The determination of moisture, silica, iron, aluminum, calcium, and magnesiuim in the lungs of normal
and lunger sheep
by Elwood P Wilson
A THESIS Submitted to the Graduate Committee in partial fulfillment of the requirements for the
Degree of Master of Science in Chemistry
Montana State University
© Copyright by Elwood P Wilson (1936)
Abstract:
Both the experimental data for the testing of the methods of analysis and the actual results when these
methods were applied have been included in this problem.
1. The "sliming", the nitric and perchloric acids, and the ashing methods of removing the organic
matter have all been tried and the ashing method used because of its applicability to the problem.
Occasionally the nitric and perchloric acids method was used to check the silica content as procured
when the ash was dehydrated with HCl.
2. It was found that the phosphate had to be removed in order to make accurate analysis. Both the
ammonium molybdate and the tin methods of removing the phosphate were tried, and the latter found
the more applicable as well as giving the better results.
3. The tin method of removing phosphate has for the first time been shown satisfactory for quantitative
work. Table VI shows its reliability in presence of relatively large amounts of phosphate and small
amounts of Al, Ca, Mg and Fe. The molybdate method does not seem to be satisfactory (see Table IV)
for the material in question where small amounts of Al, Ca, Mg and Fe must be determined accurately.
4. One of the most outstanding differences observed between normal and "lunger" lungs is that of the
heavier weight of the latter's dried material. Although the percentage moisture is about the same in both
cases, the "lunger" dry weight is 3.7 times heavier than that of the normal.
5. In the "lunger" lungs the calcium is about 2 times, magnesium about 1 1/2 times, and iron about 2
times as much as in normal lungs of the same size dry sample, while the silica and aluminum content
seem to be about the same in both cases. When the weights of the mineral content of the entire dried
lungs were calculated they were much greater in the diseased than in the normal lungs. The average of
the mineral content of the total dried "lunger" lungs contains approximately the following over that in
the normal lungs: silica 4 times, iron 5 times, aluminum 3 times, calcium 6 times, and magnesium 5
times.
6. Since blood contains considerable iron and calcium, the quantity of these two found in the analysis
of the lungs would vary with the amount of blood present. The excess blood in the lungs was kept very
low by observing precautions when removing the lungs from the sheep so as to keep its content as low
as possible, 7. It is beyond the scope of the present problem to make any conclusions as to whether or
not the increase in mineral content of the "lunger" over normal lungs could be the cause of the "lunger"
disease of sheep, To determine whether the increased mineral content is due to inhaled dusts or
pathological deposits is also excluded from the present investigation.
THE DETERMIIfATIOH OF MOISTURE, SILICA, IROH
ALUHHUM, CALCIUM, MD MAGHESIUM IU THE
LUHGS OF HORMAL AHD "LUHGERv SHEEP
■b y
EIwood P . W ils o n , J r ,
A THESIS
S u b m itte d t o t h e G ra d u a te Comm ittee 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 t h e D egree o f M a s te r o f S c ie n c e
i n C h em istry a t M ontana
S t a t e C o lle g e
A pproved:
____
? JisfA
I n Charge o f M ajor Wor.
/C h a irm a n G raduate^C om m ittee
Bozeman, M ontana
Ju n e , 1936
H 31 B
Urj, 3
-
2-
TABLE OF COIJTElTTS
Page
I.
II.
III.
IV .
V.
V I.
In tro d u c tio n
3
Review o f t h e L i t e r a t u r e
6
C o n d itio n s U nder TJhich Sheep Were R a ise d
17
P r e p a r a t io n o f Sam ples
A.
M ethod o f Removing Lungs
17
B.
D ry in g and G rin d in g
17
C.
Sam pling and P r e s e r v in g
18
M ethods o f A n a ly se s
A.
Q u a l i t a t i v e A n a ly s is
18
B.
Q u a n t i t a t i v e A n a ly s is
18
T a b u la tio n and D is c u s s io n o f D ata
A.
Q u a l i t a t i v e A n a ly s is
24
B.
Q u a n t i t a t i v e Ash D e te rm in a tio n
24
C.
M ethods o f D e s tro y in g O rg an ic M a tte r
26
D.
M ethods o f Removing P h o sp h ate
26
E.
R e s u lts o f Lung A n aly ses
33
F.
G e n e ra l D is c u s s io n
45
I
CO
r 5
M
i-i
V II.
Summary
49
Acknowledgment
51
L i t e r a t u r e C ite d
52
54884
*
3”
THE DETERMNATION OF HOISTUEE, SILICA, IEOH"
ALUEDTOH, CALCIDM, AED MAGEESIHIvl I E THE
LHEGS OF EOEMAL AED ' iLHEGEEh SHEEP
I.
IETEODECTIOE
"L unger" d is e a s e o f s h e e p , w h ich h a s b e e n d e s c r ib e d a s c h ro n ic p r o ­
g r e s s i v e p neu m o n ia, i s w e l l known, t o sh eep men o f th e E o r th w e s t, and t o
th em t h e a f f e c t e d sh eep a r e . known a s " lu n g e r s " o r "h eav ey " s h e e p « As th e
name i n d i c a t e s , t h e d i s e a s e i s one a f f e c t i n g t h e lu n g s , p ro d u c in g a r a p i d ,
j e r k y , p a n t in g ty p e o f r e s p i r a t i o n w h ich r e s u l t s i n l o s s o f c o n d i t i o n ,
e m a c ia tio n , e x h a u s tio n , an d f i n a l l y th e d e a t h o f t h e an im al*
T h is i s a d is e a s e o f ra n g e sh eep an d so f a r a s i s known a t p r e s e n t
does n o t o c c u r i n sh eep r a i s e d and m a in ta in e d i n sm a ll f l o c k s on i r r i g a t e d
p a s tu re s .
I t i s o b s e rv e d m o st commonly i n ewes o v er f o u r y e a r s o f a g e ,
b u t i t o c c u rs i n tw o , t h r e e , and f o u r - y e a r - o l d s h e e p , and i n b u ck s and
w e a th e rs a s w e l l a s i n ew es.
O c c a s io n a lly t h e symptoms a r e s e e n i n y e a r ­
lin g s .
O b s e rv a tio n s show t h a t th e d is e a s e o c c u rs i n U ta h , Wyoming, W ashing­
t o n , O regon, Id a h o , and E o r th D a k o ta , a s w e ll a s i n M o n tan a.
I n Montana,
th e l o s s from t h e d is e a s e i n t h e a v e ra g e i n f e c t e d b an d ru n s a b o u t two p e r
c e n t a n n u a l ly .
A lth o u g h t h e r e a r e some sh eep o u t f i t s t h a t a r e n o t known
t o h av e any " lu n g e r s " among t h e i r s h e e p , t h e d is e a s e i s q u i t e g e n e r a l l y
d i s t r i b u t e d an d th e t o t a l l o s s e a c h y e a r r e a c h e s a h ig h f i g u r e .
D r. H a d le ig h M arsh and D r. Howard W elch o f th e M ontana V e te r in a r y
4
— —
R esearch. L a b o r a to r y hav e c a r r i e d o u t s t u d i e s i n c o n n e c tio n w i t h th e
" lu n g e r " d is e a s e f o r f i f t e e n y e a r s .
They w orked on t h e p ro b le m l a r g e l y
from th e p a t h o l o g i c a l a n d b a c t e r i o l o g i c a l s ta n d p o in ts and i s o l a t e d from
t h e lu n g s o f " lu n g e r " sheep t h r e e s p e c ie s o f b a c t e r i a w h ich w ere a P a s t e u r e l l a , a d i p h t h e r o id b a c i l l u s , and M icro co ccu s c a t a r r h a l i s ( P f e i f f e r ) .
They s u c c e e d e d i n r e c o v e r in g i n p u re c u l t u r e t h e d ip h t h e r o id b a c i l l u s
and M icro co c cu s c a t a r r h a l i s ( P f e i f f e r ) .
On in o c u l a t i n g n o rm al sh eep w ith t h e b a c t e r i a o b ta in e d i n p u re c u l­
t u r e fro m th e lu n g s o f " lu n g e r " s h e e p , t h e d is e a s e c o u ld n o t a p p a r e n tly
b e re p r o d u c e d .
D r. M arsh an d D r. W elch fou n d fro m t h e i r o b s e r v a tio n s t h a t th e
n o rm al sh eep lu n g im m e d ia te ly a f t e r d e a th i s b r i g h t p in k i n c o l o r , te n d s
t o c o l la p s e i n t h e c h e s t c a v i t y , i s s o f t an d e l a s t i c , an d i s e a s i l y com­
p r e s s e d i n th e h a n d .
The w h o le lu n g i s ev en i n t e x t u r e an d c o n ta in s no
lum ps o r v a r i a t i o n s i n s t r u c t u r e .
The lu n g o f a " lu n g e r " sh eep i s g ra y
i n a p p e a ra n c e , f ir m , h e a v y , and f i l l s t h e c h e s t c a v i t y .
te n d t o c o l la p s e and i s n o t e l a s t i c .
I t d o es n o t
I n th e s m a lle r a n t e r i o r - lo b e s and
a lo n g t h e lo w e r m a rg in s o f t h e m ain l o b e s , an d o c c a s io n a lly , i n o th e r
p a r t s o f t h e lu n g , a r e s o l i d d a r k e r a r e a s .
The g r a d u a l e n la rg e m e n t o f
t h e s e s o l i d a r e a s i n t h e lu n g s an d th e g r a d u a l th i c k e n in g o f t h e w a ll s
o f t h e a i r s a c s th ro u g h o u t t h e lu n g s d e te rm in e th e c o u rs e o f t h e d i s ­
ease.
As th e c o n s o l i d a t i o n p r o g r e s s e s , t h e a v a i l a b l e a i r sp a c e d e c r e a s e s
u n t i l f i n a l l y t h e r e i s n o t enough a i r - c o n t a i n i n g t i s s u e t o s u p p ly th e
n e c e s s a r y am ount o f oxygen t o t h e b lo o d and t h e sheep d i e s .
The w hole p a t h o l o g i c a l p i c t u r e i s t h a t o f a lo n g c o n tin u e d i r r i t a -
-S t i o n , e i t h e r b a c t e r i a l o r m e c h a n ic a l.
M ic r o s c o p ic a l e x a m in a tio n o f th e
lu n g t i s s u e shows t h a t t h e r e a r e p r e s e n t i n th e v a r io u s p a r t s o f th e
d is e a s e d o rg a n c e r t a i n p a r t i c l e s w h ich g iv e a g r e e n is h y e llo w r e f r a c t i o n .
I t i s th o u g h t t h a t th e s e p a r t i c l e s may b e f o r e i g n m a t e r i a l , p r i n c i p a l l y
d u s ts o f in o r g a n ic n a t u r e , an d b y e i t h e r m e c h a n ic a l o r c h e m ic a l i r r i t a ­
t i o n o f th e lu n g t i s s u e so r e a c t a s t o c a u s e t h e " lu n g e r ” d i s e a s e .
B ecause o f th e f a c t s t h a t th e " lu n g e r ” d is e a s e o f sh eep c o u ld n o t
b e re p ro d u c e d i n n o rm al a n im a ls by i n o c u l a t i n g them w i t h t h e b a c t e r i a l
s p e c ie s i s o l a t e d fro m " lu n g e r " s h e e p , and t h a t h i s t o l o g i c a l o b s e r v a tio n
showed th e p r e s e n c e o f p a r t i c l e s o f g r e e n i s h y e llo w r e f r a c t i o n i n
" lu n g e r " lu n g t i s s u e , th e p r e s e n t s tu d y was u n d e rta k e n w i t h t h e o b je c t
o f d e te rm in in g th e c o n t e n t o f c d r t a i n m in e r a ls i n lu n g s o f " lu n g e r "
sh eep a s com pared w ith t h o s e i n no rm al lu n g s .
—6™
II.
REVIEW OF THE LITERATURE
The w ork o f p r e v io u s i n v e s t i g a t o r s w h ich a p p l i e s t o th e p r e s e n t p r o ­
b le m may h e d iv id e d in to - f o u r g r o u p s : f i r s t , p r e v io u s w ork done on
” lu n g e r" d i s e a s e y s e c o n d , f o r e ig n m in e r a ls t h a t a r e l i k e l y t o o c c u r i n
t h e lu n g s o f " lu n g e r " s h e e p ; t h i r d , t h e m ethod o f d e s tr o y in g o r g a n ic m at­
t e r p r e s e n t ; and f o u r t h , t h e m ethod o f a n a l y s i s .
AL
P re v io u s Work Done on "L u n g er" D is e a s e
P re v io u s w ork on t h e "L u n g er" d is e a s e o f sh eep h a s b e e n a lo n g th e
b a c t e r i o l o g i c a l and p a t h o l o g i c a l l i n e s .
W elch and M arsh (4 2 ) have w orked
f o r a number o f y e a r s on t h e b a c t e r i a l i n f e c t i o n i n t h e lu n g s o f d i s e a s ­
ed sh eep and fo u n d no d e f i n i t e c a u s e .
M arsh (S I and 32) h a s d e s c r ib e d
t h e d i s e a s e a s a p r o g r e s s iv e pneum onia o f s h e e p .
He i s o l a t e d two s p e c ie s
o f b a c t e r i a w h ich h e b e l i e v e s t o b e f a c t o r s i n th e e t i o l o g y o f th e d i s ­
e a s e b u t c o u ld n o t re p ro d u c e th e d is e a s e b y in o c u l a t i n g n o rm al sheep w ith
t h e i s o l a t e d o rg a n is m s .
■Be
F o r e ig n M in e ra ls T h at A re L ik e ly To O ccur
I n Lungs o f "L u n g er" Sheep
V ery l i t t l e w o rk h a s b e e n done on th e m in e r a l a n a l y s i s o f sheep t i s ­
s u e , e s p e c i a l l y o f t h e lu n g s .
Laws and G i l b e r t (29) h av e made a n in o r g a n ic
c h e m ic a l a n a l y s i s o f th e e n t i r e body o f t h i n , h a l f - f a t , f a t an d v e r y f a t
s h e e p , and r e c o r d e d th em i n 'p e r c e n t a g e o f l i v e w e ig h t.
F o r t h i n slieep
th e y fo und 1 .1 1 8 p e r c e n t HgPO^, 1*321 p e r c e n t baO, 0 .0 5 6 p e r c e n t MgO,
0 .1 7 3 p e r c e n t KgO, 0 .1 2 0 p e r c e n t HagO, 0 .0 3 7 p e r c e n t FegOg, an d 0 .0 2 1
p e r c e n t S iO g; f o r f a t sh eep 1 .0 4 0 p e r c e n t HgPO^, 1 .1 8 4 p e r c e n t CaO,
0 .0 4 8 p e r c e n t MgO, 0 .1 4 8 p e r c e n t KgO, 0 .0 9 7 p e r c e n t HagO, 0 .0 3 4 p e r
-
c e n t F e 2Og, an d 0 .0 2 6 p e r c e n t SiO2 .
