ENEE TOLUENE AND 3ILENE IN
AIR BY 9ASS CHROMATO9RAFY
Intr$!"cti$n
The a#omati! h2d#o!a#"o$s ha<e a$ e3t#ao#di$a#2 i$d&st#ia impo#ta$!eI the2
a#e #a' mate#ia %o# mo#e tha$ 5+; o% the to$$a)e o% pasti!s4 eastome#s a$d
s2$theti! %i""e#s a$d aso %o# d2es4 i$se!ti!ides4 medi!i$es4 et!. I$ additio$ the2 a#e
aso p#ese$t i$ %&es o"tai$ed %#om the pet#oe&m. D&e to that 'e !a$ %i$d them as
atmosphe#i! po&ti$) a)e$ts i$ <e#2 di<e#se !o$!e$t#atio$s. The most #ee<a$t a#e
"e$7e$e4 to&e$e a$d di%%e#e$t 32e$es (mi3ed isome#s*4 %oo'ed "2 the $aphthae$e
a$d the a$th#a!e$e.
The mai$ so&#!es o% the "e$7e$e a#e the si)ht %#a!tio$s o% the so%t !oa ta#
a$d the e3t#a!ted a#omati! %#a!tio$s %#om )asoi$e. The to&e$e is o"tai$ed %#om the
same so&#!es that the "e$7e$e a$d !a$ "e sepa#ated %#om it a$d %#om 32e$e "2
s&!!essi<e distiatio$s. Mo#e o% 1+; o% the p#od&!ed to&e$e "e!omes "e$7e$e "2
h2d#o0dea/2atio$. A pa#t o% the %#a!tio$s 'ith hi)h !o$te$ts o% to&e$e is added to
)asoi$e to i$!#ease the o!ta$e de)#ees. These #easo$s ead to %i$d "e$7e$e4 to&e$e
a$d 32e$e i$ the atmosphe#e i/e po&ti$) a)e$ts. No#ma2 the2 a#e a$a27ed Loi$t2
"2 )as !h#omato)#aph2 a$d the tota amo&$t is e3p#essed as =T_ (U) o% "e$7e$e4
to&e$e a$d 32e$e m
06
*.
Gas !h#omato)#aph2 (GC* is a te!h$iE&e o% sepa#atio$ <e#2 &sed to so<e
p#o"ems i$ di<e#se %ieds4 amo$) them the e$<i#o$me$ta a$a2sis. At the mome$t it
is &sed #o&ti$e2 as a !o$t#o te!h$iE&e p#ese$ti$) <e#2 )ood #eso&tio$ %o# <oatie
o#)a$i! !ompo&$ds.
The theo#eti!a p#i$!ipes o% the sepa#atio$ "2 mea$s o% GC do $ot di%%e# "asi!a2
%#om those o% the !h#omato)#aph2 i$ )e$e#a. I$ the GC the mo"ie phase is a )as4
'he#eas the statio$a#2 phase !a$ "e a soid adso#"e$t o# a iE&id #etai$ed o$ a soid
s&ppo#t (pa!/ed !o&m$* o# imp#e)$ati$) the 'as o% a !apia#2 !o&m$ (ope$
!o&m$*.
To o"tai$ a )ood sepa#atio$ a$d )ood #eso&tio$ !h#omato)#ams it is
$e!essa#2: a pe#%e!t tempe#at&#e !o$t#o o% the !o&m$4 the !o$t#o the %o'0#ate o% the
mo"ie phase a$d a s&ita"e ee!tio$ o% the mo"ie a$d statio$a#2 phases4 as 'e as o%
the dete!tio$ s2stem.
Target
To "e t#ai$ed i$ the !o##e!t &se o% sta$da#ds a$d i$st#&me$ta te!h$iE&es o%
hi)h pe#%o#ma$!e.
To "e %amiia# 'ith the )as !h#omato)#aph a$d its pe#%o#ma$!e.
Ide$ti%2 a$d dete#mi$atio$ o% "e$7e$e4 to&e$e a$d 32e$e i$ ai# sampes.
,>
Glassware and apparatus Reagents
Sampi$) set 8e$ta$e
Adso#"e$t !a#t#id)es Sta$da#ds %o# GC:
Gas !h#omato)#aph =e$7e$e (sta$da#d*
Capia#2 !o&m$ $o$ poa# To&e$e (sta$da#d*
Dete!to# FID _2e$e (sta$da#d*
So%t'a#e %o# data p#o!ess
A$a2ti!a "aa$!e
1A mL t&"es p#o<ided 'ith stop!o!/
-1 mL <o&met#i! %as/s
14 -4 6 a$d , mL pipettes
Mi!#os2#i$)e4 1+ CL
Preparati$n $' S$l"ti$n
Sto!/ so&tio$ A. I$ a -1 mL %as/ a#e i$t#od&!ed -+ mL o% pe$ta$eI !o<e#
ti)ht2 a$d 'ei)h it. Ne3t4 add 1++ CL o% "e$7e$e4 it is !o<e#ed
immediate2 a$d 'ei)hed a)ai$. 1++ CL o% to&e$e a#e added
immediate2. Remo<e the %as/ %#om the "aa$!e a$d e<e it 'ith pe$ta$e.
Sta$da#d so&tio$ =. I$ a t&"e i$t#od&!e A mL o% pe$ta$e a$d add 1 mL o%
the sto!/ so&tio$ A.
Note: !o<e# immediate2 the %ash a$d 'o#/ a'a2s i$to a %&me hood.
Pr$ce!"re
1* Sampi$) a$d sampe p#e0t#eatme$t. =2 mea$s o% the sampi$)
eE&ipme$t4 it is %o#!ed to %o' th#o&)h the adso#ptio$ !a#t#id)e a$ ai#
<o&me e3a!t2 meas&#ed (1+ L*4 so that the "e$7e$e !o$!e$t#atio$4
to&e$e a$d 32e$e a#e 'ithi$ the !ai"#ated st#ai)ht i$ea# #a$)e. I% it is
$e!essa#2 pe#%o#m the s&ita"e adL&steme$ts.
-* E3t#a!tio$. I$ the a$a2ti!a a"o#ato#2 the !o$te$t o% the sampi$) t&"e
is p&t i$ !o$ta!t 'ith 1++ mL o% pe$ta$e to e3t#a!t the o#)a$i! !ompo&$ds.
6* Fit#atio$. The e3t#a!t is %ite#ed th#o&)h a $2o$ %ite# o% +.,1 Cm.
,* Cai"#atio$. Sta$da#d so&tio$s o% "e$7e$e4 to&e$e a$d 32e$e. Usi$)
!o<e#ed t&"es4 a se#ies o% sta$da#d so&tio$s o% "e$7e$e4 to&e$e a$d
32e$e is p#epa#ed "2 di&ti$) +.14 1.+4 -.+4 6.+ a$d ,.+ mL o% so&tio$ = i$
pe$ta$e &p to a 1+ mL.
Note: !o<e# immediate2 the %ash a$d 'o#/ a'a2s i$to a %&me hood.
1* A$a2sis. The !h#omato)#aphi! a$a2sis 'i "e pe#%o#med a!!o#di$) to
%oo'i$) !o$ditio$s:
I$Le!tio$ <o&me: 1CL
,A
Co&m$: H81 t2pe phe$2meth2sii!o$e (6+m 3 +.6- mm 3 +.-1 Cm*
Gas !a##ie# : Hei&m4 %o'0#ate 1.1 mL mi$
01
I$Le!to# tempe#at&#e -6+ WC
Dete!to# tempe#at&#e -1+ WC
8#o)#ammed Co&m$ tempe#at&#e: Isothe#m at 61 @C d&#i$) 1 mi$.
F#om 61 @C to 1,+ @C at 11 @C mi$
01
Isothe#m at 1,+ @CI - mi$
Fi$a tempe#at&#e: 61 @C
1.1.* Ide$ti%i!atio$ o% "e$7e$e4 to&e$e a$d 32e$e. A s&ita"e so&tio$ o%
"e$7e$e a$d the$ o% to&e$e a#e i$Le!ted to assi)$ the #ete$tio$ times
'hi!h ao' ide$ti%2 s&!h !ompo&$ds eithe# i$ the !ai"#atio$ )#aph a$d
sampes.
1.-.* Sta$da#d i$Le!tio$s. The di%%e#e$t sta$da#d so&tio$s o% "e$7e$e4
to&e$e a$d 32e$e a#e i$Le!ted i$ the same !o$ditio$s o"tai$i$) the
!o##espo$di$) !h#omato)#ams a$d !o##e!t i$te)#atio$ o% the a#eas o% the
pea/s %#om these !ompo&$ds.
1.6.* Sampe I$Le!tio$. 1 CL o% the e3t#a!t is aso i$Le!ted i$ the same
!o$ditio$s o"tai$i$) the !o##espo$di$) !h#omato)#am a$d the i$te)#atio$
a#eas o% the "e$7e$e4 to&e$e a$d 32e$e pea/s 'hi!h 'i "e ide$ti%ied "2
its #ete$tio$ time.
Re"lt
1* Ide$ti%2 the #ete$tio$ times (t
R
* %o# the th#ee !ompo&$ds.
-* Ca!&ate the !o$!e$t#atio$ i$ C) L
01
o% "e$7e$e4 to&e$e a$d 32e$e
&si$) a dots i$ the !ai"#atio$ )#aph.
A 0t$c8 $l"ti$n1 )riteA
Vo&me o% "e$7e$e (CL* 1++
m) o% 'ei)hed "e$7e$e
C) L
01
o% "e$7e$e
Vo&me o% to&e$e 1++
m) o% 'ei)hed to&e$e
C) L
01
de to&e$e
Vo&me$ de 32e$e 1++
m) o% 32e$e (e3a!t 'ei)ht*
C) L
01
o% 32e$e
Stan!ar! n 1 ( , . /
Vo&me o% = so&tio$ (i$ mL*
C) L
01
o% "e$7e$e
C) L
01
o% to&e$e
1+
C) L
01
o% 32e$e
Write in the ta'les the area o& the pea$s in the cali'ration (raph
=e$7e$e ).L
`#ea
To&e$e ).L
`#ea
_2e$e ).L
`#ea
6* D#a' the !ai"#atio$ i$es. Rep#ese$t the a#eas o"tai$ed i$ the
!h#omato)#ams vs. the !o$!e$t#atio$ <a&es o% "e$7e$e4 to&e$e a$d 32e$e
a$d appied the !o##espo$di$) i$ea# #e)#essio$ a$a2sis.
,* O"tai$ %#om the !ai"#atio$ i$es the !o##espo$di$) !o$!e$t#atio$s o%
"e$7e$e4 to&e$e a$d 32e$e i$ the simpe so&tio$s.
Ide$ti%2 the p#ese$!e o% these th#ee !ompo&$ds "2 Lea$s o% the #ete$tio$ times.
O"tai$ the !o##e!t pea/ a#eas %#om the !h#omato)#amI the$ !a!&ate the
!o$!e$t#atio$s i$ C) m
06
.
Relate! -"eti$n
1* E3pai$ the p#o"ems #eati<e to the !o$tami$atio$ d&e to o#)a$i! !ompo&$ds4 its
o#i)i$s4 e%%e!ts a$d possi"e so&tio$s.
-* List the di%%e#e$t sampi$) s!hemes %o# o#)a$i! !ompo&$ds.
6* D#a' a s!heme o% the )as !h#omato)#aph.
,* Bhat e%%e!t has the tempe#at&#e o% the !o&m$ o$ the sepa#atio$ o% these
!ompo&$dsO
11
1-
I. 1,. DETERMINATION OF THE ?OLATILE OR9ANIC
COMPOUNDS FROM THE ATMOSPHERE USIN9 A
PORTABLE 9AS CHROMATO9RAPH
Maria7Critina BRATU
Intr$!"cti$n
The mo$ito#i$) o% the <oatie o#)a$i! !ompo&$ds (VOC* i$ the
e$<i#o$me$ta sampes is do$e eithe# "2 the !oe!ti$) o% ai# sampes a$d thei#
s&"seE&e$t a$a2sis i$ the a"o#ato#24 o# "2 dete#mi$atio$ at the sampi$) site &si$) a
po#ta"e )as !h#omato)#aph M (p#o!ed&#e des!#i"ed i$ this 'o#/*.
The a$a2sis o% the <oatie o#)a$i! !ompo&$ds %#om the atmosphe#e is do$e4
&s&a24 eithe# "2 thei# adso#ptio$ o$ !a#t#id)es 'ith spe!i%i! %ii$)s4 %oo'ed "2
the#ma deso#ptio$ a$d thei# a$a2sis 'ith the aid o% a !h#omato)#aph p#o<ided 'ith a
dete!to# MS4 FID4 o# ECD4 o# "2 !oe!ti$) ai# sampes i$ metai! <esses %oo'ed
"2 thei# p#ess&#i7atio$ 'ith a$ i$e#t )as a$d the ate# )as !h#omato)#aphi! a$a2sis o%
the mi3t&#e.
=oth a$a2sis p#o!ed&#es #eE&i#e the i$te#<e$tio$ o% the h&ma$ ope#ato#
i$!#easi$) the possi"iit2 o% appea#a$!e o% s2stemati! e##o#s that a#e #e%e!ted i$ the
p#e!isio$ o% the a$a2sis.
The &se o% a po#ta"e )as !h#omato)#aph p#o<ided eithe# 'ith a FID4 8ID o#
