The Effect of Various Ointment Bases on the

Percutaneous Absorption of Salicylates I*

b

Effect of Type of Ointment Base

By MOISE E. STOLARt, G. VICTOR ROSSI, and MARTIN BARR
An ap roach to the evaluation of the percutaneous absorption of drugs from oint-
ment gases is described. Salicylic acid and sodium salicylate were selected as test
drugs for incorporation into ointment bases representing each of four physical types
of ointment vehicles. These ointments were applied to the intact shaved skin of the
rabbit and confined by a specially designed bandage. Concentration of salicylate
in the blood was determined at hourly intervals following a plication of the oint-
ment. Salicylic acid was most effectively absorbed from gydrophilic ointment.
The extent of absorption from the other ointments studied, in decreasing order, was:
hydrophilic petrolatum containing water, etrolatum, and polyethylene glycol oint-
ment. Sodium salicylate was also absorged to the greatest extent from hydro-
philic ointment, although the degree of absorption was considerably less than that
observed with salicylic acid. On the basis of salicylate blood levels, no significant
absorption of sodium salicylate occurred from ointments prepared with petrolatum,
hydrophilic petrolatum containing water, or polyethylene glycol ointment.

a significant degree of percutaneous hydrophilic petrolatum U. S. P. XV in which water

A LTHOUGH
absorption is essential to the therapeutic
usefulness of relatively few drugs applied topi-
was incorporated, a water-in-oil emulsion base;
hydrophilic ointment U. S. P. XV, an oil-in-water
base; and polyethylene glycol ointment U. S. P.
cally in the form of ointments, i t is nevertheless XV, a water-soluble base. Salicylic acid ointments
desirable t o know the extent of such absorption were prepared to contain 6% of the active ingredient
from the various types of ointment bases. Such and sodium salicylate ointments were prepared to
information is especially important in those cases contain 6.95% sodium salicylate, which is equivalent
to 6% salicylic acid.
in which absorption of drugs through the skin Aualysis of Salicylic Acid Content of Oint-
may lead to untoward reactions, sensitization, etc. ments.-An accurately weighed sample (2 Gm.) of
It was the primary aim of this study to develop salicylic acid ointment was transferred to a 100-ml.
a standardized procedure for determining the rel- beaker to which was subsequently added 25 ml. of
ethyl alcohol warmed to 65" and previously neutral-
ative degree of absorption of drugs through the
ized with sodium hydroxide to phenolphthalein.
skin, and for evaluating the various factors which The mixture was stirred until the ointment first
influence percutaneous absorption. melted and then congealed. The supematant liquid
This report is concerned with the influence of was decanted and the procedure repeated. In those
the type of ointment base on the percutaneous cases where the ointment was miscible with alcohol,
the sample was dissolved in 50 ml. of neutralized
absorption of salicylic acid, a predominantly oil- alcohol. The salicylic acid content of the super-
soluble drug, and sodium salicylate, a predomin- natant liquids from both extractions, or solution of
antly water-soluble drug. -4 subsequent report the ointment base in alcohol, was determined accord-
will consider the effect of various surface-active ing to the U. S. P. XV assay for salicylic acid. To
compensate for possible interference from the various
agents on the absorption of these drugs from four
components in the ointment base, a blank composed
different physical types of ointment bases. of the vehicle without salicylic acid was analyzed
concurrently.
EXPERIMENTAL Analysis of Sodium Salicylate Content of Oint-
Preparation of Ointments -Salicylic acid and ments.-A 2-Gm. sample of sodium salicylate oint-
sodium salicylate, previously reduced to a fine pow- ment, accurately weighed, was transferred to a
der in a ball mill and sifted through a So. 80 sieve, 100-ml. beaker. Sodium salicylate was either ex-
were incorporated in ointment bases representing tracted with two 12.5-ml. portions of water warmed
each of four physical types. The bases selected to 65'. or dissolved along with the base in 25 ml. of
were: petrolatum U. S. P. XV, an oleaginous base; warmed water. The extraction was achieved by al-
ternatc melting and congealingof the ointment while
* Received July 16, 1959, from the Philadelphia College of being constantly stirred. The sodium salicylate
Pharmacy and Science. Philadelphia, Pa.
Abstracted from a thesis submitted by Moise E. Stolar to content of the supernatant liquids from both extrac-
the faculty of the Philadelphia College of Pharmacy and tions, or the aqueous solution of the ointment, was
Science in partial fulfillment of the requirements for the de-
gree of Doctor of Philosophy. determined according to the U. S. P. XV assay for
Presented to the Scientific Section, A. Prr. A,. Cincinnati sodium salicylate. To compensate for possible in-
meeting, August 1959.
t Present address: Warner Lambert Research Institute, terference from the ingredients of the ointment base,
M o m s Plains. N. J. a blank composed of the vehicle without sodium sali-
The authors wish to thank Chas. Pfizer and C o . for their
financial support of this study. cylate was analyzed concurrently.
144
March 1960 SCIENTIFIC
EDITION 145

