EXPERIMENTAL CRITICISM OF RECENT RESULTS IN

TESTING ADRENALIN

W. H. SCHULTZ

From the Division of Pharmacology, Hygienic Laboratory, Public Health an4 Marine
Hospital Service, Washington, D. C.

Received for publication, June 26, 1909

In Bulletin No. 55 of this Laboratory (8) the relative vaso-constrictor

activity, toxicity, and mydriatic action of certain catechol deriva-
tives of the adrenalin type’ is discussed and in connection there-
with is a review of the literature extending up to 1909. Recently,
however, there have appeared several articles the results of which
it seems well to mention and discuss.
Abderhalden and Thies (2), 1909, worked with a preparation of
1-adrenalin having an optical activity of (a) - 50.40, and with
d-adrenalin having an optical activity of (a)’ + 50.49. The enu-
ceated eyes of frogs were supported in small glass funnels and studied
1 The results in brief are as follows:
1-The blood pressure method with dogs under morphine ether anasthesia, with
the vagi cut and small doses of curare administered, is the most accurate pharmacolog-
ical essay for catechol derivatives of the adrenalin type.
2-The pupil method as modified by the author is a reliable essay for adrenalin,
but is a method less delicate and more tedious than that of blood pressure.
3-Synthetic dl-adrenalin is less active as a vaso-constrictor and as a mydriatic
than natural 1-adrenalin, the ratio being 2: 3.
4-The relative vaso-constrictor activity of the catechol derivatives in the order
named, 1- and dl-orthodioxyphenylethylanolmethylamin, ortho-dioxyphenyl-ethanol-
amin, an(l ortho-ethyl aminodioxyacetophenon, are to each other as the inverse ratios
1: 1.5: 1: 80.
5-The toxicity of the substances in the order named in 4 are to each other as the
inverse ratios of 1: (1.5 to 2): 5 : (71 to 80).
6-The relative physiological activity of these catechol derivatives seems to depend
upon the substance partaking of the properties of a secondary alcohol or of a ketone,
upon the nature and number of groups displacing the hydrogen of the amino group,
and upon the arrangement of the asymmetric carbon atom in space.

\
This OnS

CD22_479_X96G
292 W. H. SCHULTZ

in pairs. After first observing the eyes the solution to be tested was
dropped upon the corneal surface of one eye of a given pair and upon
its companion an equivalent amount of salt solution, whereupon the
eyes were again observed for one or two hours. As the result of six
experiments it is concluded that d-adrenalin does not affect the ex-
cised eye of the frog, but that the 1- product does, and since dl-adrena-
lin dilates the pupil, it does so by virtue of the lavo component in
the racemic product. Furthermore, d-adrenalin does not produce
glycosuria, the 1avo does, hence when it is thus produced the 1-,
not the d- product, is responsible.
Abderhalden and Slavu (1), 1909, studied the effect of subcutaneous
injections of adrenalin upon mice. In animals weighing from 11 to
14 grams an injection of 0.1 to 0.2 milligram of 1-adrenalin caused
the temperature to fall as much as 15.5#{176}C., the animal usually dying
when the temperature fell to 22#{176}
or 23#{176}
C. If, however, d-adrenalin
was used, a 13-gram mouse could be injected with as much as 5 milli-
grams with but slight change in temperature and no perceptible after
effects. An 11-gram mouse was injected daily with doses of increas-
ing size, starting with 1 milligram until a 5-milligram dose of d-adren-
alin was reached after which the mouse recovered from an 0.8-milli-
gram dose of 1-adrenalin, which, according to these writers, is eight
times the lethal dose. From this they infer that the d- product
causes more or less tolerance towards 1-adrenalin, and that the differ-
ence in their action is the result of physical differences present in the
two substances.
Comessatti (3), 1909, used the excised eyes of 1?. temporaria and
Hyla arborea in a comparative study of the mydriatic substances in
urine and serum and of adrenalin solutions. One eye was placed in
Ringer’s solution, the other in serum, urine, or the substance to be
tested and both illuminated by electric light. Observations were
made for a period of two hours.
TJndialyzed urine brought about opaquing of the lens and so it
was dialyzed for two or three hours in a Philippson tube, which re-
moved the substance responsible for the opalescence. Comessatti also
experimented with 5, 2.5, 1.25, 1, 0.6, 0.50, 0.30, and 0.25 per cent.
sodium chloride solutions and found that saline solutions do not
cause prompt dilation of the pupil, but 1.25 per cent. and over cause
 CRITICISM OF RECENT RESULTS IN TESTING ADRENALIN 293