ECD does $ot #eE&i#e the i$te#<e$tio$ o% the ope#ato# i$ the sampi$) a$d a$a2sis
steps4 )#eat2 simpi%2i$) the 'o#/i$) p#o!ed&#e a$d a<oidi$) the e##o#s i$he#e$t to
the i$te#<e$tio$ o% the ope#ato# i$ the sampe p#epa#i$) step.
Reagent an! EF"ip#ent
Ca##ie# )as ($it#o)e$ o% !h#omato)#aphi! p&#it24 mi$ AA.AAA;*.
C2i$de# %o# the !ai"#atio$ o% the po#ta"e GC: FSCOTTYG !2i$de# o% ,> L a$d
6++ psi (=TE_ Gas Mi3*4 !o$sisti$) o% si3 a$a2tes 'ith the !o$!e$t#atio$ i$
$it#o)e$ o% 1+ ppm ea!h4 as %oo's: "e$7e$e4 eth2 "e$7e$e4 to&e$e4 m032e$e4
o032e$e4 p032e$e.
The dete#mi$atio$s o% <oatie o#)a$i! !ompo&$ds 'e#e do$e &si$) the
po#ta"e )as !h#omato)#aph F8hoto<a!G4 mode Vo2a)e#4 'ith the %oo'i$)
!o$%i)&#atio$: t'o the#mostated i$Le!tio$ s2stems (a$ a&tomated )as i$Le!tio$ <a<e4 a
)as i$Le!tio$ po#t &si$) a )as s2#i$)e*4 a !o&m$ o<e$ that e$s&#es the the#mostati$)
o% %o&# !apia#2 !o&m$s mo&$ted at the %a!to#2 i$side it (a F"a$/G !o&m$ %o# the
a$a2sis o% the tota E&a$tit2 o% <oatie o#)a$i! !ompo&$ds4 a !o&m$ %o# the a$a2sis
o% the i)ht <oatie o#)a$i! !ompo&$ds4 a !o&m$ %o# the a$a2sis o% the medi&m
16
<oatie o#)a$i! !ompo&$ds a$d a !o&m$ %o# the a$a2sis o% the hea<2 <oatie
o#)a$i! !ompo&$ds*4 t'o )as !h#omato)#aphi! dete!to#s !o$$e!ted i$ pa#ae 'ith
sim&ta$eo&s data a!E&isitio$ (8ID0 photoio$i7atio$ dete!to# a$d ECD0 ee!t#o$
!apt&#e dete!to#* a$d the o$"oa#d s2stem %o# data p#o!essi$).sto#a)e. The po#ta"e )as
!h#omato)#aph is p#o<ided 'ith a$ ee!t#o$i! %o' #ate !o$t#oe# %o# the !a##ie# )as
a$d !o$$e!tio$ i$te#%a!e %o# the e3te#$a 8C %o# the !o$t#o o% the )as !h#omato)#aph4
t#a$s%e# o% the a!E&i#ed data a$d thei# #e0p#o!essi$) 'ith the !h#omato)#aphi!
so%t'a#e SiteCha#t.
I$ the %i)&#e "eo' is #ep#ese$ted the s!heme o% the po#ta"e )as
!h#omato)#aph F8hoto<a!G mode Vo2a)e# i$ 'hi!h a#e i$di!ated a its !ompo$e$t
pa#ts.
The s!heme o% the po#ta"e )as !h#omato)#aph G8hoto<a!G mode Vo2a)e#
)$r8ing pr$ce!"re
=e%o#e sta#ti$) the ope#atio$ 'ith the po#ta"e )as !h#omato)#aph its "atte#2
m&st "e !ha#)ed.
The i$te#$a !2i$de# o% the !h#omato)#aph is oaded 'ith !a##ie# )as ($it#o)e$
'ith the p&#it2 AA.AAA;* "2 !o$$e!ti$) the i$p&t po#t o% the i$st#&me$t th#o&)h a
p#ess&#e #e)&ato#4 to a $it#o)e$ !2i$de# o% -++ atm. The %ii$) 'ith !a##ie# )as o% the
1,
i$te#$a !2i$de# o% the !h#omato)#aph e$s&#es its a&to$om2 %o# D ho&#s4 d&#i$) this
time o$ site )as !h#omato)#aphi! meas&#eme$ts !a$ "e do$e.
A%te# the !ha#)i$) 'ith !a##ie# )as4 the !h#omato)#aph is sta#ted "2 p&shi$)
the O$.O%% "&tto$4 "ei$) #eE&i#ed a$ i$te#<a o% a"o&t ha% a$ ho&# %o# its sta"ii7atio$
a$d #ea!hi$) o% the optima 'o#/i$) pa#amete#s4 mome$t i$di!ated "2 the )#ee$ !oo#
o% the state ed o% the )as !h#omato)#aph (FRead2 stat&s i$di!ato#G*.
=e%o#e the sta#ti$) o% the dete#mi$atio$4 a sampi$) o!atio$ #e!o)$itio$
$ami$) is e$te#ed4 &si$) the /e2"oa#d o% the !h#omato)#aph. Fo# this4 the %oo'i$)
s!heme is %oo'ed:
I.. is e$te#ed ENTER MENU &si$) o$e o% the %o&# %i3ed /e2s o!ated o$
the e%t side o% the dispa2 (it #e)a#ds the &p a$d do'$ positio$i$)
o#de#4 the /e2s O$.O%%4 Sta#t.Stop4 E$te# Me$&4 E3it* the$4 &si$) the
/e2s "eo' the dispa24 is p#essed the /e2 "eo' SETU84 the$ the /e2
"eo' TAG. I$ this mome$t is e$te#ed the $ami$) o% the sampi$)
o!atio$ %o# the ate# #e!o)$itio$ o% !h#omato)#am #e!o#ded at that
site.
Aso4 a#e !hose$ the !o&m$ a$d the dete!to#4 'ith the aid o% 'hi!h the
a$a2sis 'i "e made. Fo# Fi)htG !ompo&$ds (%o# e3ampe: a!eto$e4 meth20eth20
/eto$e4 !ho#o%o#m* is !hose$ the !o&m$ C4 a$d %o# the Fmedi&mG o$es ("e$7e$e4
to&e$e4 32e$es4 meth20iso"&th20/eto$e4 t#i!ho#eth2e$e* is see!ted the !o&m$ =.
The a$a2sis time %o# the !o&m$ C is 11 mi$&tes4 a$d %o# the !o&m$ = is ha% a$
ho&#. Bith #e)a#d to the dete!to#s4 the data a!E&isitio$ !a$ ta/e pa!e sim&ta$eo&s2
o$ "oth dete!to#s4 o# o$2 o$ the photoio$i7atio$ dete!to# (i$ the !ase o% the
!ompo&$ds ha<i$) the io$i7atio$ pote$tia o'e# tha$ 1+.5 eV* o# o$2 o$ the ee!t#o$
!apt&#e dete!to# (i$ the !ase o% !ompo&$ds !o$tai$i$) atoms o% !ho#i$e o# "#omi$e*.
Fo# the see!tio$ o% the !o&m$4 is e$te#ed ENTER MENU 0 SETU8 M CONFIG M
COLUMN a$d 'ith the aid o% the /e2 E$te# Me$& is see!ted o# desee!ted a
!h#omato)#aphi! !o&m$.
A%te# the esta"ishi$) o% these pa#amete#s4 the ai# sampe to "e a$a27ed is
sampi$) "2 see!ti$) the /e2 START.STO8. Fo# -+ se!o$ds a$ ai# sampe 'i "e
d#a'$ i$ 'ith the aid o% a p&mp (d&#i$) sampi$) the i$di!ato# ed o% the sampi$)
sta)e4 $amed FSampi$) stat&s i$di!ato#G0 'i ha<e a #ed !oo#*. A%te# the -+ se!o$ds
sampe sampi$) i$te#<a4 the sampe is Ft#a$spo#tedG "2 the !a##ie# )as %#om the e$t#2
poi$t i$ the !h#omato)#aph4 th#o&)h the p#e<io&s2 see!ted sepa#atio$ !o&m$
to'a#ds the dete!to#.
Ea!h !ompo$e$t o% the sampe 'i "e #etai$ed 'ith a di%%e#e$t time i$side the
!o&m$ a$d 'i "e e&ated %#om the !o&m$ 'ith a !ha#a!te#isti! #ete$tio$ time. I% the
#espo$se o% the dete!to# is #epo#ted to the dead time4 this #espo$se 'i "e p#ese$ted as
a se#ies o% !h#omato)#aphi! pea/s sepa#ated "2 time4 'ith o$e pea/ %o# ea!h
!ompo$e$t o% the sampe. The #ete$tio$ time se#<es %o# the ide$ti%i!atio$ o% the
!ompo$e$ts o% the a$a27ed ai# sampe4 a$d the a#eas o% the !h#omato)#aphi! pea/s
a#e &sed %o# the E&a$titati<e a$a2sis.
11
The E&aitati<e a$d E&a$titati<e a$a2sis made 'ith the Vo2a)e# i$st#&me$t4
a%te# the a$a2sis o% a$ ai# sampe is "ased o$ the #ete$tio$ times a$d the a#eas o% the
pea/s !o##espo$di$) to the p#e<io&s2 a$a27ed sta$da#ds. The !h#omato)#aph
p#ese$ts i$ its data"ase a i"#a#2 o% !ompo&$ds4 'ith a sta$da#d o% ea!h i$te#esti$)
!ompo&$d "ei$) sto#ed i$ its memo#2 "2: #ete$tio$ time4 pea/ a#ea a$d !o$!e$t#atio$.
The #atio pea/ a#ea.!o$!e$t#atio$ #ep#ese$ts the se$siti<it2 %o# a !e#tai$ !ompo&$d
(e3p#essed i$ mVS.ppm*4 a$d this i$%o#matio$ is sa<ed i$ a pa#t o% the memo#2 o% the
Vo2a)e# i$st#&me$t /$o'$ as the i"#a#2 'hi!h 'i "e a&tomati!a2 &pdated d&#i$)
the !ai"#atio$ o% the i$st#&me$t.
I$ the !ase o% ea!h a$a27ed sampe4 the #ete$tio$ times !o##espo$di$) to the
#e!o#ded !h#omato)#aphi! pea/s a#e !ompa#ed a&tomati!a2 to the #ete$tio$ times o%
the !ompo&$ds %#om the !h#omato)#aph i"#a#2. I% a$ o<e#appi$) 'ithi$ the
esta"ished imits o% the pea/ #e!o)$itio$ 'i$do' is o"se#<ed4 tha$ that pea/ is
ide$ti%ied as o$e o% the !ompo&$ds %#om the i"#a#2. Fo# the !a!&atio$ o% the
!o$!e$t#atio$ !o##espo$di$) to the ide$ti%ied pea/4 its a#ea is di<ided to the se$siti<it2
o% the !o##espo$di$) sta$da#d %#om the i"#a#2 (the se$siti<it20 #atio
a#ea.!o$!e$t#atio$*. The #ete$tio$ times a$d the #atios a#ea.!o$!e$t#atio$
!o##espo$di$) to the sta$da#ds o% the !ompo&$ds %#om the i"#a#2 a#e &pdated e<e#2
time the Vo2a)e# !h#omato)#aph is !ai"#ated.
Fo# the !ai"#atio$ o% the po#ta"e )as !h#omato)#aph 'as !hose$ the optio$
o% !o$$e!ti$) it th#o&)h a %o' #e)&ato#4 to a p#ess&#i7ed !2i$de# that !o$tai$s a
mi3t&#e o% !ai"#atio$ )ases (i$ the !ase o% this a#ti!e 'as &sed %o# the !ai"#atio$ a
!2i$de# =TE_ !o$tai$i$) "e$7e$e4 eth2 "e$7e$e4 to&e$e4 m032e$e4 o032e$e4 p0
32e$e 'ith a !o$!e$t#atio$ o% 1+ ppm ea!h*.
The see!tio$ o% a o$)e# s&!tio$ i$te#<a %o# the !ai"#atio$ )as is #eE&i#ed4 to
ao' the p&#)i$) o% the 'hoe #o&te "et'ee$ the )as !h#omato)#aph a$d the
!ai"#atio$ !2i$de# "e%o#e the sta#t o% the !ai"#atio$. I$ o#de# to !ai"#ate the
!h#omato)#aph4 the !ai"#atio$ !2i$de# is ope$ed a$d the <a<e o% the %o' #e)&ato#
is so'2 ope$ed. D&#i$) the s&!tio$ o% the !ai"#atio$ )as th#o&)h the p&mp4 the
i$di!ato# o% the %o' #e)&ato# 'i "e mai$tai$ed at a "aa$!e <a&e %o&$d "et'ee$ the
ma3im&m a#ea (#ed !oo#ed a#ea* a$d the mi$im&m o$e o% the %o'4 this ao'i$) the
