830 CARDOSO & SCHAPOVAL: JOURNAL OF AOAC INTERNATIONAL VOL. 82, NO.
4, 1999
DRUGS, COSMETICS, FORENSIC SCIENCES
UV Spectrophotometry and Nonaqueous Determination of
Terbinafine Hydrochloride in Dosage Forms
CARDOSO & SCHAPOVAL: JOURNAL OF AOAC INTERNATIONAL VOL. 82, NO. 4, 1999
SIMONE GONÇALVES CARDOSO
Universidade Federal de Santa Maria, Departamento de Farmácia Industrial, Santa Maria, CEP 97119-900-RS, Brazil, and
Universidade Federal do Rio Grande do Sul, Faculdade de Farmácia, Curso de Pós-Graduação em Ciências Farmacêuticas,
Av. Ipiranga 2752, Porto Alegre, CEP 90.610-000-RS, Brazil
ELFRIDES E.S. SCHAPOVAL
Universidade Federal do Rio Grande do Sul, Faculdade de Farmácia, Curso de Pós-Graduação em Ciências Farmacêuticas,
Av. Ipiranga 2752, Porto Alegre, CEP 90.610-000-RS, Brazil
An ultraviolet spectrophotometric and a
nonaqueous volumetric method for determining
terbinafine hydrochloride (TH) in pharmaceutical
formulations are presented. The UV spectrophoto-
metric procedure was developed for assay of TH in
raw materials, tablets, and creams. The method
was tested for linearity (0.8–2.8 µg/mL, r = 0.9997),
recovery (102.00% for creams and 99.90% for tab-
lets) and precision (101.3%, CV = 0.96%, n = 9, for
creams; 100.25%, CV = 1.08%, n = 9, for tablets).
The volumetric method involves titration of TH with
0.05M perchloric acid with crystal violet as indica-
tor. This method was used for quantitative determi-
nation of TH in raw materials and tablets. Mean re-
covery and precision were, respectively, 100.41
and 101.18% (CV = 1.64%, n = 9) for TH in tablets.
There were no significant differences between the
proposed methods and a previously described
high-performance liquid chromatographic method.
The UV spectrophotometric and titrimetric meth-
ods are potentially useful for a routine laboratory
because of their simplicity, rapidity, and accuracy.
erbinafine hydrochloride, (E)-N-(6,6-dimethyl-2-hepten-
T 4-ynyl)-N-methyl-1-naphthalene methanamine hydro-
chloride (TH), is a new potent antifungal agent of the
allylamine class with broad-spectrum activity against yeasts,
dimorfic fungi, molds, and dermatophytes (1–4). It is avail-
able as tablets and creams.
No analytical procedures are reported in pharmacopoeias
for its determination in pharmaceutical preparations. TH has
been determined in biological fluids by high-performance liq-
uid chromatography (HPLC; 5–7) and microbiological
bioassays (8, 9). An HPLC method has been used to deter-
mine TH in tablets and creams (10). A UV spectrophotometric
method was proposed for TH in tablets (11), but determination
Received October 19, 1998. Accepted by JM February 17, 1999.
of TH in creams has not been reported. Spectroscopic (UV
and IR) and titrimetric techniques are used routinely in quality
control because of their simplicity and rapidity. The objective
of the present study was to develop a simple, sensitive, and ac-
curate UV spectrophotometric method to quantitate TH in
creams. Nonaqueous titration for determination of TH in raw
materials and tablets is also described.
Experimental
Chemicals and Reagents
All reagents were analytical grade: 0.05M perchloric acid,
1% crystal violet, and 6% mercuric acetate in glacial acetic
acid. TH reference (assigned purity, 99.7%) was obtained
from Galena (São Paulo, Brazil). Pharmaceuticals containing
terbinafine were obtained commercially. Tablets were
claimed to contain 125 mg terbinafine (as base), and creams
were claimed to contain 1% TH.
Apparatus
A Shimadzu (Kyoto, Japan) UV-160 double-beam
UV-visible spectrophotometer with a fixed slit width (2 nm)
and its recorder were used.
Spectrophotometry Standardization
The method was standardized with a methanolic solution
containing 1.0 mg TH reference/mL. From this solution, dilu-
tions in the range 0.2–5.0 µg/mL were made in methanol.
Absorbances of the resulting solutions were measured at the
wavelength of maximum absorbance (224 nm) with methanol
as blank. Compliance with Burger–Lambert Beer’s law was
observed in the range 0.8–2.8 µg TH/mL. Results were used to
calculate the equation of the line by linear regression
(least-squares method), and the data were evaluated by analy-
sis of variance.
Sample Preparations
(a) Creams.—An accurately weighed amount of cream
equivalent to 25 mg TH was placed in a 50 mL volumetric
flask and ca 40 mL methanol was added. After the flask was
 CARDOSO & SCHAPOVAL: JOURNAL OF AOAC INTERNATIONAL VOL. 82, NO. 4, 1999 831

