EFFECT OF ATROPINE ON CHLOROFORM

HYPERGLYCEMIA

ELLISON L. ROSS

Department of Physiology and Pharmacology, Northwestern University Medical

School

Received for publication January 5, 1920

The administration of atropine was found to reduce the hyper-

glycemia produced by ether anesthesia (1). It was thought of
value to determine the influence of atropine on the increase in
blood sugar produced by chloroform anesthesia. Since the gross
action of general anesthesia is a prominent and common one
of both ether and chloroform, differences in actions of the two
drugs are in danger of being overlooked.

EXPERIMENTAL WORK

Dogs were used as subjects. The diet of the animals was

uncontrolled except that they did not receive food within twelve
hours previous to anesthesia. Two groups, each consisting of
ten dogs, were used.
The animals of the first group were treated in the following
manner: Each dog was tied to a dog board and about 8 cc. of
blood was taken into a syringe from the jugular vein. This blood
was mixed with a small amount of powdered sodium oxalate.
The anesthetic was administered by inserting the animal’s head
into a large cylinder into which was forced air that had passed
over chloroform. A large amount of air was used in order to
reduce asphyxiation to a minimum. The manipulations were
done as quietly as possible and the animal handled carefully, to
prevent excitement. Since it had been found that atropine had
accomplished the major part of its action in the first fifteen
minutes of ether anesthesia, the animals were kept under ch.loro-
135
136 ELLISON L. ROSS

form only fifteen minutes. A second sample of blood was taken

in the same manner as the first and at the end of the fifteen
minutes of anesthesia The dextrose in the samples of blood
was determined by Benedict’s method (2). Triplicate deter-
minations were made in all cases. The results are given in
table 1.
The second group of dogs was treated in exactly the same way
as the first except that fifteen minutes before the above described
procedure was started the animals were bled and atropine given
intravenously. Atropine sulphate one to one thousand parts of
TABLE 1

Effect of chloroform anesthesia alone

NORMAL FIPTEEN MINUTES OP CELORO-

DOG
(PER CENT DEXTROSE) FORM ANESTHESIA

1 0.087 0.104
2
0.095 0.114
3 0.099 0.135
4 0.091 0.120
5 0.095 0.123
6 0.108 0.170
7 0.101 0.136
8 0.105 0.136
9 0.099 0.108
10 0.090 0.116

Average 0.097 0.1262

Per cent 100 1)

water was used to the extent of 0.1 cc. per kilogram of body
weight. This is twice the amount necessary to paralyze the vagus
nerve of the average dog (3). The results are given in table 2.
The group of animals which received chloroform only averaged
0.097 per cent of dextrose in their blood before anesthesia. After
fifteen minutes of surgical anesthesia the blood contained 0.126
per cent of dextrose. This procedure then produced an increase
of glycemia of 30 per cent.
The group of animals which received atropine before the
administration of chloroform, averaged 0.103 per cent of blood
 EFFECT OF ATROPINE ON HYPERGLYCEMIA 137

sugar before any drugs were given. Fifteen minutes action of

atropine had very little effect, the glycemia being 0.1038 per cent.
The administration of chloroform for fifteen minutes brought up
the blood sugar to 0.1334 per cent. This is an increase above
normal of 29 per cent. The increase in this case agrees so closely
both relatively and absolutely, one is forced to conclude that
atropine does not reduce chloroform hyperglycemia as it does
ether hyperglycemia.
Ether anesthesia caused an increase of blood sugar of 41 per
cent. Atropine reduced this increase to 9 per cent. Ether and
TABLE 2

Effect of chloroform anesthesia after atro pine

NORMAL FIFTEEN MINUTES FIFTEEN MINUTES OP

DOG
(PER CENT DEXTROSE) AFTER ATROPINE CHLOROFORM

1 0.094 0.095 0.126

2 0.089 0.097 0.118
3 0.109 0.111 0.135
.
4 0.112 0.106 0.157
5 0.092 0.099 0.138
6 0.109 0.105 0.138
7 0.085 0.086 0.090
8 0.123 0.124 0.192
9 0.105 0.105 0.120
10 0.112 0.110 0.120