e$s&#i$) o% a p#ope# %o' %o# the !ai"#atio$ )as. A%te# the i$t#od&!tio$ o% the
!ai"#atio$ )as i$ the !h#omato)#aph4 the !ai"#atio$ !2i$de# is !osed a$d the$ the
%o' #e)&ato# is !osed.
Bhe$ the !h#omato)#aph has to "e !ai"#ated4 the desi#ed !o&m$ is see!ted4
set as %oo's: ENTER MENU0SETU80CONFIG0COLUMN. =2 p#essi$) i$ this
mome$t the /e2 ENTER MENU is a!ti<ated the !o&m$ A o# = o# C o$ 'hi!h the
a$a2sis 'i "e !a##ied o$4 the possi"iit2 o% see!ti$) o$e dete!to# o# "oth dete!to#s i$
the same time "ei$) a<aia"e (the 8ID dete!to# o# .a$d ECD*. I$ this mome$t a#e
see!ted the 8ID o# . a$d the ECD i"#a#ies depe$di$) o$ the !hose$ dete!to#s.
Foo'i$) the s!heme ENTER MENU M LI=RARY M LIST a$d &si$) the e%t0#i)ht
a##o's is see!ted the i"#a#2 !o##espo$di$) to the see!ted !o&m$. Fo# the a!ti<atio$
15
o# the dea!ti<atio$ o% a !ompo&$d4 is %a))ed the $ame o% the !ompo&$d a$d a#e
p#essed the /e2s "eo' EDIT M CALCM8D. I% a !ompo&$d is $ot see!ted4 it is
a&tomati!a2 #emo<ed %#om the i"#a#2. Fo# the e$te#i$) o% the !o$!e$t#atio$ o% a
!e#tai$ !ompo&$d4 a#e p#essed the /e2s "eo' EDIT M CALCONC. =2 &si$) the
a##o' /e2s a#e e$te#ed the !o$!e$t#atio$s o% the !ompo&$ds %#om the !ai"#atio$ )as.
The !ai"#atio$ )as m&st !o$tai$ a the !ompo&$ds that 'e#e a!ti<ated i$ the see!ted
i"#a#2. =2 p#essi$) the /e2 START.STO8 is sta#ted the a$a2sis4 the$ a#e p#essed the
/e2s "eo' DIS8LAY M GC M CM8D to <ie' the ist o% the !ompo&$ds4 a$d at the
%i$ishi$) o% the a$a2sis it 'i "e !he!/ed i% a the !ai"#atio$ !ompo&$ds a#e p#ese$t
i$ the ist. =2 p#essi$) ENTER MENU M LI=RARY0 CALL is a!!epted the ast
e3e!&ted a$a2sis as "ei$) p#ope# %o# the !ai"#atio$ o% the po#ta"e )as
!h#omato)#aph.
1D
I.1.. SEPARATION AND DETERMINATION OF
TRIHALOMETHANES FROM )ATERS BY
9ASCHROMATO9RAPHY
Intr$!"cti$n
The t#ihaometha$es (CHC
6
4 CHC
-
=#4 CHC=#
-
4 CH=#
6
* a#e %o#med i$ 'ate#
d&#i$) the !ho#i$atio$ p#o!ess. Se<e#a %o#matio$ me!ha$isms a#e possi"e4 %#om
'hi!h 'e me$tio$ the o$e sta#ti$) %#om po2phe$oi! de)#adatio$ p#od&!ts o% the
%&<i! a!id %#om the !ompositio$ o% the $at&#a h&m&s s&"sta$!es.
The !o$!e$t#atio$ o% t#ihaometha$es i$ 'ate# is mo$ito#ed d&e to the #oe
att#i"&ted to them i$ the !a#!i$o)e$esis p#o!esses4 the ma3im&m ao'ed
!o$!e$t#atio$ "ei$) 1++ ).L (a$d #espe!ti<e2 6+ ).L CHC
6
*.
)$r8ing Pr$ce!"re
Preparing the filling of the chro"atographic colu"n
The statio$a#2 phase &sed is the sE&aa$ (14541+41141A4-60he3ameth2e$
tet#a!osa$*4 ha<i$) the "oii$) poi$t at 6D1 WC a$d the de$sit2 o% +.>+1 ).!m
6
.
CH
CH
6
CH
6
(CH
-
*
6
CH
CH
6
(CH
-
*
6
CH
CH
6
(CH
-
*
,
CH
CH
6
(CH
-
*
6
CH
CH
6
(CH
-
*
6
CH
CH
6
CH
6
The i$e#t s&"st#ate is Ch#omoso#" B4 AB0DMCS 'ith )#a$&e dime$sio$s
>+.1++ mesh. The Ch#omoso#" B is a diatomite ea#th4 that !o$tai$s sii!i! a!id i$ the
h2d#ated amo#pho&s phase4 'ith a po#o&s st#&!t&#e4 the#ma2 t#eated at A++ WC 'ith a
sma E&a$tit2 o% Na
-
CO
6
(6 ;* a$d has a spe!i%i! s&#%a!e o% 1 M 6.1 m
-
.). The AB
i$de3 desi)$ates the p#e<io&s a!id 'ashi$) o% the !omme#!ia2 dei<e#ed p#od&!t.
The i$de3 DMCS desi)$ates the modi%i!atio$ o% the s&#%a!e 'ith dimeth2!ho#osia$e
as i$ the s!heme %#om "eo'.
The !o$!e$t#atio$ o% the statio$a#2 phase o$ the mi3 s&ppo#t is o% 1+ ;.
1>
Fo# the p#epa#atio$ o% 1+ ) o% %ii$) the %oo'i$) p#o!ed&#e is empo2ed:
0 1 ) sE&aa$ is disso<ed i$ 11+ mL $0he3a$e o% !h#omato)#aphi! p&#it2
0 A.+++ ) o% Ch#omoso#" B4 AB0DMCS a#e 'ei)hed 'ith a p#e!isio$ o% [1 m)4
0 the sE&aa$ so&tio$ is po&#ed o<e# the Ch#omoso#" a$d the mi3t&#e is
homo)e$i7ed 'ith the aid o% a ma)$eti! sti##e# %o# 11 mi$&tes
0 the $0he3a$e is so'2 e<apo#ated4 p#e%e#a"2 i$ a distiatio$ i$staatio$ 'ith
#otati$) sti (the heati$) is do$e to at the most >10A1 WC*
0 the %ii$) p#epa#ed i$ this ma$$e# is e%t %o# -, ho&#s i$ a 8et#i dish.
A !h#omato)#aphi! !o&m$ %#om )ass 'ith the i$te#$a diamete# o% 6.1 mm
a$d the e$)th o% 1., m is &sed. The %ii$) o% the !o&m$ is do$e i$ the %oo'i$)
ma$$e#:
0 o$e o% the e$ds o% the !o&m$ is "o!/ed 'ith a )ass 'addi$) p&)
0 that e$d o% the !o&m$ is !o$$e!ted th#o&)h a #&""e# hose to a <a!&&m p&mp
0 th#o&)h a %&$$e o% p#ope# dime$sio$s the %ii$) is i$t#od&!ed i$ the !o&m$ i$
sma po#tio$s. A%te# ea!h additio$4 the 'as o% the !o&m$ a#e tapped 'ith a
'oode$ #od to o"tai$ a &$i%o#m setti$) o% the %ii$).
Para#eter '$r t*e Separati$n
0 Mo"ie phase: N
-
0 Statio$a#2 phase: 1+ ; sE&aa$ o$ Chemoso#" B4 AB0DMCS
0 Co&m$: i$te#$a diamete# 6.1 mm4 e$)th 1., m
0 N
-
p#ess&#e th#o&)h the dete!to#: +.6 atm
0 E$t#2 p#ess&#e o% the mo"ie phase i$ the !o&m$: +.> atm
0 Dete!to#: %ame io$i7atio$ t2pe
0 Tempe#at&#e o% the !o&m$: D+ WC
0 Tempe#at&#e o% the dete!to#: -1+ WC
0 Tempe#at&#e o% the i$Le!to#: 1-+ WC
0 Vo&me o% the i$Le!ted sampe: 6 L
0 Sta$da#ds &sed: CHC
6
4 CH=#
6
4 CHC=#
-
o% !h#omato)#aphi! p&#it2.
1A
Si O Si
O
H
-
O4 H
?
O Si Si
OH OH
Si
CH
6
CH
6
C C -
?
Si
Si CH
6
CH
6
C
Si
Si CH
6
CH
6
C
O
Met*$! '$r t*e Preparing $' t*e Sa#ple an! Stan!ar!
a* The method %o# the p#epa#i$) o% the sta$da#ds is:
0 I$ a <o&met#i! !ai"#ated %as/ o% 1+ mL4 'ith a$ impe#<io&s p&)4 a#e 'ei)hed at
the a$a2ti!a "aa$!e E&a$tities o% sta$da#d s&"sta$!e "et'ee$ -+ a$d A+ m). The
%as/ is %ied to the ma#/ 'ith metha$o. The so&#!e so&tio$ o"tai$ed i$ this
ma$$e# /ept at ? , WC !a$ "e &sed %o# t'o 'ee/s.
0 Bo#/i$) #e%e#e$!e so&tio$ o"tai$ed "2 the di&tio$ 1.-1 o% the so&#!e so&tio$
'ith metha$o. I$ this ma$$e# a#e o"tai$ed !o$!e$t#atio$s o% t#ihaometha$es at
the o#de# o% te$s o% ).L.
0 I$ a <esse 'ith the !apa!it2 o% 1+ mL4 ma$&%a!t&#ed %#om po2p#op2e$e a$d
ha<i$) the id p#o<ided 'ith a sii!o$e #&""e# sept&m4 a#e i$t#od&!ed , mL o%
distied 'ate#4 1 L o% 'o#/i$) #e%e#e$!e so&tio$ ('ith the aid o% a
mi!#os2#i$)e* a$d 1 mL pe$ta$e o% !h#omato)#aphi! p&#it2.
0 The <esse is sti##ed %o# , mi$&tes.
0 The phases a#e ao'ed to sepa#ate.
0 Bith the aid o% a mi!#os2#i$)e4 a pa#t o% the o#)a$i! a2e# is #emo<ed a$d i$Le!ted
i$ the !o&m$.
"* Method %o# the sepa#atio$ o% the t#ihaometha$es %#om 'ate#s:
0 , mL o% sampe a#e i$t#od&!ed i$ the p#e<io&s2 me$tio$ed e3t#a!tio$ <esse. 1
mL o% pe$ta$e o% !h#omato)#aphi! p&#it2 is added a$d the e3t#a!tio$ is !a##ied
o&tI 6 L a#e #emo<ed %#om the o#)a$i! a2e# a$d a#e i$Le!ted i$ the !o&m$.
0 I% the sampe !o$tai$s %#ee !ho#i$e a %e' )#a$&es o% sodi&m thios&%ate a#e
added4 a$d the$ the t#ihaometha$es a#e e3t#a!ted i$ pe$ta$e.
Pr$ceing t*e Re"lt
The aspe!t o% a !h#omato)#am is p#ese$ted i$ the $e3t %i)&#e:
Ch#omato)#am o"tai$ed at the sepa#atio$ o% the t#ihaometha$es %#om 'ate#s. 10
e3t#a!tio$ so<e$t (pe$ta$e*I -0 CHC
6
I 60CHC
-
=#I ,0CHC=#
-
I CH=#
6
.
5+
A
0U? ((@ n#1
ti#p 0#in1
+ 1 1+ 11
1
-
6
,
1
A
0U? ((@ n#1
ti#e 0#in1
+ 1 1+ 11
1
-
6
,
1
A
0U? ((@ n#1
ti#p 0#in1
+ 1 1+ 11
1
-
6
,
1
A
0U? ((@ n#1
ti#e 0#in1
+ 1 1+ 11
1
-
6
,
1
The pea/s a#eas a#e app#e!iated %#om the !h#omato)#am o% the #e%e#e$!e
sampe. The %oo'i$) <a&es a#e o"tai$ed:
e
CH=#
e
CHC=#
e
=# CHC
e
CHC
6 - - 6
A I A I A I A
The pea/s a#eas a#e app#e!iated %#om the !h#omato)#am o% the &$/$o'$
sampe. The %oo'i$) <a&es a#e o"tai$ed:
3
CH=#
3
CHC=#
3
=# CHC
3
CHC
6 - - 6
A I A I A I A
The a#ea o% a pea/ is o"tai$ed "2 m&tip2i$) the hei)ht o% the pea/ 'ith its
'idth at +.5+D %#om the hei)ht.
The &$/$o'$ !o$!e$t#atio$s a#e %o&$d %#om the %o#m&as:
e
THM
e
THM
3
THM
3
THM
e
THM
e
THM
A
! A
.......... .......... A
.........! .......... A
= .
.
'he#e: THM M t#ihaometha$e
e
THM
! M !o$!e$t#atio$ o% the #e%e#e$!e i$ the #espe!ti<e t#ihaometha$e
51
I.1/. DETERMINATION OF THE PHENOLIC COMPOUNDS IN
)ATER BY HI9H PERFORMANCE LI-UID
CHROMATO9RAPHY