Figure 1. Absorption spectrum of terbinafine hydrochloride in methanol (1.6 mg/mL): (A) reference, (B) tablets, and
(C) creams.

shaken for 30 min, the volume was made up to 50 mL with
methanol. This solution was filtered through filter paper
(Schleicher & Schull, Germany). The first 10 mL of the fil-
trate was discarded. After filtration, dilutions were made with
methanol to give a final concentration of 1.6 µg/mL. Nine de-
terminations were done in triplicate, and the absorbances of
the solutions were determined at 224 nm.
(b) Tablets.—Tablets were treated in the same manner as
creams, except that the weight of pulverized tablets used was
equivalent to 100 mg TH. This amount was transferred to a
100 mL volumetric flask, and 60 mL methanol was added. Af-
ter the flask was shaken, the volume was made up to 100.0 mL
with methanol.
Recovery Tests
Recovery of added reference TH was studied at 3 levels.
Portions of sample solutions (creams and tablets) containing
1.2 µg TH/mL were transferred to a 50 mL volumetric flask.

Table 1. Analysis of terbinafine hydrochloride in tablets and creams

Dosage form N Linear range, µg/mL CV, % Amount found, % Recovery, %

UV spectrophotometry

Creams 9 0.8–2.8 0.96 101.30 102.0

Tablets 9 0.8–2.8 1.08 100.25 99.90

Titrimetry

Tablets 9 — 1.64 101.18 100.41

HPLC

Creams 6 10.0–20.0 1.18 99.26 94.14

Tablets 6 10.0–20.0 0.62 101.15 99.25
832 CARDOSO & SCHAPOVAL: JOURNAL OF AOAC INTERNATIONAL VOL. 82, NO. 4, 1999
1.0, 2.0, and 3.0 mL TH reference solution (20 µg/mL) were
added to separate flasks to give final concentrations of 1.6, 2.0,
and 2.4 µg/mL, respectively. Recovery was calculated with the
formula proposed by AOAC INTERNATIONAL (12).
Titrimetry
To determine the average weight of a tablet, 20 tablets were
weighed and powdered. In a 100 mL flask containing a mag-
netic bar, 100 mg of accurately weighed TH was dissolved in
30 mL acetic acid, and 10 mL 6% mercuric acetate solution
and 0.5 mL crystal violet solution (indicator) was added. The
mixture was titrated with 0.05M perchloric acid until the color
changed to bluish green. The same procedure was used for
raw material. A blank determination was performed.
Recovery Test
Accurately weighed amounts of powdered tablets equiva-
lent to 100 mg TH were placed in 3 flasks. To each was added
12.5, 25, or 37.5 mg TH reference. Final concentrations were
equivalent to 90, 100, and 110%, respectively, of the declared
amounts of TH in tablets. Recovery was calculated with the
formula proposed by AOAC INTERNATIONAL (12).
Results and Discussion
Good planning is essential in the selection and develop-
ment of drug analysis methods. It is necessary to be able to
quantitate with acceptable precision, accuracy, and reliabil-
ity within the budget and other considerations of the labora-
tory, such as complexity of the sample, intent of use, nature
of the drug, availability of apparatus, and specialist skills
(13). Despite analytical methods based on chromatography,
spectrophotometric and nonaqueous volumetric methods
are still widely used because they are inexpensive and easy
to perform.
One UV spectrophotometric method exists for TH deter-
mination in tablets. The UV spectrophotometric method used
in the present study has an advantage over the reported
method because it can be used to quantitate TH in creams and
tablets without complicated extraction or sample pretreat-
ment. The method is simple, giving reproducible, reliable, and
accurate results within a short period. TH is freely soluble in
methanol and methylene chloride, soluble in ethanol, and
slightly soluble in water. Its UV absorption spectrum in meth-
anol shows an intense absorption at 224 nm (Figure 1), with an
apparent molar absorptivity of 86 978/mol·cm.
Method linearity, precision, and accuracy were evaluated. In
the UV procedure, absorbance was linearly correlated with con-