7The r e s u l t s f o r h a l f - f a t and very-
f a t sh eep w ere v e r y c l o s e t o th o s e a l r e a d y g iv e n a b o v e. ,
W eiske (4 3 ) h a s a n a ly z e d th e b lo o d o f lam bs i n c o n n e c tio n w ith a
s tu d y o f t h e e f f e c t s o f i n g e s t i o n o f a c i d s .
p e r c e n t d r y w e ig h t:
He fou n d th e f o llo w in g i n
0 .1 5 0 p e r c e n t P , 1 5 .2 5 5 p e r c e n t H, 0 .4 1 5 p e r c e n t
K2O, 2 .2 2 0 p e r c e n t Ha2O, 0 .0 6 6 p e r c e n t Ca, an d 0.029. p e r c e n t MgO.
Any d i f f e r e n c e s i n th e lu n g s o f " lu n g e r " and n o rm al sh eep o th e r th a n
p a t h o l o g i c a l d e p o s it s m ig h t b e th e r e s u l t o f f o r e i g n m a t e r i a l .
W ith t h i s
f a c t i n m in d , a s w e l l a s know ing so m eth in g o f t h e m e ta b o lism o f th e a n i ­
m a l, th e c o m p o s itio n o f t h e p o s s i b l e in o r g a n i c d u s t b r e a t h e d b y th e slieep
s h o u ld b e a d e te r m in in g f a c t o r i n t h e c a u s e o f t h e " lu n g e r " d i s e a s e .
I n t h e a n a ly s e s o f t h e s o i l s o f M o n tan a, aluminum an d m agnesium s i l i ­
c a t e s p l a y a n im p o r ta n t p a r t . .. A lso v o lc a n ic a s h i s p r e s e n t i n a c o n s id e r ­
a b le a r e a i n M ontana s o i l s .
T h is a s h a c c o r d in g t o Em erson (1 2 ) i s l a r g e l y
com posed o f a n g u la r b i t s o f g l a s s and h a s a b r a s iv e p r o p e r t i e s w h ich c la y s
do n o t p o s s e s s . Em erson ( 1 2 ) . i n a n exam ple o f th e c o m p o s itio n o f la v a
s t a t e s t h a t i n s o l u t i o n i t in c lu d e s q u a r t z , f e l d s p a r , m ic a , and h o rn ­
b le n d e .
T h is w ould g iv e la v a s o i l s th e i n g r e d i e n t s q u a r t z , alum inum ,
p o ta s s iu m , so d iu m , an d c a lc iu m s i l i c a t e s .
H ilg a r d (1 7 ) g iv e s th e f o llo w in g t a b l e f o r th e o x id e s i n t h e e a r t h 's
c ru s t:
SiO2
5 7 .7 8
MgO
3 .8 1
A l2O3
1 5 .6 7
CaO
5 .1 8
^ 20S
3 .3 1
Ha2O
3 .8 8
FeO
3=84
,K 2O
3 .1 3
P2Os
0 .3 7
MtiO
0 .2 2
FeS2
1.03
HgO ( a c i d )
0 .3 6
HgO ( b a s e )
1=42
-
8-
From t h i s c o m p o s itio n o f th e s o i l and fro m t h e m e ta b o lic a b i l i t y o f a n i ­
m a ls t o th ro w o f f v a r io u s m in e r a l m a t t e r , th e m ost l i k e l y f o r e i g n m in e r­
a l e le m e n ts t h a t one w ould e x p e c t t o f i n d i n l a r g e r q u a n t i t i e s i n t h e
” lu n g e r ” lu n g s t h a n p r e s e n t i n n o rm al lu n g s a r e s i l i c a , alum inum , i r o n ,
m agnesium , an d c a lc iu m .
T h is p ro b le m seems v e r y c l o s e l y r e l a t e d t o c e r t a i n m i n e r a l d is e a s e s
i n t h e lu n g s o f human b e i n g s .
im p o r ta n t.
Of t h e s e , s i l i c o s i s i s one o f t h e m ost
Fow w eather (1 3 ) s t a t e s t h a t t h e s i l i c a c o n te n t o f n o rm al human
lu n g s v a r i e d fro m 0 .1 2 p e r c e n t t o 0 .2 0 p e r c e n t (a v e ra g e 0 .1 6 3 p e r c e n t)
an d i n lu n g s o f a r a ilw a y man w ith m arked a n t h r o c o s i s t h e r e was 0 .1 7 p e r
cent s ilic a .
He s a i d t h e r e i s no r e l a t i o n s h i p b e tw e e n t h e am ount o f
s i l i c a p r e s e n t an d t h e am ount o f pu lm o n ary f i b r o s i s and g iv e s tw o p o s­
s i b l e an sw ers t o t h e q u e s ti o n o f s i l i c o s i s :: f i r s t , s i l i c o s i s i s d e t e r ­
m ined n o t b y t h e am ount o f s i l i c a p r e s e n t b u t b y i t s n a t u r e an d seco n d ,
s i l i c o s i s i s d e te rm in e d n o t s o l e l y b y th e am ount o f s i l i c a b u t b y c e r t a i n
c o n d i tio n s p r e s e n t i n t h e b o d y o f th e p e r s o n .
Jo n e s (2 2 ) i n h i s a n a l y s e s o f s i l i c o t i c lu n g s fo u n d t h a t th e s i l i c a
was n o t t h e r e i n a f r e e s t a t e b u t t h e r e a s p o ta s s iu m and alum inum s i l i ­
c a te s .
H is a n a ly s e s o f s i l i c o t i c lu n g s a r e 9 .5 9 p e r c e n t A l2O5 , 4 8 .0 2
p e r c e n t SiO2 , 8 .4 3 p e r c e n t F e 2O5 , an d 2 .1 6 p e r c e n t CaO, 1 .6 4 p e r c e n t
MgO, 6 .4 2 p e r c e n t Ha2O, 1 6 .4 7 p e r c e n t P2O5 , and 6 .0 7 p e r c e n t K2O.
S te w a r t, L e e d s, an d F a u ld s (3 9 ) fo u n d i n t h e i r a n a ly s e s o f H a e m a tite
M in ers Lung t h a t t h e a v e r a g e p e r c e n t o f s i l i c a i s 1 .6 6 p e r c e n t and i n
n o rm a l lu n g s 0 .1 6 p e r c e n t o f th e d r y w e ig h t.
They fou n d q u a r tz and
s e r i c i t e p a r t i c l e s up t o th e s i z e o f t e n m ic ro n s and b e l i e v e s i l i c a t o
-
be th e r e a l cause o f f i b r o s i s .
9-
I t i s a ls o s ta te d in t h i s a r t i c l e th a t
h a e m a tite i s a c o m p a r a tiv e ly h a rm le s s d u s t w h ic h s t i m u l a t e s p h a g o c y to ­
s i s , and i s r a p i d l y rem oved fro m t h e l u n g s .
T h is e v id e n c e s h o u ld i n ­
d i c a t e t h a t i r o n w ould p r o b a b ly n o t b e a n im p o r ta n t f a c t o r i n t h e cau se
o f th e " lu n g e r" d is e a s e i n sh e e p .
The d i f f e r e n c e i n t h e s i l i c a c o n te n t o f th e lu n g s o f s i l i c o n w o rk e rs
.
and t h a t i n n o rm al lu n g s, h a s b e e n shown a l s o b y L ieb an d S c h a d e n d o rff
(3 0 ).
They fo u n d t h e SiOg c o n te n t o f p a t h o l o g i c a l lu n g s t o b e 4 .6 p e r
c e n t t o 2 2 .8 p e r c e n t o f t h e a s h t i s s u e w h ile t h e v a lu e s f o r n o rm al t i s s u e
w ere 0 .0 8 p e r c e n t t o 4 .0 p e r c e n t .
M cU ally (3 4 ) i n an a r t i c l e cm th e s i l i c o n d io x id e c o n t e n t o f lu n g s
i n h e a l t h and i n d is e a s e a s s e r t s t h a t th e s i l i c a c o n te n t o f lu n g s i n ­
c r e a s e s w i t h a g e ev e n i n n o rm al p e r s o n s .
He p o i n t s o u t t h a t 1 2 .8 p e r
c e n t o f th e e a r th * s c r u s t i s f r e e s i l i c a and i n th e com bined fo rm i t i s
p r e s e n t t o t h e e x t e n t o f 4 2 .5 p e r c e n t .
He fo u n d th e s i l i c a c o n te n t o f
n o rm al lu n g s t o be 1 .1 3 mg. p e r gram o f d r i e d lu n g and i n lu n g s o f e i g h t
p e rs o n s w o rk in g i n d u s ty atm o sp h ere t h e m illig r a m s o f s i l i c a p e r gram
t i s s u e ra n g e d fro m 2 .4 t o 2 6 .0 m g ., d e p e n d in g upon th e o c c u p a tio n .
Cummins and G ladden (5 ) i n a n a r t i c l e on c o a l m in e rs lu n g found
2 .3 6 p e r c e n t s i l i c a i n lu n g s o f c o a l m in e rs who d ie d o f p u lm o n ary d i s ­
e a s e a f t e r lo n g d is a b le m e n t, and i n c o a l m in e rs n o t d y in g o f pulm onary
d i s e a s e and n o t known t o have any p ulm onary d i s a b i l i t y — 1 .2 4 p e r c e n t .
I n n o n - c o a l m in e rs t h e y fo u n d 0 .1 3 p e r c e n t s i l i c a .
I t is a ls o s ta te d
i n t h i s a r t i c l e t h a t an. e x c e s s o f i r o n i s p a r t i a l l y . a t t r i b u t e d t o th e
e x c e s s o f a c t u a l t i s s u e m a t e r i a l i n t h e f i b r o t i c lu n g s .
They s t a t e t h a t
-
10 -
i r o n i n t h e lu n g s i s due t o t h r e e c a u s e s : f i r s t , "n o rm al” t i s s u e i r o n
( i n c l u d in g i r o n o f t h e M o o d ) ; se c o n d , i r o n i n p ig m en t o f endogenous
o rig in ,- and t h i r d , i r o n o f e x t e r n a l o r i g i n fro m in h a le d d u s t .
They
fo u n d t h a t th e c o a ly m a t e r i a l in c r e a s e d a l l o u t o f p r o p o r t i o n w ith th e
i r o n and t h e r e f o r e t h e l a t t e r h a s v e ry l i t t l e
s i g n i f i c a n t v a lu e i n th e
c a u se o f t h e d i s e a s e .
S i l i c o s i s h a s b e e n p ro d u c e d e x p e r im e n ta lly b y G ard n er (1 4 ) b y
e x p o s in g g u in e a p ig s and r a b b i t s t o a n a tm o sp h e re c o n ta in in g q u a r tz
d u s t e i g h t h o u rs a day f o r s i x d ay s a w eek and o v e r a p e r io d o f one y e a r
o r m ore.
He p ro d u c e d l e s i o n s c h a r a c t e r i s t i c o f s i l i c o s i s i n man by
u s in g a d u s t c o n ta in in g 9 0 .7 5 p e r c e n t SiO2 and whose p a r t ic l e t,s iz e r a n
fro m I t o 10 m ic r o n s .
To q u o te fro m G ard n er (1 4 ) i n r e g a r d t o i n h a l a t i o n o f v a r io u s d u s t s :
,
"A co m p a riso n o f t h e r e a c t i o n t o q u a r tz d u s t w ith t h a t
o f o th e r ty p e s o f d u s t , l i k e carborundum , s o f t o v a l ,
a s b e s t o s , and g r a n i t e , h a s i n d i c a t e d t h a t t h e r e a r e
d e f i n i t e d i f f e r e n c e s i n th e re s p o n s e t o d i f f e r e n t
ty p e s o f d u s t .
I n t h e c a s e o f q u a r tz th e a c t i v i t y o f th e p h a g o c y te s
i s r e s p o n s i b l e f o r t h e c o n c e n tr a ti o n o f a d e q u a te quan­
t i t i e s o f a n i r r i t a t i n g c h e m ic a l i n d i r e c t c o n t a c t
w ith t h e c o n n e c tiv e t i s s u e s . G ra n ite ; a l s o c o n t a in s
f r e e s i l i c a , b u t t h e e le m e n ts i n i t s c o m p o sitio n a p ­
p e a r t o m o d ify t h e e f f e c t o f th e s i l i c a upon th e
p h a g o c y te s an d p e rh a p s upon th e c o n n e c tiv e t i s s u e a s
w e l l ."
I t i s G ardner* s o p in io n t h a t t h e d i f f e r e n c e i n t h e r e a c t i o n t o
q u a r tz and g r a n i t e i s n o t e n t i r e l y due to th e low er c o n c e n tr a ti o n s o f
s i l i c a i n g r a n i t e d u s t , b u t t h a t th e n o n - s i l i c e o u s com ponents o f t h i s
d u s t m o d ify t h e b e h a v io r o f t h e p h a g o c y te s so t h a t th e y do n o t co n c e n t
t r a t e th e i r r i t a t i n g s i l i c a w ith "the same r a p i d i t y t h a t th e y do i n th e
-
11 -
case o f q u a rtz .
I s z a r d (2 1 ) i n r e g a r d t o th e in f l u e n c e o f an i n s p i r e d d u s t on
s p e c i f i c i n f e c t i o n o f th e lu n g s fo u n d t h a t c a lc iu m h y d r a te d u s t s a r e
■not h a rm fu l t o lu n g s o f hum ans.
'T h is a r t i c l e s t a t e s t h a t t h e s i l i c a
c o n te n t o f a n in o r g a n ic d u s t b e a r s d i r e c t r e l a t i o n s h i p t o i t s h a rm fu l­
n e s s and t h a t n o n a b s o rb a b le d u s ts a r e t h e m ost i n j u r i o u s .
To q u o te fro m C o l l i s and Greenwood (3 ) i n r e g a r d t o i n h a l a t i o n o f
in o r g a n ic p a r t i c l e s :
"O nly p a r t i c l e s w h ich a r e in s o l u b l e i n th e f l u i d s o f
th e body when c a r r i e d i n t o th e a i r p a s s a g e s re m a in
a s f o r e i g n b o d ie s e i t h e r t o s t i m u l a t e t h e c i l i a t e d
e p e th e liu m t o o v e r a c tio n f o r t h e i r e x p u ls io n , o r , ,
i f t h e y g a i n a c c e s s t o t h e lymph c h a n n e ls , i t g iv e s
r i s e t o a p r o l i f e r a t i o n o f c o n n e c tiv e t i s s u e s . Thus
d u s ts o f i v o r y , h o r n , b o n e , and o th e r an im a l s t r u c ­
t u r e s , and o f c a lc iu m s u l p h a t e , o f lim e s to n e , and o f
o x id e o f i r o n , a r e n o t a s s o c i a t e d w ith p n eu m o n o co n io sis
i n t h e way d u s t s o f v e g e ta b le h u s k s , em ery, g l a s s ,
s a n d s to n e , an d f l i n t a r e . "
L a n d is (2 7 ) w ith r e f e r e n c e t o in o r g a n ic d u s ts s t a t e s t h a t in o r g a n ic
d u s t fkom a n y s o u rc e p ro d u c e s d e f i n i t e c h a n g e s i n th e lu n g s .
