Analysis of fish tissue for trace amounts of lead by furnace atomic absorption spectroscopy
by Dennis Riley Neuman
A thesis submitted to the Graduate Faculty in partial fulfillment of the requirements for the degree of
MASTER OF SCIENCE in Chemistry
Montana State University
© Copyright by Dennis Riley Neuman (1972)
Abstract:
Trace amounts of lead were determined in fish bone, gill, liver and muscle tissue by a wet digestion
and furnace atomic absorption. Sensitivity is 0.27 ng/g. The detection limit is 0.15 ng/g. The accuracy
of the method, determined by standard addition, is 99% ± 6%. Matrix interferences were minimized as
evidenced by similar slopes of three analytical standard curves prepared from three different chemical
matrices.
Fish samples were collected from the West Gallatin River and the Federal Fish Hatchery. The lead
levels are in the p.p.m. range. Tissue from fish within the Gallatin Canyon exhibit the highest lead
levels. It is believed that this is the result of leaded vehicle exhaust emissions. S ta te m e n t o f P e rm is s io n ' t o
Copy
In p r e s e n tin g t h i s t h e s i s in. p a r t i a l f u l f i l l m e n t o f th e r e q u ir e ­
m ents f o r an advanced deg ree a t Montana S ta te U n iv e r s ity , I a g ree t h a t
th e L ib ra ry s h a l l make i t f r e e l y a v a il a b le f o r in s p e c tio n .
I fu rth e r
a g re e t h a t p e rm is s io n f o r e x te n s iv e copying o f t h i s t h e s i s f o r s c h o la rly
p u rp o s e s may be g ra n te d by my m ajo r p r o f e s s o r , o r , i n h i s a b se n c e , by
th e D ir e c to r o f L i b r a r ie s .
I t i s u n d e rsto o d t h a t any. copying o r p ub­
l i c a t i o n o f t h i s t h e s i s f o r f i n a n c i a l g a in s h a l l n o t be allo w ed w ith o u t
my w r itt e n p e rm is s io n .
ANALYSIS OF FISH TISSUE FOR TRACE AMOUNTS OF LEAD
BY FURNACE ATOMIC ABSORPTION SPECTROSCOPY
by
DENNIS RILEY NEUMAN
V
A t h e s i s su b m itte d t o ■th e G raduate F a c u lty i n p a r t i a l
f u l f i l l m e n t o f th e re q u ire m e n ts f o r th e d eg ree
of
MASTER OF SCIENCE
in
C hem istry
Approved:
Head, M ajor D epartm ent
iitte e
GrhdHate Dean
MONTANA STATE UNIVERSITY
Bozeman, Montana
A ugust, 1972
iii
ACKNOWLEDGMENT
. The a u th o r would l i k e t o e x p re ss h i s th a n k s t o th e fo llo w in g p e o p le
f o r t h e i r a s s i s t a n c e d u rin g th e co u rse o f t h i s re s e a r c h .
To Dr. Gordon K. P agenkopf, f o r h i s good a d v ic e and guidance and
f o r c a tc h in g m ost o f th e f i s h used i n th e stu d y .
To D r. Ray W oodriff and h i s r e s e a r c h group f o r t h e i r te c h n i c a l
a d v ic e in v o lv in g th e fu rn a c e .
To Montana S ta te U n iv e rs ity C hem istry D epartm ent f o r th e f i n a n c i a l
a s s i s t a n c e p ro v id e d by a G raduate T eaching A s s is ta n ts h ip .
To h i s fa m ily f o r t h e i r m o ral su p p o rt d u rin g th e c o u rse o f t h i s
stu d y .
TABLE OF CONTENTS
Page.
L i s t o f T a b le s „ „
L i s t o f F ig u re s
.
v
.................. . . . . . . . . . . . . . . . . . . . . .
A b s tra c t . . . . . . . . . . . . . . . . . . . .
......................
vi
. . . .
v ii
PART
I
INTRODUCTION.........................................................................................................I
S ta te m e n t o f th e P r o b le m ................................... .................................. 10
II
EXPERIMENTAL PROCEDURES ............................................ . . . . . . . . .
12
Sample S e le c tio n . . . . . . . . . . . . . . . . . . . . .
12
G lassw are P r e p a r a tio n
..........................
R eagent and Sample P r e p a r a tio n . . .
A p p aratu s
III
. . . . .
14
15
RESULTS AND D ISCU SSIO N ............................... ............................................... . 18
^
...............................
18
Lead L ev els i n F is h T issu e . . . . . . . . . . . . . . . .
28
. SUMMARY AND CONCLUSIONS . . . . . . . . . . . . . . . . . . . .
38
A n a ly tic a l Method
Lead in F is h T issu e
V
......................
..................................................... ..................................
A n a ly tic a l Method
IV
„ 12
LITERATURE CITED
.
. . . . . . . . . . . . . . . . . . . .
38
. . . . . . . . . . . . . . . . . . .
38
41
L IS T OF TABLES
Table
Page
I
Furnace T em perature v s A bsorbance
.
„ 18
II
D rying Tem perature-Tim e v s A bsorbance . .
.................. ....
20
III
Recovery o f L ead,Added to F is h Samples
.................. ....
25
IV
F is h Age v s Average Length
. . . . . . . . . . . . . . . . .
28
V
Lead L ev els in F is h T issu e
............................
29
VI
Average Lead L ev els in T is s u e s by
V II
Lead in Bone T issu es.
V III
Lead in G i l l T i s s u e s ..........................................................
35
IX
Lead i n L iv e r T is s u e s
36
X
Lead L ev els in R iv e r and H atchery W ater . . . . . . . . . . .
.
. . . . . . . . . .
L o c a tio n
. ...............................................................................
.. ...............................
.
31
33
37
L IS T OF FIGURES
F ig u re
1
Page
A ir-L ead V alues a s a F u n c tio n o f T r a f f ic Volumes and
D is ta n c e from th e Highway . . . . . .
...................................
. . . .
3
2 ■ Map o f West G a lla t in R i v e r ......................
13
3
The F u r n a c e ............................... .... ...............................
16
4
S tan d ard Curve f o r Lead . . . . . . . . . . . . .
5
C a lib r a tio n Curves f o r th e Lead Added to R eagent Bland
and from S tan d ard A d d itio n Data ........................................ . . . . . .
...........................
22
23
ABSTRACT
T race am ounts o f le a d were d eterm in ed i n f i s h b o n e, g i l l , l i v e r
and m uscle t i s s u e by a w et d ig e s tio n and fu rn a c e atom ic a b s o rp tio n .
S e n s i t i v i t y i s 0.27 n g /g . The d e te c tio n l i m i t i s 0.1 5 n g /g . The
a c c u ra cy o f th e m ethod, d eterm in ed by s ta n d a rd a d d itio n , i s 99% ± 6%.
M atrix in t e r f e r e n c e s were m inim ized a s ev id en ced by s im ila r s lo p e s o f
th r e e a n a l y t i c a l s ta n d a rd c u rv e s p re p a re d from th r e e d i f f e r e n t chem­
i c a l m a tr ic e s .
F is h sam ples were c o lle c te d from th e West G a lla t in R iv e r and th e
F e d e ra l F is h H a tch e ry . The le a d l e v e l s a re i n th e p .p .m . ra n g e .
T issu e from f i s h w ith in th e G a lla t in Canyon e x h ib it th e h ig h e s t le a d
l e v e l s . I t i s b e lie v e d t h a t t h i s i s th e r e s u l t o f le a d e d v e h ic le
e x h a u s t e m issio n s.
INTRODUCTION
Since th e I n d u s t r i a l R ev o lu tio n th e r e h a s been an in c r e a s in g use
o f - le a d .
Today th e U. S. consum ption o f le a d i s o v e r one m illio n to n s
a n n u a lly .
Lead i s a n a t u r a l component i n s o i l , w a te r, an im al l i f e ,
v e g e ta tio n and th e a i r .
I t h a s been mined and used s in c e e a r ly tim e s
f o r th e making o f b ro n z e , b r a s s , p ip e s t o c a rry w a te r and f o r "sw e e tin g
w in es.
D ated snow sam ples from G reenland in d ic a te t h a t th e le a d con­
t e n t i s in c r e a s in g and h a s become an e x p o n e n tia l f u n c tio n o f tim e sin c e
1940.
There was a b a se l e v e l o f 20 /Lig P b /to n o f ic e up t o 1750.
1860 i t was 50 /ig /to n .
By
In 1940, 80 /Ltg/ton; in 1950 120 /ig /to n ; and
i n 1965 th e le a d l e v e l was 210 /Ltg/ton.
T h is sharp in c r e a s e in th e Pb
snow l e v e l h a s been a t t r i b u t e d t o th e u se o f le ad e d g a s o lin e s .
In th e p a s t 30 y e a r s th e in c r e a s in g u se o f le a d h a s cu lm in ated i n
an e n v iro n m e n ta l l e v e l f a r above t h a t which would n o rm a lly e x i s t , w ith
3 4
co n seq u en t h a rm fu l e f f e c t s on h e a l t h . ’
The m ajor s o u rc e s o f le a d
c o n ta m in a tio n a re le a d e d g a s o lin e s and p e s t i c i d e s , th e u se o f le a d in
m a n u fa c tu rin g , th e com bustion o f c o a l, i n c in e r a t io n o f r e f u s e , le ad e d
p a i n t s and earth en w a re p o t t e r y t h a t h a s been im p ro p erly g la z e d .
A m ajo r study o f le a d i n u rb a n a i r ’ was conducted i n 1961 and
1962.
3400 a i r sam ples i n th r e e m ajo r U. S. c i t i e s were a n aly z e d . The
g
a n n u a l a v erag e c o n c e n tra tio n o f le a d was 1-3 /Ltg/m . In a r e a s o f heavy
t r a f f i c th e le a d l e v e l ranged from 14 t o 44 /ig/m^.
In a c i t y w ith a
O
p o p u la tio n o f o v e r 2 .5 m illio n th e mean v a lu e was 5 jxg/m°, w h ile th e
l e v e l dropped t o 1.7 Mg/m^ in c i t i e s w ith l e s s th a n 100,000 p e o p le .
It
2
was d eterm in ed t h a t th e le a d l e v e l in u rb an a i r i s r i s i n g a p p ro x im a te ly
5%.a n n u a ll y „
Lead in g a s o lin e i s th e m ajo r so u rce o f a ir b o r n e le a d in
u rb a n a r e a s . ^
Lead a s th e t e t r a e t h y l and te tr a m e th y l a Ucyls i s added t o g a s o lin e s
t o in c r e a s e t h e i r o c tan e r a t i n g s .
The n a t io n a l l e v e l i s 2 .4 0 grams o f
T E L /g allo n o r 1 .5 4 grams le a d p e r g a l l o n , w ith a ran g e o f 1 .2 8 t o 2 .5 6
grams P b /g a llo n o f g a s o li n e .
O rganic s c a v e n g e rs , e th y le n e d ic h lo r id e
and e th y le n e d ib ro m id e , a r e a l s o added t o g a s o li n e .
The e x h a u s t
e m is sio n s c o n ta in m o stly PbBrCl and s m a lle r amounts o f PbC lg, PbBr2 ,
Pb(OH)Cl and Pb(OH)Br.
The amount o f le a d d isc h a rg e d t o th e atmosr-
p h ere in e x h a u s t g a se s v a r i e s , depending on d r iv in g c o n d itio n s , from
I 8
25% t o 75% o f th e le a d in ta k e i n th e f u e l . 5 As a consequence th e
a tm o s p h e ric c o n c e n tr a tio n s n e a r highw ays a r e an e x p o n e n tia l fu n c tio n
o f d is ta n c e from th e roadway and a l i n e a r f u n c tio n o f t r a f f i c volum e.
