Published:Journal of Chromatographic Science,
ISSN 0021-9665Volume
38, Number 6, June 2000, pp. 246-254
Validation
of a High-Performance Liquid Chromatography Method for the Assay of and Determination
of Related Organic Impurities in Nevirapine Drug SubstanceQ.
Chan Li1,*, T. Tougas1, K. Cohen1, R. Lee1, P. Meagan1, M. Corson2, and T. Muchnick3
1Analytical Sciences Department, Research and Development Center, Boehringer
Ingelheim Pharmaceuticals, Inc., Ridgefield, CT 06877; 2Pharmaceutics Department,
Research and Development Center, Boehringer Ingelheim Pharmaceuticals, Inc.,
Ridgefield, CT 06877; and 3Purdue Pharma, L.P., 444 Saw Mill River Rd., Ardsley,
NY 10502
Nevirapine
(Viramune™), a dipyridiodiazepinone, is a potent and highly specific nonnucleoside
inhibitor of HIV-1 reverse transcriptase. This paper describes the validation
of a specific, sensitive, and stability-indicating high-performance liquid chromatography
method for the assay and determination of related organic impurities in nevirapine
drug substance. This method uses a Supelcosil LC-ABZ column, a mobile phase
of 20:80 (v/v) acetonitrile–25mM NH4H2PO4 (pH 5.0), and ultraviolet detection
at a wavelength of 220 nm. This method was validated for specificity, linearity,
accuracy, repeatability, detection limit, quantitation limit, stability of analyte
solutions, robustness, and intermediate precision. Nevirapine is completely
separated from all impurities. The method is shown to be linear with coefficients
of determination r2 greater than 0.999. Average accuracy is 100.4% with a relative
standard deviation of 0.7% for the assay. Accuracy ranges from 100.1 to 102.6%
for related organic impurities. Repeatability is good, with relative standard
deviations not more than 1.4%. The detection limit and the quantitation limit
are determined to be 0.001 and 0.003%, respectively. Relative response factors
of known organic impurities are determined, permitting the use of nevirapine
at the 0.1% level as an external standard for the quantitation of these impurities.
Analyte solutions are shown to be stable for at least 2 days at ambient temperature.
The method is validated as robust, and intermediate precision is high. A system
suitability test is developed and validated, and requirements are set.
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