They d i f f e r
o n ly i n t h e s e v e r i t y o f th e le s io n s w h ich depend upon th e h a r d n e s s , s h a r p ­
n e s s , and c h e m ic a l c o m p o s itio n .
I s z a r d (2 1 ) fo u n d t h a t c a lc iu m i s p a r t i a l l y removed b y th e b lo o d
s tre a m and t h a t th e d is s o lv e d c a lc iu m i s i n th e b lo o d s tre a m alw ays a s
b ic a r b o n a te a n d p h o s p h a te > t h e e x c e s s , in tr o d u c e d b y c a lc iu m d u s t , b e in g
d e p o s ite d i n t h o s e o rg a n s n o r m a lly c h a r a c t e r i z e d b y h ig h c a lc iu m c o n te n t
su c h a s b o n e s , k id n e y s , and lu n g s .
L a n d is (2 7 ) s t a t e s t h a t o rg a n ic d u s ts p ro d u c e no l e s i o n s , b u t w here
l e s i o n s a r e e n c o u n te re d i n th e lu n g s o f th o s e who have b e e n ex p o sed to
-
12 -
o rg a n ic d u s t , t h e l e s i o n s a r e t o he a s c r i b e d t o in o r g a n ic m a t e r i a l s —
i n o th e r w o rd s , a m ixed d u s t .
He s t a t e s t h a t th e symptoms o f in o r g a n ic
d u s t a r e s n e e z in g , c o u g h in g , t i c k l i n g s e n s a t i o n i n th e t h r o a t , and
v o m itin g .
I n a p a p e r i n r e g a r d t o r e l a t i o n o f o r g a n ic d u s ts t o p n e u m o n o c o n io sis,
L a n d is (2 8 ) s t a t e s t h a t o r g a n ic d u s ts do n o t h av e t h e pow er - t o e s t a b l i s h
t h e c o n d i t i o n known a s p n e u m o n o c o n io sis.
Q u o tin g fro m L a n d is (28) j
" D is r e g a r d in g t h e p r e d is p o s in g in f l u e n c e i t may h av e
on t h e d ev elo p m en t o f t u b e r c u l o s i s , i t s h o u ld b e
c l e a r l y r e c o g n iz e d t h a t in o r g a n ic d u s t o f i t s e l f c a n
and d o es c a u se s e r io u s and d i s a b l i n g damage t o t h e lu n g s ."
C.
M ethods o f D e s tro y in g O rg an ic M a tte r
T h ere h a s b e e n a g r e a t d e a l o f w ork done on t h e v a r i o u s ways o f d e s ­
t r o y i n g o r g a n ic m a tte r i n p l a n t and a n im a l m a t t e r .
From t h e s u rv e y o f
t h e l i t e r a t u r e t h e r e seem:.- t o b e s i x m ethods w h ich a r e u s e d t o a c o n s id e r ­
a b le e x t e n t a t t h e p r e s e n t .
As Koch (2 4 ) p o i n t s o u t , th e m ethod u sed
depends upon th e m in e r a l m a t t e r t h a t i s t o b e d e te rm in e d .
He s u g g e s ts
s u lp h u r ic a c i d p l u s a c a t a l y s t f o r n i t r o g e n d e t e r m in a tio n s ; s u lp h u r ic
a c i d and n i t r i c a c i d f o r p h o s p h o ru s , b u t n o t s a f e f o r n i t r o g e n d e te rm in a ­
t i o n s j sim p le i n c i n e r a t i o n o f o rg a n ic m a t e r i a l i n a m u f f le w h ich i s n o t
s a f e f o r s u lp h u r , p h o s p h o ru s , c h l o r i n e , o r n itr o g e n ,, b u t i s u s e f u l, f o r
m ost m e t a l s ; f u s i o n w ith sodium c a r b o n a te and sodium h y d ro x id e w ith o r
w ith o u t o x id i z in g a g e n ts a s p o ta s s iu m n i t r a t e o r sodium p e r o x id e f o r s u l ­
p h u r d e t e r m in a tio n s .
H i t r i c and p e r c h l o r i c a c id s w e r e 'u s e d by G ie s e k in g , S n id e r and G etz
(1 5 ) a s w e l l a s S m ith ( 3 7 ) .
They c la im l e s s lo s s o f m in e r a l m a tte r by
t h i s m ethod t h a n b y i n c i n e r a t i o n .
They a l s o s t a t e t h a t t h e s i l i c a i s com­
p l e t e l y d e h y d ra te d w ith o u t ta k i n g th e s o l u t i o n t o d ry n e s s an d t h a t th e
-
13 -
p e r c h l o r a t e o f t h e m e ta ls a r e much more s o lu b le th a n t h e c h l o r i d e s o r
o x id e s, i n a sh e d m a t e r i a l .
Al " s lim in g " m ethod w h ich c o n s i s t e d o f t r e a t ­
in g t h e w e t o r d ry a n im a l t i s s u e w i t h c o n c e n tr a te d n i t r i c a c i d f o r a
/.p erio d o f s e v e n t o t e n days was u se d b y Jo n e s ( 2 2 ) .
The in s o l u b l e m a t e r i a l
i s f i l t e r e d and th e o r i g i n a l c y r s t a l s t r u c t u r e o f th e s i l i c e o u s m a tte r
may b e exam ined u n d e r a m ic ro sc o p e s in c e t h e n i t r i c a c i d i s b e l ie v e d t o
have no e f f e c t on th e r e s i d u e m a t e r i a l »
K ing (2 3 ) and many o th e r a u th o r s a d v is e th e u s e o f p la tin u m c r u c i ­
b l e s f o r t h e a s h in g p r o c e s s an d u s u a l l y u s e d a sam ple o f a b o u t 2 g ram s.
A n i c k e l c r u c i b l e w as u s e d b y K ro ese an d K ieuw enhuyzen (2 5 ) an d th e y
fo u n d t h a t th e n i c k e l i s n o t a p p r e c ia b ly o x id iz e d on h e a t in g a n d t h a t i t
i s o n ly v e ry s l i g h t l y a t t a c k e d b y t h e c h a r r i n g o f t h e o r g a n ic m a t t e r .
They p u t th e d r i e d g ro u n d o r g a n ic m a t e r i a l i n a c lo s e d n i c k e l c r u c i b le
and i n c i n e r a t e d i n an e l e c t r i c oven a t 300° t o 350° C. w h ile a s tre a m
o f oxygen Tias p a s s e d i n t o th e n i c k e l v e s s e l .
They o b ta in e d a c h a lk
w h ite a s h i n 3 t o 4 ho u r s .
D.
M ethods o f A n a ly s is
T h ere i s much l i t e r a t u r e on th e m ethods o f a n a l y s i s f o r an im a l and
p l a n t m a t e r i a l s a s w e ll a s f o r c e r t a i n in o r g a n i c e le m e n ts .
Many w o rk ers
h av e g iv e n t h e m ethod o f a n a l y s i s f o r c e r t a i n o f th e e le m e n ts so u g h t i n
t h e p r e s e n t s tu d y , b u t few have g iv e n a m ethod f o r t h e c o m p lete a n a l y s i s .
Cummins and S la d d e n (5 ) i n a n a r t i c l e on C oal M in ers Lung s e l e c t e d
300 t o 400 grams o f w e t lu n g ,, c u t t h e m a t e r i a l i n t o s m a ll p i e c e s w ith
s c i s s o r s , and p la c e d th e t i s s u e i n a p r o c e l a i n d i s h .
T h is w as d r ie d
o v e r a w a te r b a t h u n t i l t h e w e ig h t was c o n s ta n t and th e n th e d r ie d p ro ­
-
d u c t "was g ro u n d i n a c o f f e e m i l l .
14 -
The a n a l y s t d e te rm in e d a s h , s i l i c a ,
" c o a ly " m a t e r i a l , and i r o n o n th e sam p le.
K roese an d N ieuw enhuyzen (2 5 ) i n t h e i r e s t i m a t i o n o f s m a ll am ounts
o f s i l i c a and c a lc iu m i n t h e lu n g s o f humans a sh e d th e t i s s u e i n a n i c k e l
c ru c ib le .
The a s h was t r a n s f e r r e d t o a p la tin u m c r i c u b l e a n d f u s e d w ith
a NaOH-KOH m i x tu r e .
T h is was t h e n e v a p o r a te d t h r e e tim e s w ith co n cen ­
t r a t e d HC1, d r i e d a t H O 0 and f i n a l l y i g n i t e d .
The r e s i d u e was t r e a t e d
w i t h d i l u t e HOI , t h e s i l i c a f i l t e r e d o f f , w ashed w ith h o t w a t e r , and
f i n a l l y i g n i t e d i n a p la tin u m c r u c i b le t o c o n s t a n t w e ig h t.'
The p u r i t y o f
SiOg was ch eck ed b y e v a p o r a tio n w ith s u lp h u r ic an d h y d r o f l u o r i c a c i d s
and once more i g n i t e d t o a c o n s ta n t w e ig h t.
I n th e f i l t r a t e fro m th e
s i l i c a , th e p h o sp h o ru s was p r e c i p i t a t e d b y a d d i t i o n o f n i t r i c a c i d , am­
monium n i t r a t e , and an e x c e s s o f ammonium m o ly b d a te .
The p h ospho-m olyb-
d a te was rem oved b y f i l t r a t i o n an d t h e e x c e s s molybdenum rem oved from
th e f i l t r a t e b y s a t u r a t i o n w i t h HgS, a f t e r f i r s t b o i l i n g o f f t h e n i t r i c
a c id .
The n i c k e l w as p r e c i p i t a t e d w ith ammonium s u lp h id e a f t e r p r e ­
li m in a r y n e u t r a l i z a t i o n .
The c a lc iu m w as th e n p r e c i p i t a t e d a s th e oxa­
l a t e and w eig h ed a s CaO.
M organ an d K ing (3 5 ) h av e g iv e n a m ethod f o r m icro d e te r m in a tio n o f
s ilic a .
They a s h e d th e sam ple and u s e d p e r c h l o r i c a c i d t o d e h y d ra te t h e
s i l i c a i n p la c e o f HCl
They f i l t e r e d o f f t h e s i l i c a , i g n i t e d i n a
p la tin u m d i s h , an d w e ig h e d .
K u z ir ia h (2 6 ) e s tim a te d t h e c a lc iu m i n t h e a s h o f p l a n t and an im al
c a rc a sse s.
He rem oved th e p h o sp h o ru s w ith ammonium m o ly b d a te , p r e c i p i t a t ­
ed t h e c a lc iu m a s th e o x a l a t e , an d w eig h ed t h e l a t t e r a s CaO.
The c a lc iu m
-1 5 o x id e -was t h e n d i s s o lv e d , i n d i l u t e H Cl, f i l t e r e d , ammonium c h l o r i d e and
ammonium h y d ro x id e a d d e d , b o i l e d u n t i l o d o r o f ammonia i s f a i n t , and
i r o n and alum inum h y d ro x id e s rem oved.
a s th e o x a l a t e .
I h e c a lc iu m i s t h e n p r e c i p i t a t e d
He recommended n o t rem o v in g th e e x c e s s molybdenum s in c e
i t d id n o t i n t e r f e r e w i t h t h e p r e c i p i t a t i o n o f th e c a lc iu m .
S t a n f i e l d (5 8 ) i n h i s m ethod f o r d e te r m in in g th e molybdenum i n
p l a n t s and s o i l s a d v is e s t h e p r e c i p i t a t i o n o f t h e molybdenum a s th e s u l ­
p h id e .
T h is i s a c c o m p lish e d by s a t u r a t i n g a s tr o n g am m oniacal s o l u t i o n
w ith h y d ro g en s u lp h id e and p o u rin g th e s o l u t i o n i n t o an e x c e s s o f 6 H.
s u lp h u r ic o r h y d r o c h lo r ic a c i d , b o i l i n g t o c o a g u la te t h e p r e c i p i t a t e ,
and f i l t e r i n g o f f t h e MoSg.
McCrudden (3 5 ) h a s g iv e n e x p e r im e n ta l d a t a fo r d e te r m in a tio n o f
c a lc iu m and m agnesium i n th e p r e s e n c e o f p h o sp h o ru s an d i r o n .
H is r e ­
s u l t s w i t h th e m ethod f o r c a lc iu m a g re e w i t h th o s e g iv e n b y O f f i c i a l
M ethods o f A g r i c u l t u r a l C h em ists ( I , X E Ij 9 ) * , H a lv e rs o n an d B ergeim
( 1 6 ) , C o rle y and D en is ( 4 ) , H ille b r a n d and L u n d e ll ( 1 9 ) , an d B re a z e a le
(2 ).
The c a lc iu m i s p r e c i p i t a t e d a s t h e o x a l a te i n a s l i g h t l y a c i d s o lu ­
t i o n and th e n b ro u g h t t o s l i g h t a l k a l i n i t y w ith sodium a c e t a t e .
The
m agnesium i s d e te rm in e d i n th e f i l t r a t e fro m c a lc iu m b y e v a p o r a tin g t o
d r y n e s s , t a k i n g up w i t h HCl and f i n a l l y p r e c i p i t a t i n g i t a s t h e am­
monium p h o s p h a te .
I f i r o n i s p r e s e n t 0 .5 t o 1 .0 c c . o f a 5 p e r c e n t
sodium c i t r a t e s o l u t i o n i s ad d ed b e f o r e t h e m agnesium i s p r e c i p i t a t e d .
The u s e o f t h e c i t r i c a c i d to fo rm com plex io n s w ith t h e i r o n and
alum inum h a s a l s o b e e n v e r i f i e d b y C urtm an ( ? ) and S t . Jo h n (4 0 ) a s
w e l l a s C urtm an an d D ubin ( 8 ) .
* D en o tes th e d i v i s i o n and d e t e r m in a tio n n u m b er.
I-, ■> II V
D ien es (9 ) h a s u s e d t h e same e s s e n t i a l m ethods f o r c a lc iu m and
m agnesium &s d i d McCrudden ( 3 3 ) , "but t h e f o r m e r 's w ork was w ith m ic ro ­
c h e m ic a l a n a l y s i s w h ich in v o lv e d some m in o r c h a n g e s .