9
The le a d l e v e l was m easured a t v a r io u s d is ta n c e s (10 t o 1750 f e e t )
from highw ays s u p p o rtin g t r a f f i c lo a d o f betw een 1 9 ,8 0 0 c a r s and
9
58,000 c a r s d a i l y . F ig u re I shows th e a ir b o r n e le a d l e v e l s a s fu n c ­
t i o n s o f t r a f f i c volume and d is ta n c e from th e highw ay.
The le a d l e v e l
n e a r th e highway i s r e l a t i v e l y h ig h , b u t d ro p s o f f r a p i d l y d u rin g th e
f i r s t few f e e t (150) from th e highw ay.
The c o n c e n tr a tio n o f le a d was
red u ced 50% betw een 10 and 30 f e e t from th e highway s u p p o rtin g 58,000
v e h ic le s d a i l y .
I n a s i m i l a r study"*"® le a d in s o i l s and p la n ts was
r e l a t e d t o t r a f f i c volume and p ro x im ity t o highw ays.
S o il s and p la n ts
a lo n g h e a v ily tr a v e le d highw ays show t h a t le a d c o n c e n tr a tio n s in c re a s e
3
(1) S3,OOO vehicles daily
(2) 4 7 , 0 0 0
(4)
H
Ii
I 9,00 0
2 &3
DISTANCE (feet)
F ig u re I .
S o u rc e :
A ir-L ead V alues a s a F u n c tio n o f T r a f f ic Volume and D ista n c e
from th e Highway
E n v iro n m en tal S cien ce and T echnology,
320 (1 9 7 0 ).
4
w ith t r a f f i c volume and d e c re a se w ith d is ta n c e from th e highw ay.
o f th e le a d was a rem ovable s u rfa c e c o n tam in an t on th e p l a n t s .
Much
The
m ajor e f f e c t o f t r a f f i c was lim ite d t o th e s u r f a c e s o i l and t o a
narrow zone w ith in 100 f e e t o f th e h ig h w ay .
The a v e ra g e Pb l e v e l in
n on-washed ro a d s id e g ra s s a t 0 f e e t from th e highway was 2 5 5 .0 p .p .m .
A t 255 f e e t from th e highway th e Pb l e v e l was 4 6 .3 p .p .m .
G rass sam ples
t h a t were washed p r i o r t o th e a n a ly s is c o n ta in e d l e s s l e a d .
T h e ir
a v e ra g e v a lu e s a t 0 f e e t and 255 f e e t w ere 136 p .p .m . and 4 0 .6 p .p .m .
le a d , re s p e c tiv e ly .
The a v erag e le a d c o n te n t o f s o i l sam ples ta k e n a t
25 f e e t from th e highw ays were 1 5 9 .2 p .p .m . (0 -6 in c h e s d e p th ) , and
1 0 5 .0
p .p .m . le a d (6-12 in c h e s d e p t h ) .
A t 225 f e e t th e a v e ra g e v a lu e s
w ere 9 2 .5 p .p .m . and 7 1 .2 p .p .m . r e s p e c t i v e l y .
The h ig h e r le v e l s o f
le a d i n th e a i r , s o i l and f o l i a g e n e a r highw ays have le a d t o d is a s tr o u s
r e s u l t s f o r some o f th e a n im a ls in New York C ity z o o s.
d e a th was d i r e c t l y a t t r i b u t e d t o le a d p o is o n in g .
11
A l i o n ’s
G ra s s , le a v e s and
s o i l sam ples ta k e n from w ith in th e zoo cag es ranged up t o 3900 jig l e a d /
gram .
R alph S t r e b e l o f New York M ed ical C o lle g e , a t t r i b u t e d th e h ig h
le a d c o n c e n tr a tio n s t o a tm o sp h eric f a l l o u t .
C hildhood le a d p o is o n in g i s a v e ry r e a l d is e a s e o f th e m e tro p o lita n
g h e tto .
I t a f f e c t s p r im a r ily p re s c h o o l age c h ild r e n who l i v e in
d e t e r i o r a t e d h o u sin g p re v io u s ly p a in te d w ith le ad e d p a i n t .
A pproxim ately
8% o f a l l g h e tto c h ild r e n s u f f e r from v a ry in g d e g re ss o f plum bism .
A
c h ip o f p a i n t th e s i z e o f an a d u l t f i n g e r n a i l may c o n ta in betw een 50
5
and 100 mg o f le a d
A c h ild e a tin g a few o f th e s e a day w i l l e a s i l y
In a d u l t s a le a d -b lo o d l e v e l o f 80 fJg
exceed th e t o x i c l e v e l o f l e a d .
le a d /1 0 0 ml o f blood i s c o n s id e r e d . t o be th e minimum t o x i c l e v e l .
C h ild re n h a v in g c o n c e n tr a tio n s o f 60-80 ^g /1 0 0 ml may show c l i n i c a l
s ig n s o f plumb ism .
Above 80 j^g th e r i s k o f a c u te le a d p o is o n in g r i s e s
s ig n ific a n tly .
I n a m ajo r s tu d y
12
o f le a d and i t s t o t a l r e l a t i o n s h i p t o man, le a d
was found in a l l human o rg a n s .
A ccum ulation o f le a d w ith age in t i s ­
su e s o f A m erican human b e in g s re a c h e s a maximum in th e f i f t h o r s i x t h
decade.
Some t i s s u e s showed a 4 t o 10 tim e s in c r e a s e i n c o n c e n tra tio n
o v er v a lu e s a t b i r t h ..
was 20 p .p .m .
In lung t i s s u e a t b i r t h th e a v e ra g e le a d l e v e l
T his v a lu e in c re a s e d t o a h ig h a v erag e v a lu e o f over
70 p .p .m . a t age 40-50 y e a r s .
Bone t i s s u e a l s o showed a n a v erag e low
l e v e l o f l e s s th a n 5 p .p .m . a t b i r t h , w ith a h ig h a v e ra g e v a lu e o f
o v er 30 p .p .m . a t age 50-60 y e a r s .
S im ila r tr e n d s w ere a l s o in d ic a te d
i n th e k id n e y , p a n c r e a s , l i v e r and a o r t a .
le a d l e v e l s i n v a r io u s sea fo o d s .
w e ig h t) .
The a u th o rs d eterm in ed th e
These v a lu e s a r e a l l i n /Lig/g (w et
F ro zen s h rim p , 0 .3 1 ; f r e s h sh rim p , 0 .4 5 , l o b s t e r 2 .5 0 ; c r a b -
m e a t, 0 .0 6 ; clam s 0 .1 5 ; o y s t e r s , 0 .1 3 ; s c a l l o p s , 0 .1 5 ; s a r d i n e s , 0 .7 5 ;
and haddock, 0.27 ^ g /g .
In a more r e c e n t work
t i o n s i n f i s h were d e te rm in e d .
th e le a d c o n c e n tra ­
Salmon sam ples .c o n ta in e d 1 .3 jug P b /b ;
clam chow der, 2 .2 jug/g; tu n a , 1 .2 p g /g ; o y s t e r , 4 .3 ^ g /g ; s a r d in e ,
1 .3 y g / g ; and crabm eat 0 .8 5 y g / g .
The v a lu e s were e s ta b lis h e d on th e
6
b a s i s o f d ry w e ig h t.
The e n tra n c e o f Pb i n t o th e body i s by two r o u t e s , th e a lim e n ta ry
c a n a l and th e r e s p i r a t o r y t r a c t .
The amount o f le a d a b so rb ed from a
X 8
g iv e n a i r c o n c e n tr a tio n v a r ie s a c c o rd in g t o p a r t i c l e s i z e . 5
P a rtic le s
IjLt o r l e s s w i l l be in h a le d w ith 50% b e in g r e ta in e d in th e lu n g s .
tic le s
P a r­
and l a r g e r w i l l im pinge on mucous l i n i n g s and e i t h e r be
sw allow ed o r coughed o u t.
A pp ro x im ately 75% o f p a r t i c u l a t e Pb in
v e h ic le e x h a u s t i s l e s s th a n 0 .9 ^ in mean d ia m e te r , a s i z e e a s i l y r e ­
ta in e d in th e l u n g s .
a v e ra g e s 20-30 ^ g / d a y .
300 jtg /d a y . 5 ’
a b s o rb e d .
A b so rp tio n o f Pb by r e s p i r a t i o n i n u rb a n a re a s
The mean d i e t a r y in ta k e o f le a d i s a p p ro x im a te ly
A p p roxim ately 6-7% o f th e m e ta l in g e s te d i s a c t u a l l y
T h is r e p r e s e n ts 18 t o 21 Mg/d a y . The t o t a l d a i l y c o n c e n tra ­
t i o n o f le a d w hich i s r e ta in e d from b o th s o u r c e s , i s 38 t o 50 jug.
Lead i s a l s o e x c r e te d from th e body i n u r i n e , sw e a t, h a i r and n a i l s .
The g e n e r a l p o p u la tio n seems t o be in " le a d b a l a n c e " T h a t i s , th e
d a i l y amount o f le a d a b so rb ed (38-r50 jug) i s m atched by a p p ro x im a te ly
th e same amount e x c r e t e d .
More th a n 90% o f th e le a d i n th e body i s s to r e d i n th e s k e l e t o n .
In tim e s o f f e v e r i s h i l l n e s s o r h ig h c alc iu m m e ta b o lism , le a d may be
r e le a s e d from th e bone i n t o th e s o f t t i s s u e s .
Lead i n i t s m o b ile ,
s o lu b le form in s o f t t i s s u e s a f f e c t s c h i e f l y th e b lo o d , k id n e y s and
n e rv o u s s y s te m .
E x c e ssiv e amounts o f le a d in th e b lo o d can cause
15
anem ia and id io p a th ic hem o ch ro m ato sis.
C hronic n e p h r i t i s and in
-
7
s e v e re c a s e s k id n e y f a i l u r e may r e s u l t from h ig h le a d c o n c e n tra tio n s
in th e k id n e y s .
J u v e n ile m e n ta l r e t a r d a t i o n i l l d e fin e d n e u r o s is ,
and e n ce p h a lo p ath y can r e s u l t from th e e f f e c t o f a c u te o r p ro lo n g e d ,
c h r o n ic , ex p o su res t o l e a d .
I t h a s been e s ta b lis h e d t h a t th e in c r e a s in g e n v iro n m e n ta l l e v e l
o f le a d i s a s e r io u s p ro b lem .
A lthough th e m easured l e v e l s in b io ­
l o g i c a l system s a r e i n th e p a r t p e r m illi o n ra n g e , any f u r t h e r in c re a s e
in th e e n v iro n m e n ta l le a d l e v e l w i l l r e s u l t in a f u r t h e r c o n c e n tra tio n
in th e s e s y s te m s .
Lead ab so rb ed in t r a c e q u a n t i t i e s o v e r a lo n g p e rio d
o f tim e can accu m u late t o exceed th e th r e s h o ld l e v e l s and produce
v d e lay e d to x ic e f f e c t s .
In g e o g ra p h ic a l lo c a tio n s w here th e e n v iro n ­
m e n ta l l e v e l i s n o t e x p ec te d t o be s i g n i f i c a n t , i t sh o u ld be p o s s ib le
t o d e term in e b a s e lin e le a d le v e l s i n b i o l o g i c a l s y s te m s .
The im p o rtan ce
o f e s t a b l i s h i n g th e s e " n a t u r a l " l e v e l s c an n o t be u n d e r s t a t e d .
f u tu r e such q u e s tio n s a s ,
I n th e
have th e " n a t u r a l " l e v e l s in c r e a s e d ? , w hat
h as caused th e l e v e l s t o in c r e a s e ? , and w hat s,hould be done a b o u t
them ?, can be more f u l l y u n d e rsto o d and more r e s p o n s ib ly an sw ered .