opalescence of the lens. He thinks that the opalescence observed in

eyes immersed in undialized urine is likewise caused by a high per-
centage of inorganic salts usually found in urine. When the bulbi
were tested with dilute solutions of catechol and of adrenalin, the
latter proved to be at least ten times the more active
Blood serum of persons suffering from various diseases was also
tested but only one case gave a positive reaction and even this one
was unsatisfactory. In another set of experiments, he used 15 cc.
of fresh human serum to which was added three to four drops of
commercial adrenalin solution and allowed it to stand for twenty
to thirty hours, testing it from time to time upon the frog’s excised
eye and also with his sublimate reaction. In each test of the adrenalin
solution the reaction was prompt and positive whereas the control
serum tests yielded uniformly negative results, thus showing that
adrenalin offers considerable resistance to the action of human serum
when outside of the body. As a result of the experiments he concludes
that the enucleated frog’s eye immersed in blood serum does not
show a specific reaction towards adrenalin and that it has in this
Particular case limited yet undeniable value, especially when results
are obtained with blood serum, diluted 10 to 20 times, of persons
suffering from disease. He concludes that Schur and Wiesel (9)
must then have been testing serum containing adrenalin, since of the
substances known to exist in the blood that produce mydriasis none
of them cause a distinct dilation in qtiantities so small as would exist
in serum diluted 10 to 20 times. But should the eye fail to dilate
while in the blood serum there still remains more or less the feeling
that adrenalin may be present.
Finally, Comessatti thinks the frog’s bulbus is only in a measure
an indicator for adrenalin. It is not an infallible test object, and
even a positive reaction for adrenalin can only be relied upon when
the transverse diameter of the pupil increases in length so as t cause
an undoubted rounding within 20 to 30 minutes after treatment with
the liquid. He considers, on the other hand, the frog’s bulbus to be a
test object of the greatest value when employed in trying the strength
of pure adrenalin solutions of unknown concentration.
Recent articles by Meltzer (6) and Ehrmann (5) are for the most
part polemic in nature, the controversy arising from Ehrmann’s using
294 W. H. SCHULTZ

a modification of the pupil method. as published by Meltzer without

sufficiently acknowledging Meltzer’s priority. It is indeed true that
Meltzer cites only a few experiments. Nevertheless they were per-
formed and attention was called to the advantage of using the excised
frog’s eye for testing adrenalin solutions. In his article on the effect
of suprarenal extracts upon the frog’s eye (1904) (7) it is stated that
“in carefully excised eyes, dropping of adrenalin on the corneal sur-
face brought on promptly a dilation of the pupil which lasted many
hours.” Just how much more sensitive the excised eye is toward
adrenalin than is the intact eye seems not to have been determined
at the time; nevertheless complete excision of the bulbus had, in
Meltzer’s opinion, advantages, else why should he have concluded by
saying, “The frog’s eye, excised or in situ, might prove to be a better
reagent than the blood pressure to demonstrate the efficiency of a
suprarenal preparation”?
Meltzer (6) 1909, shows the error of calling the pupil method “Ehr-
mann’s Method,” and also describes a more recent one employed by
himself. It consists in excising the eye and placing over its ante-
rior surface a disc of rubber (previously greased with some inert
substance) in such a manner that the opening of the rubber ring
forms a receptacle over the corneal surface. Into this receptacle is
dropped the solution to be tested and by comparing these eyes with
controls, the receptacle of which contained normal saline or Ringer
solution, the limit of dilution ws determined at which the eye showed
positive mydriasis. Eyes studied under these conditions dilated so
that the pupil was apparently round when in solutions containing
not less than one part in 10,000 of Ringer.2 Meltzer thinks the
limit of sensitiveness towards adrenalin is one part of adrenalin to
10,000 parts of solvent; Ehrmann claims to be able to detect adrenalin
in body fluids if present in so small an amount as 1 to 10 million.
Ehrmann (7) considers this more recent method of Meltzer inferior
to the method of complete immersion used by other recent investi-
gators-Ehrmann (4), Cornessatti (3), Schultz (8)-and is perhaps
justified in thinking so. Yet the method recently described by Melt-
zer ought to prove valuable as a preliminary test. He thinks that
Meltzer and Auer’s (7) observations show nothing further than that
2 Catechol was found to be one-tenth as active as adrenalin.
 CRITICISM OF RECENT RESULTS IN TESTING ADRENALIN 295