E lv eh jem an d H a r t ( l l ) c o n firm ed t h e m ethod f o r i r o n g iv e n i n O f f i ­
c i a l M ethods o f A g r i c u l t u r a l C h em ists ( I , X I I , 7) and H ill e b r a n d and Lund e ll (1 8 ).
T re a d w e ll an d H a ll (4 1 ) g iv e a m ethod f o r .th e d e te r m in a tio n
o f i r o n and alum inum when th e y a r e p r o c u r e d a s com bined o x i d e s .
.W inter
and P a t t e n (4 4 ) p r e c i p i t a t e d th e i r o n and alum inum a s p h o s p h a te s and
l a t e r d e te rm in e d t h e i r o n s e p a r a t e l y b y a d j u s t i n g th e h y d r o x y l c o n c e n tr a ­
t i o n b y means o f ammonium a c e t a t e .
M yersy M u ll, and M o rris o n (36) f o l -
I
lowed a s i m i l a r scheme b u t u s e d a c o l o r i m e t r i c m ethod f o r alum inum .
■
E dw ards• (1 0 ) fo u n d t h e amount o f i r o n and alum inum i n p h o s p h a te s b y
rem oving th e p h o s p h a te w ith c i t r o m a g n esia and p r e c i p i t a t i n g t h e i r o n
and alum inum i n a b a s i c s o l u t i o n w i t h h y d ro g en s u lp h id e .
The f e r r o u s
s u lp h id e was w a sh e d , d i s s o l v e d i n d i l u t e s u lp h u r ic a c i d , b o i l e d , c o o le d ,
and t i t r a t e d w i t h s ta n d a r d p e rm a n g a n a te .
T h e ' alum inum i s c a l c u l a t e d ..
by th e d iffe re n c e .
C urtm an ( 6 ) h a s g iv e n a m ethod f o r rem o v in g th e p h o s p h a te by u se
o f t i n i n q u a l i t a t i v e a n a l y s i s and fo u n d i t q u i t e s a t i s f a c t o r y .
The
m ethod i s s im p ly a d d in g t i n t o a c o n c e n tr a te d n i t r i c a c id s o l u t i o n f r e e
fro m c h l o r i d e s and rem oving th e p h o sp h a te b y f i l t e r i n g .
From t h e l i t e r a t u r e p e r t i n e n t t o t h e p ro b le m , i t a p p e a r s t h a t con­
s i d e r a b l e w o rk h a s b een done i n a n a ly z in g t h e v a r io u s in o r g a n ic m a te r ia ls
p r e s e n t i n human lu n g s an d t h a t t h e m ethod u se d v a r i e s w i t h t h e p a r t i ­
c u l a r d e t e r m in a tio n s s o u g h t.
T here i s no l i t e r a t u r e on t h e d e te r m in a tio n
-
17 -
o f t h e m in e r a l c o n te n t o f lu n g s o f " lu n g e r " s h e e p ,
III.
COEDITIOES UEDEE WHICH THE' SHEEP WERE RAISED
A ll o f th e sh eep whose lu n g s w ere a n a ly z e d i n t h i s e x p e rim e n t w e r e
r a i s e d u n d e r t h e s u p e r v is io n o f th e M ontana E x p erim en t S t a t i o n .
The
sh eep whose lu n g s w e re c o n s id e r e d n o rm al w ere r a i s e d e i t h e r u n d e r f lo c k
c o n d i t i o n s , on t h e r a n g e , o r p a r t l y u n d e r "both.
The " lu n g e r " lu n g s
came fro m sh e e p who u s u a l l y h ad b e e n on t h e ra n g e f o r some p e r io d o f
tim e .
IT .
PRERARATIOE OF SAMPLES
The p ro c e d u re o f sam p lin g f a l l s i n t o t h r e e g ro u p s and w i l l be con­
s i d e r e d i n t h i s m anner.
A.
M ethod o f Removing Lungs
D r. M arsh o f th e V e te r i n a r y R e s e a rc h L a h o r a to r y p r o c u r e d a l l o f th e
lu n g s w h ich w ere a n a ly z e d i n t h i s p ro b le m .
I n a l l b u t one c a s e , i n
w h ich t h e sh eep d i e d , t h e y w ere k i l l e d b y b le e d in g fro m t h e j u g u l a r
v e i n and b o th lu n g s upon rem o v a l from t h e a n im a l p la c e d i n a p o r c e l a i n
d is h .
The t r a c h e a a s w e l l a s a l l o th e r t i s s u e o th e r t h a n t h a t o f lu n g
was c u t aw ay.
B.-
D ry in g an d G rin d in g
The d ry in g was done i n a n e l e c t r i c a l l y h e a te d d r y e r p r o v id e d w ith
a f a n f o r c ir c u la tin g th e a i r .
fro m 55°
t o 65° C ..
The te m p e r a tu r e o f t h e d r y e r ra n g e d
A f t e r fro m f i v e t o s e v e n d ay s i n t h i s d r y e r , d e ­
p e n d in g upon t h e tim e n e c e s s a r y f o r t h e m a t e r i a l t o come t o c o n s ta n t
w e ig h t, th e sam ple was rem oved an d g ro u n d i n a W iley m i l l .
—
C.
18 -
S am pling and P r e s e r v in g
The g ro u n d lu n g was t h e n " ta b l e d " b y p la c in g on a p ie c e o f p a p e r
a b o u t two f e e t s q u a re and p u l l i n g e a c h c o r n e r o f th e p a p e r i n s u c ­
c e s s io n o v e r th e d ia g o n a l c o r n e r , t h u s c a u s in g th e p ie c e s t o r o l l o v er
one a n o t h e r .
T h is was c o n tin u e d u n t i l e a c h c o r n e r h a d 'b e e n p u l l e d
a c ro ss f o r te n tim e s .
The t o t a l d r i e d m a t e r i a l was th e n b o t t l e d up i n
a n e i g h t ounce b o t t l e o r q u a r t c a n n in g j a r d ep en d in g upon t h e s i z e o f
t h e lu n g s .
S in c e t h e s i z e o f t h e lu n g s o f some o f t h e sh eep was. s m a ll
i t n e c e s s i t a t e d t h e k e e p in g o f th e c o m p lete lu n g s .
T h is p r a c t i c e was
c a r r i e d o u t w ith a l l t h e lu n g s .
V.
METHODS OF ANALYSES
A.
Q u a l i t a t i v e A n a ly s is
The g e n e r a l m ethods a s g iv e n i n C urtm an' s Q u a l i t a t i v e A n a ly s is
w ere u s e d t o i d e n t i f y t h e v a r io u s io n s p r e s e n t .
Be
Q u a n t i t a t i v e A n a ly s is
The m ethods o f a n a ly s e s w ere m o d ifie d t o s u i t t h e d e te r m in a tio n s
•
d e s i r e d i n a fe w c a s e s , b u t i n g e n e r a l t h e p ro c e d u re s o f t h e A s s o c ia tio n
o f t h e O f f i c i a l A g r i c u l t u r a l C h em ists ( l ) w ere f o llo w e d .
T h is r e f e r ­
en ce w i l l b e r e f e r r e d t o a s " O f f i c i a l M ethods" i n f u r t h e r c i t a t i o n s .
U s u a lly a 12 gram sam ple o f t h e d r i e d lu n g m a t e r i a l was ta k e n and c a r ­
r i e d th r o u g h th e e n t i r e a n a l y s i s , a lth o u g h i n some c a s e s o th e r s i z e s
w ere u s e d . I n a l l c a s e s t h e d e te r m in a tio n s w e re ru n i n d u p l i c a t e .
l a r g e sam ple h a d t o b e a s h e d i n o r d e r t o r e c e iv e s u f f i c i e n t l y la r g e
am ounts o f in o r g a n ic m a t e r i a l s i n t h e a s h .
A
-
19 -
MoI s t u r e i n th e Lungs
The lim g s a s r e c e iv e d i n ' a p o r c e l a i n e v a p o r a tin g d is h w ere w eighed
and t h e w e ig h t o f t h e d is h s u b t r a c t e d so a s t o o b ta in th e w e ig h t o f th e
w e t lu n g s .
A f te r t h e lu rig s h ad b e e n d r i e d a t a te m p e r a tu r e o f 55 t o
65° c . t o c o n s ta n t w e ig h t, t h e w e ig h t o f t h e d i s h was a g a in s u b tr a c t e d
fro m t h e t o t a l w e ig h t i n o r d e r t o a r r i v e a t th e d ry w e ig h t o f th e lu n g s .
The p e r c e n ta g e o f m o is tu r e was fo u n d on t h e w et b asis®
T o ta l Ash
T o ta l a sh f o r c o m p u ta tio n s was c a r r i e d o u t i n fo u r s e p a r a t e
s a m p le s , tw o o f w h ich w ere n o rm al and two o f w h ich w ere " lu n g e r " lung
s a m p le s .
A 6 gram sam ple was w eig h ed i n t o a s i l i c a d is h an d th e n p la c e d
in an e l e c t r i c m u ffle .
The te m p e r a tu r e o f th e m u ffle was q u i t e low a t
t h e s t a r t o f t h e a s h in g p r o c e s s i n o r d e r t o p r e v e n t l o s s o f m a t e r i a l b y
fla m in g an d a lso , l o s s due t o " p u f f in g u p " o f c h a r r e d m a t e r i a l .
The
a s h in g was th e n c o n tin u e d a t a te m p e r a tu r e o f 393° t o 488° C. f o r 48
t o 96 h o u rs w h ile a s tre a m o f a i r p a s s e d th r o u g h th e m u f f l e .
T h is tem ­
p e r a t u r e was th e lo w e s t a t w h ich a s h in g w ould ta k e p la c e c o m p le te ly .
A t a h ig h e r te m p e r a tu r e t h e a s h had a te n d e n c y t o f u s e .
S i l i c a d is h e s w ere u s e d i n p la c e o f p la tin u m d is h e s s in c e a la r g e
sam ple had t o b e a s h e d i n o r d e r t o p ro c u re a s u f f i c i e n t l y l a r g e a s h and
t h a t su c h l a r g e sam p les when ash e d may hav e d e s t r u c t i v e a c t i o n on th e
.p la tin u m .'
L arge p la tin u m d is h e s w ere n o t a v a i l a b l e .
The a s h in g a s c a r r i e d o u t f o r th e in o r g a n ic a n a l y s i s was done i n a
n i c k e l c r u c i b l e a s s u g g e s te d b y K roese an d H ieuw enhuyzen (2 5 ) b y p la c in g
t h e v e s s e l w i t h t h e o r g a n ic t i s s u e s i n a n e l e c t r i c m u ffle w hose te m p e ra -
-
20 -
•fcure was j u s t h ig h enough t o c h a r th e o r g a n ic m a tte r and low enough t o
p r e v e n t th e " p u f f in g up" o f t h e m a t e r i a l a s much a s p o s s i b l e .
A f te r
th e m a t e r i a l had s to p p e d sm oking t h e te m p e r a tu r e o f th e m u f f le was r a i s e d
t o 393° t o 488° C. f o r t h e re m a in d e r o f t h e a s h in g p r o c e s s .
T o ta l a s h
c o u ld n o t be d e te rm in e d b y t h i s m ethod s in c e a c e r t a i n am ount o f n i c k e l
o x id e and s a l t s w ere fo rm ed b y t h e i g n i t i o n o f th e a s h i n t h e n i c k e l
c ru c ib le .
S i l i c a d is h e s c o u ld n o t b e u s e d f o r a s h in g th e sam ple f o r i n ­
o rg a n ic a n a l y s i s b e c a u se some s i l i c a may be removed from t h e d i s h and
g iv e f a l s e r e s u l t s i n th e s i l i c a d e t e r m in a tio n .
S i l i c a D e te r m in a tio n
The s i l i c e o u s m a tte r was d e te rm in e d i n t h e ash e d m a t e r i a l a c c o rd ­
in g t o H ill e b r a n d (2 0 ) and O f f i c i a l M ethods ( I , Z I I , 5 ) , th e a sh e d
m a t e r i a l b e in g p u t in to , a b e a k e r and e v a p o r a te d t h r e e tim e s w i t h con­
c e n t r a t e d HCl and f i n a l l y t a k i n g up t h e r e s i d u e w ith d i l u t e H C l.
A f te r
f i l t e r i n g , t h e f i l t r a t e was e v a p o r a te d t o d ry n e s s b u t r a r e l y e v e r .gave
any s i l i c a .
The s i l i c e o u s m a tte r was f i l t e r e d th r o u g h a Whatman 42
f i l t e r p a p e r and i g n i t e d i n a p la tin u m c r u c i b l e t o c o n s ta n t w e ig h t.
The s i l i c a c o n te n t w as p ro c u r e d b y e v a p o r a tin g th e s i l i c e o u s m a tte r
w ith s u lp h u r ic and h y d r o f lu o r ic a c i d s u n t i l c o n s ta n t w e ig h t was r e a c h e d .
The d i f f e r e n c e o f w e ig h t o f t h e s i l i c e o u s m a tte r b e f o r e an d a f t e r t h i s
tr e a tm e n t gave th e : w e ig h t o f th e SiOg p r e s e n t .
I r o n D e te r m in a tio n
The f i l t r a t e r e c e iv e d a f t e r a l l th e s i l i c e o u s m a tte r h ad b een r e ­
moved was made up t o a Volume o f 100 c c . .
A 90 c c . p o r t i o n was p la c e d
i n a b e a k e r f o r t h e d e te r m in a tio n o f alum inum , c a lc iu m , an d m agnesium .
—
21 “
The re m a in in g 10 c c . w ere u s e d f o r th e ir o n d e te r m in a tio n a c c o r d in g t o
th e m i c r o c o lo r i m e t r i c p ro c e d u re o f th e O f f i c i a l M ethods ( I , % I I , 7)
w ith th e p r e c a u t i o n ta k e n o f ad d in g th e same amount o f n i t r i c a c id t o
b o th unknown and known s o l u t i o n s o f i r o n .
P h o sp h a te Removal
S in c e c a lc iu m a s w e l l a s m agnesium p h o s p h a te w ould p r e c i p i t a t e
when a s o l u t i o n c o n t a in in g t h e i n g r e d i e n t s i s made a l k a l i n e , t h e p h o s­
p h a te h a s t o be rem oved.
The m ethod o f rem oving th e p h o s p h a te b y am­
monium m o ly b d ate a c c o r d in g t o K roese and K ieuw enhuyzen (2 5 ) and O f f i­
c i a l M ethods ( I , I I j 7) was t r i e d , b u t d i f f i c u l t y was fo u n d i n rem oving
b o th t h e p h o s p h a te and e x c e s s molybdenum.