The d e te r m in a tio n o f le a d i n b i o l o g i c a l sam ples h a s been accom­
p lis h e d by c o l o r i m e t r i c ^ a n d
s p e c tro p h o to m e tric m eth o d s"^’' ^ 5"^
9 10 13 17 20
and more r e c e n tly by c o n v e n tio n a l flam e ato m ic a b s o r p tio n > > > >
and fla m e le s s a to m ic a b s o r p tio n .
21 22 23 24
? » >
B efore th e a n a ly s is can
be made i t i s n e c e s s a ry t o o x id iz e th e o rg a n ic m a tte r t o CO^ and HgO,
8
le a v in g o n ly th e in o rg a n ic s a l t s a s r e s i d u e .
In g e n e r a l, th e r e a r e
t h r e e a p p ro a ch e s t o t h i s problem , a c id d i g e s t i o n , h ig h te m p e ra tu re
d ry a s h in g , and a r e l a t i v e l y new method o f low te m p e ra tu re oxygen
plasm a a sh in g
A l l t h r e e m ethods w i l l acco m p lish th e o x id a tio n
o f th e o rg a n ic m a t e r i a l s .
In th e d ry a s h in g te c h n iq u e , o x id a tio n i s
acco m p lish ed by h e a tin g th e sam ple t o r e l a t i v e l y h ig h te m p e r a tu r e s ,
400° t o 7 SO0G.
A tm ospheric oxygen s e r v e s a s th e o x id iz in g a g e n t.
The tim e o f a s h in g , depending upon sam ple s i z e , can e a s i l y re a c h 12 t o
16 h o u r s .
In a c id d ig e s t io n
p ro c e d u re s th e te m p e ra tu re s a r e much
low er and l i q u i d c o n d itio n s a r e m a in ta in e d .
by o x id iz in g a g e n ts in s o l u t i o n .
O x id a tio n i s c a r r ie d o u t
N i t r i c , p e r c h l o r i c , s u l f u r i c and
h y d ro c h lo ric a c id s and c o m b in atio n s o f th e s e a r e u s u a ll y used t o d i g e s t
th e s a m p le s .
D is s o lu tio n tim e s a g a in v a ry w ith sam ple s i z e , b u t th e y
a r e c o n s id e ra b ly l e s s th a n i n d ry a s h in g .
I n th e low te m p e ra tu re
oxygen plasm a te c h n iq u e , o x id a tio n i s acco m p lish ed by p la c in g th e sam ple
i n a n oxygen plasm a w hich s e r v e s a s th e o x id iz in g a g e n t.
The plasm a i s
a c h ie v e d by i r r a d i a t i n g oxygen w ith r a d i o fre q u e n c y r a d i a t i o n which
io n iz e s th e oxygen c r e a tin g a p la sm a .
300°C, a r e m a in ta in e d .
Low te m p e r a tu r e s , l e s s th a n '
O x id a tio n tim e s a r e u s u a lly lo n g e r th a n a c id
d ig e s t io n y e t l e s s th a n th e dry a s h in g te c h n iq u e .
Once th e sample i s
c o m p le te ly o x id iz e d th e r e s id u e i s u s u a ll y d ilu te d w ith w a te r and
d e te r m in a tio n s o f t r a c e m e ta ls m ade.
C o lo rim e tric and s p e c tro p h o to m e tr ic te c h n iq u e s depend upon th e
9
developm ent o f p r e c is e c o lo r s u n d er c a r e f u l l y c o n tr o lle d pH c o n d itio n s .
In th e a n a ly s is o f le a d a t th e t r a c e l e v e l in b i o l o g i c a l sam p les, i t i s
u s u a lly n e c e s s a ry t o p r e c o n c e n tra te and e x t r a c t th e m e ta l u sin g d i t h i zone o r ammonium 1 - p y r r o lid in e c a r b o d ith io n a te (APDC) and m ethyl i s o ­
b u ty l k e to n e (MIBK).
n e c e s s a ry .
The u se o f a la r g e number o f r e a g e n t b la n k s i s
L arge sam ple s i z e s a r e r e q u ir e d and th e t o t a l a n a ly s is
tim e , b e ca u se o f th e e x te n s iv e sam ple p r e tr e a tm e n t, i s q u ite lo n g .
The m ost p o p u la r method o f t r a c e m e ta l a n a ly s is i n b i o l o g i c a l
sam ples i s c o n v e n tio n a l flam e atom ic a b s o r p tio n .
E x tr a c tio n and
p r e c o n c e n tr a tio n s te p s a r e u s u a lly used t o a l l e v i a t e ch em ical and
s p e c t r a l in te r f e r e n c e s and t o a c h ie v e b e t t e r d e te c tio n l i m i t s .
Large
sam ple s i z e s a r e u s u a lly n e c e s s a ry w hich can r e p r e s e n t a r e a l h a n d ica p
f o r c l i n i c a l sam ples such a s b lo o d , u r i n e , s p in a l and lymph f l u i d s .
N on-flam e ato m ic a b s o r p tio n r e p r e s e n ts th e m ost p ro m isin g method
f o r t r a c e m e ta l a n a l y s i s .
The h ig h s e n s i t i v i t i e s and low d e te c tio n
l i m i t s co u p led w ith v e ry s m a ll sam ple s iz e s make t h i s te c h n iq u e th e
m ost v e r s a t i l e method f o r a n a ly s is o f t r a c e m e ta ls i n b i o l o g i c a l s y s tm es o C hem ical and s p e c t r a l in t e r f e r e n c e s r e s u l t i n g from th e com­
p l i c a t e d sam ple m a trix can be e lim in a te d o r a t l e a s t red u ced t o an
a c c e p ta b le l e v e l .
10
S ta te m e n t o f th e Problem
A irb o rn e le a d from v e h ic le e x h a u s ts i s th e main so u rc e o f le a d
c o n ta m in a tio n i n u rb a n a r e a s .
The le a d l e v e l n e a r highw ays i s depen­
d e n t upon th e p ro x im ity o f th e roadway and th e t r a f f i c volum e.
Vege­
t a t i o n and s o i l sam ples n e a r highw ays show t h a t le a d c o n c e n tr a tio n s
in c r e a s e w ith t r a f f i c volume and d e c re a s e w ith d is ta n c e from th e h ig h ­
way.
U. S . Highway 191 p a r a l l e l s th e W est G a lla t in R iv e r v e ry c lo s e ly
f o r i t s e n t i r e r o u te w ith in G a l l a t i n Canyon.
volume i s a p p ro x im a te ly 1000 v e h i c l e s .
27
The a v e ra g e d a ily t r a f f i c
A lthough th e canyon can
h a rd ly be d e s c rib e d a s u rb a n , th e f i s h w ith in th e r i v e r have been e x ■■
posed t o le a d c o n ta in in g e x h a u s t e m is s io n s .
The d e te r m in a tio n o f th e le a d c o n te n t o f t i s s u e s from th e s e f i s h
may in d ic a te t o w hat e x te n t le a d h as accu m u lated a s a r e s u l t o f t h i s
e x p o s u re .
In u n iq u e lo c a t i o n s where f i s h have n o t been exposed t o le a d
c o n ta in in g e x h a u s ts i t sh o u ld be p o s s ib le t o e s t a b l i s h b a s e lin e le a d
le v e ls in t h e i r t i s s u e s .
F is h from th e F e d e ra l F is h H a tc h e ry , n o r th ­
e a s t o f Bozeman and from th e W est G a l l a t i n R iv er w ith in Y ello w sto n e
P a rk have n o t b een d i r e c t l y exposed t o v e h ic le e x h a u s ts .
A com parison o f th e le a d t i s s u e c o n c e n tr a tio n s o f th e s e f i s h t o
th o s e from w ith in t h e canyon may e s t a b l i s h t o w hat e x t e n t th e le v e l s
v a ry in a b i o l o g i c a l system a s a r e s u l t o f ex posure t o le a d c o n ta in in g
v e h ic l e .e x h a u s t e m is s io n s .
11
The scope o f t h i s r e s e a r c h p r o j e c t was f o u r f o l d :
1)
To e s t a b l i s h th e le a d c o n te n t in t i s s u e s o f f i s h t h a t have
been exposed t o le ad e d e x h a u s ts ;
2)
To e s t a b l i s h b a s e lin e le a d t i s s u e le v e l s in f i s h t h a t have n o t
b e e n exposed t o le a d e d e x h a u s ts ;
3)
To d e term in e w hat e f f e c t , i f a n y , th e le a d e m issio n s have had
on t h i s b i o l o g i c a l s y s te m ;
4)
To d e v elo p a s e n s i t i v e , a c c u r a te and r e l i a b l e method o f a n a ly s is
f o r le a d in f i s h b o n e , g i l l , l i v e r and m uscle t i s s u e s u s in g an a c id
d ig e s t io n p ro c e d u re and fu rn a c e ato m ic a b s o r p tio n .
EXPERIMENTAL PROCEDURES
Sample S e le c tio n
F is h w ere c a u g h t by th e s ta n d a rd hook and l i n e method by G. K.
Pagenkopf .and D. R. Neuman from s i x lo c a tio n s a lo n g th e West G a lla t in
R iv e r.
F ig u re 2 i l l u s t r a t e s a map o f th e West G a l l a t i n R iv e r.
sam ple c o l l e c t i o n s i t e s a r e w ith in G a l l a t i n Canyon:
Four
s o u th o f Cascade
C reek ; n e a r th e mouth o f Cascade C re e k ; s o u th o f G reek C reek and n e a r
th e mouth o f P o rcu p in e C reek .
Samples w ere a l s o ta k e n in Y ellow stone
P ark n e a r th e mouth o f Fan C reek and n o r th o f th e town o f G a lla t in
G atew ay.
A l l f i s h ta k e n from th e r i v e r and a n aly z e d w ere n a t i v e .
P la n te d f i s h have had t h e i r d o r s a l a d ip o se f i n c lip p e d p r i o r t o b e in g
p la n te d .
T h is a d ip o se f i n w i l l n o t grow b a c k .
28
F is h sam ples were
a l s o c o lle c te d from th e F e d e ra l F is h H atch ery in B rid g e r Canyon,
n o r t h e a s t o f Bozeman.
Of a t o t a l o f 39 f i s h , 30 w ere Rainbow T ro u t (Salmo g a r d u e r i ) ,
4 w ere C u tth r o a t T ro u t ( Salmo c l a r k i ) , 3 were Brown T ro u t (Salmo t r u t t a )
and two w ere W h ite fis h ( Coregonus w i l l i a n s o n i ) .
The f i s h sam ples w ere
la b e le d a c c o rd in g t o t y p e , d a te and l o c a t i o n o f c a tc h and fr o z e n a s soon
a s p o s s ib le a f t e r b e in g c a u g h t.
W ater sam ples were c o lle c te d in p o ly ­
e th y le n e b o t t l e s , from th e same l o c a t io n s a s th e f i s h w ere ta k e n .
The
w a te r sam ples were a c i d i f i e d w ith n i t r i c a c id t o a s s u r e th e s o l u b i l i t y
o f any le a d s a l t s .
G lassw are P r e p a r a tio n
I t was e s s e n t i a l t h a t a l l g la ss w a re b e im m aculately c le a n t o a v o id
13
FIGURE 20 W0 Gallatin River
/ G r e e k Cr.
Park Boundry
9 P orcu p in e Cr.
W0 Gallatin River
Sample Sites o
Taylor Fk.
14
any c o n ta m in a tio n .
Nine 1 0 0 .0 ml v o lu m e tric f l a s k s , tw e lv e 50 ml p y rex
b e a k e rs and an a s s o rtm e n t o f p i p e t t e s were b o u g h t new and used e x c lu ­
s iv e l y f o r le a d a n a l y s i s .
P r i o r t o i t s i n i t a l use a l l th e g la ssw a re
was washed in doubly d i s t i l l e d w a te r and soaked in h o t c o n c e n tra te d
HNOg.
