A STUDY OF THE ANTISEPTIC AND THE PHARMA-

COLOGIC PROPERTIES OF META-CRESOL ACETATE

ISIDOR GREENWALD

Prom the Laboratory of Biological Chemistry of Columbia University, at the College

of Physicians and Surgeons, New York

Received for publication June 8, 1911

I. INTRODUCTION

Phenol and the cresols have been used as antiseptics for many
years. They are not suitable intestinal antiseptics, however,
because, being quite soluble, they are readily absorbed from the
stomach and, in any but very diKite solutions, are extremely
corrosive. Their external use, also, is dangerous. Absorption
of phenol or a cresol from a large area is apt to result in severe
intoxication and the application of aqueous solutions in dressings,
etc., to the extremities is frequently followed by gangrene. Con-
sequently, these substances are used externally to a much smaller
extent than was formerly the case and are employed internally
chiefly in the form of compounds such as salol. Such compounds
are but sparingly soluble in water and are not materially decom-
posed in the stomach. They are gradually broken down in the
intestine, where they yield the active constituents. These are
produced in such low concentrations as to be almost non-irritant.
Of the three isomeric cresols and phenol, meta-cresol is the
least toxic. This has been reported by a number of observers.
Thus, Tollens gives the data summarized on the next page.

‘Tollens: Arch. f. Exp. Path. and Pharm., 1905, lii, p. 220.

513
514 ISIDOR GREENWALD

LETHAL DOSE PER GRAM LETHAL DOSE PER EILO

Frog Mouse Cat

mg. ,ng. gm.

Phenol 0.10 0.35 0.09

Para-cresol 0.15 0.15 0.08
Ortho-cresol 0.20 0.35 0.09
Meta-cresol 0.25 0.45 0.12

Meili2 reported that meta-cresol was less toxic to rabbits than

either phenol, ortho-cresol or para-cresol. Binet,3 who used rats
and guinea pigs, found that meta-cresol is less toxic than either
of its isomers.
Although less toxic to animals, meta-cresol has a greater bac-
tericidal action than either of its isomers or phenol. This has
been established by the researches of many investigators, among
whom may be mentioned Hammer,4 Seybold5 and Hammer!.6
The great toxicity and corrosive action of the phenols seem to
be influenced by the hydroxyl group attached to the benzol
ring. If a phenol is modified by the introduction of a radical
which eliminates the hydroxyl group, the toxicity is very much
diminished. It appears, therefore, that if meta-cresol could be
modified in a manner that would diminish its toxic and corrosive
properties without interfering with its germicidal activity, the
product might be of considerable therapeutic value.
Several years ago, Dr. Nathan Sulzberger, of this city, began an
effort to produce, from meta-cresol, a compound that would
possess the strong germicidal and analgesic action of the phenols,
but which would lack their irritating and escharotic properties.
Meta-cresol acetate was the outcome of that investigation. Dr.
Sulzberger submitted the results of his work to Dr. Gies, with
whose advice I have performed the experiments described in this
paper.
2Meili: Vergleichende Bestimmung der Giftigkeit der drei isomeren Kresole
und des Phenols. Berne, 1891.
3Binet: Rev. med. de la Suisse romande, 1895, p. 561.
‘Hammer: Arch. f. Hyg., 1891, xii, p. 3.59.
6Seybold: Z. f. Hyg., 1898, xxix, p. 377.
llammerl: Hyg. Rundschau, 1899, ix, p. 1017.
 STUDY OF THE PROPERTIES OF META-CRE5OL ACETATE 515

II. EXPERIMENTAL

General Physical and Chemical Properties of Meta-Cresol Acetate

Physical properties. Meta-cresol acetate is a colorless, oily

liquid, which boils at 212#{176}
(uncorrected) and possesses an agree-
able odor. Its specific gravity at 26#{176}
(1120 at 4#{176} 1) is 1.048.
=