an adrenalin solution of 1:1,000 or 1:10,000 after injection or instilla-

tion causes mydriasis in the frog’s eye, which, he states, has already
been observed by Lewandowsky. He also maintains that the remark-
able increase in sensitivity of the frog’s eye by enucleation of the
bulbus was observed neither by Lewandowsky nor by Meltzer but by
himself, who alone has used this method to test blood or body fluids.
The matter stands thus: it was Meltzer who first used the excised
frog’s eye for testing the mydriatic action of adrenalin, and it was
he who called attention to its convenience and delicacy as a pharma-
cological test object, though without attempting to show how much
more sensitive is the excised than the intact eye. To Ehrmann credit
is due for bringing the method into prominence by reason of his experi-
ments with body fluids, and for exciting interest in the subject by
claiming for the eye as a result of enucleation a thousandfold greater
sensitivity towards adrenalin.
With reference to Abderhalden and Thies’s (2) work it will be
noticed that they do not mention having guarded against errors
resulting from changes of temperature, light value, and the possibility
of mechanical stimuli. As a result considerable oscillation in the
lengths of the pupillary axes is a natural consequence. Furthermore,
they used solutions too concentrated to prove the relative activity of
synthetic dl- and natural 1-adrenalin. It is possible that maximum
mydriatic effects can be obtained with a 1:5000 solution, although
the small number of experiments cited by them does not bring out
this point so clearly as is shown in tables 1 and 2 of the present arti-
cle. Any method like that last used by Meltzer (6) and by Abder-
halden and Thies (2) in which it is necessary to drop upon the surface
of the cornea the solution to be tested is open to serious objections
and cannot be expected to yield quantitative results.
In Comessatti’s work with the pupil he does not seem to have
guarded against changes of temperature, but he immersed the bulbi
in the solution tested and also studied them in artificial light. Except
for not taking into account the temperature, his technique is good
and his conclusions ought to bear with them considerable weight.
Yet I cannot entirely agree with him that in order to be sure that
the mydriasis resulting from a given solution is a purely adrenalin
effect the pupil must dilate to roundness in from 20 to 30 minutes.
296 W. H. SCHULTZ