Between usage th e g la ssw a re was washed and r in s e d i n doubly
d i s t i l l e d w a te r and soaked in h o t c o n c e n tra te d n i t r i c a c i d .
R eagents and Sample P r e p a r a tio n
R e d i s t i l l e d HNO3 and HClO^ were used t o w et a sh th e t i s s u e s a m p le s .
Lead n i t r a t e , t h a t had been d r ie d f o r two h o u rs 3 was used t o p re p a re
th e s ta n d a rd le a d s o l u t i o n s .
d i s t i l l e d c o n d u c tiv ity w a te r .
A ll d i l u t i o n s were made w ith doubly g la s s
Sample f i s h were removed from th e f r e e z e r
and a llo w ed t o thaw a t room te m p e ra tu re .
le n g th were re c o rd e d .
The t o t a l f i s h w e ig h t and
G i l l s 5 l i v e r t i s s u e and s e c tio n s o f ' backbone
and m uscle t i s s u e were removed by d i s s e c t i o n and f r e e z e d rie d by a
Thermovac l y p h o l i z e r .
A pproxim ately one gram o f th e d r ie d t i s s u e was
weighed and th e n d ig e s te d in a m ix tu re o f 7 .0 0 ml o f c o n c e n tra te d
HNO3 and 5 .0 0 ml o f c o n c e n tra te d HClO^.
The s o lu tio n s w ere slo w ly
h e a te d on a h o t p l a t e u n t i l a l l foam ing had sto p p ed and d is s o l u t i o n
was a c h ie v e d .
A t t h i s p o in t th e te m p e ra tu re was in c re a s e d so a s t o
re d u c e th e volume t o a b o u t I m l.
in g o f HClO^.
T h is was accom panied by copious fum­
The maximum te m p e ra tu re was 88°C.
The u s u a l p re c a u tio n s
were m a in ta in e d when u s in g p e r c h lo r ic a c id t o d i g e s t o rg a n ic m a t e r i a l .
The c o l o r l e s s sam ples were th e n a n a l y t i c a l l y tr a n s f e r r e d t o c lea n e d
15
1 0 0 .0 ml v o lu m e tric f l a s k s and d i lu te d t o volum e.
A r e a g e n t b la n k
s o l u tio n and s ta n d a rd curve check s o l u t i o n were c o n c u r r e n tly d ig e s te d
w ith a l l s a m p le s .
T o ta l a n a ly s is tim e , in c lu d in g l y p h o l i z a t i o n , f o r
8 sam ples was a p p ro x im a te ly 8 h o u r s .
A p p aratu s
F urnace a to m ic a b s o r p tio n s p e c t r o s c o p y ^ -3 ^ was used t o a n a ly z e
th e f i s h t i s s u e sam ples f o r t r a c e s o f l e a d .
shown in F ig u re 3 .
The fu rn a c e d e s ig n i s
I t i s h e a te d by a G .E. a r c w e ld e r t o an i n t e r n a l
te m p e ra tu re o f 1800°C .
Argon g a s , a t a flo w r a t e o f 35 m l/m in , was
c o n tin u o u s ly flu s h e d th ro u g h th e fu rn a c e t o p re v e n t th e o x id a tio n o f
th e g r a p h ite and t o c a r r y o u t h o t r e s i d u a l g a s e s .
R a d ia tio n from th e h ollow c ath o d e tu b e , s e t a t 12 ma, was chopped
a t 400 c y c le s /s e c u s in g a PAR model B Z-I l i g h t c h o p p e r.
lo c k - i n a m p l i f i e r i s tu n ed on t h a t fr e q u e n c y .
A PAR HR-8
The l i g h t p assed th ro u g h
th e h e a t e r tu b e o f th e fu rn a c e and was fo c u s se d on th e e n tra n c e s l i t
(H O fjpi) o f a Spex 3 /4 m e te r C ze rn y -T u m e r s p e c tro p h o to m e te r.
The
a m p lifie d s i g n a l from th e p h o to m u ltip lie r was m o n ito red by a T e c k tro n ic s
ty p e RM503 o s c illo s c o p e and d is p la y e d on a H oneyw ell 6 -in c h r e c o r d e r .
o
The Spex m onochrom ator was s e t a t t h e 2170 A le a d l i n e . An E ppendorf
p i p e t t e was u sed t o t r a n s f e r 20 p i o f th e sample i n t o th e g r a p h ite
cups.
The cups w ere p la c e d u n d er a n IR l i g h t and h e a te d u n t i l th e
s o lv e n t had e v a p o r a te d .
The cup was in tro d u c e d i n t o th e fu rn a c e th ro u g h
th e sam ple p o r t , and r e s t e d a t th e in n e r end o f th e s id e tu b e .
Its
■RUBBER INSULAtlON
3 0 CM.
FLUSH VENT
r—INSULATOR
GRAPHITE INSULATION
ivV.V;
v-: "
.
‘
=. .
v.
4-
y
/H EA TER TU O E
:
T
Z l SHIELD
Tl
TUBE
%
j
S I
*:4
I I
—
W
SPIRAL HEATER
TUBE CONTACT
Q
s t a in l e s s
st e e l
0
RUBBER
C 3 G R A P H IT E
E 3 CO PPER
Q
□
QUARTZ
g r a p h it e
IN SU L A T IO N
F ig u re 3 .
The Furnace
6
17
c o n te n ts were im m ed iately atom ized i n t o th e h e a te r tu b e .
The peak
h e ig h t was re a d a s a f u n c tio n o f a b so rb an c e and compared t o s ta n d a rd
c u rv e s .
fu rn a c e .
P r io r t o u se each g r a p h ite cup was h e a te d t o 1 8 50OC in th e
T h is e lim in a te d any a b so rb an c e due t o c o n ta m in a tio n .
RESULTS and DISCUSSION
A n a ly tic a l Method
B efo re th e a c t u a l a n a ly s is f o r le a d i n th e t i s s u e . sam ples could
be made, i t was n e c e s s a ry t o d e te rm in e th e i n t e r n a l fu rn a c e te m p e ra tu re
t h a t would p ro v id e th e g r e a t e s t s e n s i t i v i t y f o r a c o n s ta n t c o n c e n tra tio n of le a d .
35
T ab le I shows th e r e l a t i o n s h i p betw een fu rn a c e tem p­
e r a tu r e and a b so rb an c e f o r a g iv e n c o n c e n tr a tio n o f l e a d .
The t a b l e
i n d ic a te s t h a t th e maximum s e n s i t i v i t y i s a c h ie v e d n e a r ISOO0C.
T able I .
F urnace T em perature vs A bsorbance f o r
-1 0
8 x 10
grams o f le a d
F urnace T em perature
A verage A bsorbance
IlBO0C
.!SB
1300°C
„370
1450°C
.SOS
, 1600°C
.645
ITBO0C
.680
ISSO0C
„695
1900°C
.663
E a rly in th e e x p e rim e n ta l work th e a n a ly s is o f t i s s u e sam ples w ere
e x h ib itin g seven® ch em ica l i n t e r f e r e n c e s .
The HClO^ w hich was used in
th e a c id d ig e s t io n p ro c e d u re gave r i s e t o n o n - s p e c if ic a b s o r p tio n .
If
th e g r a p h ite cups were n o t h e a te d u n d er th e IR lamp f o r a lo n g enough
p e rio d o r a t a s u f f i c i e n t te m p e ra tu re , th e HClO^ would n o t be c o m p le tely
d riv e n o f f .
When th e cups were in s e r t e d i n t o th e fu rn a c e th e HClO^
19
would "fu m e," m asking th e t r u e ato m ic a b s o r p tio n s i g n a l .
h ig h , i n c o n s i s t e n t a b so rb an c e v a lu e s .
T his caused
T ab le I I shows t h a t th r e e
d i f f e r e n t ch em ica l m a tric e s were used t o o p tim iz e th e tim e and te m p e ra ­
t u r e o f d r y in g .
90°C t o 190°C .
The te m p e ra tu re u n d e r th e d ry in g lamp was v a rie d from
The d ry in g tim e was v a r ie d from 15 t o 30 m in u te s .
H ig h er te m p e ra tu re s were a c h ie v e d u s in g a m u ffle f u r n a c e .
A s ta n d a rd
aqueous s o l u t i o n o f le a d was p re p a re d so t h a t 20 ^ l gave a c o n c e n tra tio n
o f 2 .0 x 10
g P b.
The a b so rb an c e v a lu e o f t h i s c o n c e n tr a tio n , .1 7 0 ,
was u n a ffe c te d by d ry in g a t v a rio u s te m p e r a tu r e s - tim e s .
i l l u s t r a t e d in column th r e e of. th e t a b l e .
T h is is
20 ^ l o f th e r e a g e n t b la n k ,
w hich c o n ta in e d th e d ig e s t in g a c id HNO3 and HClO^, gave th e ab so rb an ce
v a lu e s shown i n column f o u r .
A t low te m p e r a tu r e s - s h o r t d ry in g tim es
th e s e v a lu e s a r e h ig h , i n d ic a tin g i n t e r f e r e n c e from th e HClO^.
Lead
was added t o th e a c id c o n ta in in g r e a g e n t b la n k so t h a t 20 ^ l p ro v id ed a
c o n c e n tr a tio n o f added 2 .0 x 10
v a lu e s shown i n th e l a s t colum n.
t u r e s - s h o r t d ry in g t i m e s .
P b.
T h is s o lu tio n gave a b so rb an c e
The v a lu e s a re h ig h a t low tem p era­
A t v e ry h ig h te m p e ra tu re s th e ab so rb an ce
v a lu e s a r e low er th a n w hat i s e x p ec te d from th e added le a d a l o n e ; t h i s
in d ic a te s t h a t th e m e ta l i s b e in g l o s t due t o v o l a t i l i z a t i o n a t th e s e
te m p e r a tu r e s .
A t a te m p e ra tu re o f 190°C and a d ry in g tim e o f 30 min­
u te s th e a b so rb an c e v a lu e o f th e r e a g e n t b la n k s o l u tio n was reduced t o
.1 7 7 .
A lthough t h i s v a lu e was r e l a t i v e l y h ig h , i t was c o n s i s t e n t ,
h av in g a d e v ia tio n o f ± .003 a t th e 95% c o n fid e n ce l e v e l .
The optimum
T ab le I I .
D rying Tem perature-T im e v s A bsorbance
A bsorbance
T em perature u n d er
d ry in g lamp
Time cups d rie d
u n d er lamp
2 .0 x 10 "^g P b ^
R eagent blarik^3 ^
2 .0 x IO"10
g Pb, r e a g e n t
b la n k (a )
90°C
IOO0C
MO0C
190 C
15 min
15
15
15
.170
.170
.170
.170
.958
.886
.782
.769
90°C
IOO0C
MOOC
190°C •
20 min
20
20
20
.170
.170
.170
.170
.732
.688
.647
. .408
.853
. .782
.699
.602
90°C
IOO0C
MO0C
190°C
30 min
30
30
30
.170
.170
.170
.170
.508
.356
.214
.177
.699
.515
.414
.347
295°C
340°C
400°C
450°C •
15 min
15
15
15
.031
.022
.017
.044
.026
.004
.031
.004
cups p la c e d i n m u ffle fu rn a c e
^ a l l v olum es, 20
— -------
—
—
'
.1.000
.920
• .744
.657
21
c o n d itio n s o f d ry in g te m p e ra tu re and tim e were s e t a t 190°C and 30
m in u te s.
. The a n a l y t i c a l s ta n d a rd curve i s shown in F ig u re 4.
The s ta n d a rd
le a d s o lu tio n s f o r t h i s p l o t ran g e in c o n c e n tra tio n from 0.2 5 to 5.00
x 10“5 g / l .
A 20 / l l a l i q u o t o f each p ro v id e d th e c o n c e n tra tio n ra n g e ,
0 .5 0 t o 10.0 x IO- -^ g le a d .