It is insoluble in water and glycerol, but is freely miscible with

common organic solvents, such as alcohol, ether, chloroform,
petroleum ether and benzo!.
Chemical stability. Meta-cresol acetate is quite stable. A
sample that had been about the laboratory for over five months
had the same boiling point as a fresh preparation. It gives only
those tests for meta-cresol in which strong hydrolyzing reagents
are used.
Resistance to hydrolysis. In order to estimate the rapidity
with which meta-cresol acetate may be hydrolyzed under various
conditions, 10 cc. were added to each of three bottles containing
500 cc. of 0.5 per cent hydrohloric acid solution, 1 per cent
sodium carbonate solution and distilled water, respectively, all
at 40#{176}
C. Ten cubic centimeter samples of the acid and alkalin
solutions were removed immediately after the addition of the
meta-cresol acetate and at intervals thereafter. Each was titrated
with am.monium hydroxid or sulfuric acid solution, with
Congo red as the indicator. The distilled water was tested for
acidity and with ferric chiorid solution for free cresol. The ferric
chlorid reaction was negative throughout, probably because
the meta-cresol formed was kept in solutior in the excess of the
acetate. The figures obtained in these experiments are given
in Table I.
In the acid and alkalin solutions, meta-cresol acetate was grad-
ually decomposed. Hydrolysis by distilled water proceeded very
slowly. Apparently, then, meta-cresol acetate would be decom-
posed very slowly under physiological conditions.
Action on protein. When thoroughly shaken with a fairly
concentrated solution of egg-white, meta-cresol acetate coagu-
lated some of the protein. Under these conditions the droplets
516 IBIDOR GREENWALD

TABLE I

Hydrolysis of meta-cresol acetate in 0.5 per cent HC1 solution, 1 per cent Na2CO3
solution and di8tilled water
Cubic centimeters of acid or alkalin solution required to neutralize 10 cc. of
the liquid

0.5 PER CENT HC1 1 PER CENT Na,COs H,0 (ACIDITY)

hours
0 (Control) 7.1 11.0 none
1 7.1 10.6 none
2 7.4 10.4 none
3 7.3 10.2 none
5 7.3 10.1 none
8 7.5 9.8 none
24 7.7 9.6 none
31 8.05 9.6 none
51 8.3 9.3 none
75 8.6 9.0 none
120 8.8 9.2 none

Days
6 9.0 9.2 none
8 9.4 8.8 none
10 9.6 8.7 none
14 12.8 8.5 none
17 13.1 8.3 none
22 8.3 slight
30 0.3
35 0.3
193 0.8
232 0.8
254 1.8

4Completely dissolved.

of meta-cresol acetate were coated by coagulated protein, which

prevented their coalescence. The greater part of the protein,
however, was not coagulated even after contact with the acetate,
with frequent shaking, for four days.

Antiseptic Action

Meta-cresol acetate acts as a powerful antiseptic. In one

experiment 0.02 cc. in 100 cc. of peptone broth kept. at 38#{176}
prevented putrefaction for over seventy-two hours. In two flasks,
 STUDY OF .THE PROPERTIES OF META-CRESOL ACETATE 517

each containing 100 cc. of such broth, to which had been added,
respectively, .0.04 and 0.08 cc. of meta-cresol acetate, there was
no appreciable bacterial growth after two months in the incubator.
In another series of experiments 0.06 cc. added to 100 cc. of pep-
tone broth checked the multiplication of bacteria to such a degree
that prior to the fifth day of incubation, there was no evidence
of film formation. Under these conditions 0.08 cc. prevented
putrefaction indefinitely.
Comparative tests were made with meta-cresol. It was found
that 0.07 cc. in 100 cc. of peptone broth checked putrefaction for
five days and 0.1 cc. prevented it indefinitely. In this respect,
therefore, meta-cresol was found to be as active as the free cresol.
The broth that had been preserved with meta-cresol acetate
did not give a test for cresol with ferric chloride. Apparently,
then, the antiseptic properties of the acetate were not due to its
decomposition into acetic acid and meta-cresol.
Urine to which a little meta-cresol acetate had been added did
not decompose. The acetate, however, was slowly hydrolyzed,
the acidity of the urine increasing slightly from day to day.