All salicylic acid ointments assayed to contain

5.90-6.05YGsalicylic acid ; all sodium salicylate oint-
ments assayed to contain 6.80-7.10% sodium salicy-
late, equivalent to 5.86-6.12% salicylic acid.
Test Animal.-Male New Zealand white rabbits
weighing between 3.0 and 3.6 Kg. were used throngh-
out the study. The animals were maintained on
Purina rabbit pellets and water ad libitum and
housed individually in temperature and humidity
controlled quarters. All experiments were per-
formed in these same quarters.
Application of Ointment.-Hair was removed1
from the skin of the dorsal area between the forelegs
and hind-legs on both sides of the spine twenty-four
hours prior to application of the ointment.
A bandage was devised t o restrict and control the
area of contact between the ointment and the skin of Fig. 1.-Components of the bandage used in applica-
the rabbit. The edges of an 8.35 X 14.7 cm. piece of tion of test ointrneuts t o the shaved rabbits skin.
aluminum foil were doubled over and flattened 1cm.
on each side to produce a rectangular plate measur-
ing 6.35 X 12.7 cm. with a 1-cm. reinforced margin.
An accurately weighed 7.5-Gm. sample of ointment
was uniformly spread over one surface of the plate,
whose opposite side was centered on a 7.5 X 20 cm.
strip of adhesive tape. The entire assembly was
then applied to the shaved skin of the rabbit and
carefully adjusted to conform to the contours ol the
area. To minimize contamination and insure ade-
quate contact between the ointment and the skin,
this assembly was covered with an Ace elastic band-
age and fastened with strips of gauze bandage around
the forelegs and hind-legs of the animal. The various
components of the bandage are illustrated in Fig. 1,
and the appearance of the bandage applied t o the
rabbits skin is illustrated in Fig. 2. The ointment
remained in contact with the skin for the nine-hour Fig. 2.-Appearance or bandage assembly in position
on the rabbit.
experimental period.
Procedure.-Due to time limitations, not more
than seven animals could be utilized during any one
experimental day. Therefore, on each of ten con- presented in Fig. 3. The results of a similar study
secutive experimental days, ointments containing conducted with sodium salicylate are illustrated in
either salicylic acid or sodium salicylate prepared Fig. 4. Each point on the graph represents an aver-
with the various type ointment bases were studied age of the determinations on groups of ten rabbits.
together with a salicylic acid ointment prepared On the basis of the mean hourly blood levels of
from hydrophilic ointment U. S. P. XV, as a control. salicylate, as reported in Figs. 3 and 4, it is apparent
One and one-half milliliters of blood were with- that the type of ointment base exerts a definite in-
drawn from the marginal ear vein of the rabbit prior fluence on the extent of percutaneous absorption of
to, and a t hourly intervals for nine hours after appli- both salicylic acid and sodium salicylate. I t may
cation of ointment. The blood samples were al- also be noted, however, that the effect of the oint-
lowed t o clot a t room temperature and were then ment base is considerably greater in regard t o the ab-
chilled in a refrigerator. After centrifugation a t sorption of salicylic acid as compared to sodium
2,500r. p. m. for thirty minutes, the serum was an- salicylate.
alyzed for salicylate according to the method de- Salicylic acid was absorbed through the skin to a
scribed by Ungar, et al. (1). The results obtained greater extent from hydrophilic ointment than from
from a group of ten animals treated with each oint- the other types of bases investigated. The extent of
ment base were analyzed by analysis of variance and absorption of sa.licylic acid from the ointment bases
the t test (2). studied, in decreasing order, was found to be: hy-
drophilic ointment, hydrophilic petrolatum, and
petrolatum. Statistical analyses indicated a signifi-
RESULTS cant difference among the average blood salicylate
concentrations obtained after application of each of
The relative extent of absorption through the in-
these ointments. As measured by blood salicylate
tact rabbit skin of salicylic acid incorporated in hy- level, the degree of absorption of salicylic acid from
drophilic ointment, petrolatum, hydrophilic petrola- polyethylene glycol ointment was negligible.
tum containing water, and polyethylene glycol oint- According to the criterion employed in this study,
ment, as determined by blood salicylate concentra- percutaneous absorption of salicylic acid and sodium