A cco rd in g t o O f f i c i a l Me­
th o d s ( I , I I , 7) 70 c c . o f m o ly b d ate s o l u t i o n h a s t o be added f o r e v e ry
d e c ig ra m o f P 2O5 p r e s e n t i n a n HCl f r e e s o l u t i o n .
T h is w o u ld n e c e s s i ­
t a t e th e a d d in g o f a b o u t 300 t o 400 c c . o f m o ly b d ate s o l u t i o n i n o r d e r
t o p r e c i p i t a t e a l l o f th e p h o s p h a te p r e s e n t .
T h is d i l u t i o n seemed t o
i n t e r f e r e w ith t h e p r e c i p i t a t i o n o f th e p h o s p h a te .
A lso much d i f f i ­
c u l t y was e n c o u n te r e d on t r y i n g t o r i d t h e s o l u t i o n o f t h e e x c e s s mo­
lybdenum b y t h e m ethod s u g g e s te d b y S t a n f i e l d ( 3 8 ) .
I n a n e f f o r t , t o remove t h e p h o s p h a te th e q u a l i t a t i v e m ethod o f
Curtm an ( 6 ) was t r i e d and fo u n d t o g iv e good r e s u l t s .
T h is m ethod con­
s i s t e d o f e v a p o r a tin g t h e a l i q u o t t o d ry n e s s a f t e r th e i r o n sam ple
had b e e n rem oved and t a k i n g i t up w ith 10 c c . o f c o n c e n tr a te d n i t r i c
a c id .
T h is was ta k e n to d ry n e s s and 10 c c , o f n i t r i c a c i d a g a i n ad d ed .
A f t e r t a k i n g t h i s t o d ry n e s s t h e r e s id u e was ta k e n up w ith co n c e n ­
t r a t e d n i t r i c a c i d , h e a te d u n t i l a l l m a t e r i a l h ad gone i n t o s o l u t i o n .
22 —
—
an d a b o u t 2 gram s o f t i n i n th e fo rm o f CP f o i l was a d d e d .
T h is was
b o i l e d and e v a p o r a te d t o a b o u t 0 .5 o f a c c . , 25 o c . o f w a te r w ere
a d d e d , t h e s o l u t i o n s t i r r e d , a llo w e d t o s t a n d , and f i n a l l y f i l t e r e d .
The p r e c i p i t a t e w as w ashed fro m t h e f i l t e r p a p e r i n t o a b e a k e r and 10
c c . o f h o t w a te r a d d e d .
T h is was a g a in f i l t e r e d , w ashed w i t h w a te r ,
ad d ed t o t h e o r i g i n a l f i l t r a t e , and t h e t o t a l volume e v a p o r a te d to
a b o u t 25 c c .
S in c e some o f th e o r i g i n a l p r e c i p i t a t e te n d s t o go
th r o u g h th e p a p e r , t h e s o l u t i o n i s a g a in f i l t e r e d .
The f i l t r a t e , a l ­
th o u g h f r e e fro m p h o s p h a te , u s u a l l y c o n t a in s s m a ll q u a n t i t i e s o f le a d
and c o p p e r o r i g i n a l l y p r e s e n t a s i m p u r i t i e s i n t h e t i n f o i l .
To g e t
r i d o f th e s e and an y t i n t h a t m ig h t b e p r e s e n t , th e s o l u t i o n i s made
a l k a l i n e w i t h ammonium h y d r o x id e , made s l i g h t l y a c id w ith HC1, 5 grams
o f ammonium c h l o r i d e a d d e d , an d f i n a l l y s a t u r a t e d w ith h y d ro g e n s u l ­
p h id e .
The p r e c i p i t a t e i s ' f i l t e r e d , , w ashed w i t h ammonium n i t r a t e ,
and f i l t r a t e made s tr o n g ly a c i d w ith H C l.
T h is i s b o i l e d t o g e t r i d
o f e x c e s s h y d ro g en s u lp h id e and t h e f r e e s u l f u r w hich form s i s f i l t e r ­
e d o f f and w ashed w ith h o t w a te r .
The f i l t r a t e i s e v a p o r a te d t o a b o u t
75 c c .
D e te r m in a tio n o f Aluminum
The c o n c e n tr a te d f i l t r a t e o b ta in e d fro m above a f t e r t h e p h o sp h a te
re m o v a l i s t r e a t e d f o r FegOg and AlgOg a c c o r d in g t o O f f i c i a l M ethods
( I , XXXVII, 5 6 ) .
The i r o n c o n te n t i n t h e com bined o x id e s i s found b y
u s in g t h e p ro c e d u re i n O f f i c i a l M ethods ( I , XXXVII, 5 7 ) .
o f . FegOg i s c a l c u l a t e d fro m t h e am ount o f i r o n fo u n d .
The amount
The q u a n t i t y o f
AlgOg i s fo u n d b y s u b t r a c t i n g th e am ount o f FegOg fro m t h e w e ig h t o f
-
t h e com bined o x id e s=
23 -
The amount o f th e alum inum i s found b y u s e o f
c h e m ic a l f a c t o r s =
D e te r m in a tio n o f C alcium
The am m oniacal f i l t r a t e
from t h e R20S d e te r m in a tio n i s s a t u r a t e d
■with h y d ro g en s u lp h id e t o remove th e n i c k e l .
T h is i s f i l t e r e d o f f and
•washed w i t h a I p e r c e n t ammonium n i t r a t e s o l u t i o n .
The f i l t r a t e i s
a c i d i f i e d w ith HCl an d b o i l e d . The f r e e s u lp h u r i s f i l t e r e d o f f , w ash­
ed w ith h o t w a te r , and t h e r e s u l t i n g f i l t r a t e
200 c c .
c o n c e n tr a te d t o ab o u t
The c a lc iu m was d e te rm in e d a c c o rd in g t o O f f i c i a l M ethods
( I , X I I , 9) e x c e p t t h a t th e c a lc iu m o x a l a te was p r e c i p i t a t e d i n a c o ld
s o l u t i o n w h ic h w as v e r y s l i g h t l y a c i d i n r e a c t i o n t o m e th y l o r a n g e .
D e te r m in a tio n o f M agnesium
M agnesium was d e te rm in e d i n a s i n g l e p r e c i p i t a t i o n a c c o rd in g t o
O f f i c i a l M ethods ( I , XXI, 3 7 ) .
-
V I.
24
'
TABULA.!IOU AUD DISCUSSIOIT OF DATA
An im p o r ta n t f a c t o r i n g e t t i n g good r e s u l t s i n t h i s p ro b le m i s
t h a t o f o b ta in in g a m ethod w h ich w i l l a c c u r a t e l y and u n if o r m ly a p p ly
t o a l l o f t h e a s h e d sam p les o f t h e lu n g m a t e r i a l .
W ith t h i s f a c t i n
m ind t h e v a r io u s m ethods w ere t r i e d i n o r d e r t o d e te rm in e w h ich w ould
g iv e th e m ost a c c u r a te r e s u l t s .
A.
Q u a l i t a t i v e A n a ly s is
A q u a l i t a t i v e a n a l y s i s was r u n on tw o n o rm al and two " lu n g e r "
lu n g sam p les w h ic h w ere a s h e d i n s i l i c a d i s h e s .
The m ethods g iv e n i n
C urtm an*s Q u a l i t a t i v e C hem ical A n a ly s is w ere u s e d and i n d i c a t e d th e
p r e s e n c e o f S iO g, alum inum , p h o s p h a te , i r o n , c a lc iu m , an d m agnesium i n
b o th n o rm al and " lu n g e r " lu n g s .
g o t t e n u s in g t h e s e m e th o d s.
Uo o th e r q u a l i t a t i v e t e s t s c o u ld be
Much tim e w as s p e n t t r y i n g t o g e t th e
o n ly a v a i l a b l e s p e c tr o s c o p e i n p r o p e r w o rk in g c o n d itio n i n o r d e r t o
o b t a i n t h e s p e c tru m p h o to g ra p h o f b o th th e n o rm al and " lu n g e r ” lu n g
s a m p le s .
I t was f i n a l l y d e c id e d theCt t h e in s tr u m e n t c o u ld n o t g iv e
f i n e enough l i n e s f o r u s e , i n s p e c tru m s t u d i e s , and t h e r e f o r e c o u ld
n o t b e u s e d i n t h e p r e s e n t p ro b lem .
B.
Q u a n t i t a t i v e Ash D e te rm in a tio n
T ab le I shows t h e r e s u l t s o b ta in e d i n a s h in g a 6 gram sam ple o f
tw o n o rm al and tw o " lu n g e r ” ew es.
The " lu n g e r ” a v e ra g e a s h e d w e ig h t
w as 0 .1 1 5 6 gram s h e a v i e r o r 4 1 .8 p e r c e n t h ig h e r th a n th e a v e ra g e
w e ig h t o f t h e n o rm al lu n g a s h .
T h is d e t e r m in a tio n shows t h a t t h e r e
i s 'n e a r l y l|h ti m e s a s much m in e r a l m a tte r i n " lu n g e r " a s t h e r e i s i n
n o rm a l lu n g s a m p le s .
-
25 -
T a b le I .
W eight o f T o ta l A sh i n 'lL ungerw and i n n o rm al Lungs
Sheep
number
• S iz e o f
sam ples
gms
W eight o f ash
gms
L unger Lungs
GlTD
6
0 .3 7 3 1
5 5 TB
6
0 .4 1 2 4
A verage
6
0 .3 9 2 8
Borm al Lungs
430
6
0 .2 6 1 8
314
6-
0 .2 9 2 6
A verage
6
0 .2 7 7 2
-
C.
26 M ethods o f D e s tro y in g O rg a n ic M a tte r
T hree m ethods vrere t r i e d f o r d e s tr o y i n g th e o r g a n ic m a t t e r p r e s e n t
and a r e n am ely t h e a s h m ethod a c c o r d in g t o K roese and Ziem venhuyzen
( 2 5 ) j s e c o n d , d i g e s t i o n o f lu n g w ith n i t r i c and p e r c h l o r i c a c i d s a c c o rd ­
in g t o S m ith (3 7 ) } a n d t h i r d , " s lim in g ” m ethod o f d i g e s t i o n "by means o f
n it r ic a c id .
S in c e t h e l a t t e r a p p l ie d more t o m ic r o s c o p ic a l s t u d i e s
o f t h e s i l i c e o u s m a tte r p r e s e n t th a n t o c h e m ic a l a n a l y s i s , i t was n o t
u s e d a f t e r p r e li m i n a r y e x p e r im e n t.
The n i t r i c and p e r c h l o r i c a c id s
o f f e r e d a q u ic k and e a s y m ethod o f d e s tr o y i n g o rg a n ic m a tte r a s w e ll
a s a c c u r a te r e s u l t s f o r s i l i c a s in c e th e p e r c h l o r i c a c i d d e h y d ra te s
t h e s i l i c a w ith o u t h a v in g t o e v a p o r a te t h e s o l u t i o n t o d r y n e s s .
How­
e v e r , s in c e t h e p e r c h l o r a t e s form ed b y t h i s tr e a tm e n t i n t e r f e r e d w ith
t h e t i n m ethod o f rem o v in g th e p h o s p h a te th e m ethod was u s e d o n ly t o
ch ec k t h e r e s u l t s on s i l i c a a s o b ta in e d b y th e a s h and HCl d e h y d r a tio n .
D.
M ethods o f Removing P h o sp h ate
Tn o r d e r t o e s t a b l i s h a p p r o x im a te ly t h e am ounts o f s i l i c a , a lu m i­
num, i r o n , c a lc iu m , and m agnesium i n sh e e p lu n g s t h e e n t i r e m a t e r i a l
o f a n o rm a l ewe was ash e d i n a n i c k e l c r u c i b l e a t 393° t o 4 8 8 ° and
th e r e s u l t i n g a s h a n a ly z e d .
The SiOg was rem oved b y d e h y d r a tio n w ith
HCl and 5 e q u a l a l i q u o t s ta k e n fro m t h e f i l t r a t e f o r d e te r m in a tio n s
on co m p ariso n o f m e th o d s.
Two o f t h e a l i q u o t s w ere r u n on e a c h m ethod
so a s t o a c t a s ch e c k s on one a n o t h e r .
I n t h e f i r s t tw o t h e p h o sp h a te
was l e f t i n s o l u t i o n and i n th e re m a in in g two i t was rem oved b y u se o f
ammonium m o ly b d ate s o l u t i o n .
T ab le I I t a b u l a t e s t h e gram s o f e a c h
e le m e n t fo u n d w h ile T ab le I I I r e p r e s e n t s t h e p e r c e n ta g e s fo u n d .
T ab le
-
27 -
T ab le I I .
W eig h ts o f Aluminum, I r o n , C alciu m and' M agnesium a s
D eterm in ed i n a Hormal S h eep ’ s L u n g s, Vfith
and W ith o u t Removal o f P h o sp h a te
P h o sp h a te
Humber o f
d e te r m in a tio n s * rem oved
P h o sp h ate
• grams
I
no
2
no
S
yes
4
yes
Aluminum
gram s
Iro n
grams
C alciu m
gram s
M agnasium
grams
0 .1 0 8 9
0 .0 1 2
0 .0 0 3 7
0.0 0 0 8
0 .0 7 3 6
0 .0 1 1
0 .0 0 5 4
0.0008
0 .0 1 4 1
0 .0 5 0 7
0 .0 1 0
0 .0 0 6 9
0 .0 0 0 8
0.0134
0 .0 5 0 7
0 .0 1 1
0 .0 0 6 4
0 .0 0 0 8
———
■
*E ash sam ple was a n a l i q u o t c o n ta in in g 2 0 .9 gram s o f m a t e r i a l and
r e p r e s e n t e d o n e - f i f t h o f t h e t o t a l d r y w e ig h t o f t h e lu n g s .
-
28 -
T ab le I I I .
P e rc e n ta g e , o f Aluminum, I r o n , C alciu m , and Magnesium, a s
D eterm in ed i n a Uormal S h eep ’ s L ungs, W ith
and W ith o u t Removal o f P h o sp h ate
Humber o f
d e te r m in a tio n s *
P h o sp h a te
rem oved
Aluminum
per cent
Iro n ■
p e rc e n t
C alcium
per cent
M agnesium
per cent
I
no
0 .1 0 4
0 .0 1 1
0 .0 0 3 5
0 .0 0 0 7
2
no
0 .0 7 1
0 .0 1 0
0 .0 0 3 2
0 .0 0 0 7
3
yes
0 .0 4 8
0 .0 0 9
0 .0 0 6 1
0 .0 0 0 7
4
yes
0 .0 4 8
0 .0 1 0
0 .0 0 6 1
0.0 0 0 8
#S ash sam ple w as a n a l i q u o t c o n t a in in g 2 0 .9 gram s o f m a t e r i a l
and r e p r e s e n t e d o n e - f i f t h o f th e t o t a l d r y w e ig h t o f th e lu n g s .
