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Bigeleisen,
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in the development of a drug product can provide clues to such (17) M. Uedeira and H. Uedeira, Kolloid 2. 2.Polynz., 194,
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(20) S. Subramanian, D. Balasubramanian, and J. C. Ahluwalia,
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(2) G. Zografi, Ph.D. thesis, University of Michigan, 1961. Shah, and D. Smith, Trans. Furuduy Soc., 62, 1963(1966).
(3) T. J. Schoch and E. C. Maywald, Anal. Chem., 28,382(1956). (22) C. H. Giles, J. Amer. Chem. Soc., 91,759(1969).
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42, 1lO(1926). 48101

DRUG STANDARDS

Spectrophotometric Determination of
Diphenhydramine Hydrochloride in an
Antiallergic Cream

FABRIZIO De FABRIZIO

Forman ( 3 ) developed an assay for dienestrol in a

Abstract 0A specific method for the quantitative determination of cream using urea-inclusion chemistry to remove the
diphenhydramine hydrochloride in a cream formulation has been
developed. The method entails the extraction of diphenhydramine
excess of monostearin. The information obtained from
by chloroform, further purification of the extracted base by column any of these studies is valuable and may indicate a
chromatography using alginic acid, and its spectrophotometric general approach to analysis employing the two basic
determination in the eluate at 258 mp. steps of extraction and cleanup. However, each of them
Keyphrases Diphenhydramine cream-analysis 17 Column chro- is a specific case which depends upon the physical and
matography-separation UV spectrophotometry-analysis chemical properties of the active component.
Therefore, it is reasonable to assume that different
methods of separation may be required for differing
The isolation of a pharmacologically active ingredient formulations containing the same active ingredients
from a pharmaceutical formulation containing surfac- and for differing active ingredients contained in similar
tants is often difficult. Various authors have followed formulations. Diphenhydramine hydrochloride is found
different procedures for the removal of unwanted in- in various combinations in commercially available
gredients. Jones (1) described a method for the deter- pharmaceutical preparations, and various assays for
mination of diethylstilbestrol in a water-dispersible its determination have been reviewed (4-6). No pro-
suppository using column chromatography followed by cedure, however, has been reported involving the
TLC. Gottlieb (2) used refluxing with an organic sol- quantification of diphenhydramine hydrochloride in an
vent to break down the emulsion and subsequently
recovered the active drug (which was also diethyl-
The analysis was applied to a product containing 2 % of diphen-
stilbestrol) using an aluminum column. More recently, hydramine hydrochlorideand marketed as “Allergin Cream.”

1470 0 Journal of Pharmaceutical Sciences

Table I-Results of Determination of Diphenhydramine Hydro- Table II-Analysis of Diphenhydramine Hydrochloride in Eight
chloride in Eight Synthetic Mixtures” Commercial Formulations