tion (results expressed as mg. % salicylic acid), is salicylate occurred maximally between the fourth
1 Oster electric animal clipper, model A-2, Ang-Ra No. 2
and sixth hour after application of the ointment to
head. the skin of the experimental animal. It is, of course,
146 JOURNAL OF THE AMERICANPHARMACEUTICAL
1 this base. Statistical analyses of blood salicylate
1 1 I I
0 2 4 6 8 actual experiments and were not, a t any time, placed
TIME IN HOURS
Fig. 3.-The effect of the type of ointment base on
the percutaneous absorption of salicylic acid. 0,
hydrophilic ointment ; A, hydrophilic petrolatum
with water; 0, petrolatum; A, polyethylene glycol
ointment.
U i I /
0 2 4 6 8 stances are not or only poorly absorbed, since the
TIME IN HOURS
Fig. 4.-The effect of the type of ointment base on
the percutaneous absorption of sodium salicylate.
0,hydrophilic ointment; 0 , hydrophilic petrola-
tum with water; A, petrolatum; A, polyethylene
glycol ointment.
recognized that the blood salicylate level a t any
given time represents a complex interaction of such
factors as absorption, storage, biotransformation.
and excretion.
Sodium salicylate was found t o be absorbed
through tbe skin to a measurable degree when in-
corporated in hydrophilic ointment. although the ex-
tent of absorption was considerably less than that ob-
served with salicylic acid ointments prepared with
ASSOCIATION Vol. 49, No. 3
concentrations indicated that there was no signifi-
cant absorption of sodium salicylate from ointments
compounded with petrolatum, hydrophilic petrola-
tum containing water, or polyethylene glycol oifit-
ment.
DISCUSSION
Method.-The procedure developed in this in-
vestigation eliminates certain variables inherent in
previous methods for studying percutaneous absom-
tion, which may, in part, be responsible for the
nnmerous contradictory statements regarding drug
absorption to be found in the literature. For ex-
ample, inunction of the ointment base, which is
commonly employed in absorption studies, was
avoided in this investigation since it is obviously diffi-
cult to control adequately the force of application
and surface area involved.
Following a study of dermal permeability in
chronically heat- and humidity-stressed rats, Clay
and Kelson (3)suggested that the dermal permeabil-
ity of drugs may be altered by stress. To minimize
the Occurrence of a situation which might possibly be
considered as stressful, in this study all animals were
conditioned t o the experimental routine twice weekly
for a period of one month prior to performance of the
in a restraining device.
Previously reported quantitative methods for
studying percutaneous absorption have generally
consisted of a one-point determination of test com-
pound in either urine or blood collected at a specified
interval after application of the ointment. The
technique employed in this investigation, whereby
blond salicylate concentration was determined at
hourly intervals for an extended period of time fol-
lowing application of the ointment, was thought to
enable a more valid comparison of the extent of per-
cutaneous absorption of medicament from various
ointment bases.
Solubility of Drugs.---With the ointment bases
utilized in this stady it was found that salicylic acid,
a pi edorninantly oil-soluble drug, was substantially
absorbed through the skin and was absorbed to a
much greater extent than sodium salicylate, a prg-
dominantly water-soluble compound.
I t is the opinion of many investigators in this field
that oil-soluble substances are absorbed through the
skin, whereas water-soluble or oil-insoluble sub-
skin is largely impermeable to water and therefore
to water-soluble compounds.
The percutaneous absorption of oil-soluble sub-
stances is often quite rapid and the extent of absorp-
tion appears to increase in those cases where the
compound possesses a degree of water solubility in
addition to oil solubility. Despite extensive reports
in regard to the influence of oil and water solubility
on percutaneous absorption (4-7). it has not been
established that the percutaneous absorption of com-
pounds is a function of their distribution coefficients
(i. e., oil solubility/water solubility).
Bischler, et al. (8). refute the necessity of water
solubility in regard to the absorption of compounds
through the skin. They demonstrated excellent
percutaneous absorption of oleates of certain alka-
loids which they state to be insoluble in water.
March 1960 EDITION
SCIENTIFIC 147