Two a d d i t i o n a l a n a l y t i c a l c u rv e s were
o b ta in e d from s ta n d a rd a d d itio n d a ta and le a d added t o th e a c id d i ­
g e s tio n r e a g e n ts .
These a r e i l l u s t r a t e d i n F ig u re 5.
o f th e le a d - a c id d ig e s t io n curv e i s .1 7 7 .
v a lu e f o r th e re a g e n t b la n k .
The i n t e r c e p t
T h is i s th e ab so rb an ce
The i n t e r c e p t o f th e p l o t o b ta in e d from
s ta n d a rd a d d itio n d a ta i s . 388.
T h is v a lu e i s h ig h e r a s i t r e p r e s e n ts
th e re a g e n t b la n k v a lu e and a ls o th e le a d n a t u r a l l y o c c u r rin g i n th e
t i s s u e sam ple.
ra n g e .
A ll th r e e e x h ib it l i n e a r i t y i n t h i s c o n c e n tra tio n
The slo p e o f th e p l o t i n F ig u re 4 i s 0.8 8 x 10^.
o f th e o th e r two a r e 0 .8 6 and 0 .8 7 x 10^.
The s lo p e s
The le a d s e n s i t i v i t y ob­
ta in e d by fu rn a c e atom ic a b s o r p tio n i s many tim e s b e t t e r th a n t h a t ob­
ta in e d by c o n v e n tio n a l fla m e s ^
The a b s o lu te s e n s i t i v i t y , which i s th e
c a lc u la te d le a d c o n c e n tr a tio n t h a t g iv e s an ab so rb an ce v a lu e o f .004
(1%A) i s 5 .5 x 10 ^
g Pb.
A d e te c tio n l i m i t o f 0 .15 n g /g i s c a lc u ­
l a t e d a s tw ic e th e s ta n d a rd d e v ia tio n a t th e 95% c o n fid e n c e l e v e l f o r
th e r e a g e n t b la n k .
The e x c e lle n t s e n s i t i v i t y and low d e te c tio n l i m i t
r e s u l t s in p a r t from th e f a c t t h a t th e atom s a re c o n fin e d i n a long
n o n - tu r b u le n t l i g h t p a th f o r a g r e a t e r le n g th o f tim e .
ABSORBANCE
22
GRAMS
F ig u re 4.
Pb K IO
S tan d ard Curve f o r Lead
23
9
GRAMS
F ig u re 5.
Pb x IO
C a lib r a tio n Curves f o r Lead Added to R eagent Blank
and from S tan d ard A d d itio n Data
IO
24
The a c c u ra c y o f t h i s method h a s been e v a lu a te d u s in g s ta n d a rd
a d d itio n .
A known q u a n tity o f le a d was added t o a p p ro x im a te ly e q u iv a ­
l e n t s a m p le s, I g , o f d rie d m uscle t i s s u e .
d ig e s te d and a b so rb a n c e s d e te rm in e d .
T able I I I .
The sam ples were th e n
The r e s u l t s a r e p re s e n te d in
The v a lu e s a r e based on a t l e a s t 5 d e te r m in a tio n s , and a l l
d e v ia tio n s a r e a t th e 95% c o n fid e n c e l e v e l .
th e Pb found in m uscle t i s s u e in ^ g /g .
The f i r s t column r e p r e s e n ts
P re d ic te d le a d i s th e sum o f
th e n a t u r a l l y o c c u rrin g le a d ( 1 s t colum n) and th e am ount o f le a d a d d ed .
The l a s t column r e p r e s e n ts th e r a t i o o f Pb found t o Pb p r e d ic te d , o r
th e f r a c t i o n re c o v e re d .
The d a ta i n T ab le I I I in d ic a te t h a t v i r t u a l l y
no le a d i s b e in g l o s t d u rin g th e p ro c e d u re .
The d a ta a l s o show one
p a r t i n t r i n s i c le a d can be d eterm in ed i n th e p re se n c e o f 49 p a r ts
added Pb.
The le a d p r e s e n t n a t u r a l l y i n th e s e f i s h m uscle t i s s u e s
ra n g e s from 0.12 t o 1 .8 1 jyg/g.
The p e r c e n t re c o v e ry ra n g e s from 95
t o 102 w ith a, c e n t r a l te n d en c y o f 99%.
fid e n c e l e v e l i s a b o u t ± 6 % .
le a d i n m uscle t i s s u e i s ± 5% .
The d e v ia tio n a t th e 95% con­
The " r e l a t i v e s ta n d a rd d e v ia t io n f o r th e
Over t h i s ran g e o f le a d c o n c e n tra tio n
i t i s concluded t h a t th e a c c u ra c y o f th e method ex ceed s th e o b ta in a b le
p re c is io n .
u s in g
212
These re c o v e ry r e s u l t s a r e c o n s is te n t w ith e x p erim en ts
36
Pb w ith th e HClO^-HNOg w et a s h in g te c h n iq u e o f c o c o a .
The method p re s e n te d i n t h i s work h a s th e a d v an ta g e t h a t th e
q uoted s e n s i t i v i t y and th e d e te c tio n l i m i t can be o b ta in e d w ith r e a l
a s w e ll a s s y n t h e t i c s a m p le s .
The l i n e a r i t y and s lo p e s o f th e th r e e
25
T able I I I .
Recovery o f Lead Added t o F is h Samples
Pb i n Muscle
T iss u e
Pb Added3
1 .8 1 ± .14
0 . 0 0 ± .00
1 .8 1 ± .14
0.43 ± .01
2 .2 4 ± .14
2 .2 8 ± .08
1 .0 1 7 ± .072
1 .8 1 ± .14
0 .9 0 ± .01
2 .7 1 ± .14
2 .7 2 ± .13
1 .0 0 4 ± .071
1 .8 1 ± .14
1 .6 0 ± .01
3 .4 1 ± .14
3.3 3 ± .05
0.977 z .043
0 .7 7 ± .07
0 . 0 0 ± .00
0.77 ± .07
3.33 ± .01
4 .1 0 ± .07
3 .9 9 ± .01
0.973 ± .017
0 .7 7 ± .07
1 .3 6 ± .01
2 .1 3 ± .07
2 .1 0 ± .16
0.986 ± .082
0.12 ± .10
0 . 0 0 ± .00
0 .1 2 ± .10
2 .2 7 ± .01
2 .3 9 ± .10
2 .3 8 ± .22
0 .9 9 5 ± .10
0.12 ± .10
4 .8 8 ± .01
5 .0 0 ± .10
5.00 ± .16
1 .0 0 0 ± .038
0 .1 2 ± .10
8 .8 4 ± .01
8 .9 6 ± .10
8.52 ± .18
0.952 ± .023
a
Pb P r e d ic te d 3
Pb Found3
Pb Found/
Pb P r e d ic te d
1 .8 1 ± .14
0.7 7 ± .07
0.12 ± .10
le a d in fJg/g d ry w e ig h t, d e v ia tio n s a t 95% c o n fid e n c e l e v e l
26
a n a l y t i c a l cu rv es in d ic a te t h a t t h i s sy stem i s r e l a t i v e l y f r e e o f any
m a trix e f f e c t s .
S im ila r a n a l y t i c a l c u rv e s u sin g th e c arb o n rod a to m iz e r
and th e ta n ta lu m b o a t assem bly do n o t e x h i b i t com parable l i n e a r i t y .
These te c h n iq u e s a r e a l s o s e n s i t i v e t o th e volume o f a n a ly te u se d .
T h is i s n o t a problem w ith th e fu rn a c e s in c e th e s o lv e n t i s e v a p o ra te d
p r i o r t o a to m iz a tio n .
The d e te c tio n l i m i t f o r le a d by th e carb o n ro d method i s 20 x
10
—12
g Pb.
37
The ta n ta lu m b o a t p ro c e d u re
38
r e p o r ts a d e te c tio n l i m i t
o f 1 .1 9 (jg/100 ml and an a b s o lu t e ’ s e n s i t i v i t y o f 0 .1 x IO- ^g Pb.
"
23
atom r e s e r v o i r te c h n iq u e h as a d e te c tio n l i m i t o f 0 .3 n g /g .
The
Many o f th e s e o th e r non-flam e te c h n iq u e s e x h i b i t s e v e re ch em ical
and s p e c t r a l in te r f e r e n c e s
22 23 24
9 9
w hich a r i s e p a r t l y from th e com­
p l i c a t e d c h em ica l m a trix o f th e ash ed b i o l o g i c a l sam ple and from th e
method o f d ry in g and a s h in g o f th e sam p le.
In th e c arb o n ro d a to m iz e r
method d ry in g 9 a s h in g and a to m iz a tio n o f th e sample a l l ta k e p la c e on
th e c arb o n rod i n s u c c e s s iv e programmed s t e p s .
T h is i s a n a tte m p t t o
re d u c e a n a ly s is tim e by u s in g n o n - p r e tr e a te d sam ples o f v e ry s m a ll s i z e ,
5-20 j j l .
I n th e c a se o f Pb d e te r m in a tio n in re d b lo o d c e l l s
22
th e
m a trix e lem en ts c an n o t be removed c o m p le te ly d u rin g th e a sh in g c y c le
and th u s a weak nonatom ic a b s o r p tio n s i g n a l i s superim posed on th e
a to m ic a b s o r p tio n p e a k .
A d e u teriu m background c o r r e c t o r has been
s u c c e s s f u lly used t o a l l e v i a t e t h i s p ro b le m .
In th e atom r e s e r v o i r te c h n iq u e
23
th e m a trix e f f e c t s were so
27
s e v e re t h a t a s o lv e n t e x t r a c t i o n was n e c e s s a ry t o a c h ie v e th e e x c e l l e n t
d e te c tio n l i m i t .
The ta n ta lu m b o a t p r o c e d u r e ^ a l s o in c lu d e d a s o lv e n t
e x t r a c t i o n s te p b e fo re th e a n a ly s is co u ld be s u c c e s s f u lly c a r r ie d o u t.
In t h i s p ro c e d u re we have s e p a ra te d th e d i s s o l u t i o n , d ry in g and
a to m iz a tio n s te p s and i t i s b e lie v e d t h a t t h i s r e s u l t s in a m in im iz a tio n
o f any m a trix i n t e r f e r e n c e s .
The a d v a n ta g e s o f t h i s method in c lu d e
s u p e r io r s e n s i t i v i t y , a c c u ra c y and r e p r o d u c i b i l i t y .
T here i s no need
f o r la r g e sa m p le s, te d io u s e x t r a c t i o n o r p r e c o n c e n tr a tio n s t e p s .
S in ce a c id d ig e s t io n i s u s e d , th e method i s r e a d i l y am endable t o th e
a n a ly s is o f any ty p e o f t i s s u e o r bone sam p le .
28
Lead L ev els i n F is h T is s u e s
The r e l a t i o n s h i p betw een f i s h age and le n g th was o b ta in e d from th e
S ta te F is h and Game D e p a rtm e n t^ and i s i l l u s t r a t e d i n T able IV.
T able IV.
F is h Age v s Average Length
F is h Age
1
2
3
4
Average Length
year
y ears
y ears
y e a r s and o ld e r
5 .2 ”
7 .7 "
9 .4 "
1 2 .0 "
These v a lu e s a r e b ased on f i s h sam ples from th e West G a lla t in R iv er
w ith in G a l l a t i n Canyon.
The f i s h sam ples from th e F e d e ra l F ish H atchery
and from th e G a l l a t i n R iv e r i n Y ellow stone Park were grouped by t h e i r
le n g th a lo n e .
The v a lu e s o f th e le a d c o n c e n tra tio n s in th e f i s h t i s s u e s a re
p re s e n te d i n T able V.
f i s h ag e.
The d a ta i s a rra n g e d a c c o rd in g t o in c re a s in g
The t o t a l f i s h w e ig h t, le n g th , lo c a t io n o f c o lle c tio n and
ty p e a re a ls o g iv e n .