Found for Eight Claim for

Mixture Containing Taken, g. Determinations, g.b Diphenhydramine
Preparation Hydrochloride, g. % Found, % of Claim
Diphenhydramine hydro- 2.00 1.97, 2.01, 2.10, 1.98,
chloride 2.04, 1.96, 2.01, A 2 105.30
2.10 B 2 98.73
C 2 100.23
a The preparation also contains emulsifying wax, liquid paraffin, D 2 99.45
parabens, glycerin, water, color, and menthol. b Mean percentage re- E 2 102.70
covery of diphenhydramine hydrochloride = 100.5 %. F 2 100.68
_..
G 2 98.io
emulsified cream base. The procedure described in this H 2 99.79
paper is a modification of an analysis method for the
determination of diphenhydramine hydrochloride in a absorbance of the solution was then determined at 258 mp, using
cough mixture (6). The method entails the recovery 4-cm. cells and 0.1 N hydrochloric acid as blank.
of the diphenhydramine by chloroform extraction and RESULTS AND DISCUSSION
further purification through an alginic acid column,
followed by quantitative spectrophotometry at 258 mp. The cream examined was of the oil-water type. The oil phase
contained menthol as a coolant. A nonionic emulsifying wax was
used as the emulgent, as well as providing a cream of the required
EXPERIMENTAL consistency. The aqueous phase contained diphenhydramine hydro-
chloride, parabens, red color, and glycerin as a humectant. In a
Apparatus-A Beckman DB spectrophotometer and 4-cm. preliminary study, attention was focused on the possibility of find-
square, fused silica cells were used. A glass column, 30 X 1.8 cm., ing a combination of solvents which would help liquify the cream
with a stem, 5 cm., was fitted with a buret key. and at the same time facilitate the diphenhydramine extraction.
Reagents-The following were used : cation-exchange resin, The most suitable system found was chloroform-water. By initially
alginic acid, 40-100 mesh (available from British Drug Houses); acidifying the cream, the difficulty, which is normally encountered
2 N hydrochloric acid in water; 0.1 N hydrochloric acid in water; in alkaloidal extractions using chloroform-water systems, of extrac-
5 % hydrochloric acid in water; 95% ethanol in water; and 80% tion of the active ingredient favoring the aqueous phase instead
ethanol in water. Except where otherwise specified, all reagents of the chloroform phase is overcome. In addition to diphenhydra-
were of British Drug Houses’ Analar quality. mine, the chloroform extract contains menthol, liquid paraffin,
Standard Solutions-The following solutions were prepared with parabens, and the emulsifying wax. For this reason, the residue
suitable reference standards: (a) diphenhydramine hydrochloride, after the evaporation of chloroform was dissolved in 95 % ethanol
2 g./100 ml. in 5 % hydrochloric acid; and ( b ) diphenhydramine instead of water which would have given a turbid solution. Al-
hydrochloride, 3.2 mg.1100 ml. in 0.1 N hydrochloric acid. though the alcoholic solution used does not dissolve the oily par-
Column Preparation-Alginic acid, about 4 g., was slurried in ticles in the residue, these are easily eliminated by filtration of the
water and allowed to soak for 4 hr. The slurry was poured into a solution through a pledget of cotton wool. When the procedure
glass column which had been fitted with a cotton wool plug and was followed to analyze eight synthetic mixtures of the cream
allowed to settle. The column was washed with 2 N hydrochloric (prepared in a manner similar to commercial formulation), the
acid, until the absorbance of the eluate (pathlength 4 cm.) was results in Table I were obtained.
less than 0.005 at 258 mp, and then with distilled water until the When the method was applied to eight commercial products,
eluate was neutral to litmus. Finally, 25 ml. of 80% ethanol was the results in Table I1 were obtained.
passed through the column. The reasonable results obtained with commercial and empirical
Sample Treatment-An amount corresponding to approximately products establish the validity of this procedure.
2 g. of cream (about 40 mg. diphenhydramine hydrochloride)
was accurately weighed into a 150-ml. separator. The cream was REFERENCES
suspended in 6 ml. of 5 % hydrochloric acid. The suspension was
extracted with four successive portions of 15 ml. each of chloro- (1) L. N. Jones, M. Seidman, and B. C. Southworth, J. Phurm.
form; each extraction was filtered through a pledget of cotton wool Sci., 57, 646(1968).
into a 100-ml. volumetric flask, and the solution was brought to (2) S. Gottlieb,J. Amer. Pharm. Ass., Sci. Ed., 37, 147(1948).
volume. An aliquot of 20 ml. was pipeted into a 25-ml. volumetric (3) B. J. Forman and L. T. Grady, J . Pharm. Sci., 58,1262(1969).
flask. The chloroform was evaporated to dryness on a water bath (4) “The United States Pharmacopeia” 17th rev., Mack Publishing
with the aid of a current of air. The residue was dissolved in 95% Co., Easton, Pa., 1965, p. 207.
ethanol, and the solution was brought to volume. An aliquot, 10 ( 5 ) J. Levine, J. Pharm. Sci., 54,485(1965).
ml., was pipeted onto the prepared column, and the solution was (6) F. De Fabrizio, J . South African Chem. Znst., XX, 194(1967).
allowed to pass through the column at a rate of 1 ml./min. The
column was then washed with 50 ml. of 80% ethanol divided ACKNOWLEDGMENTS AND ADDRESSES
into two portions, also at a rate of 1 ml./min., and finally with 200
ml. of water at a rate of 4 ml./min. Diphenhydramine was subse- Received February 16, 1970, from the Control Pharmaceutical
quently eluted with 0.1 N hydrochloric acid at a rate of 1 ml./min. Laboratory of Biopharm (Pry.) Limited, Johannesburg, Republic of
The first 5 ml. of eluate was discarded and the balance collected South Africa.
into a 100-ml. volumetric flask until 100 ml. was collected. The Accepted for publication April 3, 1970.

Vol. 59, No, 10, October 1970 0 1471