On the other hand, it has been shown by the use of tistically significant blood salicylate levels. The
immunologic procedures that oil-insoluble antigens possibility remains, however, that some degree of
are absorbed from either ointments or wet-packs sodium salicylate absorption did occur from the lat-
(9-12), although the degree of absorption is ad- ter three ointments, which could not be detected on
mittedly small. the basis of blood salicylate level. Evidence for this
Considerable emphasis has been placed on the im- assumption may be based on the known biologic
portance of oil solubility, since i t is believed that this properties of salicylates, and the indicative but not
property tends to enhance percutaneous absorption statistically significant blood salicylate concentra-
by the transfollicular route (13). In contrast, rela- tions obtained with petrolatum and hydrophilic pe-
tively little attention has been directed to the trolatum ointments.
influence of the solubility of the drug in the ointment Since the literature in regard to percutaneous ab-
vehicle on the extent of percutaneous absorption. sorption of sodium salicylate is meager, it was not
This factor will be considered in a subsequent publi- possible t o corroborate the results obtained in this
cation. study. However, Kionka (20) and Miyazaki (21) re-
It remains equivocal as t o whether sodium ported the absorption through the skin of sodium
salicylate is absorbed through the intact skin as such salicylate incorporated in lanolin, a water-in-oil
or only after conversion to salicylic acid. A study emulsion type base, and petrolatum containing olive
by Schwenkenbecher (14) in 1904 indicated that oil. These authors did not note any significant dif-
salicylates per se were not absorbed percutaneously. ference in the extent of absorption from the two
This conclusion was drawn from an experiment in types of ointment bases. Valette and Cesar (22)
which salicylate but not lithium ion was found in a demonstrated the superiority of an oil-in-water type
urine sample collected twenty-four hours after the base to other liquid vehicles with respect to the per-
topical application of a 2% lithium salicylate solu- cutaneous absorption of sodium salicylate.
tion. Schwenkenbecher, thereFore hypothesized
that salicylate salts were converted in the skin to
salicylic acid which was subsequently absorbed and SUMMARY
excreted in the urine. More recent renal excretion
studies (15) indicate, however, that the lithium ion 1. A procedure, utilizing t h e intact rabbit
does not appear in the urine until two to three days skin, is described for the study of percutaneous
after the oral ingestion of lithium chloride. absorption of drugs.
Aqueous solutions of sodium salicylate (6.95%),as 2. The relative extent of percutaneous absorp-
well as sodium salicylate ointments having a con- tion of salicylic acid and sodium salicylate from
tinuous aqueous phase, were found to have a p H
range of 6.2 to 6.5. The pH of the skin of the rabbit four different physical types of ointment bases
was determined t o range from 6.0 t o 7.4, values was investigated.
which are in agreement with tbose previously re- 3. The influence of the type of ointment base