Maria7Critina BRATU
Intr$!"cti$n
The phe$o a$d s&"stit&ted phe$os #ep#ese$t !ommo$ "20p#od&!ts i$ ma$2
i$d&st#ia p#o!esses. At !o$!e$t#atio$s o% the o#de# o% pp"4 i$ e$<i#o$me$ta
!o$ditio$s depe$di$) o$ tempe#at&#e a$d pH4 these !ompo&$ds ma2 pe#sist %o# se<e#a
da2s o# 'ee/s. It 'as demo$st#ated that the phe$os a#e to3i! %o# most aE&ati!
o#)a$isms a$d4 e<e$ i$ o' !o$!e$t#atio$s4 the2 ha<e a "ad i$%&e$!e o$ the taste a$d
sme o% 'ate#. =e!a&se o% this4 i$ the !ase 'he#e di%%e#e$t t2pes o% 'ate#s 'i "e
&sed as 'ate# so&#!es %o# h&ma$ !o$s&mptio$4 is imposed the !o$tami$atio$ !he!/ %o#
these o#)a$i! po&ta$ts. I$ the E&#opea$ U$io$4 the ma3im&m ao'ed !o$!e$t#atio$
%o# di%%e#e$t phe$os i$ the d#i$/i$) 'ate# is e3t#eme2 sma. I$ this sit&atio$4 is
imposed the &se o% a$ a$a2ti!a method that 'i e$s&#e a hi)h see!ti<it2 a$d
se$siti<it2.
I$ this 'o#/ is p#ese$ted a method %o# the dete#mi$atio$ o% phe$os %#om
'ate# "2 &si$) a p#e0!o$!e$t#atio$ p#o!ed&#e "2 see!ti<e e3t#a!tio$ o% these
!ompo&$ds %#om 'ate# o$ adso#"e$t !a#t#id)es a$d dete#mi$atio$ o% the e3t#a!ted
phe$os %#om the !a#t#id)e "2 hi)h pe#%o#ma$!e iE&id !h#omato)#aph2 'ith UV
dete!tio$.
0 A!eto$it#ie o% H8LC )#ade (0ie!el !e 1aen*I 'ate# o% H8LC )#ade (/lu$a*4
metha$o o% H8LC )#ade %/lu$a*4 a!eti! a!id (Al!rich*.
0 Ca#t#id)e S8E %o# p#e0!o$!e$t#atio$: S&pe!ea$ ENVI0Ch#om 8 %Supelco",
!opo2me# st2#e$e0di<i$2 "e$7e$e (the pa#ti!e si7e is: >+015+ CmI the po#e
diamete#s a#e: 11+01D1 a*.
0 Sta$da#d o% phe$os: E8A 5+,0M 8he$os Mi3 %Supelco", %o#med %#om a mi3t&#e
p#epa#ed i$ metha$o o% 11 a$a2tes4 at di%%e#e$t !o$!e$t#atio$s: phe$o (1++
C).mL*I ,0$it#ophe$o (-1++ C).mL*I -4,0di$it#ophe$o (11++ C).mL*I -0
!ho#ophe$o (1++ C).mL*I -0$it#ophe$o (1++ C).mL*I -4,0dimeth2phe$o (1++
C).mL*I ,450di$it#o0o0!#eso (-1++ C).mL*I -4,0di!ho#ophe$o (1++ C).mL*I ,0
!ho#o0m0!#eso (-1++ C).mL*I -4,450t#i!ho#ophe$o (11++ C).mL*I
pe$ta!ho#ophe$o (-1++ C).mL*.
0 Sta$da#d o% phe$os: E8A 5+,0S 8he$os Mi3 %Supelco" , %o#med %#om 11
i$di<id&a phe$o so&tio$s4 'ith the !o$!e$t#atio$ o% 1++ m).L p#epa#ed i$
5-
metha$o4 'ith the <o&me o% 1 mL4 phe$oI ,0$it#ophe$oI -4,0di$it#ophe$oI -0
!ho#ophe$oI -0$it#ophe$oI -4,0dimeth2phe$oI ,450di$it#o0o0!#esoI -4,0
di!ho#ophe$oI ,0!ho#o0m0!#esoI -4,450t#i!ho#ophe$oI pe$ta!ho#ophe$o* a$d
a <ia 'ith a mi3t&#e o% a the a"o<e me$tio$ed !ompo&$ds.
0 Soid phase e3t#a!tio$ s2stem t2pe Va!&&m Ma$i%od FS&pe!oG 'ith 1- positio$s.
0 LiE&id t#ap t2pe S&pe!o S8E Va!&&m 8&mp T#ap Rit.
0 Va!&&m p&mp Heidoph.
The dete#mi$atio$s 'e#e made &si$) a hi)h pe#%o#ma$!e iE&id
!h#omato)#aph F8e#/i$ Eme#G se#ies -++4 eE&ipped 'ith:
0 E&ate#$a#2 p&mpI <a!&&m de)asse# %o# so<e$tsI
0 a&to0sampe# (%o# the a&tomated i$Le!tio$ i$ the !h#omato)#aphi! !o&m$ o% a
p#e<io&s2 see!ted E&a$tit2 o% sampe a$d %o# the a&tomated 'ashi$) o% the
s2#i$)e*I
0 !o&m$ o<e$ %o# the#mostati$) the !h#omato)#aphi! !o&m$I
0 UV0VIS dete!to# t2pe FDiode A##a2G 'ith the possi"iit2 o% sim&ta$eo&s spe!t#a
a!E&isitio$I
0 !omp&te# (so%t'a#e*.
)$r8ing Pr$ce!"re
Sa"ple preparation
The 'ate# sampe is a!idi%ied to a pHH- to p#e<e$t the io$i7atio$ o% the
phe$o !ompo&$ds.
/he preconcentration of the sa"ple on a SP0 cartridge
The !a#t#id)e is a!ti<ated "2 passi$) th#o&)h it 5 mL o% metha$o at a %o' #ate o%
1 mL.mi$.
The !a#t#id)e is !o$ditio$ed "2 passi$) th#o&)h it 5 mL o% a!idi%ied 'ate# (pHH-*
at a %o' #ate o% 1 mL.mi$.
Th#o&)h the !a#t#id)e a#e passed at a %o' #ate o% -.1 mL.mi$4 1++ mL o% sampe
pa!ed i$ a$ additio$a <esse.
The e&tio$ %#om the !a#t#id)e is do$e 'ith - mL o% so<e$t mi3t&#e metha$o.
a!eti! a!id 1;4 at a %o' #ate o% 1 mL.mi$.
A%te# this the e&ate is a$a27ed !h#omato)#aphi!a2 i$ o#de# to sepa#ate the
!ompo$e$ts a$d dete#mi$e them.
Chro"atographic anal%sis
I$ the <esses (<ias* o% the a&tomated sampe !ha$)e# (a&to0sampe#* a#e
i$t#od&!ed the sta$da#ds a$d sampes to "e a$a27ed (a"o&t - mL*..
The s2stem H8LC F 8e#/i$ Eme# F se#ies -++ 'i "e &sed.
A !o&m$ 'ith #e<e#sed phase F=#o'$eeG t2pe 8e!osphe#e C 1>4 is &sed4
ha<i$) the po#e diamete# o% 6 Um4 the e$)th o% the !o&m$ "ei$) >6 mm a$d the
diamete# ,.5 mm.
56
A$ e&tio$ 'ith i$ea# )#adie$t o% the mo"ie phase 'i "e &sed: %#om 61 ;
a!eto$it#ie.'ate# (pHH-.-* &p to 1++ ; a!eto$it#ie4 o<e# a$ i$te#<a o% , mi$&tes.
The mo"ie phase %o's th#o&)h the !h#omato)#aphi! !o&m$ 'ith a %o' #ate
o% -.1 mL.mi$. The !h#omato)#aphi! a$a2sis do$e i$ these !o$ditio$s ta/es %i<e
mi$&tes. The e&tio$ o#de# %#om the !h#omato)#aphi! !o&m$ is the %oo'i$): phe$oI
,0$it#ophe$oI -0!ho#ophe$oI -4,0di$it#ophe$oI -0$it#ophe$oI -4,0dimeth2phe$oI
,0!ho#o0m0!#esoI -4,0di!ho#ophe$oI ,450di$it#o0o0!#esoI -4,450t#i!ho#ophe$oI
pe$ta!ho#ophe$o.
I$ the !ase o% 'ate# sampes 'ith a mo#e !ompe3 !ompositio$4 %o# a "ette#
sepa#atio$ o% the phe$oi! !ompo&$ds4 !a$ "e &sed a$othe# !h#omato)#aphi! !o&m$:
t2pe S&pe!osi LC0>4 'ith the dime$sio$s 11 !m e$)th 3 ,.5 mm. i.d. 'ith the
pa#ti!e diamete# o% 1 Cm. The same e&tio$ 'ith i$ea# )#adie$t o% the mo"ie phase
is &sed (A: metha$o.1; a!eti! a!idI =: H
-
O.1 ; a!eti! a!id* as %oo's: 101++ ; A ?
A1 M +; = i$ 6+ mi$&tes. The mo"ie phase %o's th#o&)h the !h#omato)#aphi!
!o&m$ 'ith a %o' #ate o% 1 mL.mi$. The a$a2sis time to sepa#ate the 11 phe$oi!
!ompo&$ds is 11 mi$&tes4 the e&tio$ o#de# %#om the !h#omato)#aphi! !o&m$ is the
%oo'i$): phe$oI ,0$it#ophe$oI -4,0di$it#ophe$oI -0!ho#ophe$oI -0$it#ophe$oI
-4,0dimeth2phe$oI ,450di$it#o0o0!#esoI -4,0di!ho#ophe$oI ,0!ho#o0m0!#esoI
-4,450t#i!ho#ophe$oI pe$ta!ho#ophe$o.
I$ "oth sit&atio$s4 the tempe#at&#e o% the !o&m$ is set at 6+ WC4 the
meas&#eme$t "ei$) do$e 'ith a UV0VIS dete!to# t2pe D2ode A##a2 'ith
sim&ta$eo&s spe!t#a a!E&isitio$4 the phe$oi! !ompo&$ds "ei$) sim&ta$eo&s2
mo$ito#ed at t'o 'a<ee$)ths: -1, $m a$d ->+ $m.
I$ o#de# to esta"ish the #ete$tio$ time !o##espo$di$) to ea!h phe$oi!
!ompo&$d %#om the mi3t&#e is &sed the phe$o sta$da#d: E8A 5+,0S 8he$os Rit
%Supelco" , %o#med %#om 11 i$di<id&a phe$o so&tio$s4 'ith the !o$!e$t#atio$ o% 1++