Effects on Animals

Experiments on frogs. Subcutaneous injection. Meta-cresol

acetate being insoluble in water and separating from it very read-
ily, it was impossible to use an aqueous or saline emulsion in
this work. In the experiments upon frogs a mixture of glycerol
and water having the same density as meta-cresol acetate was
used. Such a mixture contains about 20 per cent of glycerol
and, when shaken vigorously with meta-cresol acetate, forms a
fairly stable emulsion. A control injection of the maximum vol-
ume of the water-glycerol solution which was used as a carrier in
these experiments was without effect. The injected emulsion
contained 2.01 grams of meta-cresol acetate in 100 cc. Since the
lethal dose of meta-cresol acetate was found to be considerably
greater than that calculated from the figures given by Tollens
(p. 2), 1.635 gram of meta-cresol was made up to 100 cc. with
20 per cent glycerol solution and portions of this liquid were
518 ISIDOR GREENWALD

injected into a number of frogs. When injected into a dorsal

lymph sac both meta-cresol acetate and meta-cresol, in such
media, produced the usual symptoms of phenol poisoning. Over-
excitability and convulsions were more marked in the frogs that
were given the acetate, and paralysis was the chief symptom in
those that received the free cresol. The essential data are sum-
marized in Table II.
TABLE II

Comparative toxicity of meta-cresot and its acetate introduced subcutaneou8ly.

Experiments on frogs

META-CRESOL ACETATE META-CRESOL

Amount Administered Metacresol Equivalent Amount Administered

per Gram per Gram per Gram

mg. mg. ing.

Maximum dose not fol-
lowed by symptoms 0.207 0.149 0 147
Minimum dose followed
by symptoms 0.258 0.186 0.199
Maximum dose followed
by symptoms but not
terminating in death 0.614 0.442 0.486
Lethal dose 0.469* 0338 0.294*
Lethal dose 0.588f 0.423 0.287t
Lethal dose 0.714 0.514 0.419
Lethal dose 0.637#{176} 0.458 0.427#{176}

*Death occurred between nine and twenty-four hours after injection.

tDeath in two hours.
tDeath in eight hours.
§Death in one and one-half hours.
#{182}Deathin four hours.
#{176}Deathwithin one hour.

Considerable variation in the resistance of frogs to the influ-

ence of these substances was observed but the results show that
the poisonous action of meta-cresol tcetate on frogs is approxi-
mately equal to that of its meta-cresol equivalent.
Experiments on dogs. Administration per os. A number of
dogs received meta-cresol acetate and also meta-cresol by mouth.
Each dose was given in gelatin capsules concealed in balls of hashed
meat. The animals were found to vary greatly in their resistance
as is shown by the data summarized in Table III.
 STUDY OF THE PROPERTIES OF META-CRESOL ACETATE 519

TABLE III

Comparative toxicity oj meta-cresol and its acetate given by mouth. Experiments

on dogs
META-CRE8OL ACETATE META-CRESOL

Amount Administered Meta-cresol Amount Administered

per Kg. Equivaient per Kg. per Kg.

gin. gin. gin.

Maximum dose not fol-
lowed by twitching of
muscles 0.449 0.323 0.377
Minimum dose followed
by twitching of mus-
cles 0.442 0.318 0.377
Maximum dose followed
by twitching of mus-
cles but not resulting
in death 0.733 0 528 0 810
Minimum lethal dose 0.836* 0.602 0.794f
Lethal dose 1.096 0.800 0.907
Lethal dose 1. 126f 0.810
Lethal dose 0.998 0.718

*Death in forty-eight hours.

fDeath in two hours.

tDeath in twenty-six hours.
§Death in six hours.
#{182}Death between nine and twenty-four hours.