My own experience has been that the length of the pupillary axis
ought to increase by at least 0.4 to 0.5 mm. and maintain this length
within a few hundredths of a millimeter in order to eliminate the
possibility of the eye having dilated in response to mechanical or
thermal stimuli. In other words, if an eye in adrenalin solution
dilates to maximum in 30 minutes and immediately begins to con-
strict, returning to normal within two or three hours, it is almost
certain that the dilation noted during the first thirty minutes was
not necessarily due tO adrenalin.
A 1:125,000 solution of natural 1-adrenalin may cause the pupil to
dilate more widely than does a 1:1000 solution. And in the course
of dilation it is not always the short axis that dilates most rapidly,
though this may seem to be the case when observed by the naked
eye. Measurements show that the long axis often increases in length
more rapidly than does the short one, and upon reaching a (see 42b
and 48b) maximum length it then comes to a standstill or decreases
in length while the short axis gradually reaches its greatest length.
Then, like the long axis, it usually decreases in length and finally
comes to rest, whereupon both axes remain stationary or oscillate in
length slightly. At this time the pupil may be round, oval, or irregular,
depending upon the eye used.
It would seem that the degree of mydriasis is not proportional to
the strength of the adreralin solution. Data with reference to this
are clearly brought out in tables I and II, in which experiments 37b,
48b, 52b, 56b, and 59b are all with eyes of approximately the same
size, yet they do not show that a definite relation exists between
the amount of dilation and the concentration of the solution used.
The degree of adrenalin reaction should instead be sought for in what
may be called the maximum dilation time. For although solutions
of adrenalin do not necessarily cause a degree of mydriasis in propor-
tion to their concentration, the more concentrated solutions require
a shorter time in which to bring about maximum mydriasis.3 Even
By maximum mydriasis is meant the greatest length of one or both pupillary
axes acquired under existing experimental conditions, not the greatest possible
dilation for the eye to attain under normal conditions. Thus, with a given drug the
axes may increase in length up to a certain point and no further, after which they
either shorten or remain stationary. This greatest length is almost invariably less
than that attained in the normal eye when dilated to its greatest possible size, as
in very dim light.
 CRITICISM OF RECENT RESULTS IN TESTING ADRENALIN 297

this unit is not to be relied upon when eyes are chosen promiscu-
ously as in tables I and II. It is, however, very reliable when eyes
of freshly caught frogs are studied in pairs so that the right and left
eye of a given frog are immersed, say, in 2 cc. of a 1:20,000 or in a
1:40,000 solution of adrenalin at 20 or 26#{176}
0.
It would seem that there is a certain optimum concentration in
which adrenalin acts most uniformly upon the enucleated frog’s eye.
This optimum concentration, though varying somewhat with indi-
viduals, is approximately one part of adrenalin base to 20,000 of
Ringer solution. With such a solution at 20#{176}
the dilation time is
about 27 minutes. If more concentrated solutions be used there will
be a gradual shortening of this time until a certain per cent. (again
depending upon the individual) of adrenalin content is reached, after
which further increase of concentration fails to shorten the time
required for the axes to reach their greatest length. Thus it is pos-
sible to choose a set of solutions of such concentration that all produce
a maximum mydriatic effect, though some members of the set con-
tain ten times as much adrenalin per cc. as do others. Naturally,
the dilation time of eyes immersed in such solutions will be about
the same. If, on the other hand, weak solutions are used which con-
tain less than 1:625,000, the dilation time at 20#{176}
to 23#{176}
becomes variable
and the degree of mydriasis is usually so small as to be of uncertain
origin. For comparative work, whether quantitative or qualitative
in nature, it would seem then advisable to limit the experiments, at
23#{176}
C., to solutions containing not more than one part of adrenalin
base to 10,000 parts of Ringer and not less than one of adrenalin to
125,000 of Ringer and certainly not less than 1: 625,000.
Finally, to those interested in a clinical method for determining
the presence of adrenalin in body fluids the pupil method is, according
to Ehrmann and others, an exceedingly delicate and reliable test-
object. Yet in the light of my own experiments I cannot recommend
it in such strong terms as do these writers. If the excised eye yields
uncertain results when immersed in a 1:625,000 Ringer adrenalin
solution of known composition, what must one expect with a patho-
logical serum the adrenalin content of which is undoubtedly less than
this, not to mention the possible influence of other less well-known
substances that by their combined action may cause the slight
mydriasis often observed with such sera?
298 w. H. SCHULTZ

TABLE I
Mydriatic Action of Natural i-Adrenalin and of Synthetic di-Adrenalin
(Excised eyes of Rana halecina, Kalm)

NATURAL L-ADRENALIN SYNTHETIC DL-ADRENALIN

.# .a ia
CONCEN-

Temp. Increase in TRATION Temp.