Average 0.1030 0.1038 0.1334

Per cent 100 100 129

chloroform are usually considered to have essentially the same

action. That atropine reduces ether hyperglycemia and does
not that of chloroform suggests that the group of factors giving
rise to the two hyperglycemias differ. Of the things that may
be mentioned as possible causes of increase in blood sugar are,
first, asphyxia (4), second, reduced cardiac efficiency which pro-
duces asphyxia, and third, direct action of ether or chloroform
on glycogen in the liver to set dextrose free. The possibility of
one of these factors being more influential with the one anesthetic
than the other and the influence of atropine on this factor was
thought to be worth investigating.
138 ELLISON L. ROSS

Asphyxia will produce an increase in the sugar content of the

blood (4). In the process of inducing anesthesia asphyxia of a
greater or lesser degree may be produced. By the usual cone
method of administering ether or chloroform sufficient air is not
allowed to enter the patient’s respiratory passages and varying
degrees of asphyxia are bound to result. In our experimental
work this factor was practically eliminated. The animal’s head
being inserted into a large cylinder into which was forced a
large amount of air which had passed over the anesthetic, insured
an abundance of oxygen.
TABLE 3

Cessation of respiration as produced by ether and chloroform with and without

atro pine

ETHER CHLOROFORM
DOG

Without atropine With atropine Without atropine With atropine

1 17 11 35 32
2 13 14 27 30
3 16 19 50 60
4 13 9 41 43
5 13 20 40 26

Average 14.4 14.6 38.6 38.2

The irritating action of ether or chloroform on the respiratory

passages is considerable. This irritation leads to more or less
suspension of respiration. Asphyxia so resulting must give rise
to some hyperglycemia. The question arose whether atropine
might not reduce this factor for ether and not for chloroform. To
test this point 20 anesthesias were done on 10 dogs. Ether was
used on half the number of dogs and chloroform on the remainder.
The first anesthesia was induced without any premedication.
The second was preceded by an intravenous injection of 0.1 cc.
of 0.1 per cent of atropine per kilogram of body weight. Pneu-
mographic tracings were taken of each case. The time of sus-
pended breathing was measured in seconds in all cases. The
results are given in table 3.
 EFFECT OF ATROPINE ON HYPERGLYCEMIA 139

The data presented in the preceding table indicates a wide

variation in animals as far as suspension of respiration is con-
cerned. Ether alone caused an average stoppage of breathing
of 14.4 seconds and with atropine a cessation of same of 14.6
seconds. Atropine did not seem to have any influence on this
factor. Chloroform caused a stoppage averaging 38.6 and with
atropine a stoppage of 38.2 seconds. Here also, atropine had
no effect. The striking difference between ether and chloroform
is indicated. Chloroform caused very much greater cessation of
respiration than ether.
TABLE 4 -

Change in heart rate from ether as affected by atro pine

NO ATROP1NE BEFORE ANESTHF.BIA WITH ATROPINE BEFORE ANESTHESIA

DOG

Before During Difference Before During Difference

1 134 132 -2 180 180 00

2 96 108 +12 213 204 -9
3 136 152 +16 159 147 -12
4 126 132 + 6 138 177 +39
5 130 138 +8 150 180 +30
6 228k 162*

Average 124.4 132.4 +8.0 178.0 175.0 -3.0

* Without these values included in average

168.0 177.6 +9.6

Asphyxia may be produced by a reduction of the efficiency

of the heart. This is strikingly evident in heart disease. It is
well known that atropine increases the heart rate through reduced
vagus action. Irritating substances like anesthetics in the
respiratory passages are known to reduce reflexly the heart rate
(5). It was thought well to see if atropine affected the heart
rate the same for the two anesthetics. If atropine should prevent
a slowing of the heart in ether anesthesia and not in chloroform
anesthesia, we would have an answer to our problem. To
determine the facts on this phase of the question, a series of
five dogs was anesthetized with ether and the heart rates deter-
mined before and during the anesthesia. Later the same dogs
were given the same dose of atropine as above mentioned and
140 ELLISON L. ROSS

then the same observations as before were made. Another series

of five dogs was treated in the same way except that chloroform
was used in the place of ether. The heart rates were determined
by dotting on paper every heart beat heard through the stetho-
scope for a period of twenty seconds. Three determinations
were made in each case and the average taken as the value. The
results are given in tables 4 and 5.
The heart rates of dogs treated as described are subject to
many strong influences. The temperament of the animal, the
manner in which he is handled, the training of the dog, and the
surroundings are all factors which affect the heart rate before
the anesthetic. During the first fifteen minutes of anesthesia,