From the figures in Table III it seems that meta-cresol acetate

is somewhat more toxic than its chemical equivalent of meta-
cresol. At the same time it should be noted that some of the
results do not warrant this conclusion. Probablj the number
of dogs used (eight for the acetate and seven for meta-cresol)
was not sufficient to rule out the element of uncertainty.
The symptoms usually observed after administration of meta-
cresol acetate were drowsiness, followed in about one hour, if
the dose was sufficiently large, by twitching of the muscles and
by convulsions. When a non-lethal dose was administered, these
effects disappeared in a few hours and recovery was rapid. Other-
wise the animal remained in a rather apathetic state until death
ensued. At autopsy the chief findings were several small fresh
520 ISIDOR GREENWALD

ulcers in the small intestine, the mucosa of which was usually

congested; and possibly somewhat more marked congestion and
ulceration of the mucosa of the large intestine.
The dog that died forty-eight hours after the administration of
meta-cresol acetate presented a hemorrhagic area, about two
inches in diameter, on the outside of the fundic portion of the
stomach. The liver resembled the so-called “nutmeg liver;”
kidneys and spleen showed a passive congestion.
The dog that received 1.096 gram of meta-cresol acetate per
kilo, was found dead the following morning. A number of small
subcutaneous and mesenteric hemorrhagic areas were discerned.
The liver and kidneys were dark brown and the gross markings
were unusually distinct.
It was rather surprising to find that in no case was the gastric
mucosa congested or affected in any perceptible manner.
Two dogs were made the subjects of continued dosage per os. One,
weighing 14 kilos, received 15.2 grams of rneta-cresol acetate in
three days, none for five days, and then a total of 44 grams dur-
ing the next ten days. In the first period the doses were increased
from day to day. In the second period, which was ten days in
length, the meta-cresol acetate was given daily except on the
fourth, fifth and ninth days. No ill effect was noted other than a
disposition to vomit any considerable volume of food given less
than seven or eight hours after administration of the meta-cresol
acetate.
Another dog, weighing 4.2 kilos received 2.561 grams of meta-
cresol acetatejn five days and 8.181 grams in the next seven days.
In each period the daily doses were almost uniform. The day
after the acetate was last given the dog was killed with chloro-
form. There were five small, old, healed ulcers such as are fre-
quently seen in dogs, in the small intestine. Otherwise nothing
abnormal was found.
Intravenous injection. Under cocaine anesthesia the right
femoral vein of a fox terrier weighing 7 kilos was exposed and
connected by means of a cannula and rubber tube, to a
burette containing physiological salt solution. At intervals por-
 STUDY OF THE PROPERTIES OF META-CRESOL ACETATE 521

tions of a solution of meta-cresol acetate in olive oil7 (26.2 grams

of meta-cresol acetate per 50 cc. of solution), were injected into the
rubber tube near the cannula and then washed into the circula-
tion with 5 or 10 cc. of the saline solution. Injection of 0.5 cc.
of the oily liquid caused the heart to beat more rapidly almost
immediately, the pulsations increasing to over 150 per minute
and decreasing slowly to 130. Three minutes later 0.5 cc. more
were injected. This was followed immediately by marked muscle
twitching, which passed away in three minutes. The injection
was then repeated with the same effec1. One cubic centimeter
was then given. The symptoms were severe and continued fifteen
minutes. Then 1.25 cc. were injected, Twitching and convul-
sive movements became very marked and continued so for a few
minutes, then disappeared slowly and were entirely absent in
thirty minutes. Recovery was rapid and, within an hour of the
last injection, the dog was quite normal and ate eagerly. The
total amount injected was 3.75 cc. containing 1.965 gram of
meta-cresol acetate. This was equivalent to 1.415 gram of
meta-cresol or 0.202 gram per kilo. Gibbs and Hare8 found that
0.15 gram of meta-cresol per kilo., injected intravenously at
one time, was fatal.
Intraperitoneal injection. The dog used in the experiment just
described was kept under observation for four days and in all re-
spects behaved like a normal dog. On the fifth day 4.2 grams of
meta-cresol acetate in 20 cc. of olive oil were injected into the per-
itoneal cavity. Muscle twitching soon followed but ceased in four
hours. The next day the dog was quite apathetic and ate very
little. On the following morning he was dead. At autopsy a
mesenteric hemorrhage was found opposite the site of injection.
Otherwise everything was apparently normal.
Local action. The local action of meta-cresol acetate was. next
studied. A dog had two feet bound with cotton compresses
saturated with the acetate. The compresses were removed after