m).L. Ea!h phe$oi! !ompo&$d is di&ted 1:1+ 'ith metha$o. =2 i$Le!tio$ i$ the
!h#omato)#aphi! s2stem o% 5+ CL %#om ea!h di&ted phe$oi! !ompo&$d so&tio$
!o##espo$di$) to +.5 C)4 is dete#mi$ed the !o##espo$di$) #ete$tio$ time %o# ea!h
!ompo&$d. As a #es&t4 the ide$ti%i!atio$ o% the phe$oi! !ompo&$ds sepa#ated %#om
the a$a27ed sampe 'i "e do$e a&tomati!a2 th#o&)h the so%t'a#e o% the
!h#omato)#aph4 "2 !ompa#i$) 'ith the #ete$tio$ times o% the sta$da#ds p#e<io&s2
i$t#od&!ed i$ the H8LC s2stem.
The E&a$titati<e a$a2sis o% the phe$oi! !ompo&$ds is do$e "2 the
i$te#poatio$ o% the a#ea o% the !h#omato)#aphi! pea/ !o##espo$di$) %o# ea!h sepa#ate
!ompo&$d4 o$ the !ai"#atio$ !&#<e o"tai$ed &si$) mi3t&#es o% di%%e#e$t
!o$!e$t#atio$s o% sta$da#d so&tio$s o% phe$oi! !ompo&$ds p#epa#ed i$ metha$o
(<.<*.
To e$s&#e the #ep#od&!i"iit2 o% the #ete$tio$ times !o##espo$di$) to the
phe$oi! !ompo&$ds4 a$d %o# a hi)h p#e!isio$ o% the dete#mi$atio$s4 "e%o#e the sta#t o%
the !h#omato)#aphi! a$a2ses4 is #eE&i#ed a$ i$te#<a %o# the sta"ii7atio$ o% the
!h#omato)#aphi! s2stem o% at east o$e ho&#.
5,
/a$ing the calibration curve
The !ai"#atio$ !&#<e is t#a!ed i$ %o&# poi$ts. Fo# this4 the phe$o mi3t&#e
sta$da#d E8A 5+,0M FS&pe!oG is di&ted to a #atio o% 1:-1 (<.<*4 1:1+ (<.<*4 1:1++
(<.<*4 #espe!ti<e2 1:1-1 (<.<*. The !o$!e$t#atio$s !o##espo$di$) to the phe$oi!
!ompo&$ds mi3t&#e %#om the %o&# o"tai$ed so&tio$s4 e3p#essed i$ m).L a#e p#ese$ted
i$ ta"e A. The E&a$tities o% phe$o e3p#essed i$ C) %#om the sta$da#d so&tio$s
p#epa#ed %o# the t#a!i$) o% the !ai"#atio$ !&#<e a#e p#ese$ted i$ ta"e =.
I$ %o&# <esses o% the a&tosampe# o% the !h#omato)#aph a#e i$t#od&!ed
app#o3imate2 - mL %#om ea!h o% the %o&# so&tio$s4 %#om 'hi!h 'i "e a&tomati!a2
i$Le!ted 5+ CL.
Ta%le A. Co$!e$t#atio$s (e3p#essed i$ m).L* o% the phe$oi! !ompo&$ds &sed %o# the
t#a!i$) o% the !ai"#atio$ !&#<e.
P*en$lic c$#p.
in t*e tan!ar!
EPA [email protected]
C$nc. $'
p*en$lic
c$#p. in t*e
tan!ar!
0S"pelc$1
C$nc. $'
p*en$lic
c$#p. in t*e
tan!ar!
0S"pelc$1
!il"te! ':12
<it*
#et*an$l
C$nc. $'
p*en$lic
c$#p. in t*e
tan!ar!
0S"pelc$1
!il"te! ':2(
<it*
#et*an$l
C$nc. $'
p*en$lic
c$#p. in t*e
tan!ar!
0S"pelc$1
!il"te! ':'((
<it*
#et*an$l
C$nc. $'
p*en$lic
c$#p. in t*e
tan!ar!
0S"pelc$1
!il"te! ':'12
<it*
#et*an$l
8he$o 1++ m).L -+ m).L 1+ m).L 1 m).L , m).L
,0Nit#ophe$o -1++ m).L 1++ m).L 1+ m).L -1 m).L -+ m).L
-4,0
Di$it#ophe$o
11++ m).L 5+ m).L 6+ m).L 11 m).L 1- m).L
-0Cho#ophe$o 1++ m).L -+ m).L 1+ m).L 1 m).L , m).L
-0Nit#ophe$o 1++ m).L -+ m).L 1+ m).L 1 m).L , m).L
-4,0
Dimetiphe$o
1++ m).L -+ m).L 1+ m).L 1 m).L , m).L
,450Di$it#o0o0
!#eso
-1++ m).L 1++ m).L 1+ m).L -1 m).L -+ m).L
-4,0
Di!ho#ophe$o
1++ m).L -+ m).L 1+ m).L 1 m).L , m).L
,0Cho#o0m0
!#eso
-1++ m).L 1++ m).L 1+ m).L -1 m).L -+ m).L
-4,450
T#i!ho#ophe$o
11++ m).L 5+ m).L 6+ m).L 11 m).L 1- m).L
8e$ta!ho#ophe
$o
-1++ m).L 1++ m).L 1+ m).L -1 m).L -+ m).L
51
Ta%le B. Amo&$ts o% phe$os e3p#essed i$ C) %#om sta$da#d so&tio$s p#epa#ed %o#
the !ai"#atio$ )#aph (i$Le!ted <o&me H 5+ CL*
P*en$lic c$#p.
in t*e tan!ar!
C$nc. $'
p*en$lic
c$#p. in t*e
tan!ar!
0S"pelc$1
C$nc. $'
p*en$lic
c$#p. in t*e
tan!ar!
0S"pelc$1
!il"te! ':12
<it*
#et*an$l
C$nc. $'
p*en$lic
c$#p. in t*e
tan!ar!
0S"pelc$1
!il"te! ':2(
<it*
#et*an$l
C$nc. $'
p*en$lic
c$#p. in t*e
tan!ar!
0S"pelc$1
!il"te! ':'((
<it*
#et*an$l
C$nc. $'
p*en$lic
c$#p. in t*e
tan!ar!
0S"pelc$1
!il"te! ':'12
<it*
#et*an$l
8he$o 1++ m).L 1.- C) +.5 C) +.6 C) +.-, C)
,0Nit#ophe$o -1++ m).L 5.+ C) 6.+ C) 1.1 C) 1.- C)
-4,0
Di$it#ophe$o
11++ m).L 6.5 C) 1.> C) +.A C) +.D- C)
-0Cho#ophe$o 1++ m).L 1.- C) +.5 C) +.6 C) +.-, C)
-0Nit#ophe$o 1++ m).L 1.- C) +.5 C) +.6 C) +.-, C)
-4,0
Dimetiphe$o
1++ m).L 1.- C) +.5 C) +.6 C) +.-, C)
,450Di$it#o0o0
!#eso
-1++ m).L 5.+ C) 6.+ C) 1.1 C) 1.- C)
-4,0
Di!ho#ophe$o
1++ m).L 1.- C) +.5 C) +.6 C) +.-, C)
,0Cho#o0m0
!#eso
-1++ m).L 5.+ C) 6.+ C) 1.1 C) 1.- C)
-4,450
T#i!ho#ophe$o
11++ m).L 6.5 C) 1.> C) +.A C) +.D- C)
8e$ta!ho#ophe
$o
-1++ m).L 5.+ C) 6.+ C) 1.1 C) 1.- C)
A%te# the o"tai$i$) o% the !h#omato)#ams !o##espo$di$) to ea!h i$Le!ted
phe$o mi3t&#e4 the appa#at&s ide$ti%ies the !ompo&$d "ased o$ the #eside$!e time
a$d a&tomati!a2 i$te)#ates the a#eas !o##espo$di$) to ea!h ma3im&m. The %o&#
!h#omato)#ams o"tai$ed a#e !o$side#ed !ai"#atio$ sta$da#ds4 a$d a)ai$ i$ a$
a&tomated mode the so%t'a#e o% the appa#at&s )e$e#ates the !ai"#atio$ !&#<es %o#
ea!h !ompo&$d at the %o&# !o$!e$t#atio$s. Di%%e#e$t t2pes o% !ai"#atio$ !&#<es ma2
"e o"tai$ed. The most pop&a# (aso !hose$ i$ o&# !ase* is the I
st
o#de# i$e 'ith the
i$te#se!tio$ o% the o#i)i$. I% the F!ai"#atio$ !&#<eG $eeds to "e e3te$ded to hi)he#
!o$!e$t#atio$ <a&es it is #e!omme$ded the &se o% a II
$d
o#de# eE&atio$ )i<e$ that the
ma3im&m i$ea#it2 de<iatio$ 'i $ot e3!eed -1 ;.
It !a$ "e $oti!ed that "2 the adso#ptio$ o$ the !a#t#id)e o% 1++ mL o% sampe
!o$tami$ated 'ith phe$oi! !ompo&$ds4 %oo'ed "2 the e&tio$ 'ith - mL o% mi3t&#e
metha$o.a!eti! a!id 1;4 most phe$oi! !ompo&$ds !a$ "e easi2 dete#mi$ed at
!o$!e$t#atio$s "eo' the <a&e o% +.1 pp" &si$) the UV dete!tio$.
55
I.12. DETERMINATION OF ALDEHYDES AND DETONES IN
AMBIENT AIR
Adeh2des a$d /eto$es samped as -.,0di$it#ophe$2h2d#a7o$es i$
imp#e)$ated t&"es !a$ "e a$a2sed i$ e3t#a!ts 'ith hi)h pe#%o#ma$!e iE&id
!h#omato)#aph2 (H8LC*.
Reagents
0 A!eto$it#ie4 H8LC0E&ait24 Rath"&#$ Chemi!as Ltd.4 No. RH 1+15.
0 Metha$o4 Me#!/.
0 Tet#ah2d#o%&#a$4 Me#!/.
0 Bate#4 E&a#t7 distied a$d io$ e3!ha$)ed %#om a Miipo#e FMiiKG 'ate#
p&#i%i!atio$ s2stem.
0 8otassi&m iodide4 p.a4 F&/a.
0 S&%&#i! a!id4 Me#!/
0 Etha$o.
0 Ca#"o$2!ompo&$ds $eeded.
0 -4,0Di$it#ophe$2h2d#a7i$e4 F&/a.
-nstru"entation
The %oo'i$) i$st#&me$ts a$d eE&ipme$t ma2 "e &sed:
0 A He'ett08a!/a#d 1+1+ mod&a# s2stem4 !o$sisti$) o% a DA>1-A E&ate#$a#2
so<e$t s&pp2 s2stem4 a G16+5A diode a##a2 dete!to#4 a$d a$ a&tosampe#.
0 No<a08a/ C
1>
4 , Cm pa#ti!es (No. >56,,4 Bate#s Asso!iates*4 11+ mm X 6.A mm
i.d.*.
0 Bate#s i$0i$e 8#e!o&m$ Fite#4
0 S2#i$)es a$d <o&met#i! )ass'a#e.