No. OF SOLN.
No. Increase in
of length of of length of . S
Sn. axes i GM. PER of Soln
Exp. Exp. axes
s. C.#{149}
.- .-.
0 os
s E
z z
long short long short
48l 23.2 0.780 0.600 13 1000cc. 48a 23.2 0.780 0.725 18
22.8 0.685 0.605 15 Ringer 22.8 0.785 0.660 24

49b 23 1.045 1.380 14 49a 23.0 1.145 1.425 22

1.170 1.255 36

51b 23 1.005 1.365 16 2000c.c. 51a 23.4

Ringer 23 0.850 0.930 21

50a 23 1.135 1.525 21 SOb 23.3

1.085 1.580 34 1.130 1.440 21

35a 22.1 0.620 1.390 18

0.890 1.245 20

37b 23 0.650 1.160 26 37a 20.6 1.240 1.485 21

#{149} 0.530 1.260 13 22 1.220 1.560 25

39a 23 0.845 1.090 20 39b 23.4 1.060 1.325 15

22.6 0.825 1.090 13 23.8

40a 1.125 1.600 22 5000cc. 40b 23 0.750 0.970 17

1.000 1.685 42 Ringer 23.5

41a 23.8 0.760 1.150 14 41b 23.8 1.010 1.080 17

0.540 1.190 31 0.910 1.205 26

52b 23.2 0.830 0.630 1 52a 23.2 1.100 0.935 23

23.8 0.715 0.780 45 23.4 1.090 0.990 21

53a 23.8 0.800 0.925 24 53b 23.8 0.970 1.125 25

0.790 0.965 21 0.955 1.190 36

54b 23.3 1.115 1.755 14 56a 0.865 1.310 34

1.335 1.740 17 0.880 1.220 23

42b 22.5 1.100 0.720 67 42a 22.5 0.595 1.045 70

22.8 1.080 0.735 60 25000c.c. 0.595 1.055 84
Ringer
43b 21.8 0.940 1.480 66 43a 21.2 1.225 1.685 61
22 0.930 1.490 74 21.8 1.145 1.685 54

55b 23.8 0.925 0.805 31 55a 22.2 0.760 0.480 56

23.6 0.905 0.830 24 22.4 0.620 0.550 31

56b 22.6 1.220 1.165 61 56a 22.6 1.080 .285 46

23 1.215 1.225 26 1.075 1.295 43
 CRITICISM OF RECENT RESULTS IN TESTING ADRENALIN 299

TABLE II
Mydriatic Action of Nalural i-Adrenalin and of Synthetic di-Adrenolin
(Excised eyes of Rana halecina, Kalm)

NATURAL L-ADRENALIN SYNTHETIC DL-ADRENALIN

I
I9u.cI 9,C
-e

CONCEN- Increase in
No Temp Increase n TRATION
OF SOLN. No. Temp.
of of length of of of length of d
Exp. 5n. axes 1 GM. PER Exp. :
Soln axes
e4
‘3,_sI oi.
.E, 2S

long short long short

44b 22 1.080 1.375 55 125000cc. 44a 22 0.980 1.375 332
22.2 i 0.950 1.505 114 Ringer 0.985 1.335 359

57b 22.8 0.910 1.040 95 57a 22.6 0.870 0.795 153

23 0.835 1.065 187 23 0.835 0.825 203

59b 22.6 0.990 1.085 84 59a 22.4 0.815 0.990 117

22.8 0.990 1.100 127 23.8 0.765 1.155 69
-

45a 22.2 0.410 0.430 114 625000cc. 45b 22.2 0.180 0.040 174
22.4 0.310 0.440 118 Ringer 22.4 0.060 0.060 51