TABLE 5

Change in heart rate from chloroform as affected by afro pine

NO ATROPINE BEFORE ANESTHESIA WiTH ATROPINE BEFORE ANESTHESIA

DOG

Before DUIing Difference Before During Difference

1 132 112 -20 161 192 +31

2 110 112 +2 188 183 -5
3 . 132 110 -22 196 185 -11
4 128 128 00 198 178 -20
5 141 134 -7 194 174 -20
6

Average 128.6 119.2 -9.4 187.4 182.4 -5.0

the degree of struggle during the administration of the anesthetic,

the method of giving the drug, and the rate of intake of the drug,
are factors which alter the heart rate. So it is not surprising
to find the wide individual variations recorded in the above
tables. With no premedication the heart beat averaged 8.4
beats more when the animal was under the ether than before.
When atropine was given before anesthesia the heart rate averaged
a loss of three beats due to anesthesia. The data on one of the
cases was so different from the others it was thought justifiable
to omit it from the calculation of the average. When’ this is
done the average gain amounted to 9.6 beats. The differences
in heart rate seem to be little or not at all affected by atropine.
 EFFECT OF ATROPINE ON HYPERGLYCEMIA 141

Chloroform alone caused a decrease in heart rate of 9.4 beats

per minute. When atropine was administered before chloroform
the decrease averaged 5 beats per minute. The influence of
atropine on this heart rate decrease is considered negligible.
The comparison of the effects of ether and chloroform on the
heart rate does show a decided difference, as is commonly known
to exist. Ether increased the heart rate approximately the same
amount that chloroform decreased it. It is known that chloro-
form injures heart muscle and blood pressure is reduced (6).
With the muscle injury, lowered blood pressure and the decreased
heart rate, the efficiency of the heart must be considerably
reduced. It is thought sufficient asphyxia may result in this
way for chloroform anesthesia to cause appreciable amounts of
hyperglycemia.
The most likely source of the blood dextrose which is increased
by anesthesia is the glycogen of the muscles and especially of
the liver. It is conceivable that ether or chloroform might have
the power to cause a liberation of dextrose from glycogen in
living liver cells. If such an action could be demonstrated and
the difference of ether and chloroform measured, the influence
of atropine on the action of each could be determined. A number
of attempts were made in this direction.
A dog was bled from the jugular vein as long as possible and
then tfte dog was kified by injecting ether into the fourth ventricle
of the brain. The blood was defibrinated, filtered through gauze
and diluted with an equal volume of physiological salt solution.
This mixture was divided into three equal parts. To one ether
was added to the strength of 0.4 per cent, the concentration
found in the blood of anesthetized animals. To another volume
enough chloroform was added to make the concentration 0.05
per cent, which is the strength of chloroform in the blood of
anesthetized animals. The three solutions were kept in stoppered
bottles and heated to body temperature. The portal vein was
isolated and a cannula inserted. The rest of the hepatc cord was
ligated. The vena cava just below the entrance of the hepatic
veins was ligated. A cannula was put in the vena cava toward
the liver just above the entrance of the hepatic veins. The

THE JOUR. OF PHARM. AND EXPER. THERAP., VOL. XV, NO. 2

142 ELLISON L. ROSS

body of the dog was kept warm with an electric pad. The liver
was then perfused three times with untreated blood, three times
with ether blood, three times with untreated blood, and three
times with chloroform blood. Then the series was repeated.
Determinations of the dextrose content of ether and chloroform
bloods were made before and after each triplicate perfusion.
The results are given in table 6.
A second test of the glycolytic powers of ether and chloroform
was made in a manner similar to the preceding. It differed in
that the blood was diluted one to three of normal salt solution.
The perfusions differed. They were in the following order, first,
normal salt solution; second, undrugged blood; third, ether
blood; fourth, normal salt solution; fifth, undrugged blood;
TABLE 6

Liver perfusions with ether and chloroform bloods

Results expressed in perceets of dextrose

PREPARATION INCRESSE
PERFUSION PERFUSION

First ether 0.068 6.50 6.43

First chloroform 0.080 7.20 7.12
Second ether 6.50 12.02 5.52
Second chloroform 7.20 11.11 3.91

sixth, chloroform blood; seventh, normal salt solution, and eighth,

undrugged blood. No solution was used for more than one
perfusion. The results are given in table 7.
The data given in table 7 confirms the suggestion given by the
results recorded in table 6, i.e., that the liver glycogen gives up
dextrose to a certain point independent of the presence of ether
or chloroform in the blood mixture which washes it away.
The power of chloroform or ether to liberate dextrose from
glycogen, was tested by another experiment. A dog was bled
to death. The blood was defibrinated and divided into three
parts. Ether was added to one part to the strength used above,
and chloroform was added to another part to the strength used
before. The liver was removed and several lobes were ground
to a pulp. To each portion of blood 25 grams of liver preparation
 EFFECT OF ATROPINE ON HYPERGLYCEMIA 143