Fi a 1 mL s2#i$)e 'ith a!eto$it#ie. (Coe!t the sampe e3t#a!t i$ a 6 mL
$a##o' $e!/ %as/*. E&ate the de#i<ati<es "2 so'2 (app#o3imate 1.1 mL.mi$*
p&shi$) a!eto$it#ie th#o&)h the !a#t#id)e. Stop the e&tio$ 'he$ the 6 mL ma#/ is
#ea!hed. T#a$s%e# app#o3imate2 +.1 mL o% the sampe so&tio$ to a - mL a&tosampe#
<ia a$d sea the <ia. The sampe is $o' #ead2 %o# H8LC a$a2sis.
1+ CL o% a sampe o# sta$da#d so&tio$ is sepa#ated "2 &si$) a E&ate#$a#2
mi3t&#e o% metha$o.'ate#.a!eto$it#ie.tet#ah2d#o%&#a$. The %oo'i$) ta"e sho's the
)#adie$t p#o%ie 'hi!h is &sed.
5D
K&ate#$a#2 )#adie$t 'hi!h sepa#ates the !a#"o$2h2d#a7o$es o% i$te#est (EME8* at a
%o' #ate o% +.> mL.mi$.
Ti#e G Tetra*&!r$'"ran G Acet$nitrile G )ater G Met*an$l
+.+ 1>.+ --.+ 5+.+ +.+
+.1 1>.+ --.+ 5+.+ +.+
-+.+ >., 6D., 1,.- +.+
-,.+ +.+ +.+ 6,.+ 55.+
,+.+ +.+ +.+ 11.+ >1.+
,1.+ +.+ +.+ 11.+ >1.+
,1.+ +.+ 1++.+ +.+ +.+
,>.+ 1>.+ --.+ 5+.+ +.+
The dete!tio$ a$d E&a$ti%i!atio$ is !a##ied o&t at 65A $m ("a$d 'idth -- $m*
&si$) ,D, $m ("a$d 'idth 1+ $m* as the #e%e#e$!e 'a<ee$)th. The dete!tio$ a$d
E&a$ti%i!atio$ o% di!a#"o$2s is !a##ied o&t at ,,+ $m ("a$d 'idth -- $m* &si$)
66D $m ("a$d 'idth 1+ $m*.
Foo'i$) !a#"o$2!ompo&$ds sho&d "e meas&#ed: metha$a4 etha$a4
p#ope$a4 p#opa$a4 p#opa$o$e4 -0meth20p#ope$a4 "&ta$a4 -0"&ta$o$e4 60"&te$0-0
o$e4 pe$ta$a4 he3a$a4 "e$7e$e!a#"adeh2de4 etha$dia4 o3op#opa$a.
Blan8
Ea!h da2 a$a2ses o% !a#"o$2!ompo&$ds a#e pe#%o#med4 a a"o#ato#2 "a$/
sho&d "e p#epa#ed. 8e#iodi!a2 %ied "a$/s sho&d "e o"tai$ed o$!e e<e#2 'ee/. The
"a$/ e<es o% metha$a4 etha$a4 a$d p#opa$o$e 'i p#o"a"2 !ha$)e 'ith !a#t#id)e
"at!h $&m"e# a$d the "at!h $&m"e# o% a!eto$it#ie. The "a$/ e<e sho&d $ot
e3!eed +.+1 C).m
6
o% the !a#"o$2 !ompo&$d i$ a sampe <o&me o% D1+ it#es.
Preparati$n $' H&!raH$ne
1. Disso<e ,++ m) o% -4,0di$it#ophe$2h2d#a7i$e i$ - mL A5; s&ph&#i! a!id. This
so&tio$ is the$ added4 6ith stirrin(, to 16 mL D1; etha$o. A$2 &$disso<ed
soid is #emo<ed "2 aid o% a 8aste&# pipette.
-. A <o&me !o##espo$di$) to 1++ m) o% the !a#"o$2!ompo&$d is t#a$s%e##ed to -+
mL etha$o.
5>
6. The !a#"o$2so&tio$ (step -* is tha$ t#a$s%e##ed to the DN8H so&tio$ (step 1*
'ith sti##i$). Let the so&tio$ sta$d %o# 11 mi$&tes to !ompete the #ea!tio$.
,. Fite# the so&tio$ i$ step 64 a$d #e!#2stai7e the h2d#a7o$e %#om aE&eo&s
etha$o. (H2d#a7o$es %#om &$sat&#ated.a#omati! !a#"o$2s sho&d "e
#e!#2stai7ed %#om aE&eo&s a!eto$it#ie.*
1. D#2 the h2d#a7o$e a$d do a p&#it2 test (H8LC0UV*.
Cali%rati$n
8#epa#e a sto!/ so&tio$ %#om ea!h !a#"o$2h2d#a7o$e "2 disso<i$)
app#o3imate2 1 m) (?.0 1;* i$ 1++ mL a!eto$it#ie. (These sto!/ so&tio$s 'i "e
#ead2 %o# &se.* Cai"#atio$ so&tio$s a#e p#epa#ed "2 di&tio$ o% the sto!/ so&tio$s
(1 C).mL to - C).mL is s&ita"e %o# most a$a2ses*.
-"anti'icati$n
The !o$!e$t#atio$ o% !a#"o$2!ompo&$ds i$ the ai# sampe e3p#essed as ).m
6
4
is )i<e$ "2:
C 0 Co$!e$t#atio$ o% the !a#"o$2 !ompo&$d i$ the ai# sampe SC).m
6
T
! 0 Co$!e$t#atio$ o% the !a#"o$2 !ompo&$d i$ the sta$da#d SC).mLT
H(s* 0 8ea/ hei)ht.a#ea o% the !a#"o$2 !ompo&$d i$ the sta$da#d S!o&$tsT
H(p* 0 8ea/ hei)ht.a#ea o% the !a#"o$2 !ompo&$d i$ the sampe S!o&$tsT
/ 0 Co$<e#sio$ %a!to# (e.). %#om h2d#a7o$ to !a#"o$2* metha$a:+.1,-A4
etha$a:+.1A5,4 p#ope$a:+.-6D6 et!.
V 0 Sampe <o&me Sm
6
T
< 0 Vo&me o% the p#epa#ed sampe SmLT
Inter'erence
Fai&#e to #emo<e o7o$e "2 the o7o$e s!#&""e# 'i #es&t i$ se#io&s &$de#0
estimati$) o% some !a#"o$2h2d#a7o$es.
Re'erence
1. Vai#a<am&#th24 A.4 Ro"e#ts4 N.M. a$d Ne'ma$4 L. (1AA-* Methods %o#
dete#mi$atio$ o% o' moe!&a# 'ei)ht !a#"o$2 !ompo&$ds i$ the atmosphe#e: a
#e<ie'. Atmos. 4nviron., 2;A, 1A5101AA6.
-. Sem#4 N. (1AA1* Dete#mi$atio$ o% <oatie !a#"o$2 !ompo&$ds i$ !ea$ ai#.
/resenius J. Anal. Chem., 7904 5D-05DD.
5A
6. D2e4 C. a$d Oehme4 M. (1AA-*. Comme$ts !o$!e#$i$) the H8LC sepa#atio$ o%
a!#oei$ %#om othe# C
6
!a#"o$2 !ompo&$ds as -4,0di$it#ophe$2h2d#a7o$es:
a p#oposa %o# imp#o<eme$t. J. 1i(h 0es. Chrom., 154 10>.