ported (16). Therefore, it is doubtful that the con- on the absorption of salicylic acid and sodium
centration of free salicylic acid present at such p H
values could account for the amounts detected in the salicylate through t h e skin is considered.
blood of rabbits examined during this investigation.
Effect of Type of Ointment Base.-The results ob- REFERENCES
tained in the study of percutaneous absorption of
(1) Ung?, G., Damgaard, E., and Wong, K. W., Proc.
salicylic acid from various types of ointment bases, SOC. Expfl. Bad. Med. 80.45(1952).
agree, in general, with the findings of Monocarp ( 2 ) Croxton, F. E?, Elementary Statistics with Applica-
(17, 18) who demonstrated that salicylic acid was 1953.in Medicine. Prentice-Hall Inc.. New York, N. Y.,
tion
absorbed to a greater extent from a n oil-in-water 13) Clav. M. M.. and Nelson.. 1. _ W... THISJOURNAL.. 43..
emulsion base than from a water-in-oil emulsion type 230(1954). -
(4) Burgi, E.,Die Durchlassigkeit der Haut fur Arzne-
base or an oleaginous base. These data are futher ien und Gifte Springer, Berlin, Germany 1942 p. 110.
substantiated by the results of a clinical study con- (5) Rothman, S., J . Lab. Clin. Md., 28, 1305(1943).
(6) Calvery, H. 0..Drake. J. H., and Laug, E. P.. Phys-
ducted by Polano, et al. (19), who demonstrated by a iol. Rm., 26,495(1946).
series of patch tests that the degree of skin irritation (7) Valette, G., and Cavier, R., J . Physiol., 39,137(1947).
181 Bischler. A.. Favre. M.. Frommel. E.. and Valette.
produced by 20% salicylic acid ointments was sub- G., hits Pharmacol: Toxico).. 4,122(1948):
stantially greater with an oil-in-water emulsion ve- (9) Kimura, G., Orient, J . Dis. Infanfs, 28, 15(1940).
(10) Walzer, A., and Sack,S. S., Arch Dermafol. and Syphi-
hicle than with a water-boil emulsion base. These , 101. 49 427(1944).
investigators found no evidence of irritation follow- (i1)Walzer A. ibid. 41 692(1940).
(12) Golova&f; M.,Co;pl. rene. soc. bjol., 94. 6(1926).
ing topical application of salicylic acid incorporated (13) Rothman. S..J . SOC.Cosmcfac Chcmasls. 6. 193(1955).
in polyethylene glycol ointment. (14j Schwenkenb&her. A., Arch. Anal. u. Physiol., 121,
- - ~
fiFill9M)
The influence of the ointment base on the extent of (15) Foulks, J.. Mudge, G. H.. and Gilman, A., A m . J .
absorption of sodium salicylate was less distinct than Phrsrol., 168, 642(1952).
(16) Draize, J. H.. J . Inoesf. Dermalol., 5, 77(1942).
in the case of salicylic acid, inasmuch as a measurable (17) Monocarp. C.. Arch. e x p f l . Palhol. Pharmakol.. 141,
degree of absorption of sodium salicylate was ob- 50(1929). (18) Monocarp, C., ibid.. 141, 87(1929).
served only when hydrophilic ointment was used as (19) Polano, M. K.,Bonsel, J., and Van Der Meer, B. J.,
the vehicle. Application to the rabbits skin of so- Dermafologica. 101, 69(1950).
(20) Kionka, H., K l m . Wochschr.. 10, 1570(1931).
dium salicylate ointments prepared with petrolatum, (21) Miyazaki. K., Japan J . Dermafol. Urol., 31, 113
hydrophilic petrolatum containing water, or poly- (1931). (22) Valette, G., and Cesar, R., Ann. $harm. franc., 6 ,
ethylene glycol ointment, was not followed by sta- lG(1948).