A ll v a lu e s were d eterm in ed on th e b a s is o f dry
sample w e ig h t and a re quoted i n [ig P b /g sam ple.
t i o n and f i s h ty p e a re coded.
The lo c a t io n o f c o l l e c ­
The key i s a t th e bottom o f th e t a b l e .
The ran g e o f le a d v a lu e s i s q u ite l a r g e ; from 0.0 7 /ig /g i n th e l i v e r
t i s s u e o f a Rainbow T ro u t from th e H atch ery t o 7.65 /ig /g i n th e l i v e r
t i s s u e o f a Rainbow T ro u t from th e West G a lla t in so u th o f Greek C reek.
29
T able V.
L ength
In c h e s
E s t.
Age
W eight
grams
6 .0
I
7 3 .4
6 .5
I
7 .0
Lead L e v e ls in F is h T iss u e
L o c a tio n
Type
Jig P b/g sam ple
Bone
G ill
L iv e r
Muscle
C3/R
. 1 .3 6
1.3 2
2 .2 5
—
7 0 .0
C2/R
3 .9 6
1 .2 4
1-2
70.2
C l/R
7 .5
1-2
95.6
C2/R
4 .0 3
7 .5
1-2
9 0 .4
C3/R
8 .0
2
8 9 .0
8 .0
2
8 .0
-
6 .6 1
3.07
'
1 .9 4
0 .2 0
0 .57
0 .5 0
7 .6 5
C2/R
4 .0 4
3.0 4
2 .9 5
1 0 6 .0
C3/R
3 .4 9
5.72
2 .1 0
2
9 9 .6
H/R
0.3 3
0.42
0.0 7
8 .5
2
1 23.2
h/ r
2 .4 5
0 .3 1
0 .9 6
8 .5
2
1 1 6 .5
C3/R
1 .5 3
1 .1 6
0 .97
9 .0
2-3
1 2 3 .4
C4/CT
2 .6 7
2.33
0 .8 0
9 .2 5
2 -3 ,
P5/CT
2 .2 5
1 .7 8
1 .6 3
1 5 5.7
—
0 .7 4
9 .5
3
1 6 0.3
C l/R
3 .5 6
2 .1 2
1 .9 6
9 .5
3
1 5 4 .6
C l/R
3 .3 3
2 .5 6
2 .0 2
9 .7 5
3
1 5 2 .4
C l/R
6.6 2
---- —
1 .1 1
1 0 .0
3
2 0 1 .0
C2/R
■ 6 .2 9
1 0 .0
3
1 8 0 .0
H/R
0 .4 3
0 .7 1
0 .1 0
•------ -
1 0 .0
3
1 9 2 .4
H/R .
0 .6 1
1 .0 5
1 .3 1
0.12
1 0 .5
3
2 6 0 .0
P5/CT
2 .5 0
. 1 .6 5
1 .4 2
. 3-4
2 2 0 .6
C3/R
4 .8 1
0.76
2 .1 4
3-4
2 9 4 .5
C3/R
2 .7 2
2 .1 1
1 .2 7
1 1 .0
1 1 .0
.
'-------
------ -
---- ----
30
T able V. ( c e n t . )
A*g Pb/g sample
L ength
In ch es
E s t.
Age
W eight
grams
L o c a tio n
Type
Bone
G ill
L iv e r
Muscle
1 1 .5
3-4
2 9 4 .1
C3/R
2 .7 6
2.0 7
1 .3 0
1 1 .5
3-4
3 07.1
C4/R
3 .3 0
4 .2 1
1 .7 0
1 .3 4
1 1 .5
3-4
172.2
C2/R
2 .5 5
2 .4 3
1 2 .0
4
390.6
C4/B
5.1 1
2 .5 3
1 .4 3
1 .4 0
1 2 .0
4
3 05.0
P5/CT
3 .1 5
3.0 6
2 .8 9
1 2 .5
4
4 5 3 .4
C2/R
4 .9 7
2 .5 5
1 2 .5
4
343.5
C l/R
5.5 6
1 3 .0
4-
540.0
P5/W
2 .5 6
2 .0 0
2 .4 3
1 4 .0
4-
4 3 6 .4
C3/W
4 .3 5
1.4 7
0 .2 1
1 4 .0
4-
4 6 0 .5
G/B
4 .4 5
3 .1 1
3.2 8
1 4 .0
4-
4 4 3 .0
C l/R
6 .0 8
1 .0 8
1 4 .5
4-
7 50.0
P5/B
3 .2 0
2 .1 2
2 .0 0
1 5 .0
4-
4 15.6
C4/R .
6 .2 0
7 .4 0
2 .5 0
1 5 .5
4-
558.2
C3/R
2 .7 8
2 .7 8
0 .9 7
1 8 .0
4-
7 40.0
G/R
4 .9 8
6.55
C
P
H
G
-
Canyon
P ark
H atchery
N o rth o f G a l l a t i n Gateway
R - Rainbow T ro u t
B - Brown T ro u t
CT - C u tth r o a t T ro u t
W - W h ite fis h
1
2
3
4
5
------0.77
-
1 .8 1
South o f C ascade C reek
Near Mouth o f Cascade Creek
South o f G reek C reek
Near Mouth o f P o rcu p in e C reek
N ear Mouth o f Fan Creek
31
T ab le VI p r e s e n ts th e a v e ra g e le a d l e v e l s f o r a l l t i s s u e sam p les,
a rra n g e d by l o c a t i o n and t i s s u e t y p e .
t i s s u e s a n a ly z e d a r e a l s o g iv e n .
The ra n g e s and th e number o f
S in c e th e le a d v a lu e s a r e grouped
t o g e t h e r , r e g a r d le s s o f th e age o f th e f i s h , th e ra n g e s a r e a p p re c ia b ly
la rg e .
In m ost c a s e s th e s ta n d a rd d e v ia tio n i s l a r g e r o r a t l e a s t com­
p a ra b le t o th e a v e ra g e v a lu e .
S e v e ra l o f th e in d iv id u a l le a d .v a lu e s
d e v ia te from t h e i r means by more th a n tw ic e th e s ta n d a rd d e v ia tio n .
T h is c l e a r l y in d ic a te s t h a t f u r t h e r d i f f e r e n t i a t i o n on th e b a s is o f
age m ust be made.
T able V I.
A verage Lead L e v e ls i n T iss u e s by L o c a tio n
Canyon F is h
T iss u e
A v e ra g e ^
Range
Number o f Samples
Bone
3 .8 6 ± 1 . 6 4
0 .5 7 - 6.62
24
G ill
2 .4 7 ± 1 .6 6
0 .5 0 - 7 .4 0
22
L iv e r
2 .1 1 ± 1 .9 7
0 .2 0 - 7 .65
19
P a rk F is h
T iss u e
A verage ^a ^
Range
Number o f Samples
Bone
2 .7 3 ± .41
2 .2 5 - 3 .2 0
5
G ill
2 .1 2 ± .55
1 .6 5 - 3 .0 6
5
L iv e r
2 .0 2 ± .59
1 .4 2 - 2 .8 9
5
H a tch e ry F is h
T iss u e
Bone
G ill
L iv e r
-
A v e rag e ' '
Range
Number o f Samples
0 .9 5 ± 1 .0 0
0 .3 3 - 2 .4 5
4
0.62 ±
.33
0 .3 1 - 1 .0 5
4
0 .6 1 ±
.62
0 .07 - 1 .3 1
4
^lead v a lu e s in jjg /g , dry w e ig h t
32
However, some g e n e r a l te n d e n c ie s a r e in d ic a te d .
The h ig h e s t le a d
l e v e l o c cu rs i n bone t i s s u e o f f i s h fro m .w ith in th e Canyon.
T his
v a lu e , 3 .8 6 ± 1 .6 4 jUg/g, i s h ig h e r th a n th e le a d bone c o n te n t o f f i s h
from th e P a rk , 2 .7 3 ± A l jLZg/g and sam ples from th e H a tc h e ry , 0 .9 5 ±
1 .0 0 p g /g .
T h is same g e n e r a l tr e n d i s a l s o in d ic a te d i n th e g i l l and
liv e r tis s u e s .
T h at i s , th e a v e ra g e le a d c o n c e n tr a tio n in g i l l t i s s u e
i s g r e a t e s t i n Canyon sam ples a t 2 .4 7 ± 1 . 6 6 IJlgfg3 w h ile th o s e from
th e P ark show a g i l l le a d c o n te n t o f 2 .1 2 ± .55 [Jg/g.
The a v erag e
le a d l e v e l in g i l l t i s s u e o f H atch ery f i s h i s 0.62 ± .33 y g / g .
In l i v e r
t i s s u e th e a v e ra g e le a d c o n c e n tr a tio n s a r e 2 .1 1 ± 1 . 9 7 , 2 .0 2 ± .59 and
0 .6 1 ± .62 y g f g f o r Canyon, P ark and H atch ery sam ples r e s p e c t i v e l y .
Two o th e r im p o rta n t tr e n d s a r e in d ic a te d from th e d a t a .
F irs t,
t h e a v e ra g e v a lu e s o f th e le a d t i s s u e c o n c e n tra tio n s o f H atch ery f i s h
a r e on th e o rd e r o f t h r e e t o f o u r tim e s lo w er th a n v a lu e s in s im ila r
t i s s u e s o f f i s h from th e G a l l a t i n R iv e r .
S eco n d ly , w ith in each d e s ig ­
n a te d l o c a t i o n th e le a d c o n c e n tr a tio n in bone t i s s u e i s th e h i g h e s t ,
fo llo w e d by g i l l , and th e lo w e s t v a lu e s a r e e x h ib ite d by l i v e r t i s s u e .
T h is im p lie s t h a t more o f th e m e ta l i s b e in g s to re d in th e s k e le to n
th a n i n th e o th e r o rg a n s .
These le a d c o n c e n tr a tio n s a r e in good a g re e ­
m ent w ith o th e r v a lu e s o f l e a d . i n f i s h sa m p le s, in t h a t th e y a re a l l i n
th e p .p .m . r a n g e .
V alues r e p o rte d f o r salm on. Tuna, o y s te r and s a r d in e
a r e 1 . 3 , 2 . 2 , 1 .2 , and 4 .3 ^ g /g r e s p e c t i v e l y .
13
O th e rs a r e l o b s t e r ,
2 .5 0 y g / g 3 f r e s h s h rim p , 0 .4 5 y g / g and s c a l l o p s , 0 .1 3 y g f g .
12
These
33
l a s t th r e e v a lu e s a r e on th e b a s is o f w et w eig h t and r e f l e c t low er le a d
c o n c e n tr a tio n s .
In o rd e r t o lo o k more c lo s e ly a t th e d a ta , T a b le s V I I , V I I I , and
IX p r e s e n t th e a v e ra g e le a d t i s s u e c o n c e n tra tio n s grouped a c c o rd in g to
l o c a t i o n , t i s s u e ty p e and age b r a c k e t .
W ithin each age g ro u p , f i s h
from th e Canyon a re a e x h i b i t th e h ig h e s t le a d c o n c e n tr a tio n s in bone
t i s s u e , T able V I I .
F o r exam ple, in th e 2-3 y e a r - o ld b r a c k e t th e a v e r ­
age le a d l e v e l i s 4 .4 9 ± 2 .1 0 jjg /g , w h ile bone t i s s u e o f P ark sam ples
show a 2 .3 7 pig/g l e v e l .
low er a t 0 .5 2 j jg /g .
The H atch ery f i s h bone t i s s u e c o n te n t i s much
The d a ta a l s o i n d i c a t e t h a t th e r e i s a n accum ula­
t i o n o f le a d w ith a g e .
T ab le V I I .
