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African Journal of Pure and Applied Chemistry Vol.
2 (2), pp. 010-017, February, 2008
ISSN 1996-0840
© 2008 Academic Journals
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Development and validation
of LC method for the determination of leflunomide in
pharmaceutical formulations using an experimental design
Srinivas Rao V2, Sunanda
K. K3, Narasimha Rao M.1, Allam Appa
Rao1, Maheswari IL1 and Srinubabu G1*
1Center
for Biotechnology, Andhra University College of Engineering
(A), Visakhapatnam- 530003, India.
2St
John's Pharmacy College, Vijayanagar, Bangalore-40, India.
3Department
of Microbiology, Andhra University College of Science and
Technology, Visakhapatnam-530003, India.
*Corresponding author. Email:
srinubabuau6@gmail.com.
Phone: +91-891-2844204.
Accepted
16 January, 2008 |
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Abstract |
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A rapid and sensitive
RP-HPLC method with UV detection (260 nm) for routine analysis of
leflunomide in a pharmaceutical formulation was developed.
Chromatography was performed with mobile phase containing a mixture of
acetonitrile and phosphate buffer (60:40, v/v) with flow rate of
0.8 ml min−1. Quantitation was accomplished with internal
standard method. The procedure was validated for linearity (correlation
coefficient = 0.9999), accuracy, robustness and intermediate precision.
Experimental design was used for validation of robustness and
intermediate precision. Plackett- Burman design was used to screen the
essential factors for optimization. To test robustness, three factors
were considered; percentage v/v of acetonitrile in mobile phase, flow
rate and pH; an increase in the flow rate results in a decrease of the
drug found concentration, while the percentage of organic modifier and
pH have no important effect on the response. For intermediate precision
measure the variables considered were: analyst, equipment and number of
days. The RSD value (0.93%, n = 24) indicated a good precision of
the analytical method. The proposed method was simple; highly sensitive,
precise, accurate and retention time less than 6 minutes indicating that
the method is useful for routine quality control.
Key words:
Leflunomide,
HPLC, validation, robustness testing, experimental design. |
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