•
-
29 -
I I I i s c a l c u l a t e d fro m t h e d a t a i n T a b le I I .
I t i s o b v io u s t h a t i n
t h e c a s e s w here th e p h o s p h a te was n o t rem oved t h e aluminum, c o n te n t i s
h ig h e r and t h e c a lc iu m c o n t e n t lo w er t h a n i n th o s e w here t h e p h o sp h a te
•was rem oved.
These d i f f e r e n c e s i n d i c a t e t h a t c a lc iu m p h o s p h a te m ust
p r e c i p i t a t e when t h e s o l u t i o n i s made a l k a l i n e and t h e r e f o r e i s w eighed
a s AlgOg t h u s i n c r e a s i n g t h e amount o f alum inum and d e c r e a s in g th e
am ount o f c a lc iu m .
I n o r d e r t o t e s t t h e a c c u r a c y o f th e v a r io u s m ethods o f a n a l y s e s ,
a known s o l u t i o n was made up w h ic h c o n ta in e d t h e a p p ro x im a te q u a n t i ­
t i e s o f th e e le m e n ts fo u n d i n t h e n o rm al s h e e p 's lu n g s a s t a b u l a t e d i n
T a b le I I .
The s to c k s o l u t i o n o f t h e known was made b y d i s s o l v i n g t e n
tim e s t h e am ount r e p r e s e n t e d b e lo w , m aking up t o 100 c c . an d ta k in g 10
c c . a liq u o ts .
Q u a n t i t i e s P r e s e n t i n Known 10 c c . P o r tio n
0 .0 0 5
gm.
i r o n i n form o f C .P . w ire
0 .0 5 0 0 gm.
alum inum o r .3975 gm. AlCl g . SHgO
0 .0 1 0 0 gm.
c a lc iu m o r .3065 gm. CaClg.SNgO
0 .0 1 0 0 gm.
m agnesium o r .0846 gm. MgClg.SHgO
0 .3 0 0 0 gm.
n i c k e l o r .6 6 4 0 gm. N iC lg
0 .0 1 9 9 gm.
p h o sp h o ru s o r .1 3 2 4 gm. NaNH^HPO^.4Hg0
I n t h e f i r s t d e t e r m in a tio n o f t h i s known th e p h o s p h a te w as removed
b y ammonium m o ly b d ate s o l u t i o n and t h e e x c e s s o f th e l a t t e r a s w e ll a s
n i c k e l n o t rem oved d u r in g th e c o u rs e o f t h e e n t i r e a n a l y s i s .
The r e ­
s u l t s a r e t a b u l a t e d i n T ab le IT u n d e r sam p les N os. I and 2i
A n o th e r d e t e r m in a tio n was c a r r i e d o u t on t h e same known s o l u t i o n
w ith o u t rem o v in g th e p h o s p h a te and th e r e s u l t s o b ta in e d t a b u l a t e d i n
-
30 -
T ab le I T .
C o n te n ts o f a Hnoivn- S o lu ti o n an d R e s u lts O b ta in e d
b y V a rio u s M ethods
Sample num ber
Known
P h o sp h a te
gram s
Aluminum
grams
Iro n
gram s
0.0199
0 .0 5 0 0
0 .0 0 5 0
"
M agnesium
grams
C alcium
gram s
0 .0 1 0 0
0.0 1 0 0
R e s u lts ■O b ta in e d
I
0.0197
0 .0 5 1 1
0 .0 0 5 0
0 .0 1 0 7
0 .0 1 2 0
2
0.0208
0 .0 5 0 0
----
0.O1O9
0.0 1 0 9
3
———
O olSlS
——”
0 .0 0 1 8
0 .0 0 5 1
——w
0 .0 1 2 7
4
• 0 .0 2 0 2
. 0 .0 5 1 4
5
0 .0 1 9 9
0 .0 4 4 5
0 .0 1 3 7
.
0.0 0 8 9
0 .0 1 1 4
—
31 -
T a b le IV u n d e r sam ple No. 3»
& m ethod w h ic h c o n s is te d o f rem oving .th e p h o sp h a te w ith ammonium
m o ly b d ate and rem o v in g t h e e x c e s s o f th e l a t t e r a s w e l l a s th e n i c k e l
b y p r e c i p i t a t i n g w i t h h y d ro g e n s u lp h id e i n a n am m oniacal s o l u t i o n
b ro u g h t b a c k n e a r l y t o a c i d i t y w ith HCl w as r u n on a known s o l u t i o n
i
c o n t a in in g t h e same amount o f a l l e le m e n ts e x c e p t no i r o n was p r e s e n t .
The r e s u l t s o b ta in e d a r e t a b u l a t e d i n T a b le IV u n d e r sam p les N os. 4
and 5 .
The b e s t r e s u l t s seem ed t o b e o b ta in e d when th e p h o s p h a te was
rem oved b u t th e e x c e s s m o ly b d ate s o l u t i o n n o t ta k e n o u t i n t h e e n t i r e
c o u rse o f th e a n a ly s is .
T h is m ethod was. t h e n u se d on th e sam p les o f
th e same lamb's * lu n g s p r o c u r e d a t a m eat m a r k e t.
V a r io u s ways o f
d e s tr o y i n g th e o r g a n ic m a tte r w ere t r i e d and th e r e s u l t s o f th e a n a ly - '
s e s t a b u l a t e d i n T a b le V .
S in c e t h e d e te r m in a tio n s w ere a l l r u n i n
d u p l i c a t e a n d t h e w e ig h ts o f th e v a r io u s e le m e n ts d id n o t seem t o
c h e c k , o th e r m ethods o f rem oving th e p h o s p h a te w ere s o u g h t.
A n o th e r known s o l u t i o n was made f o r t e s t i n g t h e a c c u r a c y o f th e
t i n m ethod o f rem o v in g t h e p h o s p h a te .
The known c o n ta in e d i n e a c h 10
cce p o r t i o n t h e f o llo w in g w e ig h ts o f e a c h e le m e n t w eig h ed fro m th e
m a te ria l as in d ic a te d .
Q u a n t i t i e s P r e s e n t i n Ehbtvn 10 c c . P o r t i o n
0 .0 1
gm.
0 .0 5
gm.
I r o n ; f r oid 'C .P . w ir e
Aluminum o r .3975 gm. AlCl g . SHgO
0 .0 1
gm.
C alcium o r .0365 gm. CaClg.SHgO
0 .0 1
gm.
M agnesium o r .0846 gm. MgClg« 6 ^ 0
0 .2 1 6 9 gm.
P h o sp h o ru s o r 1 .4 4 0 gm. NaNH^HPO^.
I t w as fo u n d t h a t t h e i r o n was p a r t i a l l y removed i n some c a s e s and
T ab le T
Grams o f S i l i c a , I r o n , Aluminum, C alciu m , M agnesium and P hosphorus
P e n to x id e Found i n Sam ples o f Lamb* Lungs "When D i f f e r e n t
M ethods o f D e s tro y in g O rg an ic M a tte r Were Used
W eight o f
sam ple
gram s
M ethod o f d e s tr o y in g
o r g a n ic m a tte r
3-
IO eOO
2
10 e 35
Sam ple
number
Ca
Mg
grams
grams
grams
P2°5
grams
0 .0 0 5 3
0*0030
0 .0057
0.0063
0.2087
0 .0 0 4 0
0 .0 0 8 9
0 .0 1 0 0
0 .0066
0 .2411
Fe
SiOg
grams
grams
ash
0 .0 0 2 0
HEiO3 and HCIO4
0*0027
Al
i
os
3
10*35
s lim in g
0 .0 0 4 0
0 .0 0 3 6
0 .0 0 2 4
none
0 .0100
0.2161
4
10.00
ash
0 .0 0 2 4
0 .0 0 7 2
0*0288
0 .0 1 0 8
0.0180
0 .2175
*T hree s e t s o f lu n g s o b ta in e d fro m t h e m eat m a rk e t, h i s t o r y unknown
r
-
e n t i r e Iy- i n o t h e r s "by t h e t i n .
33 -
T ab le T I shows th e r e s u l t s when th e
known c o n ta in e d th e q u a n t i t y o f i n g r e d i e n t s a s i n d i c a t e d above a s w e ll
a s when i t c o n ta in e d tw ic e a s much alum inum w i t h th e o th e r in g r e d i e n ts
re m a in in g th e sam e.
The v a lu e s o b ta in e d seem t o i n d i c a t e t h e a c c u ra c y
o f t h i s m eth o d .
T a b le T I ! shows th e r e s u l t s when d e t e r m in a tio n s w ere r u n on. t h e
same lu n g s u s in g ammonium m o ly b d ate and t i n m ethods o f rem oving t h e
p h o s p h a te .
As c a n b e s e e n much b e t t e r r e s u l t s w ere o b ta in e d by u s e o f
th e t i n m e th o d .
G re a t d i f f i c u l t y was e n c o u n te re d i n th e ammonium
m o ly b d a te m ethod i n g e t t i n g sam p les t o c h e c k .
The t i n m ethod i s v e ry
e a s i l y and q u ic k ly a p p l i e d a s w e ll a s g iv i n g good r e s u l t s .
B..
R e s u lt s o f Lung A n a ly se s
T a b le T i l l shows t h e p e r c e n ta g e o f e a c h in g r e d i e n t a s o b ta in e d i n
a n a ly s e s o f n o rm a l an d " lu n g e r " lu n g s .
The p e r c e n ta g e i s a n a v e r a g e ■
o f tw o c h e c k s r u n on s e p a r a t e sam ples whose s i z e i s d e s ig n a te d i n th e
ta b le .
I n m ost c a s e s t h e a n a l y s i s o f e a c h s e t o f lu n g s w as r u n th r e e
o r more tim e s and th e f i g u r e s r e a c h e d m o st f r e q u e n t l y u s e d i n T able
T ill.
T a b le s IX’ and X t a b u l a t e th e gram s o f e a c h i n g r e d i e n t u s e d t o ob­
t a i n t h e p e r c e n ta g e s i n T a b le T i l l .
The s i z e sam ple f o r i r o n and
s i l i c a was t h e o r i g i n a l w e ig h t o f t h e sam ple s in c e t h e s e d e te r m in a tio n s
w ere r u n b e f o r e an y a l i q u o t s w ere t a k e n . The s i z e sam ple f o r alum inum ,
c a lc iu m , and m agnesium was s m a lle r b e c a u s e a 90 c c . a l i q u o t o f 100 o c .
was u s e d .f o r t h e i r a n a l y s i s .
T a b le XI r e p r e s e n t s th e am ounts o f t h e v a r io u s m in e r a ls i n th e
54 -
•
. T ab le V I.
D e te r m in a tio n o f Knoim S o lu ti o n When P h o sp h ate
i s Removed b y T in * '
Sample num bers
Aluminum
grams
C alcium
grams
I
0.0443
0 ,0 1 0 0
2
0 .0 4 9 4
0 .0 1 0 0
3
0.0405
0.0100
0 .0 1 1 1
4
0 .0 4 4 0
0 .0 1 1 0
0 .0 1 1 3
5
0 .0 4 9 2
0 .0 1 0 7
0 .0 0 9 7
6
0.0490
0 .0 1 0 4
0.0098
7
0 .0 9 4 0
0 .0 1 0 0
0.0112
8
0 .0 9 8 8 •
0.0101
0 .0 1 2 6
Magnesium
gram s
0 .0 1 1 3
■0.0115 •
^ A n aly se s o f knoTOi s o l u t i o n used" o n Sam ples I t o 6 , i n c l u ­
s i v e : A l 0 .0 5 g ram s, Ca 0 .0 1 g ram s> Hg 0 .0 1 g ram s. F o r
Sam ples 7 an d 8 : A l 0 .1 0 grams an d Ca and Mg same a s a b o v e.
-
35 -
T a b le V I I .
C om parison o f T in 'and Ammonium M olybdate M ethods f o r
Removal o f P h o sp h a te i n t h e Same Lungs
Sheep
number
Sample
number
795N
I
Am. m o ly b d ate
0 .0 1 0
0 .4 8 2
0 .0 7 3
2
Am. m o ly b d ate
0.016-
*
*
. 3
tin
0 .0 1 7
0 .0 6 3
. 0 .0 6 2
4
tin
0 .0 2 0
0 .0 6 2
0 .0 6 9
I
Am. m o ly b d ate
0 .0 2 0
.0 .5 6 0
0 .0 7 0
2
Am. m o ly b d ate
0 .0 2 0
0 .1 6 0
0 .1 0 0
3
tin
0 .0 2 5
0 .0 5 3
0 .0 5 8
4
tin
0 .0 2 5
0 .0 5 4
0 .0 5 8
40F and
137?
M ethod u se d
Aluminum
p er cent
C alcium
per cen t
Magnesium
per cent
^ D e te rm in a tio n r u in e d b e c a u se molybenum o x id e fo rm ed
when c a lc iu m p r e c i p i t a t e d .
T ab le H l I
P e rc e n ta g e o f M o is tu r e , I r o n , S i l i c a , Aluminum, C alcium and Magnesium
C o n ta in e d i n Lxmgs fro m Hormal and "L unger" Sheep
S iz e
of
sample
gms
Sheep
number
444H
795H
40F and
137F**
F-48
430
516
D-83
A verage
Dry '
w e ig h t
Ixmgs M o istu re *
gms
i
10
12
64
78
12
65
70
107
86
91
7 8 .2
12
12
12
12
76 o i l
76 o i l
79.26
79 o14
80*73 .
84.59
8 1 .3 7 ‘
7 9 ,6 2 ■
_____ M ethod o f
d e s tr o y in g
rem oving
p h o s p h a te o r g a n ic m a tte r
tin
Hormal Lungs
a s h in g
It
tr
tr
U
if
tr
it
it
it
it
!I Lxmger "
2570
2578R
617D***
557B
96D
A -60
A verage
12
12
12
12
12
12
■231
201
341
315
230
360
280
82*75
81.06
82.93
81.96
80*78
82.43
81.99
Ii
11
it
ii
Ii
I!