Lead i n Bone T issu e
Bone T issu e
Canyon
E s t . Age
-Average
H atchery
P ark
# sam ples
A verage
1-2
2 .7 1 ± 1 . 4 9
7
2-3
4 .4 9 ± 2.10
5
2 .3 7
3 -4
3 .9 6 ± 1 .4 4
8
4-
4 .8 5 ± 1 .6 0
4
# sam ples
A verage # sam ples
1 .3 9
2
2
0 .5 2
2
3 .1 5
I
—
—
2 .8 8
2
SM W H
I
In sam ples from th e Canyon a re a th e a v e ra g e v a lu e o f le a d in 1-2 y e a r
o ld f i s h i s 2 .7 1 ± 1 .4 9 p g /g .
4 y e a rs and o l d e r .
T h is v a lu e i s 4 .8 5 ± 1 .6 0 ^ g /g in f i s h
A lthough th e sam ples in th e 2 -3 y e a r old group a r e
h ig h e r th a n th o s e i n th e 3 -4 y e a r old b r a c k e t, th e s ta n d a rd d e v ia tio n
i s la r g e in d ic a tin g t h a t some o f th e v a lu e s may be o u t o f l i n e f o r t h i s
34
age g ro u p .
In bone t i s s u e s from P ark f i s h th e tr e n d o f in c r e a s in g le a d
l e v e l s w ith in c r e a s in g age i s a l s o d e m o n stra te d .
However, th e r e i s
only I bone t i s s u e sam ple i n th e 3 -4 y e a r old g ro u p .
I t s v a lu e , 3 .1 5
p g /g , does n o t f i t t h i s t r e n d , b u t s in c e i t i s only one sam ple th e
v a lu e may n o t be r e p r e s e n t a t i v e .
In H atch ery sam ples th e r e a p p e a rs
t o be a d e c r e a s in g c o n c e n tr a tio n o f le a d w ith in c r e a s in g a g e .
th e r e a r e two t i s s u e sam ples in each age g ro u p .
However,
The v a lu e o f 1 .3 9 ^ g /g
i n th e 1-2 y e a r old age b r a c k e t i s an a v e ra g e o f 0.33 and 2 .4 5 ^ g /g .
The h ig h v a lu e o f 2 .4 5 ng/g may i n d i c a t e a n anomaly i n an in d iv id u a l
fis h .
The a c c u m u la tio n o f le a d w ith age h as been w e ll e s ta b lis h e d in
hum ans,
12
w ith th e h ig h e s t v a lu e s o c c u rrin g in th e f i f t h t o s ix t h
decade o f l i f e i n human o rg a n s .
I n g i l l t i s s u e , w ith in each age g ro u p , th e le a d c o n c e n tra ­
t i o n s a r e a g a in g r e a t e s t i n f i s h from th e Canyon.
i n T able V I I I .
T h is i s i l l u s t r a t e d
T his g e n e r a l tr e n d i s b ro k en in th e 3 -4 y e a r old age
b r a c k e t , where th e g i l l t i s s u e o f one sam ple from th e P ark shows a
h ig h e r le a d l e v e l th a n th e a v e ra g e v a lu e f o r Canyon f i s h .
The le a d
l e v e l o f 3 .0 6 ^g/g i s w ith in th e ra n g e o f th e le a d c o n te n t o f Canyon
s a m p le s .
The le a d c o n c e n tr a tio n in th e g i l l o f t h i s one f i s h may: n o t
be i n d i c a t i v e o f th e t r u e l e v e l a s i t i s th e same f i s h t h a t e x h ib ite d
th e h ig h le a d bond l e v e l o f 3 .1 5 jj g /g .
The H atch ery t i s s u e s show a
much lo w er le a d c o n c e n tr a tio n a t 0 .3 6 and 0.88 jjg/g i n r e s p e c tiv e age
35
g ro u p s .
The g i l l t i s s u e s in T able V III d e m o n strate a s l i g h t
te n d en c y t o accu m u late le a d w ith a g e .
The Canyon Samples in c re a s e
from 2 .2 s ± 1 .3 9 j^g/g i n young f i s h t o 3 .1 8 ± 2 .9 0 ^ g /g in o ld e r f i s h .
The H atchery show th e same t r e n d .
There i s a d e f i n i t e in c r e a s e from
0 .3 6 t o 0.88 jrg/g in t h e i r g i l l t i s s u e s .
a p p e a r tc) in c r e a s e w ith a g e .
The le a d l e v e l s in P ark f i s h
The one v a lu e , 3 .0 6 ^ g f g , from th e 3 -4
y e a r old age group i s o u t o f l i n e a s in d ic a te d b e f o r e .
T ab le V I I I .
Canyon
Lead in G i l l T issu e
G i l l T issu e
P a rk
H atch ery
E s t . Age
A verage
# sam ples
1—2
2 .2 5 ± 1 .3 9
7
-------------
—
0 .3 6
2
2 -3
2 .3 3 ±
.21
3
1 .7 1
2
0 .88
2
3 -4
2 .3 8 ± 1 .0 1
7
3 .0 6
I
—
—
4-
3.'18 ± 2 . 9 0
4
2 .0 6
2
A verage
# sam ples
A verage # sam ples
T ab le IX i l l u s t r a t e s th e le a d c o n c e n tr a tio n s in l i v e r t i s s u e .
In
th e 2-3 y e a r o ld age b r a c k e t th e le a d l e v e l s o f Canyon f i s h and P ark
sam ples a r e n e a r ly e q u iv a l e n t, 1 .4 7 ± .6 1 p g /g and 1 .5 2 ^ g /g r e s p e c t i v e l y
H atch ery sam ples i n th e same age gro u p e x h i b i t a l e v e l o f 0 .7 0 p g /g
which i s a p p ro x im a te ly o n e - h a lf o f th e above v a lu e s .
Lead a ccu m u lates
w ith age i n H atch ery sam ples and f i s h from th e P a rk .
There a p p e a rs t o
be r e v e r s a l o f t h i s tr e n d in f i s h l i v e r t i s s u e from th e Canyon.
1 -2 y e a r o ld age group th e l e v e l i s 3 .3 9 ± 2 . 8 1 jig /g .
In th e
Two v a lu e s ,
36
6 .6 1 p g /g and 7 .6 5 p g /g a r e in c lu d e d i n t h i s a v e r a g e .
These v a lu e s 9
e s p e c ia ll y in t h i s ag e g ro u p , seem u n re a so n a b ly h ig h . . I f th e y were t o
be ig n o re d th e a v e ra g e would drop t o 1 .2 8 ± 1 .1 0 f j g / g .
T h is many
r e p r e s e n t a more r e a l i s t i c le a d l e v e l f o r f i s h l i v e r t i s s u e in t h i s
age b r a c k e t.
The o ld e r f i s h from th e Canyon fo llo w th e tr e n d more
c lo s e ly .
T a b le IX .
Lead i n L iv e r T issu e
L iv e r T issu e
Canyon
: . Age
A verage
P ark
# sam ples
1—2
3 .3 9 ± 2 .8 1
7
2 -3
1 .4 7 ±
.61
4
3 -4
1 .5 7 ±
.35
4-
1 .2 3 ± 1 .1 6
A verage
H atchery
# sam ples
Average
# sam ples
—
0 .5 1
2
1 .5 2
2
0 .7 0
2
5
2 .8 9
I
3
2 .2 1
2
—
I n o rd e r t o o b ta in more in fo rm a tio n a b o u t th e f i s h le a d e n v iro n ­
m ent, w a te r sam ples from th e R iv e r and H atch ery were a n a ly z e d f o r th e
m e ta l.
The w a te r u sed a t th e f i s h H atch ery i s from n a t u r a l h o t and c o ld
w a te r s p r i n g s .
c o v e re d .
The w a te r i s s to r e d i n la r g e h o ld in g ta n k s which a r e
T here i s a m ixing ta n k f o r th e warm and c o ld w a te r so t h a t a
c o n s ta n t te m p e ra tu re i s m a in ta in e d .
The w a te r i s p ip ed t o tro u g h s
where th e f i s h a r e m a in ta in e d and f e d .
W ater sam ples w ere c o lle c te d
from lo c a tio n s a lo n g th e W. G a l l a t i n and from d i f f e r e n t a re a w ith in
th e h a tc h e ry co m p les.
These v a lu e s a r e p re s e n te d in T ab le X.
37
T able X.
Lead L evels in R iv e r and H atchery W ater
L o catio n
Lead C o n c e n tra tio n s
ng/m l
W. G a lla t in R iv e r
N orth o f G a lla t in Gateway N orth o f Cascade Creek
South o f Cascade Creek
N orth o f Greek Creek
South o f Greek Creek
P o rcu p in e Creek Area
Fan Creek Area
2.9 3
2 .56
2 .9 4
2.65
2.96
2.81
2.65
F is h H atchery
Hot S p rin g s
Mixed Hot and Cold S p rin g s
Mixed Hot and Cold S p rin g s
F is h Tank
F is h Tank
1.38
1.4 2
1.41
1.39
1 .4 4
The le a d c o n c e n tr a tio n s in th e R iv e r a re a t th e low ra n g e o f v a lu e s
e x h ib ite d i n o th e r r i v e r s i n th e U. S .^
The n a t i o n a l av erag e le a d
l e v e l i n m ajo r r i v e r s i s 23 n g /m l, w ith <a. minimum v a lu e o f 2ng/m l.
The a v erag e v a lu e o f le a d i n th e w a te r from th e West G a lla t in i s
2 .8 1 n g /m l.
These le a d w a te r l e v e l s a re th r e e o rd e rs o f m agnitude
lo w er th a n th e le a d c o n c e n tra tio n t h a t w i l l have damaging e f f e c t s on
th e f i s h .
A le a d c o n c e n tra tio n o f 25 /Ltg/ml w i l l r e t a r d Brook T ro u t
grow th.
A t 10 Mg/ml th e r e i s no e f f e c t on t h e i r grow th.
SUMMARY AND CONCLUSIONS
A n a ly tic a l Method
The a n a l y t i c a l method p re s e n te d in th e p r o j e c t d e m o n strates t h a t
le a d can be a n a ly z e d a t t r a c e c o n c e n tr a tio n s in b i o l o g i c a l sam ples
u s in g an a c id d ig e s t io n p ro c e d u re and fu rn a c e ato m ic a b s o r p tio n .
The
a n a ly s is was d i r e c t - th e r e was no need f o r e x t r a c t i o n o r p r e c o n c e n tra ­
tio n s te p s .
From s ta n d a rd a d d itio n d a ta th e a c c u ra c y o f th e method was
e v a lu a te d a t 99% ± 6%.
M atrix in te r f e r e n c e s were a b s e n t.
T his was
in d ic a te d by th e s i m i l a r s lo p e s o f th e th r e e a n a l y t i c a l s ta n d a rd c u r v e s .
The u se o f s m a ll sam ples makes th e method a p p lic a b le t o th e a n a ly s is o f
o th e r ty p e s o f b i o l o g i c a l sam ples where s iz e may be th e m ost c r i t i c a l
c o n s id e r a tio n .
S ince a c id d ig e s t io n was u s e d , th e method i s am enable
t o th e a n a l y s i s o f o th e r m e ta ls , b o th th o s e c o n sid e re d t o x i c t o b i o -I o g ic a l system s and th o s e w hich a r e n e c e s s a ry f o r th e o p tim a l f u n c tio n
o f g ro w th , h e a lin g and th e a c t i v i t y o f many m e ta b o lic p r o c e s s e s .
The
e x c e l l e n t s e n s i t i v i t y and low d e te c tio n l i m i t makes th e fu rn a c e ato m ic
a b s o r p tio n te c h n iq u e one o f th e m ost ad v an tag eo u s m ethods f o r th e
a n a ly s is o f t r a c e m e ta ls i n b i o l o g i c a l s y s te m s .
Lead L e v e ls i n F is h T is s u e s
S e v e ra l g e n e r a l tr e n d s in th e le a d t i s s u e c o n c e n tr a tio n s o f f i s h
have been e s ta b lis h e d a s a r e s u l t o f t h i s s tu d y .
