A SIMPLE RP-HPLC METHOD FOR THE ASSAY OF CLOTRIMAZOLE-

LOADED NANOCAPSULE SUSPENSIONS

SAURIN, S, FERREIRA, L. M., SILVEIRA, F., RASCOVETZKI, R.H., CHASSOT, J. M.,
SCHAFFAZICK, S. R
2
, CRUZ, L.
3
*

(1)
PPG em Cincias Farmacuticas, UFSM, Santa Maria, Brazil- sarasaurin@gmail.com
(2)
Curso de
Farmcia, UFSM, Santa Maria, RS, Brazil.
(3)
PPG em Cincias Farmacuticas, UFSM, Avenida
Roraima, 1000, prdio 26, CEP: 97105-900, Santa Maria, RS, Brazil - leticiacruz@smail.ufsm.br
Keywords: Clotrimazole, nanocapsule, validation, HPLC

1. Introduction

Clotrimazole (CTZ) is an antifungal imidazole
derivative that can be used topically for fungal
infections. Regarding the literature, there are
some methods to determine clotrimazole in
topical formulations. However, the most part of
them employ expensive organic solvents, buffer
solutions and present a long retention time
1,2,3
.
Taking this into account, the aim of this work
was to validate a simple RP-HPLC method for
the assay of CTZ in nanocapsule suspensions.

2. Methods

The parameters evaluated to the validation were
specificity, linearity, limits of detection (LOD)
and quantification (LOQ), precision, accuracy
and robustness. The chromatographic conditions
were RP Phenomenex column (250 x 4.60 nm, 5
m particle size) mobile phase methanol-water
(90:10) at isocratic flow rate of 1mL/min,
injection volume at 20 L and the detection was
performed at 229 nm. Interfacial deposition of
preformed polymer method
4
was used to prepare
the CTZ-loaded nanocapsules (NC-CTZ) using
medium chain triglycerides as oily phase and
Eudragit RS

100 as polymer at a drug

concentration of 1mg/mL.

3. Results
In the range of 2.0-10.0 g/mL the method was
linear presenting a regression equation y =
42648x-14382 with an adequate determination
coefficient (R= 0.9974). LOD and LOQ were
0.26 g/mL and 0.88g/mL, respectively.
Repeatability (intra-day precision) was
evaluated on six injections of the same NC-CTZ
sample. The percentage relative standard
deviation (RSD %) for each well-recovered
CTZ was less than 1.5%. In its turn,
intermediate precision (inter-day) presented
RSD of 2%. Accuracy was determined using
samples of known concentrations NC
suspensions spiked with three different
concentrations of standard solution (3.0, 5.0 and
7.0 g/mL). The recovery was found within the
range of 98103%. The specificity, evaluated
using blank NC suspensions, showed no
interference from excipients. Finally, robustness
was evaluated by the variation of the flow rate
(0.9 and 1.1 mL/min) and the composition
mobile phase (85:15 and 95:05 methanol-
water). The results showed that the mentioned
variations in method conditions give a little
RSD, lower than 3 %, confirming the robustness
of the method.

4. Conclusion
The proposed method is specific, precise,
accurate, linear for the concentration range of
2.0-10.0 mg/mL and robust. It is suitable to be
applied to assay clotrimazole in nanocapsule
suspensions avoiding the use of buffer solution
in the mobile phase.
Acknowledgments
We thank FAPERGS for financial support.
References
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Talanta, 2010.

3- British Pharmacopeia, British, United Kingdom, London,
2010.

4- Fessi, H.; Puisieux, F.; Devissaguet, J.P. European
Patent 0274961 A1, 1988.