7On account of the great danger of producing oil embolism care was taken to
introduce slowly only very small doses of the oily liquid at each injection.
Gibbs and Hare: Archiv. f. Phys., Suppi. Band., 1889, p. 271.
522 ISIDOR GREENWALD

twelve hours and the feet were found to be unaffected. There was
no inflammation whatever. Two days later, under cocain anes-
thesia, two longitudinal incisions, about two inches long, were
made in the skin of the back of the neck. On the following day
one of the wounds was covered with a cotton compress saturated
with meta-cresol acetate, and the other with dry absorbent cotton.
This treatment of the wounds was repeated daily for three days.
As it was very difficult to keep the compresses in position their
use was then discontinued and the treated wound was painted with
meta-cresol acetate three or four times a day on four successive
days. At no time did the dog appear to be annoyed by the ace-
tate and the two wounds .healed with equal rapidity.
Subcutaneous injection. Subcutaneous injections of meta-cresol
acetate were very well borne by dogs. In one animal, weigh-
ing 7.5 kilos, the injection of 3.5 grams of undiluted meta-cresol
acetate, and also, later, of 5 cc. of olive oil solution containing
3.14 grams of the acetate, was followed by only slight transient
local reaction. Another dog, weighing 8.5 kilos, received 6.29
grams of meta-cresol acetate. The swelling that followed did
not disappear but softened and, after puncture three days later,
100 cc. of bloody pus were removed. The skin about the point
of puncture was very thin and an open wound soon resulted, but
the dog kept this clean and it healed rapidly.
Excretion and effects on metabolism. Jonescu9 found that of 4
grams of meta-cresol, given in four daily doses to a dog weighing
11.96 kilos, none was excreted in the feces and only 1.768 grams
or 46.5 per cent in the urine. In another experiment in which
1 gram of meta-cresol was given on each of three successive days,
the same dog excreted in the urine 1.505 gram, or 50.17 per cent
of the amount administered. Similar results were obtained in
this work with meta-cresol acetate.
A dog weighing 4.19 kilos was kept upon a daily diet of 75
grams of hashed meat (prepared and preserved as described by
GiesIo), 20 grams of cracker meal, 15 grams of lard, 5 grams of

5Jonescu: Biochem. Zeit., 1906, i, p. 399.

‘#{176}Gie.s:
Am. Journ. Physiology, 1905, xv, p. 235; Proc. Soc. Exp. Biol. and Med..
1908, v, p. 27.
 STUDY OF THE PROPERTIES OF META-CRESOL ACETATE 523

bone ash and 175 cc. of water. The dog was fed at 9 a.m., but
a little of the meat was reserved until noon when it was given
with an empty gelatin capsule (control). After the dog had been
kept on the above diet for eight days the collection of urine and
feces was begun. The excreta for four days were obtained. On
the fifth day a gelatin capsule containing 0.512 gram of meta-
cresol was given two hours after feeding. . Nothing unusual was
noticed within the next ninety minutes, but on the following
morning the urine was found to be contaminated with vomit.
This day’s urine was discarded. During the next four days the
dog received 2.0488 grams of meta-cresol acetate, equivalent to
1.475 grams of meta-cresol, in four approximately equal daily
doses, which were given in gelatin capsules in meat three hours
after feeding.
The feces of this period were treated with boiling H2S04 solu-
tion and the volatile matter distilled from the mixture. No phe-
nols could be detected in the distillate. The urines of each
period were combined and analyzed. The analytic methods
which were employed are indicated in the following summary.

Total nitrogen: Kjeldahl method.