IV
H
ii
11
ii
Iro n
%
S ilic a
%
Aluminum
%
0 ,0 7 9
0.083
0 .038
0 .0 2 7
0.075
0 .0 8 3
0 .0 8 3
0 ,0 6 3
0 ,0 9 4
0 .0 8 0
0 ,0 1 8
0 .0 0 5
0 .021
0 .2 5 0
0*170
0 .1 4 6
0 .0 8 4
0*049
0 ,0 6 3
0 ,1 2 7
0 .0 2 8
0 .0 3 7
0 .0 2 4
0 .032
0 .0 1 1
0 .0 1 5
0 .0 1 9
,
C alcium
%
M agnesium
%
0.0 1 9
0 ,0 1 9
0 .0 5 0
0 .0 6 2
0 .0 7 0
0 .0 6 5
0 .0 2 5
0 .0 2 4
0 .0 2 6
0 .0 1 7
0 .0 5 3
0 .0 5 0
0 .0 7 0
0 ,0 5 4
0 .0 6 1
0.059
0 .0 5 8
0 .0 6 2
0 .0 6 8
0 .072
0.059
0 .0 6 5
0 .0 7 0
0 .0 6 6
0 .1 2 5
0 .0 9 5
0 .0 4 4
0 .1 8 4
0.0 9 7
0 .0 7 7
0 .0 7 8
0 .0 9 2
0 .0 8 9
0 ,0 9 0
0 .0 9 2
0 .0 8 6
0 .0 1 1
0.020
Lxmgs
0 .0 1 0
0 .0 0 4
0 .0 2 3
0 .0 2 1
0.020
0.022
0 .0 1 9
0.021
0,013
0 .0 1 9
*A11 p e r c e n ta g e s e x p re s s e d o n b a s i s o f d ry w e ig h t, e x c e p t m o i s t u r e , w hich was w et b a s i s .
* * T o ta l d ry w e ig h t o f Ixmgs d iv id e d b y 2 .
***Sheep d ie d an d much b lo o d i n Ixm gs«
?
-
37 -
T ab le IX .
W eig h ts o f Aluminum, C alcium and IvIagnesium Found i n
Sam ples o f F orm al and "L u n g er1’ Lungs .
Sheep
number
S iz e o f
sam ple*
gms
D ry w e ig h t
o f lu n g s
gms
Aluminum
C a lc ium
gms
Magnesium
gms
F o rm al Lungs
9.72
444H
64
0 .0 0 1 7
0 .0 0 4 5
0 .0 0 6 2
7951
1 0 .8
78
0 .0 0 2 1
0.0067
0 .0 0 7 1
40F & 137F**
1 0 .8
129
0 .0 0 3 0
0.0064
0 .0 0 7 0
F -48
1 0 .8
70
0 .0 0 2 7
0 .0 0 5 3
430
1 0 .8
107
0 .0 0 2 7
0 .0 0 7 6
0 .0 0 7 4
516
1 0 .8
86
0 .0 0 1 8
0 .0 0 5 8
. 0.0077
D-S3
1 0 .8
91
0 .0 0 1 1
0.0066
0 .0 0 6 4
7 8 .2
0.0020
0 .0 0 6 2
0.0069
A verage
.
0 .0 0 6 7
"L u n g er" Lungs
257C
10.8
231
0 .0 0 2 3
0 .0 0 7 6
0,0 0 8 5
25791
10.8
201
0 .0 0 2 3
0 .0 0 7 1
0.0082
617D
1 0 .8
341
0.0024
' 0 .0 1 3 4
d .o io o
557B
1 0 .8
315
0 .0 6 2 0
0 .0 1 0 3
0.0 0 9 6
96D
1 0 .8
230
0.0023
0 .0 0 4 7
0.0 0 9 7
ArSO
1 0 .8
360
0.0016
0.0198
0.0100
280
0 .0 0 2 1
0.0105
0.0093
A verage
♦W eights o f alum inum , c a lc iu m an d m agnesium c a l c u l a t e d on
same b a s i s o f s i z e o f sa m p le .
♦ ♦ C o n sid e re d a s two s e t s o f lu n g s i n com puting a v e r a g e .
-
58
-
T ab le X .
W eig h ts o f S i l i c a and I r o n Found i n Sam ples o f Normal
and "L unger" Lungs
Sheep
number
S iz e o f ,,
sam ple*
gms
Dry -w eight
o f lu n g s
gms
S ilic a
mgs
Iro n
gms
Normal Lungs
444N
10
64
0.0 0 3 9
0 ,0 0 7 9
795N
.1 2
78
0.0 0 3 2
0 .0 1 0 0
40F & 137F**
12
129
0 .0 0 2 1
0 .0 0 9 0
F-48 '
12
70
0 .0 0 0 5
0 .0 1 0 0
430
12
107
0.0 0 2 5
0 .0 1 0 0
516
12
86
0.0 0 1 3
0 .0 0 7 5
D-83
12
91
0 .0 0 1 9
0 .0 1 1 3
78 c.2
0 .0 0 2 1
0 .0 0 9 3
A verage
"L unger" Lungs
2570
12
231
0 .0 0 3 4
0 .0 3 0 0
2579R
12
201
0.0 0 4 5
0 .0 2 0 0
617D***
12
341
0 .0 0 2 9
0 .0 1 7 0
557B
12
315
0 .0 0 3 9
0 .0 1 0 0
96D
12
230
0.0012
0 .0 0 5 2
A -60
12
360
0 .0 0 0 5
0 .0 0 7 5
0 .0 0 2 7
0 .0 1 5 0
A verage
280
-
■
^W eights o f s i l i c a and i r o n c a l c u l a t e d t o same b a s i s o f
s i z e o f s a m p le .
^ C o n s i d e r e d a s tiro s e t s o f lu n g s i n com puting a v e r a g e <,
***Sheep d ie d and, much b lo o d i n lu n g s .
**
39 —
t o t a l d r ie d lu n g s o f n o rm al and " lu n g e r ” s h e e p .
These r e s u l t s w ere
c a l c u l a t e d from t h e d a t a i n T a b le s IX and X .
The grams o f SiOg and alum inum a r e r e p r e s e n t e d i n F ig u r e I w h ile
F ig u re 2 r e p r e s e n t s th e gram s o f c a lc iu m , i r o n , and m agnesium i n n o r ­
m al and " lu n g e r " l u n g s .
The c u rv e s w ere made on th e b a s i s o f t h e i n ­
c r e a s i n g s i l i c a c o n t e n t , i n o r d e r t o d e te rm in e i f t h e alum inum , i r o n ,
c a lc iu m , o r m agnesium in c r e a s e d c o r r e s p o n d in g ly .
I t I s o b v io u s t h a t
th e s e m in e r a ls do n o t f o llo w t h e shape o f th e s i l i c a c u r v e .
T h e re fo re
t h e s e r e s u l t s i n d i c a t e t h a t th e alum inum , c a lc iu m , i r o n , an d m agnesium
a r e n o t p r e s e n t i n th e fo rm o f s i l i c a t e s .
The o r d e r in ' w h ich th e
lu n g s a p p e a r and t h e number u s e d t o d e s ig n a te them i n F ig u r e s I and 2
a re :
L unger
No.
No.
No e
No.
N o.
No.
I
2
3
4
5
6
£.-60
960
257-C
2579-R
617-D
557-B
Uormal
No.
No.
No.
No.
No.
. No.
No.
7
8
9
10
11
12
13
F-48
516
40—F and 137-F
D-83
795-N ■
430
444-N
F ig u re 2 shows t h a t t h e r e i s a g e n e r a l r e l a t i o n s h i p b e tw e e n th e
i r o n , m agnesium and c a lc iu m c o n te n t i n n o rm al sh e e p , w h ile i n " lu n g e r"
lu n g s t h e r e seem s t o be o n ly r e l a t i o n s h i p b etw ee n th e m agnesium and
c a lc iu m .
F ig u r e s 3 and 4 b a s e d on t h e lu n g s a s t h e y in c r e a s e i n c a lc iu m
c o n te n t a l s o show t h e r e l a t i o n s h i p o f c a lc iu m , m agnesium , and i r o n i n
t h e n o rm a ls , and m agnesium an d c a lc iu m i n th e " l u n g e r s " .
The alum inum ,
s i l i c a , and " lu n g e r " i r o n do n o t seem t o re s e m b le any o f t h e o th e r
-
40 -
T ab le XIe
T o ta l W eight o f S i l i c a , I r o n , Aluminum, C alcium and
M agnesium Found i n D rie d Lungs
Sheep
number
Dry w e ig h t
o f lu n g s
gms
S ilic a
gms
Iro n
gms
Aluminum
gms
C alciu m
gms
Magnesium
gms
Eorm al Lungs
444N
664
0 ,0 2 5 0
0 ,0 5 0 6
0 .0 1 1 2
0 .0 3 0 6
0.0 4 0 8
79 SE
78
0 ,0 2 0 8
0 .0 6 5 0
0 .0 1 5 2
0 .0 4 8 4
0.0 5 1 3
6 4 ,5
0 ,0 1 1 3
0 ,0 4 8 3
0 .0 1 7 9
0 .0 3 8 2
0 .0 4 1 9 ,
79
0 ,0 0 2 9
■ 0 .0 5 8 3
0 .0 1 7 5
0 .0 3 4 4
0 .0 4 3 4
430
107
0 .0 2 2 5
0 .0 9 0 0
0.0268
0 .0 7 5 5
516
86
0.0093
0 .0 5 3 8
0 .0 1 4 3
0.0462
0.0613
D-83
91
0 .0 1 4 4
0 .0 8 5 7
0 .0 0 9 3
0 .0 5 5 6
■0.0 5 4 0
7 8 .2
0 ,0 1 4 7
0 .0 6 2 5
0 .0 1 6 3
0 .0 4 5 9
0 .0 5 1 0
4 0? & 137F*
F -48
A verage
'
0,0 7 3 5
“L unger" Lungs
2570
231
0 .0 6 3 5
0 .5 7 7 5
0 .0 4 9 1
0 .1 6 2 3
0.1816
2579R
201
0 .0 7 5 4
0=3350
0 .0 4 2 8
0 .1 3 2 1
0.1 5 2 6
617D**
341
0 ,0 8 2 3
9 .4 8 3 0 . 0 .0 7 5 8
0 .4 2 3 0
0 .3 1 5 7
557B
315
0 ,1 0 2 4
0.2625
0 .0 5 8 3
0 .3 0 0 5
0 .2 8 0 0
96D
230
0 .0 2 3 0
0 .0 9 9 7
0 .0 4 9 0
0 .1 0 0 1
0 .2 0 6 6
A -60
360
0 .0 1 5 0
0 .2 2 5 0
0 .0 5 3 3
0.6599
- 0.3 3 3 3
0 .0 6 0 3
0 .3 3 0 4
0 .0 5 4 7
0 .2 9 6 3
0.2 4 4 9
A verage
A, 280
'
^ D iv id ed com bined d ry w e ig h t b y 2 t o g e t a v e ra g e d r y w e ig h t o f
e a c h s h e e p ’ s lu n g s , c o n s id e r e d v a lu e a s two s e p a r a t e s e t s i n
d e te r m in in g a v e r a g e s , .
**Sheep d ie d a n d much b lo o d i n l u n g s .
-
41 -
0 2 / _______ .042.________ .063_________ .084________ JOS'
G r a m s
I
/
t
i
I
I
I
7
2.
Q
3
9
4
L ung
I
I
I
I
/0
S //
N um ber
l
I
I
6
/2
/J
G ram s o f \5///ca and A /u m /n u m pe.r
Gunp in G /orm a/ a no/ L unger C/heep
/ Lunges A r r a n g e d A c c o r d in g t o
v /n c r e a tfe
in G ih c a
C o n te n t
—° ------ ° — G iO z
--------------G /
in Lungera
m Normah
f /p u r e ,
— + A/ in Lungera
------------ A! in AIormaia
/
60
./Z
2+
36
4S
\
QI
/
I
^
I
^
I
I
I
<9
j
3
-f
/<?
Lung
/Y o
I
I
I
JT
I
i
I
i
//
6
/Z"
/j
Grams o f G a fc zu m t / f f U^ezeszt/m, arzcf fr o z i
per Lung' z/z /Vozzrzof oncf V u n g e r S h e e p
(Lurpge /frrozzpecf Vfccorafzng /o
Ir) c reuse zn Szfzcu C onfenfj
--------Cu zn L u n g e r s
------------Lffg zn W orm ofs
------ C u
— H- L f g
z/ ?
/n
L Y orznuA s
2a
n g e r s
—o—o— G e
— ---- G e
G zo u rzr
S
zn
zn
L u n g e r s "
/fo r m u fs
03
06
os>______ _^£
^7X-^y//?c5
Grams Oft SiJica a n d A/U m/num p e r
Z unp/ //? /Vorm a/ anc/ d a n g e r Sheep
( L urgs A r r a n g e d
A cco rd /rrg So
Increase /n Ca/c/um Con/en/)
— -+- /9/ //7 Z angers'
SiOji in L u n g e r s
JzZ^ in Aiorma/s
----------/9/ in /Vorma/s
/ rZ G O Z te
J
44-
6ze
•
(2ram s cC Co/c/urn , Artagnes/um one/ Iron
per Lung
/n A/orm a/ one/ L unger !Sleep
( L u n g s Arranged /According /o
In crea se
/n Co/c/u/r? Con/on/)
it
m
O -O -/ e /n L u n g e rs"
Co in //orma/s
Ce /n /Vorma/s
— Hg /n Lungers
4 ^Co /n L u n g e r s ------------- Mg /n L/orma/s
H /G o /e r
45 -
—
c u rv e s.
The s i g n i f i c a n t f a c t o r r e s u l t i n g fro m a l l o f t h e s e f i g u r e s i s
t h a t t h e m in e r a l c o n te n t i s much l a r g e r i n " lu n g e r" th a n i n n o rm al
lu n g s .
J
The lu n g s w ere a r r a n g e d i n F ig u r e s 3 a n d 4 a c c o rd in g t o t h e i r i n ­
c r e a s e i n c a lc iu m an d a r e r e p r e s e n t e d a s f o l l o w s :
L unger
I)
2)
3)
4)
5)
6)
Uormal
96-D
2579-R
257-C
557-C
617-D
A-60
7)
8)
9)
10).
11)
12)
13)
'
F.
444-N
F-48
4 0 -F an d 137-F
516
795-1T
D-83
430
G e n e ra l D is c u s s io n
A la r g e p a r t o f t h e w o rk done on t h e p r e s e n t p ro b lem was c o n fin e d
t o e x p e rim e n ta l w ork on m ethods of- a n a l y s i s .
Of t h e ,v a r io u s m ethods
o f d e s tr o y i n g t h e o r g a n ic m a t t e r w h ich w ere t r i e d th e a s h in g method
was t h e one a p p l ie d i n th e a n a l y s i s o f t h e . lu n g s a m p le s.
The n i t r i c
and p e r c h l o r i c a c i d s m ethod o f S m ith (3 7 ) gave good r e s u l t s a s w e ll
a s a r a p i d m ethod o f r i d d i n g th e m a t e r i a l o f t h e o rg a n ic m a tte r p r e s e n t .
T h is m ethod i s a d v a n ta g e o u s i n t h a t f o r a s i l i c a d e t e r m in a tio n th e
s o l u t i o n does n o t hav e t o b e ta k e n to d ry n e s s i n o r d e r t o d e h y d ra te t h e
s ilic a .