were added. The amount of dextrose in the fluid filtered from

the above mixture was determined immediately after mixing,
after fifteen minutes of incubation and after thirty minutes of
incubation. It was thought that since anesthetic hyperglycemia
appeared quickly, that if ether or chloroform acted as liberators
of dextrose from glycogen, they acted within thirty minutes.
The results are given in table 8.

TABLE 7

Liver perfusion with blood preparations containing ether and chloroform

Results expressed in percents of dextrose

PREPARATION . PERVUSION
PERFUSION
INCREASE

Blood alone 0.03 3.03 3.00

Blood with ether 0.03 2.55 2.52
Blood alone 0.03 0.10 0.07
Blood with chloroform 0.02 0.10 0.08
Blood alone 0.02 0.06 0.04

TABLE 8

Liver glycolysis as affected by ether and chloroform

Results expressed in percents of dextrose

BLOOD WITH
SAMPLE BLOOD ALONE
OFC

Before incubation 0.47 0.41 0.42

Fifteen minutes incubation 0.60 0.61 0.61
Thirty minutes incubation 0.75 0.73 0.72

It is quite clear from this experiment that neither chloroform

nor ether has any effect directly on glycolysis of dead liver
glycogen. Therefore it was thought unnecessary to make any
tests in this connection with atropine.
It is common knowledge that chloroform is capable of causing
pathology in the liver. The question arose whether in the living
and intact animal chlorofotm hyperglycemia might not be the
result of chloroform injury to the liver and in this way the
mechanism of chloroform hyperglycemia differ from that of ether.
144 ELLISON L. ROSS

Davis and Whipple have shown that fasting increases the suscepti-
bility of the liver to injury by chloroform anesthesia (4). They
also have shown that a carbohydrate diet protects the liver to
a certain degree from injury by chloroform. It was thought
well to determine the chloroform hyperglycemia before and after
fasting. The animals were subjected to only a short fast of
two days. The results are given in table 9.
In attempting to interpret the meaning of the results given
in table 9 a further question seemed to be vital. Does the amount
of stored glycogen influence the amount of dextrose set free in
TABLE 9

Effect of fasting on chloroform hyperglycemia

BEFORF FASTING AFTER FASTING

DOG
Before After i Before After i
chloroform chloroform ncrease chloroform chloroform ncrease

1 0.078 0.135 0.057 0.083 0.085 0.002

2 0.096 0.129 0.033 0.087 0.111 0.024
3 0.112 0.144 0.032 0.101 0.138 0.037
4 0.097 0.132 0.035 0.088 0.100 0.012
5 0.109 0.152 0.043 0.089 0.124 0.035
6 0.100 0.115 0.015 0.076 0.093 0.017
7 0.099 0.133 0.034 0.093 0.107 0.014
8 0.109 0.147 0.038 0.109 0.112 0.003
9 0.114 0.147 0.033 0.098 0.120 0.022
7 0.106 0.133 0.027 0.093 0.105 0.012

Average 0.1020 0.1367 0.0347 0.0917 0.1095 0.0178

the blood by the action of any agency? Since ether has relatively
no injurious action on the liver, it seemed that to obtain the
data given in table 9 for ether in the place of chloroform would
go far in answering our question. Therefore a series of 10 dogs
was used to determine the ether hyperglycemia before and after
a fast of two days. The data is given in table 10.
From table 9, it is found that chloroform anesthesia before
fasting caused a hyperglycemia of 0.0347 per cent am! after
fasting 0.0178 per cent. There was a decrease in the rise of
blood sugar of nearly 50 per cent. The fasting had accomplished
two things, i.e., the store of glycogen was reduced and the liver
 EFFECT OF ATROPINE ON HYPERGLYCEMIA 145

was made more susceptible to injury by chloroform. Ether

anesthesia, according to table 10, increased the dextrose of the
blood 0.0387 per cent before fasting and 0.0336 per cent after
fasting. The fast had little or no effect on ether hyperglycemia.
With ether the liver injury factor is negligible, but the fast
reduced the store of liver glycogen the same in the ether series
as it did in the chloroform series of animals. Since the ether
hyperglycemia was not altered materially by fasting, we are led
to conclude that the reduction in the store of glycogen by two