Urea: Benedict method.’1
Total sulfur: Benedict method.”
Total and inorganic sulfates: Folin methods.’3
Glycuronic acid: Distillation with HC1 solution and precipitation
with phloroglucin, according to Tollens.’4 +

Phenols: Distillation as in Kossler and Penny’s method,’5 but, as the

action of iodine solutions upon meta-cresol has not yet been studied, a
bromination process was used. Ditz and Cedivoda, and others, have
studied the action of bromine water upon solutions of meta-cresol and
have found that a tri-brom derivative is readily formed. In our exper-
iments Lloyd’s’7 modification of the Koppesschaar method was used. It

“Benedict: J. Biol. Chem., 1910, viii,p. 405.

‘2Benedict: J. Biol. Chem., 1909, vi, p. 363.
“Folin: J. Biol. Chem., 1906, i, p. 131.
‘4Tollens: Z. Phys. Chem., 1909, lxi, p. 95; lxiv, p. 39.
“Kossler and Penny: Z. Phys. Chem., 1893, xvii, p. 117.
‘Ditz and Cedivoda: Z. f. Angew. Chem., 1899, pp. 873 and 897.
‘7Lloyd: J. A. Chem. Soc., 1905, xxvii, p. 16.
 524 ISIDOR GREENWALD

was first tried with solutions of pure meta-cresol and found to give ac-
curate results.
TABLE IV

Analytical Results. Metabolism Experiment on a dog

kilos grams grams gram gram gram gram gram per centper ceis per cent

Fore period.... 4.2 12.5 11.24 0.6& 0.492 0.462 0.247 0.014 + .89.91 71.83 93.78
Meta-cresol ace-
tate period. . 4.2 11.95 10.23 0.629 0.479 0.206 1.175 0.674 : 85.61 76.12 43.00

may be noted that in these urines, Tollens’ colorimetric method gave results
indicating a glycuronic acid content of at least 6.25 and 7.75 grams respectively.
A number of normal dog urines were found to give the reaction in correspondingly
high dilution. Apparently, then, the test may be more delicate than Tollens
supposed or, as is much more probable, some dog urines contain a substance which
either is not, precipitated by basic lead acetate and ammonium hydroxid or does
not yield furol on boiling with 12 per cent HCI solution, but which does give Tol-
lens’ test for glycuroriic acid.

If the amount of cresol excreted in the first period is subtracted

from that eliminated in the second period the difference is 0.66
gram or 44.74 per cent of that given. The ethereal sulfate
excretion for the second period was equivalent to 273 mg. of
sulfur, while in the control period it was only 30 mg. The dif-
ference, 243 mg., if present as meta-cresol sulfuric acid, would be
equivalent to 0.816 gram of meta-cresol. Similarly with the
glycuronic acid, the difference in the excretion for the two pe-
nods is 0.928 gram (calculated as the lactone), corresponding to
0.569 gram of cresol. The total amount of cresol accounted for
by the increased ethereal sulfate and glycuronic acid excretion is
1.385 gram or 93.91 per cent of the amount taken. Apparently,
then, somewhat more than half of the meta-cresol derived from
the meta-cresol acetate was oxidized, though without any consid-
erable decomposition of the aromatic nucleus; and these oxida-
. ., + ,+ . . + ,,,