F i r s t , Canyon f i s h
e x h i b i t h ig h e r le a d c o n c e n tr a tio n s in t h e i r b o n e , g i l l and l i v e r
t i s s u e s th a n f i s h from th e Park and th e F is h H a tc h e ry .
Second, H atch ery
le a d t i s s u e l e v e l s a r e a p p ro x im a te ly th r e e t o fo u r tim e s low er th a n
39
s im ila r t i s s u e s from th e P ark and th e Canyon f i s h .
th e r e s u l t o f t h e i r c lo s e ly c o n tr o lle d environm ent.
t o accum ulate w ith age i n a l l th r e e t i s s u e s s tu d ie d .
T h is i s undoubtedly
T h ird , le a d a p p e a rs
F o u rth , th e le a d
c o n c e n tr a tio n s a re h ig h e s t i n bone t i s s u e , fo llo w ed by g i l l t i s s u e and
liv e r tis s u e .
T h is i n d i c a t e s t h a t more le a d i s s to re d i n th e s k e le to n
th a n i n th e o th e r two o rg a n s.
A U o f th e v a lu e s a re i n th e p .p .m . ran g e which i n d i c a t e s t h a t
t h e r e i s n o t a s i g n i f i c a n t s t r e s s in any o f th e f i s h due to le a d .
The
d a ta from T able VI show t h a t th e le a d bone t i s s u e c o n c e n tra tio n in
Canyon f i s h i s 1.41 ± .6 3 tim e s h ig h e r th a n bone t i s s u e from f i s h i n th e
P ark .
In g i l l t i s s u e th e Canyon sam ples have a le a d l e v e l 1.16 ± .8 4
tim e s h ig h e r th a n P ark f i s h .
The Canyon f i s h have a l i v e r le a d concen­
t r a t i o n 1.05 ± 1 .0 3 tim e s h ig h e r th a n P ark sam ples.
I t i s im p o rta n t to
n o te h e re t h a t th e s ta n d a rd d e v ia tio n s a re la r g e a s t h i s d a ta was com­
p i l e d w ith o u t re g a rd t o f i s h ag e.
I t would be ad v an tag eo u s t o have a
v e ry la r g e number o f sam ples so t h a t a more com plete s t a t i s t i c a l p i c t u r e
co u ld be drawn.
However, i t i s f e l t t h a t i f th e r e were a la r g e number
o f sam p les, th e s t a t i s t i c s would ch an g e, b u t th e g e n e r a l tr e n d would
rem ain.
T hat i s , f i s h sam ples from w ith in th e Canyon have s l i g h t l y
h ig h e r le a d l e v e l s .
In two r e c e n t s t u d i e s , e x p e rim e n ta l an im als were exposed to a i r ­
borne le a d c o n c e n tr a tio n s e q u iv a le n t t o th o s e found i n u rb an a r e a s .
was d eterm in ed t h a t th e in h a la tio n o f u rb an a i r c o n ta in in g on th e
It
40
g
a v e ra g e o f 2 .5 yg Pb/m
le a d l e v e l s .
17
can r e s u l t in s i g n i f i c a n t e le v a t i o n o f bone
However, t h e i r s tu d ie s d id n o t d is c l o s e any a d v erse
42
Mice
th a t
3
were exposed t o le a d l e v e l s betw een 2 .6 and 1 5 .6 ^g Pb/m f o r a 1 5 h e a lt h e f f e c t s in th e a n im a ls b ecau se o f t h i s e x p o s u re .
month p e rio d showed le a d bone l e v e l s betw een 1 4 .3 jijg/g and 2 0 .9 jjg/g
(d ry w e i g h t ) .
These v a lu e s a r e s i g n i f i c a n t when compared t o an a v erag e
v a lu e o f 1 0 .2 fjg/g i n c o n tr o l a n im als exposed to an a ir b o r n e le a d
l e v e l o f 0.32 (jg/rn^.
The f i s h w ith in th e Canyon a r e exposed to le a d e x h a u s t e m issio n
from th e t r a f f i c lo a d o f 1000 v e h ic le s d a i l y .
The s l i g h t l y h ig h e r
le a d l e v e l s i n th e t i s s u e s o f th e s e f i s h i s b e lie v e d t o b e th e r e s u l t
o f t h i s e x p o s u re .
t h i s tim e .
The a c t u a l e f f e c t o f th e ex posure i s m inim al a t
I f th e t r a f f i c lo a d were t o in c r e a s e g r e a t l y th e n i t
would be e x p e c te d t h a t th e t i s s u e c o n c e n tr a tio n s would in c r e a s e a l s o .
■I n c o n c lu s io n , an im p o rta n t a s p e c t , p erh ap s th e m ost im p o r ta n t,
i s t h a t th e le a d l e v e l s in a b i o l o g i c a l system e s s e n t i a l l y f r e e from
th e u rb an e n v iro n m e n ta l l e v e l , have b een e s t a b l i s h e d .
These le v e l s
may be u sed a s th e b a s is o f f u r t h e r s tu d y and e v a lu a tio n o f th e le a d
e c o lo g i c a l p ro b lem .
9
■
LITERATURE CITED
1.
S. K. H a ll, E n v iro n . S c i. T e c h n o l., _6, 30 (1 9 7 2 ).
2.
P. B ry ce-S m ith , Chem. B r i t . , 7_, 56 (1 9 7 1 ).
3.
C. C. P a tte r s o n , A rch. E n v iro n . H e a lth , 11, 344 (1 9 6 5 ).
4.
C. C. P a tte r s o n , T. D. S a lv ia , S c i. C i t i z . , 10_, 66 (1 9 6 6 ).
5.
"Survey o f Lead i n th e Atmosphere o f Three Urban C om m unities,"
U. S. D epartm ent o f H e a lth , E d u c a tio n and W elfare, P u b lic H e alth
S e rv ic e P u b lic a tio n No. 999-AP-12 (1 9 6 5 ).
6.
M. T. S t e r b a r , Symposium o f E n v iro n m en tal Lead C o n tam in atio n ,
P u b lic H e a lth S e rv ic e P u b lic a tio n No. 1440 (1 9 6 6 ).
7.
G. L. T e rH a rr, M. A. B aynard, N a tu re , 232, 533 (1 9 7 1 ).
8.
A. L. M ills , Chem. B r i t . , 7_, 160 (1 9 7 1 ).
9.
R. H. D aines , H. M otto, D. M. C h ilk o , E n v iro n . S c i. T e c h n o l., 4 ,
318 (1 9 7 0 ).
10.
H. L. M o tto, R. H. D aines , D. M. C h iIk o , C. K. M otto, E nviron.
S c i. T e c h n o l., 4 , 231 (1 9 7 0 ).
11.
R. J . B a z e ll, S c ie n c e , 1 7 3 , 130 (1 9 7 1 ).
12.
H. A. S c h re o d e r, e t
13.
J . C. M eranger, E. Somers, B u ll. E n v iro n . Contam. T o x ic o l., 3_,
360 (1 9 6 8 ).
14.
R. A. K ehoe, I n s t . P u b l. H e a lth H ygiene, 24 (1 9 6 1 ).
15.
H. A. S c h re o d e r, A. P. Nason, C lin . Chem., 17_, 461 (1 9 7 1 ).
16.
E. B. S a n d e li, C o lo rim e tric D e term in atio n o f T rac e s o f M e ta ls ,
I n te r s c ie n c e Pub. I n c . , p . 388 (1 9 5 9 ).
17.
R. G. S m ith, J . S z a jn a r , L. H eck er, E n v iro n . S c i. T e c h n o l., 4 , 333
(1 9 7 0 ).
18.
O f f i c i a l Methods o f A n a ly s is , 1 0 th e d . , A ssoc. O ff. A rg. Chem.,
p . 367 (1 9 6 5 ).
a l . , - J . Chron. D i s ., 1 4 , 408 (1 9 6 1 ).
42
19.
H. A. S c h re o d e r, J . H. T ip to n , Arch. E n v iro n . H e a lth , 17
(1 9 6 8 ).
—
20.
J . B. W i llis , A nal. Chem. , 34, 614 (1 9 6 2 ).
21.
H. T. D e lv e s, A n a ly s t, 95, 431 (1 9 7 0 ).
22.
J . P. M atousek, B„ J . S te v e n s, C lin . Chem.,
23.
D. A. S e g a r, J . L. G i l i o , A b s tr a c ts , P itts b u r g h C onference on
A n a ly tic a l C hem istry, C le v ela n d , O hio, March 1972, p a p e r number 35
24«
J . Y« Hwang, S. B. S m ithe, e t . a l . , i b i d , p a p e r number 37.
25.
R. B e r s in , e t a l . , Symposium on Trace C h a r a c te r iz a tio n , N a tio n a l
Bureau o f S ta n d a rd s , O ctober 1966.
26.
C. B e rk le y , et^ a l . , T r a n s a c tio n s o f th e New York Academy o f
S c ie n c e s , _3, 331 (1 9 6 7 ).
27.
E. R e is e r , T r a f f ic Volume o f Highway 191, p r i v a t e com m unications,
Montana S ta te Highway D epartm ent, Bozeman, Montana.
28.
E. R. V in c e n t, P r o je c t B io lo g is t, F is h Length and Age, p r iv a te
com m unication, Montana S ta te F is h and Game D ep artm en t, Bozeman,
M ontana.
29.
R. W o o d riff,
G. Ramelow, S pectrochim . A c ta ., 2 3 4 , 665 (1 9 6 8 ).
30.
R. W o o d riff,
R. S to n e, AppI . O pt. , 7_, 1337 (1 9 6 8 ).
31.
R« W o o d riff,
R. S to n e , A. H eld , Appl. S p e c tro s c . , 22_, 408 (1 9 6 8 ).
32.
R. W o o d riff,
250 (1 9 7 0 ).
B. R. C u lv e r, K. W. O lse n , A ppl0 S p e c tro s c . ,
33.
R. W o o d riff, D. S h ra d e r, A nal. Chem., 43, 1918 (1 9 7 1 ).
34.
R. W o o d riff, J . Lech, A nal. Chem., 44, 1323 (1 9 7 2 ).
35.
J . Lech, F urnace Tem perature in Lead A n a ly s is , p r i v a t e communica­
t i o n s , C hem istry D epartm ent, Montana S ta te U n iv e r s ity , Bozeman,
Montana.
36.
T. F. G orsuch, A n a ly s t, 84, 135 (1 9 5 9 ).
17, 363
965
(1 9 7 1 ).
24,
43
37.
M. D. Amos, e t
a l . , A nal. Chem. , 43, 211 (1 9 7 1 ).
38.
H. T. S e lv e s , A n a ly s t, 95_, 431 (1 9 7 0 ).
39.
R. 0 . B ecker, J . A. S padaro, 'E. W. B erg, Bone and J o i n t Surg. ,
50A, 326 (1 9 6 8 ).
40.
J . F. Kopp, R. C. K ro n e r, F e d e ra l W ater P o llu tio n C o n tro l Asso­
c i a t i o n , D iv is io n o f P o ll u tio n S u r v e illa n c e , C i n c i n a t t i , Ohio
(1 9 7 0 ).
41.
D. Dorfman, W. R. W hitw orth, J . F is h . Res. Bd. Can.
(1 9 6 9 ).
42.
F. G. H e u te r, G. L. C o u te r, et^ ‘ a l . , Arch. E n v iro n . H e a lth , 12,
553 (1 9 6 6 ).
^
—
#
26, 2493
—
M
ONTiM
i ___
3 1762 10015080 2
t e n n is I
con. 2
^ n o l - n o l r’ o f f i r - h
tis su e for trace
amounts o f le a d
NAMC
HH So 98
we ^
ANC
A O O H t^ l
H * \t
'
__________