International Journal of Current Research and Review
ISSN: 2231-2196 (Print)ISSN: 0975-5241 (Online)
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IJCRR - Vol 02 Issue 01, January, 2010

Pages: 17-27

Date of Publication: 30-Nov--0001

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Author: Jawade S. K., Khanage S.G., Mohite P.B., Deshmukh V.K.

Category: Healthcare

Abstract:A simple, accurate and precise spectrophotometric method has been developed and validated for the
estimation of nevirapine from bulk drug and tablet formulations. Nevirapine shows a sharp peak at
241.0 nm in first order derivative spectrum with n =1. The drug follow Beer-Lambert?s law in the
concentration range of 4-24 \?g/ml in this method. Result of the analysis was validated statistically and found
accurate. The method was successfully applied for determination of drug in tablets, wherein no
interference from tablet excipients was observed, indicating the specificity of the developed method. Thus the
proposed method can be used successfully for routine analysis of nevirapine from capsule and tablet

Keywords: Nevirapine, Ultraviolate Spectroscopy, Derivative spectroscopy, validation, precise, accurate .

Full Text:


Nevirapine (Fig.1), 11-cyclopropyl-4- methyl-5, 11-dihydro-6H-dipyrido [3, 2-b: 2?, 3?- e][1,4] diazepin-6-one is a reverse transcriptase (RT) inhibitor of human immunodeficiency virus type 1 (HIV-1)1,2. Nevirapine inhibits replication of HIV-1 by interfering with viral RNAdirected DNA polymerase (reverse transcriptase). It binds directly to herodimeric HIV-1 reverse transcriptase and exerts a virustatic effect by acting as a specific, noncompetitive HIV-1 reverse transcriptase inhibitor; it appears to inhibit viral RNA- and DNAdependent DNA polymerase activities by disrupting the catalytic site of the enzyme3 . Literature survey reveals that there are analytical methods available for determination of nevirapine from biological matrices 4-14, bulk drug and dosage forms 15-17, and analytical methods for determination of nevirapine with combination of other antiviral drugs 18-52 Literature survey further revealed that there were very few reported RP-HPLC and spectrophotometric estimation methods for the analysis of nevirapine. Thus, an appropriate analytical procedure for the quantitative determination of nevirapine from bulk drugs is of considerable importance. Keeping this objective in mind an attempt has been made to develop and validate UV estimation method for the analysis of nevirapine which would be highly sensitive, having good resolution and reproducible. Various validation aspects of the analysis, accuracy, precision, recovery and the limits of detection and quantification etc., have been measured.

Material and Methods The working standard of nevirapine was procured from Cipla Ltd., India. analytical grade Hydrochloric acid. 0.1N HCl prepared in distilled water used as solvent. JASCO V-630 UV/VIS spectrophotometer was used with 1cm matched quartz cells. Tablets of 200 mg strength were procured from local pharmacy (Nevimune-200). Accurately about 100mg of the pure drug was weighed and dissolved in sufficient quantity of 0.1N HCl and volume made up to100ml with 0.1N HCl to give standard stock solution (1


The solvent chosen for UV must take into account the chemical nature and polarity of the drug molecule. Nevirapine is practically insoluble in water, soluble in 0.1 N hydroclodic acid, dichloromethane, dimethylsulphoxide and dimethylformamide, slightly soluble in methanol. Calibrator solutions were prepared in 0.1N HCl. The method developed was validated for limit of detection (LOD) and limit of quantitation (LOQ) in order to determine and ensure sensitivity of the developed method. The method was found to be linear over the range 4-24 μg/ml per spot with coefficient of mg/ml). Aliquots of standard stock solution were pipette out and suitably diluted with 0.1N HCl to get final concentration of 4-24μg/ml of standard solution. The solution were scanned in the spectrum mode from 400 nm to 200 nm wavelength range and the first order derivative spectra were obtained at n =1 (Method A) a sharp peak was obtained at 241nm (Figure-1). The absorbance difference at n=1 (dA/dλ) was calculated by the inbuilt software of the instrument which is directly proportional to the concentration of the standard solution .A calibration curve was plotted taking the absorbance difference (dA/dλ) against the concentration of the standard solutions. The method was applied for the sample solution of known concentration and was found be satisfactory for analysis of tablet formulation.

correlation 0.9987. (Table 1) Intra-day and inter-day precision studies showed a % RSD was less than 5.00%, indicating the method was precise. The accuracy values obtained, in the range 98.53 – 100.73 % for drug are indicative of excellent accuracy and recovery. This indicates the method is specific. Stability studies were carried out for standard. It was found to be stable in sample solution, prior to development and after development. The developed method was then validated and successfully applied for quantitation of nevirapine from the formulation. To ensure accuracy of the method, recovery studies were performed by standard addition method at 80%, 100% and 120% level,

to the pre-analyzed samples and the subsequent solutions were re-analyzed. At each level, three determinations were performed and the results obtained are shown in Table 3. The results of recovery studies were within the specified limits of ICH guidelines54. Lower values of %RSD reflect the accuracy of the method. Precision, expressed in terms of %RSD was determined in terms of intra-day and interday precisions, analyzing the drug at six different concentrations, determining each concentration thrice. The sample solutions were analyzed using the method for 3 consecutive days, repeating the process twice a day at different period. The results obtained are summarized in Table 3 and reflect high degree of precision. Two different analysts performed assay on marketed tablets of the drug, in similar operational and environmental conditions, using the developed method to determine its ruggedness. A typical absorbance spectrum of the drug is shown in Fig.1.


The developed and validated UV estimation method reported here is rapid, simple, accurate, sensitive and specific. The method was also successfully used for quantitative estimation and analysis of Nevirapine from formulation. Thus the reported method is of considerable importance and has great industrial applicability for quality control and analysis of Nevirapine from bulk drug and formulations.


Authors thank Cipla Ltd., India for supplying the authenticated standard of Nevirapine and Mr. Ajay Pise for his moral support. Also thanks to M.E.S. College of Pharmacy for providing the requirements.





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