STUDY OF THE PROPERTIES O’ META-CRESOL ACETATE 525

tion pnoducts, as well as unchanged meta-cresol, were excreted

mainly as salts of the conjugated sulfuric and glycunonic acids.
Oxidation products of meta-cresol acetate in the urine. Very early
in the course of this work it was noted that the urine of dogs that
received meta-cresol acetate became very dark on standing,
thus indicating the presence of homologues of pyrocatechol and
hydroquinon. Preusse’8 having reported that he could find no
meta-oxybenzoic acid or di-hydroxy-toluenes in the urine of a
dog to which he administered 10 grams of meta-cresol, it was
thought advisable to attempt the isolation of such oxidation
products as might theoretically be expected. These were meta-
oxybenzoic acid, homo-pyrocatechol, iso-homo-pyrocatechol and
hydrotoluquinon. Accordingly, the combined uines of four
dogs that had received a total of 50.7 grams of meta-cresol ace-
tate by mouth, and 22.7 grams subcutaneously, were evaporated
in several portions to syrups, and the latter extracted with alco-
hol. The combined extracts were evaporated to a syrup and
allowed to crystallize. The crystals were used in an unsuccessful
attempt to isolate the potassium salt of meta-cresol sulfuric acid;
nothing but urea was obtained. Some of the filtrates from the
crystals were placed in a vacuum desiccator over concentrated
sulfuric acid. After standing several days a few crystals formed
in the thick syrup obtained, but they could not be satisfactorily
separated from it.
The combined filtrates were strongly acidified with sulfuric
acid and continuously distilled in a current of steam. At the
end of a period of thirty hours considerable cresol was found to
be passing into the distillate. The distillation was interrupted
and the liquid in the distillation flask, after cooling, was thor-
oughly extracted with ether in a continuous extraction apparatus.
The ether extract was shaken with sodium bicarbonate solution.
The latter was subsequently washed with a little ether. The com-
bined ethereal solutions were distilled until all the ether was
removed. A current of steam was then passed through the resid-
ual liquid until the distillate no longer gave a precipitate with

‘8Preusse: Z. f. Phys. Chem., 1881, v, p. 57.

526 ISIDOR GREENWALD

bromine water. The liquid remaining in the distilling flask was

filtered from a tarry residue, evaporated to a syrup and set aside
in a refrigerator. No crystals appeared even after several weeks.
The syrup was finally dissolved in water and lead acetate added to
complete precipitation. Basic lead acetate was added to the
filtrate; a slight precipitate was produced.
The first lead acetate precipitate was decomposed with sul-
furic acid. The liquid was filtered and the filtrate extracted
with ether. This extract, after distillation of the ether, left a red
oil which could not be made to crystallize. Ferric chloride when
added to the aqueous solution of the oil produced a green color
which soon changed to brown. When ammonium hydroxid
solution was added after treatment with ferric chloride, the color
changed to violet. With sodium carbonate solution the color
changed to a red violet. The substance reduced ammoniacal
silver solution in the cold and Fehling solution on slight warming.
Ammonium hydroxide turned the aqueous solution of the sub-
stance green. A boiling solution in chloroform gave a blue-green
color on treatment with solid potassium hydroxide.
The precipitate produced by basic lead acetate yielded a smaller
quantity of a substance having apparently the same properties
as that obtained from the first precipitate. Exactly the same
reactions were given by a sample of homopyrocatechol, which was,
however, not quite pure.
The filtrate from the lead precipitates was freed from lead with
hydrogen sulfid, filtered and evaporated. The residue was a
brown syrup which gave no color with ferric chlorid nor with
chloroform and potassium hydroxid. When treated with anilin
and acetic acid the liquid remained clear. Apparently it was
neither toluquinon nor hydrotoluquinon.
The sodium bicarbonate extract was acidified with sulfuric
acid and extracted with ether. The ether was distilled off; it
left a brown crystalline residue. This was pressed on a porous
plate. It was then dissolved in water and treated with animal
charcoal, but with little effect. The liquid was evaporated to
crystallization. The crystals were dried on a porous plate and
dissolved in boiling toluol. On cooling, crystals were deposited.
 STUDY OF THE PROPERTIES OF META-CRESOL ACETATE 527