D+
I.15. DETERMINATION OF ALACHLOR USIN9 DIRECT ELISA
Intr$!"cti$n
This method p#o<ides a E&a$titati<e e<a&atio$ o% aa!ho# %#om 'ate#
&tii7i$) ELISA (e$72me i$/ed imm&$oso#"e$t assa2* te!h$oo)2. Aa!ho# is a
G#o&p I !ompo&$d4 'ith ad<e#se a!&te heath e%%e!ts %#om e3pos&#e to o'
!o$!e$t#atio$s. The assa2 is simpe a$d adapta"e e$o&)h that it ma2 "e &sed o$ site
i$ the %ied as a s!#ee$i$) method. Di#e!t ELISA i$<o<es &s&a2 immo"ii7atio$ o%
a$ti"od2 o$ the 'e s&#%a!e "2 simpe adso#ptio$4 %oo'ed o% additio$ o% a$a2te
(aa!ho#* a$d a"eed a$a2te de#i<ati<e (aa!ho# de#i<ati<e a"eed 'ith pe#o3idase
(HR8**4 'hi!h 'i sta#t to !ompete %o# a$ti"od2 (a%%i$it2 p&#i%ied a$ti0aa!ho#4
de<eoped i$ sheep* a!ti<e sites. Usi$) a s&"st#ate mi3t&#e spe!i%i! %o# t#a!e# ma#/e#
(i$ o&# !ase the s&"st#ate4 e. ). 646V4141V0Tet#amet2"e$7idi$e a$d H
-
O
-
i$ 6+ mL
a!etate "&%%e# 1+ mM4 pH 1.- is spe!i%i! %o# HR8*4 the #ea!ted a$a2te !o$!e$t#atio$
!a$ "e e<a&ated i$di#e!t2 "2 dete#mi$i$) the !o$!e$t#atio$ o% "o&$d %#a!tio$ o%
t#a!e#. Spe!t#omet#i!a dete!tio$ (,1+ $m* ao's to ha<e a E&a$titati<e dete#mi$atio$
o% aa!ho# "2 di#e!t ELISA. The method is appied %o# aa!ho# dete#mi$atio$ %#om
'ate# sampes (e. ). d#i$/i$) 'ate#4 s&#%a!e 'ate#4 mi3ed domesti! a$d i$d&st#ia
'aste'ate#s4 )#o&$d'ate#4 #ea)e$t 'ate#s*.
-0S(-450dieth20phe$2*(metho32meth2*ami$oT0-0o3oetha$e s&%o$i! a!id
(ESA* !#oss #ea!ts 'ith aa!ho# i$ the ELISA. ESA is $ot dete!ted "2 sta$da#d H8LC
o# GC. Aa!ho# #eated a!eta$iides ma2 aso i$te#%e#e 'ith the dete#mi$atio$ o%
aa!ho#4 s&!h as a!eto!ho#4 metoa!ho#4 a$d metaa32. I$ o#de# to p#e<e$t e<e$t&a
e##o#s i$ a$a2sis4 the !#oss #ea!ti<it2 o% &sed a$ti"od2 %o# possi"e i$te#%e#e$!e has to
"e e<a&ated.
Intr"#ent
0 Mi!#o0pate 'ith A5 'es (NUNC4 =#a$d 8#od&!ts*.
0 U$i<e#sa mi!#o0pate #eade# (EL >++4 =io0te/ I$st#&me$ts INC.*.
0 A&tomati! m&ti0%i$)e# pipette (> positio$s*.
Sa#ple C$llecti$nE Preer+ati$n an! St$rage
Coe!t a sampes i$ ,+ mL "ottes i$to 'hi!h 6 m) o% sodi&m thios&%ate
!#2stas ha<e "ee$ added to the empt2 "ottes L&st p#io# to shippi$) to the sampi$)
site. Bhe$ sampi$) %#om a 'ate# tap4 ope$ the tap a$d ao' the s2stem to %&sh &$ti
the 'ate# tempe#at&#e has sta"ii7ed ()e$e#a2 o$e to t'o mi$&tes*. AdL&st the %o' to
a"o&t 1++ mL.mi$ a$d !oe!t the sampe %#om the %o'i$) st#eam. Reep sampe
seaed %#om !oe!tio$ time &$ti a$a2sis. Bhe$ sampi$) %#om a "od2 o% 'ate#4 %i
D1
the sampe !o$tai$e# 'ith 'ate# %#om a #ep#ese$tati<e a#ea. Sampi$) eE&ipme$t4
i$!&di$) a&tomati! sampe#s4 m&st $ot &se pasti! t&"i$)4 pasti! )as/ets4 o# a$2 pa#ts
that ma2 ea!h i$te#%e#i$) a$a2tes i$to the sampe. A&tomati! sampe#s that
!omposite sampes o<e# time sho&d &se #e%#i)e#ated )ass sampe !o$tai$e#s.
Resid&a !ho#i$e i$ the sampe sho&d "e #ed&!ed "2 addi$) 1+ m).L o%
sodi&m s&%ite (this ma2 "e added as a soid 'ith sti##i$) o# sha/i$) &$ti disso<ed4 o#
as a p#epa#ed so&tio$*.
AdL&st the sampe to pH - "2 addi$) 5 N HC. It ma2 #eE&i#e &p to , mL to
a!!ompish this. It is <e#2 impo#ta$t that the sampe "e de!ho#i$ated "e%o#e addi$)
the a!id to o'e# the pH o% the sampe. Addi$) sodi&m s&%ite a$d HC to the sampe
"ottes p#io# to shippi$) the "ottes to the sampi$) site is $ot pe#mitted. HC sho&d
"e added at the sampi$) site to #eta#d a$2 mi!#o"ioo)i!a de)#adatio$ o% method
a$a2tes. Sampes m&st "e i!ed o# #e%#i)e#ated at ,@C %#om the time o% !oe!tio$ &$ti
e3t#a!tio$. 8#ese#<atio$ st&d2 #es&ts sho' that the a$a2te is sta"e %o# 1, da2s i$
sampes that a#e p#ese#<ed it. Re%#i)e#ated sampe e3t#a!ts ma2 "e sto#ed &p to 6+
da2s.
Sa#ple Preparati$n
Aa!ho# is e3t#a!ted a$d !o$!e$t#ated %#om the sampe "2 S8E (soid phase
e3t#a!tio$* &si$) soid phase e3t#a!tio$ dis/4 -1 mm4 SD=03! (6M Empo#e
TM
4 o#
eE&i<ae$t*. Remo<e the SD= dis/ %#om the sampe# a$d pa!e i$ a )ass "otte 'ith a$
ID a#)e e$o&)h to ao' the %ite# to ie %at. Add -.+ mL o% pesti!ide %#ee metha$o to
the SD= dis/4 !ap4 a$d sha/e o$ a pat%o#m o#"ita sha/e# %o# 11 mi$&tes. T#a$s%e# the
metha$o so&tio$ to a , mL sampe <ia 'ith a 8TFE0i$ed !ap. Usi$) a positi<e
dispa!eme$t pipette o# s2#i$)e4 #emo<e a$ aiE&ot o% sampe a$d di&te it a mi$im&m
o% 1:1++ 'ith deio$i7ed 'ate#. B#ap #emai$de# o% the sampes 'ith pa#a%im a$d sto#e
#e%#i)e#ated.
Note: I% !o$!e$t#atio$s a#e hi)he# tha$ +.1 C).sampe4 %&#the# di&tio$s a#e
$eeded to meet the ta#)et #a$)e o% the assa2. M&tipe di&tio$s ma2 "e do$e i% the
!o$!e$t#atio$ #a$)e is &$/$o'$.
Anal&i
0.peri"ent ': Opti"isation of tracer and antibod% concentration &3Bi4
di"ensional5 0+-SA)
1. The pate is !o<e#ed 'ith 1++ CL o% a$ti"od2 so&tio$ (i.e.4 i$ the #a$)e 1+++ 0 +
m).L i$ 1+ mM !a#"o$ate "&%%e#4 pH A.54 o$ the !o&m$ 1 to 1- "2 s&!!essi<e
di&tio$*4 a$d the adso#ptio$ is ao'ed to p#o!eed o<e# $i)ht at ,C.
-. A%te# the #esid&a so&tio$ is #emo<ed4 the pate is 'ashed 'ith 8=ST (app#o3.
-++ CL . 'e*.
6. 1++ CL o% di%%e#e$t !o$!e$t#atio$s o% t#a!e# (i.e.4 i$ the #a$)e 1.1++ 0 + t#a!e#
di&tio$ i$ 8=ST* a#e added o$ the se!o$d dime$sio$ (e.).4 #o' A to H* "2
s&!!essi<e di&tio$. The i$!&"atio$ is ao'ed to p#o!eed %o# 1 ho&#s.
D-
Optimisatio$ o% the a$ti"od2 !o$!e$t#atio$.
,. The #esid&a so&tio$ is #emo<ed4 a$d the pates a#e 'ashed a$d d#ied as
des!#i"ed i$ step -.
1. -++ CL o% the s&"st#ate so&tio$ is added i$ ea!h 'e. The #ea!tio$ is ao'ed to
p#o!eed %o# 1+ mi$&tes4 'he$ it is stopped "2 additio$ o% 1+ CL o% ,M H
-
SO
,
a$d
i$!&"ate a)ai$ %o# a$othe# -106+ mi$.
5. The a"so#"a$!e is #ead at ,1+ $m.
The optim&m !o$!e$t#atio$s o% t#a!e# a$d a$ti"od2 a#e !hose$ as the o$es that
)i<e ,+ M D+ ; "i$di$).
0.peri"ent 1: Anal%te calibration and anal%sis of real sa"ples
1. The pate is !o<e#ed as des!#i"ed i$ step 14 e3pe#ime$t 14 e3!ept that o$2 the
optim&m !o$!e$t#atio$ o% a$ti"od2 is &sed. I$!&"atio$ o<e# $i)ht at ,C.
-. The #esid&a so&tio$ is #emo<ed a$d the pate is 'ashed as des!#i"ed i$ step -4
e3pe#ime$t 1.
6. The a$a2te !ai"#atio$ (i.e.4 %#om 1 ppm to + ppm aa!ho#* is pe#%o#med o$ the
Fo$)G dime$sio$ o% the pate i$ di%%e#e$t sampe mat#i!es4 as %oo'i$).
D6