centration in the range 0.8–2.8 µg/mL at 224 nm: y = 0.2664x +
0.0036, with a correlation coefficient of 0.999 (n = 6).
Precision expresses the closeness of agreement among var-
ious measurements obtained from multiple sampling of the
same homogeneous sample under prescribed conditions. It is
usually expressed as the variance, standard deviation, or coef-
ficient of variation (CV) of a series of measurements. Preci-
sion may be considered at 3 levels: repeatability, intermediate
precision, and reproducibility (14).
Method repeatability was studied by assaying 9 samples of
creams or tablets, containing about the same concentration of
TH, during the same day, and under the same experimental
conditions. CVs were 0.96% for creams and 1.08% for tablets,
indicating good precision.
Accuracy expresses the agreement between the accepted
value (either as conventional true value or an accepted refer-
ence value) and the value found. Accuracy is reported as per-
cent recovery of a known amount of analyte added to the sam-
ple (14). Mean recoveries were 102.00% for creams and
99.90% for tablets. Results are comparable with declared
amounts and with those obtained by HPLC (Table 1). Analy-
sis of variance indicated no significant difference between UV
spectrophotometry and HPLC (p < 0.01).
The other method described here is a nonaqueous
titrimetry. Most medicinal agents are weak acids or bases and
can be assayed in a nonaqueous medium (15). Many salts of
halogen acids may be titrated in acetic acid or acetic anhydride
after addition of mercuric acetate, which removes the halide
ion as the unionized mercuric halide complex and introduces
the acetate ion (16, 17). TH is an amine salt, and the reaction
in a nonaqueous medium between TH and 0.05M perchloric
acid provides a basis for precise and accurate determination of
TH in raw materials and tablets.
Method validity was assessed by assay of tablets. The
amount of TH found in tablets were 101.18% of the label claim,
with CV = 1.64%. Mean recovery was 100.41%. Levels deter-
mined by the proposed method did not differ significantly (p <
0.01) from those determined by HPLC (Table 1).
Conclusion
The UV spectrophotometric and nonaqueous methods pre-
sented here are simple, precise, accurate, and convenient.
Therefore, they can be useful for routine analyses and qual-
ity-control assays of TH in pharmaceutical formulations.
References
(1) Petrany, G., Ryder, N.S., & Stütz, A. (1984) Science 224,
1239–1241
(2) Nussbaumer, P., Leitner, I., Mraz, K., & Stütz, A. (1995) J.
Med. Chem. 38, 1831–1836
(3) Abdel-Rahman, S., & Nahata, M. (1997) Ann. Pharmacother.
31, 445–456
(4) Balfour, J., & Faulds, D. (1992) Drugs 43, 259–284
(5) Schatz, F., & Haberl, H. (1989) Arzneim. Forsch. 39,
527–532
(6) Zehender, H., Denouël, J., Roy, M., Le Saux, L., & Schaub,
P. (1995) J. Chromatogr. B 664, 347–355
(7) Denouël, J., Keller, H.P., Schaub, P., Delaborde, C., &
Humbert, H. (1995) J. Chromatogr. B. 663, 353–359
(8) Häuser, M., Schmitt, H.J., Bernard, E.M., & Armstrong, D.
(1988) Eur. J. Clin. Microb. Infect. Dis. 7, 531–533
(9) Kan, V.L., Henderson, D.K., & Bennett, J.E. (1986)
Antimicrob. Agents Chemother. 30, 628–629
(10) Cardoso, S.G., & Schapoval, E.E.S. (1999) J. Pharmac.
Biom. Anal. 19, 809–812
 CARDOSO & SCHAPOVAL: JOURNAL OF AOAC INTERNATIONAL VOL. 82, NO. 4, 1999 833

(11) Wang, J. (1996) Zhongguo Yiyao Gongye Zazhi 27, 363–365

(12) Official Methods of Analysis (1990) 15th Ed., AOAC,
Arlington, VA
(13) Metha, A.C. (1997) Analyst 122, 83R–88R
(14) International Conference on Harmonization (ICH): Valida-
tion of Analytical Procedures (1996) Commission of the
European Communities, Switzerland
(15) Korolkovas, A., & Ferreira, E.I. (1986) Rev. Portuguesa
Farm. 36, 1–21
(16) The United States Pharmacopoeia (1995) 23rd Rev., U.S.
Pharmacopoeial Convention, Rockville, MD
(17) British Pharmacopoeia (1993) HM Stationery Office, Lon-
don, UK