These consisted of small needles and irregular hexagonal plates

which melted at 190 degrees (uncorrected). A solution of some
of the material gave a very faint green coloration with ferric
hlorid solution. A few crystals, dissolved in concentrated
sulfuric acid solution and warmed, gave an orange-red liquid.
The crystals tasted sweet. When 63.3 mg. were dissolved in
water and titrated with am.moriium hydroxid solution, using
Congo red as the indicator, 4.3 cc. of the alkalin solution were
required. Calculated for C6114 (OH) COOH, the volume is 4.6
cc. After its titration, the liquid was acidified with sulfuric acid
and extracted with ether. The ether was allowed to evaporate
spontaneously in an open dish. It left a yellow ring with pure
white crystals in the center. These gave no color with ferric
chlorid solution and melted at 197#{176} (uncorrected). Probably the
substance was meta-oxy-benzoic acid, which possesses the above
properties and melts at 200#{176}.
Unfortunately this work was begun several weeks after the
collection of the urines and was frequently interrupted by other
work, so that whatever hydroxy-toluenes were originally present
may have been, at least in part, oxidized and condensed to un-
known substances. This was indicated by the very dark color
of the urines under examination.
Attempts were made to isolate the conjugate glycuronic acid
by K#{252}lz’smethod’9 but without success. In one case, urine from
an 8.1 kilo dog that had received 8.8 grams of meta-cresol ace-
tate was used immediately after collection. The precipitate
produced by basic lead acetate in the aqueous solution of the alco-
hol-ether extract was decomposed with hydrogen sulfid and the
filtrate from the lead sulfid was evaporated. The residue was a
non-crystaffizable syrup. It was hydrolyzed with hydrochloric
acid solution. The liquid gave a very intense reaction for gly-
curonic acid with naphthoresorcin but, after neutralization, gave
no color with ferric chlorid solution. On warming, however, an
odor resembling that of quinon was noted. The rest of the
liquid was extracted with ether and this evaporated. The resi-

“ K#{252}l.z:
Z. f. Biologic, 1890, xxvii, p. 247.
528 ISIDOR GREENWALD

due gave a negative reaction with ferric chlorid solution. When

dissolved in chloroform and boiled with potassium hydroxid solu-
tion a red-brown color was obtained. Toluquinon gives the
same reaction. Possibly this substance was present.
To the filtrate from the basic lead acetate precipitate, sulfuric
acid solution was added in a quantity sufficient to effect complete
precipitation. The precipitate was filtered out and washed.
The filtrate was extracted with ether, the extract evaporated and
the oily residue dissolved in water. It gave a slight transient
green coloration with ferric chlorid solution, but no other color
appeared after the addition of ammoninm hydroxide and sodium
carbonate solutions. On warming the liquid with ferric chloride
solution, a quinon-like odor was noticed. Ammoniacal silver
solution was reduced in the cold and Fehling solution on warming.
The remainder of the liquid was evaporated to dryness. The
amorphous residue was dissolved in chloroform and this solution
boiled with solid potassium hydroxid. A reddish brown color
was produced. It seems quite probable that hydrotoluquinon
was present.
In a number of cases it was found that, after administering
meta-cresol acetate by. mouth or subcutanously, the urine did
not decompose. Such urines were kept from forty-eight to
fifty-three days without becoming alkaline. In the dogs that
received meta-cresol acetate by mouth this was the case only when
the dose was just about large enough to cause muscle twitching.
Urines voided after subcutaneous injections which caused no
geiwral symptoms were preserved without the addition of an
antiseptic for forty days. The minimum amount that was found
to do this was 4.1 grams for a dog weighing 8.5 kilos. The next
day the same dog received 6.3 grams. The urines of the follow-
ing three days, separately collected, remained acid for forty days.

III. SUMMARY OF GENERAL CONCLUSIONS

1. Meta-cresol acetate, a colorless liquid of pleasant odor, is

practically insoluble in water, is not readily hydrolyzed and is
a strong antiseptic agent.
 ,‘,..

STUDY OF THE PROPERTIES OF META-CRESOL ACETATE 529

2. It does not exhibit the marked protein-coagulating action

of free meta-cresol.
3. It is probably unaffected in the stomach but is decomposed
in the intestine.
4. It is non-corrosive and has very little harmful local effect
except in the intestine.
5. When injected into the dorsal lymph sac of frogs or given
by mouth to dogs, it is about as poisonous as the equivalent
amount of meta-cresol.
6. When taken into the body it is partially oxidized to di-
hydroxy-toluenes and meta-oxy-benzoic acid, which are in great
part excreted as the conjugate sulfuric and glycuronic acids.

It gives me great pleasure to here thank Prof. William J. Gies

for suggesting this investigation and for his interest and advice
during the course of the work.