The s i l i c a c o n te n t b y t h i s m ethod u s u a l l y ch eck ed w i t h t h a t
o b ta in e d b y a s h in g an d d e h y d r a tin g t h r e e tim e s w ith H C l.
The r e a s o n
t h a t t h e n i t r i c an d p e r c h l o r i c a c i d s m ethod was n o t u s e d i n t h e a n a l y s i s
o f a l l th e m in e r a ls s o u g h t i n t h i s p ro b lem i s t h a t t h e p e r c h l o r a t e s
form ed i n t e r f e r e d w ith t h e t i n m ethod o f rem oving t h e p h o s p h a te .
S m ith
(3 7 ) c la im s t h a t f o r su c h d e te r m in a tio n a s s u l f u r , p h o s p h o ru s , e t c . .
-
46 -
t h e n i t r i c and p e r c h l o r i c a c id s m ethod i s m ore a c c u r a te t h a n a s h in g
s in c e i n th e l a t t e r , i f p r o p e r p r e c a u tio n s a r e n o t ta k en > t h e s e e l e ­
m en ts a r e v o l a t i l i z e d and l o s t .
The e x p e r im e n ta l tvork done on th e m ethods o f rem oving th e phos­
p h a te te n d s t o f a v o r t h e t i n m ethod o v e r th e ammonium m o ly b d ate me­
th o d .
The t i n m ethod o f rem o v in g a la r g e q u a n t i t y o f p h o s p h a te i n
p re s e n c e o f sm a ll q u a n t i t i e s o f alum inum , c a lc iu m , and m agnesium g iv e s
a c c u r a te r e s u l t s a s T a b le V I show s.
Two' gram s o f t i n w ere s u f f i c i e n t
t o remove a l l o f t h e p h o s p h a te i n a 12 gramm a te ria l.
sam ple o f t h e d r i e d lu n g
The p h o s p h a te p r e c i p i t a t e m ust b e w ashed w e ll so a s n o t to
lo s e any o f th e alum inum , c a lc iu m , o r m agnesium t h a t may te n d t o a d h e r
t o th e p r e c i p i t a t e .
U s u a lly no t i n was p r e s e n t i n t h e f i l t r a t e , b u t ,
i f su c h w ere p r e s e n t , i t c a n e a s i l y and c o m p le te ly be rem oved b y
s a t u r a t i n g t h e f i l t e r e d s o l u t i o n w i t h EgS.
One o f t h e m ost im p o r ta n t f a c t o r s t h a t d e s e r v e s much a t t e n t i o n i n
rem oving t h e p h o s p h a te by th e t i n m ethod i s t h a t o f h a v in g no f r e e
c h l o r i d e io n o r HCl p r e s e n t .
These seem t o h y d ro ly z e t h e p r e c i p i t a t e
form ed and t h u s n o t r e n d e r t h e s e p a r a t i o n o f p h o sp h a te c o m p le te .
The
p h o s p h a te p r e c i p i t a t e may b e c o n s id e r e d e i t h e r a s a s t a n n i c p h o sp h a te
o r a s a m e ta s ta n n ic a c i d w i t h th e p h o s p h a te a d so rb e d w i t h i n i t .
Ex­
p e r im e n ta l e v id e n c e seems t o fa v o r th e l a s t named b e c a u se th e . amount
o f p h o s p h a te rem oved b y a c e r t a i n num ber o f grams o f t i n i s n o t con­
s ta n t .
I f i t w ere c o n s ta n t t h e e v id e n c e w ould p o in t t o a d e f i n i t e
compound, b u t s in c e i t v a r i e s i t a p p e a rs t o b e a n a d s o r p ti o n r e a c t i o n .
I r o n i n s m a ll q u a n t i t i e s a s h a s b e e n fo u n d i n t h e lu n g s a n a ly z e d seems
-
t o b e l a r g e l y rem oved b y t h e t i n .
47 A t e s t f o r t h e i r o n c o n te n t b e f o r e
and a f t e r t h e rem o v al o f t h e p h o s p h a te showed t h a t i n th e fo rm e r case
t h e r e was s u f f i c i e n t i r o n t o b e m e a su re d , w h ile many tim e s i n t h e I a t - '
t e r c a s e t h e r e w ere o n ly t r a c e s .
The alum inum d id n o t seem t o be a f ­
f e c t e d i n t h e same way a s t h e i r o n .
The v a r io u s r e s u l t s oh a knowo. s o lu
t i o n show t h i s to be t r u e .
The g e n e r a l m ethod f o r th e i r o n d e te r m in a tio n was a c c o r d in g to
O f f i c i a l M ethods ( I , X I I , 7 ) .
T h is d e te r m in a tio n was made i n th e p r e ­
se n c e o f p h o s p h a te an d no d i f f i c u l t y was e n c o u n te r e d w ith t h e c o lo r
f a d in g w h ic h E lv eh jem and H a r t ( I l ) n o t i c e d .
These i n v e s t i g a t o r s u se d
KMhO^ i n s t e a d o f HHOg t o keep th e i r o n i n a f e r r i c s t a t e , b u t b e t t e r
r e s u l t s w ere o b ta in e d i n th e p r e s e n t p ro b le m b y u s in g HHOg.
I n th e
i r o n d e t e r m in a tio n r e c o r d e d i n c o n n e c tio n w ith t h i s p ro b lem 0 .5 c c .
o f c o n c e n tr a te d n i t r i c a c id and I c c . o f ECHS w ere ad d ed t o b o th th e
known and unknown, i r o n d e t e r m i n a t i o n s .
The c o l o r o f t h e f e r r i c t h i o ­
c y a n a te form ed d id n o t fa d e and th e a c c u r a c y o f th e m ethod was p ro v e n
b y a known d e t e r m in a tio n .
The a p p l i c a t i o n s o f t h e m ethods w ere k e p t a s u n ifo rm a s p o s s ib le
th ro u g h o u t t h e a n a l y s i s o f b o th " lu n g e r " and n o rm al lu n g s a m p le s .
The
t o t a l a v e ra g e p e r c e n ta g e o f alum inum and s i l i c a re c o rd e d i n T a b le Y I I I
seem t o b e ab o u t t h e sam e, w h ile th e a v e ra g e p e r c e n ta g e s o f i r o n , c a l ­
cium and m agnesium seem t o b e h ig h e r i n th e ” lu n g e r" th a n i n th e n o r ­
m a ls .
The l a r g e s t d i f f e r e n c e seems t o be i n t h e p e r c e n ta g e o f c a lc iu m .
One o f th e m ost s i g n i f i c a n t d i f f e r e n c e s b etw ee n " lu n g e r " and n o r ­
m al lu n g s i s th e l a r g e r s i z e and w e ig h t o f th e f o r m e r .
The p e rc e n ta g e
—
48
“
o f m o is tu r e i n th e tw o ru n s a b o u t t h e sam e, b u t t h e w e ig h t o f t h e d r ie d
" lu n g e r " lu n g s, i s b e tw e e n t h r e e an d f o u r tim e s l a r g e r t h a n t h e n o rm al.
When t h e w e ig h ts o f th e v a r io u s m in e r a ls fo u n d i n t h e sam p les a r e
c a l c u l a t e d p e r w e ig h t o f t o t a l lu n g s a s T a b le X show s, t h e .c o n t e n t o f
t h e v a r io u s m in e r a ls i n th e " lu n g e r " lu n g s w e re h ig h e r t h a n i n t h e n o r ­
m al lu n g s .
The s i l i c a a p p ro x im a te ly 4 t i m e s , th e i r o n ab o u t 5 t i m e s ,
th e alum inum 3 , c a lc iu m 6 , and t h e m agnesium 5 tim e s t h e i r c o rre s p o n d ­
in g v a lu e i n t h e t o t a l n o rm al lu n g s .
T hese a l l seem t o in c r e a s e i n a
h ig h e r p r o p o r ti o n th a n d o es t h e d r y w e ig h t.
From t h e s e r e s u l t s i t c a n d e f i n i t e l y be s e e n t h a t t h e r e i s an i n ­
c r e a s e i n t h e m in e r a l c o n te n t o f " lu n g e r " o v e r t h a t o f n o rm a l lu n g s .
The d i f f i c u l t y e x i s t s i n d i s t i n g u i s h i n g w h e th e r th e h ig h e r m in e r a l con­
t e n t i s due t o in h a le d d u s ts o r p a t h o l o g i c a l . d e p o s i t s . A f u r t h e r p a th o ­
l o g i c a l s tu d y w h ich i s beyond t h e sco p e o f t h e p r e s e n t i n v e s t i g a t i o n
may d e c id e t h i s p ro b le m .
•“
TH.
49 “
SDMMaEY
B o th th e e x p e r im e n ta l d a t a f o r th e t e s t i n g o f t h e m ethods o f a n a l­
y s i s and th e a c t u a l r e s u l t s w hen t h e s e m ethods w ere a p p l ie d h av e b e e n
in c lu d e d i n t h i s p ro b le m .
1.
The " s l i m i n g " , th e n i t r i c and p e r c h l o r i c a c i d s , an d t h e a s h in g
m ethods o f rem oving t h e o r g a n ic m a tte r hav e a l l b e e n t r i e d and th e a s h ­
in g m ethod u s e d b e c a u s e o f i t s a p p l i c a b i l i t y t o t h e p ro b le m .
O c c a s io n a l­
l y t h e n i t r i c and p e r c h l o r i c a c id s m ethod was u s e d t o c h e c k th e s i l i c a
c o n te n t a s p ro c u r e d when t h e a s h was d e h y d ra te d w ith H C l.
2.
I t w as fo u n d t h a t t h e p h o s p h a te h a d to be rem oved i n o r d e r t o
make a c c u r a te a n a l y s i s .
B o th .th e ammonium m o ly b d ate and t h e t i n m ethods
o f rem oving t h e p h o s p h a te w ere t r i e d , and t h e l a t t e r fo u n d t h e more ap ­
p l i c a b l e a s w e ll a s g iv in g th e b e t t e r r e s u l t s .
3.
The t i n m ethod o f rem o v in g p h o s p h a te h a s f o r t h e f i r s t tim e b e e n
shown s a t i s f a c t o r y f o r q u a n t i t a t i v e w o rk . T a b le T I shows i t s r e l i a b i l i t y
i n p r e s e n c e o f r e l a t i v e l y l a r g e am ounts o f p h o s p h a te an d s m a ll am ounts
o f A l, Ca, Mg an d F e .
The m o ly b d ate m ethod does n o t seem t o b e s a t i s ­
f a c t o r y (s e e T ab le IT ) f o r th e m a t e r i a l i n q u e s tio n w here s m a ll am ounts
o f A l, Ca, Mg an d Fe m ust b e d e te rm in e d a c c u r a t e l y .
4.
One o f th e m o st o u ts ta n d in g d i f f e r e n c e s o b se rv e d b etw ee n n o r ­
m al and " lu n g e r " lu n g s i s t h a t o f t h e h e a v i e r w eig h t o f t h e l a t t e r ' s
d rie d m a te r ia l.
A lth o u g h t h e p e r c e n ta g e m o is tu r e i s a b o u t t h e same i n
b o th c a s e s , th e " lu n g e r " d ry w e ig h t i s 3 .7 tim e s h e a v ie r t h a n t h a t o f
t h e n o rm a l.
5.
I n t h e " lu n g e r " lu n g s th e c a lc iu m i s a b o u t 2 t i m e s , magnesium
-
50
-
a b o u t 1-|- t i m e s , an d i r o n a b o u t 2 tim e s a s much a s i n n o rm al lu n g s o f th e
same s i z e d r y sam p le, w h ile th e s i l i c a and alum inum c o n te n t seem t o be
a b o u t t h e same i n b o tn c a s e s » "When th e w e ig h ts o f th e m in e r a l c o n te n t o f
t h e e n t i r e d r ie d lu n g s w ere c a l c u l a t e d t h e y w ere much g r e a t e r i n th e d i s ­
e a s e d t h a n i n t h e n o rm a l lu n g s .
The a v e ra g e o f th e m in e r a l c o n te n t o f th e
t o t a l d r i e d " lu n g e r " lu n g s c o n t a in s a p p r o x im a te ly th e f o llo w in g o v e r t h a t '
i n th e n o rm al lu n g s : s i l i c a 4 t i m e s , i r o n 5 t i m e s , alum inum 3 t i m e s , c a lc iu m
6 t i m e s , and m agnesium 5 t i m e s .
6.
S in c e b lo o d c o n t a in s c o n s id e r a b le i r o n and c a lc iu m , t h e q u a n t ity
o f t h e s e two fo u n d i n t h e a n a l y s i s o f t h e lu n g s w ould v a r y w ith th e
am ount o f b lo o d p r e s e n t .
The e x c e ss b lo o d i n t h e lu n g s was k e p t v e r y low
b y o b s e rv in g p r e c a u tio n s w hen rem oving t h e lu n g s from th e sh eep so as t o
k eep i t s c o n te n t a s lo w a s p o s s i b l e .
7.
I t i s beyonfl t h e sco p e o f t h e p r e s e n t'p r o b le m ' t o make any c o n c lu ­
s io n s a s t o w h e th e r o r n o t th e in c r e a s e i n m in e r a l c o n te n t o f th e " lu n g e r"
o v e r n o rm a l lu n g s c o u ld b e th e c a u se o f th e " lu n g e r " d is e a s e o f s h e e p „
To
d e te rm in e w h e th e r t h e in c r e a s e d m in e r a l c o n t e n t i s due t o in h a le d d u s ts o r
p a t h o l o g i c a l d e p o s i t s i s a l s o e x c lu d e d from t h e p r e s e n t i n v e s t i g a t i o n .
• -
51
-
ACKNOWLEDGl/iEMI
The a u th o r im ishes t o ta k e t h i s o p p o r tu n i ty t o e x p r e s s h i s a p p re c ia /
t i o n t o P r o f e s s o r Edmund Burke and Mr. FrahIc D o n ald so n o f th e Montana
A g r i c u l t u r a l E x p erim en t S t a t i o n , t o D r. H a d le ig h M arsh and D r. Howard
W elch o f t h e M ontana V e te r i n a r y R e se a rc h L a b o r a to r y , and t o D r. 0 . E .
S h e p p a rd , D r. E . L . H a e n is c h , D r. B. L. J o h n so n , and P r o f e s s o r P 0 C.
G ain es o f t h e C h e m istry D epartm ent o f M ontana S t a t e C o lle g e f o r tim e ,
a s s i s t a n c e , an d a d v ic e g iv e n him d u r in g t h e y e a r w h ile t h i s w ork was
b e in g c o m p le te d .
52 —
—
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( S I)
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ReTiewed by
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.
5 , (1914) p . 1 5 3 .
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/0 0
'5
N3T8
W69d
cop. 2
54884
Wilson, E. P.
The determination of
moisture, silica,iron,
aluminu, calcium and
magnesium in the lungs