TABLE 10

Effect of fasting on ether hyperglycemia

BEFORE FASTING AFTER FASTING

DOG
Before After Before After
anesthesia anesthesia ncrease anesthesia anesthesia ncrease

1 0.082 0.136 0.054 0.079 0.125 0.046

2 0.096 0.134 0.038 0.086 0097 0.011
3 0.085 0.126 0.041 0.076 0.126 0.050
4 0.081 0.17 0.056 0 100 0.144 0.044
5 0.085 0.126 0.041 0.111 0.144) 0.029
6 0.100 0.132 0.032 0.112 0.144 0.032
7 0.085 0.145 0.060 0.072 0.096 0.024
8 0.086 0.143 0.057 0.090 0.152 0.062
9 0.108 0.114 0.006 0.096 0.114 0.018
10 0.091 0.093 0.002 0.085 0.105 0.020

Average 0.0899 0.1286 0.0387 0.0907 0.1243 0.0336

days of fasting has no influence on the increase of blood sugar.

This being determined, we are able to interpret the findings with
chloroform in relation to fasting. The injury to the liver cells
was the only factor which remained to explain the changes in
chloroform rises in blood dextrose. This injury was not only
not the cause of the freeing of dextrose into the blood but was
a positive factor in preventing its liberation into the blood
stream.
146 ELLISON L. ROSS

SUMMARY AND CONCLUSION

A group of animals was anesthetized with chloroform and the

increase in blood dextrose was determined. Another group of
animals was given atropine before chloroform anesthesia and the
change in glycemia determined. The change in the amount of
sugar in the blood was not affected by atropine.
It had previously been reported that atropine administered
before ether anesthesia reduced the increase in blood sugar. A
cause for this difference in ether and chloroform was sought.
The inhibition of respiration during the induction of anesthesia
with ether was compared with that of chloroform. It was found
that atropine had no effect on the results of either chloroform
or ether. Chloroform clearly caused very much more asphyxia
by this phase of its action than ether.
The influence of ether and chloroform anesthesia on the heart
rate as altered by atropine was measured. Atropine did not
materially change the relations with either chloroform or ether.
Chloroform decreased the heart rate about equal to the increase
caused by ether, which amounted to approximately 5 per cent.
A series of tests were made on the effect of ether and chloroform
in blood upon liver glycolysis. It was found that neither of
these anesthetics had any influence on the rate of dextrose liber-
ation from dead liver cells.
The relation of injury of liver cells to the changes of dextrose
in the blood was determined for chloroform. The means of
increasing the injury to the liver cells was by having the animals
fast for a period before the administration of chloroform. It
was found that with the increased liver injury the rise in blood
sugar was decreased. The reduced store of glycogen, which
accompanied the increased injury to the liver by chloroform,
was found to be without influence as was shown by the absence
of an alteration of ether hyperglycemia through fasting.
From the results obtained, the following conclusions may be
drawn:
1. Atropine administered before chloroform anesthesia did not
reduce the hyperglycemia.
 EFFECT OF ATROPINE ON HYPERGLYCEMIA 147

2. Atropine administered before ether or chloroform anesthesia

did not alter the changes in either heart rate or respiration.
3. Chloroform reduced heart rate while ether increased it, a
fact which has been observed by others.
4. Chloroform caused more than twice .as much respiratory
inhibition compared to that of ether.
5. A two days fast decreased chloroform hyperglycemia and
did not affect ether hyperglycemia.
6. Chloroform asphyxiates markedly more through respiratory
inhibition and reduced heart rate than ether. This asphyxiation
is the probable cause of a large part of chloroform hyperglycemia
and the cause for its not being altered by atropine.

REFERENCES

(1) Ross, E. L.: Jour. Pharm. and Exper. Therap., xii, no. 7, February, 1919.
(2) BENEDICT, S. R.: Jour. Biol. Chem., xxxiv, 203,. 1918.
(3) PILcEER, J. D., AND SOLLMANN, T.: Jour. Pharm. and Exper. Therap., v,
317, 1914.
(4) DAVIS, N. C., D WHIPPLE, G. H. : Arch. Inter. Med., xxiii, no. 5, 1919.
(5) MEYER AND GOTTLIEB: Pharmacology, p. 59, Lippincott, 1914.
(6) Soul&&rN, T.: Manual Pharmacology, pp. 566-567, W. B. Saunders Co., 1917.