46b 22.2 0.590 0.390 307 46a

22.3 0.585 0.455 325 23 0.555 0.315 367

60b 22.4 0.440 0.495 23; 60a 22.4 0.390 0.385 74

22.8 0.430 0.535 17

61b 22.4 0.205 0.225 11 61a 23 0.285 0.475 22

65b 22.2 0.215 0.210 77

22.6 0.210 0.260 95

66a 29.2 0.360 0.390 249

47b 0.315 0.565 17 3125000c.c. 47a 0.280 0.285 249

0.125 0.280 145 Ringer 0.228 0.365 275

62b 22.2 0.520 0.585 14 62a 22.8 0.380 0.360 67

0.510 0.580 219 0.355 0.400 79

63b 22.8 0.140 0.245 83 63a 0.230 0.180 288

23.6 0.170 0.195 182 0.060 0.065

64b 22.6 0.130 0.235 38

26.8 0.140 0.180 64

The figures in Tables I and II represent the changes in the length of the long and short axes
of the pupil measured by the pupilometer described in a previous article (8). The excised bulbi
were placed in glass containers of about c.c. capacity, kept at a constant temperature and a
light intensity equivalent to that of 16 c. p. incandescent light 10 inches away. The normal or
zero reading is of the constricted pupil in Ringer solution, at the temperature given in the tables.
After once bringing the eye to a given temperature the solution in which it was studied was
brought to the same temperature, especial care being taken not to disturb the eye in trans-
fering the solutions.
300 W. H. SCHULTZ

At its very best the excised frog’s eye as a pharmacological assay

for adrenalin is inferior to the blood-pressure method. As a qualita-
tive test it is perhaps one of the most sensitive test objects known,
but it is not a characteristic test (Comessatti, Meltzer) and observa-
tions convince me that too much weight ought not to be attached
to results with it in clinical diagnosis.
As has been mentioned, by taking eyes as they come and compar-
ing the relative mydriatic effect of solutions containing equivalent
amounts of natural 1- and synthetic dl-adrenalin base, not enough
difference can be detected to warrant the conclusion that di-adrenalin
is one-half as active as the 1- product. This is shown in tables I and II..
It is true there are special cases which seem to indicate that the hevo
form is a more potent mydriatic than the racemic. But the burden
of evidence in these tables supports the idea that the two are, as
mydriatics, about equal. If, however, the two substances be tested,
the one upon the left eye and the other upon the right eye of the
same frog, results are obtained which seem to agree with those recorded
by the blood-pressure method. One is safe, then, in concluding that
hevo-adrenalin is more active than the racemic product.
With reference to their article on toxicity, Abderhalden and Slavu
(1) cite some very interesting data on changes in body temperature
resulting from subcutaneous injections of 1-, d-, and dl-adrenalin. Yet
it would indeed be very remarkable if a substance that interferes
with the processes of respiration, circulation, and sugar metabolism
as does 1-adrenalin, did not also lower the body temperature. In
spite of their observations of the great difference between d- and
1-adrenalin in lowering the body temperature and of certain experi-
ments from which it is possible to calculate approximately the lethal
dose of 1-adrenalin, there is not a sufficient number of these experi-
ments cited to establish the lethal dose, much less to determine the
relative toxicity of the 1- and dl-adrenalin. Nor by the few experi-
ments cited is it possible to make sure that d-adrenalin can establish
a tolerance toward 1-adrenalin. The reason for this will become
evident after studying the tables published in Bulletin 55 of this lab-
oratory. As a result of injecting nearly four hundred animals it
was found that certain white mice withstand relatively larger doses
of adrenalin, whereas others succumb to very small doses. When
 4