1 - 6 , 1 5 D > A 1+ 11 1-
A

=

C

D

E

F

G

H
O
p
t
i
m
i
s
a
t
i
o
$

o
%

t
#
a
!
e
#

!
o
$
!
e
$
t
#
a
t
i
o
$

The t#a!e# is added to the optimised !o$!e$t#atio$ so that the %i$a <o&me i$ ea!h
'e is 1++ CL. The i$!&"atio$ is pe#%o#med %o# 1 ho&#s.
,. The #esid&a so&tio$ is #emo<ed a$d the pate is 'ashed as des!#i"ed i$ step -4
e3pe#ime$t 1.
1. -++ CL o% the s&"st#ate so&tio$ is added i$ ea!h 'e. The #ea!tio$ is ao'ed to
p#o!eed %o# 1+ mi$4 'he$ it is stopped "2 additio$ o% 1+ CL o% , M H
-
SO
,
a$d
i$!&"ate %o# -106+ mi$.
5. The a"so#"a$!e is #ead at ,1+ $m.
Rep$rt
1 E3pai$ ho' 2o& !hoose the P"estP a$ti"od2 a$d t#a!e# di&tio$ a$d 'h2 2o& did
i$ that 'a2O
- Estimate IC
1+
4 the d2$ami! #a$)e a$d the dete!tio$ imit %o# aa!ho# dete#mi$atio$
&si$) di&tio$ !&#<e i$ "&%%e# a$d sampe.
6 App#o3imate the mat#i3 i$%&e$!e i$ ea!h !ase (i.e.4 "2 !a!&ati$) the #atio
"et'ee$ IC
1+
si)$a o% !ai"#atio$ i$ "&%%e# a$d si)$a !o##espo$di$) to the same
poi$t i$ the !ase o% &#i$e di&tio$*.
, Estimate the !o$!e$t#atio$ o% aa!ho# i$ the &$/$o'$ sampe.
D,

1 - 6 , 1 5 D > A 1+ 11 1-
A

=

C

D

E

F

G

H
8=ST
8=ST
8=ST
Sampe &$di.
Sampe4 1.1+
Sampe4 1.1++
Sampe4 1.1+++
=a$/

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II.

AUTOMATIC ANALYTICAL METHODS FOR ENVIRONMENTAL MONITORING AND CONTROL

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