CRITICISM OF RECENT RESULTS IN TESTING ADRENALIN 301

injected subcutaneously with 0.008 milligram per gram body weight

these mice usually die. Very rarely they may die from 0.004 milli-
gram per gram body weight, whereas others may survive as large a
dose as 0.017 milligram per gram. Usually, however, 0.008 milligram
of natural I-adrenalin per gram body weight proves fatal, and may
be considered the lethal dose.
If, instead, synthetic di-adrenalin be used, 0.006 to 0.008 milligram per
grain body weight may cause death, but this is exceptional. Twelve thou-
sandths to 0.016 milligram per gram body weight causes a rather high
death rate and may be said to be the lethal dose, so that the relative
toxicity of natural 1-adrenalin and synthetic di-adrenalin is as the
inverse ratio of 1 :(1.5-2). This is in perfect accord with blood-
pressure experiments in which the relative vaso-constrictor activity
of these two products is in the ratio of 2:3, the natural 1-adrenalin
being the stronger.
In conclusion it may be said that:
(1) In order to detect with excised frog’s eye the difference between
natural 1-adrenalin base and synthetic dl-adrenalin base one must use
eyes as nearly alike as possible, preferably one solution upon the
right eye and the other upon the left eye of a given frog. In this
case the 1-adrenalin is shown to be more active than the dl-, the ratio
of physiological activity being approximately that indicated by the
blood-pressure method.
(2) There is an upper and a lower limit of concentration beyond
which solutions of adrenalin fail to yield graduated results. Above a
certain concentration further increase in adrenalin content influences
the dilation time so slightly as to render accurate determinations
thereof almost impossible, and by exceeding the lower limit of dilu-
tion not only is the dilation time variable for solutions of a given
concentration but the degree of mydriasis is so slight as to be of
uncertain origin. If this is correct, Ehrmann must modify his views
regarding the possibility of detecting adrenalin in solutions so dilute
as one part in ten million, and Abderhalden and Thies’s results are of
doubtful value since they worked with solutions too concentrated.
(3) In comparing two samples of adrenalin the solutions should be
diluted or concentrated as the case requires, so that one solution when
tested on the right eye shows the same dilation time as the other
solution tested on the left eye of the same frog.
302 w. H. SCHULTZ

(4) The dilation time is a better index of the relative physiological

activity of two adrenalin solutions than is the degree of mydriasis.
(5) The pupil ought to so dilate that there is an increase in the
length of its axis of not less than 0.4 to 0.5 mm. and to remain in
this condition, to within a few hundredths of a millimeter, for some
hours; otherwise the mydriasis may be of doubtful origin. It is not
necessary to assume, as Comessatti and Meltzer do, that the pupil
ought to dilate to roundness within 20 or 30 minutes after placing
the eye in the solution in order to prove the presence of adrenalin.
(6) For white mice the lethal dose of natural 1-adrenalin is 0.008
mg. per gram body weight and for dl-adrenalin about .012 to .016
mg. Or, according to Abderhalden and Thies, it is .1 g. fora medium-
sized mouse (about .007 mg.).

(1) Abderhalden, E. u. Slavu: Weitere Studien #{252}berdas physiologische Verhalten

von 1-, d-, und di-Suprarenin, Zeitschr. f. physiol. Chem., lix, 129, 1909.
(2 Abderhalden, E. u. Thies, F.: Weitere Studien #{252}berdas physiologische Ver-
halten von I-, d-, und dl-Suprarenin, II Mitteilung, Zeitschr. f. physiol. Chem., lix,
22-28, 1909.
(3) Comessatti, G.: Ueber den Wert der Froschbulbus-Reaktion und einige Eigen-
schaften des Adrenalins, Arch. f. exp. Path. u. Pharmakol., lx, 233, 1909.
(4) Ehrmann, R.: Ueber eine physiologische Wertbestimmung des Adrenalins und
semen Nachweis im Blute, Arch. f. exper. Path. u. Pharmakol., Leipz., liii, 97-111,
1905.
(5) Ehrmann, R. Zur Methode des qualitativen und quantitativen Nachweises
Kleinster Adrenalin-mengen in Blut- und KOrperfl#{252}ssigkeiten, Deutsche med. Woch-
nschr., xxxv, 676, 1909.
(6) Meltzer, S. J.: Bemerkungen zur Wirkung von Adrenin auf die Froschpupille,
Deutsche med. Wochnschr., xxxv, 575, 1909.
(7) Meltzer, S.J. and Auer, C. M.: The effect of suprarenal extract upon the pupils
of frogs, Amer. J. Physiol., xi, 449-454, 1904.
(8) Schultz, W. H.: Quantitative pharmacological studies-I. Adrenalin and
Adrenalin-like Bodies. Bulletin 55, U. S. Pub. Health and Marine-Hospital Service,
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