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IJCRR - 2(1), January, 2010

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DERIVATIVE SPECTROPHOTOMETRIC ESTIMATION OF NEVIRAPINE IN BULK DRUG AND
PHARMACEUTICAL DOSAGE FORMS

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
formulations.

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

Full Text:

INTRODUCTION

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

RESULTS AND DISCUSSION

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.

CONCLUSION

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.

ACKNOWLEDGEMENT

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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A Study by Purushottam S. G. et al. entitled "Development of Fenofibrate Solid Dispersions for the Plausible Aqueous Solubility Augmentation of this BCS Class-II Drug" is awarded Best article for Vol 13 issue 10

A Study by Kumar S. et al. entitled "A Study on Clinical Spectrum, Laboratory Profile, Complications and Outcome of Pediatric Scrub Typhus Patients Admitted to an Intensive Care Unit from a Tertiary Care Hospital from Eastern India" is awarded Best Article for Vol 13 issue 09

A Study by Mardhiah Kamaruddin et al. entitled "The Pattern of Creatinine Clearance in Gestational and Chronic Hypertension Women from the Third Trimester to 12 Weeks Postpartum" is awarded Best Article for Vol 13 issue 08

A Study by Sarmila G. B. et al. entitled "Study to Compare the Efficacy of Orally Administered Melatonin and Clonidine for Attenuation of Hemodynamic Response During Laryngoscopy and Endotracheal Intubation in Gastrointestinal Surgeries" is awarded Best Article for Vol 13 issue 07

A Study by M. Muthu Uma Maheswari et al. entitled "A Study on C-reactive Protein and Liver Function Tests in Laboratory RT-PCR Positive Covid-19 Patients in a Tertiary Care Centre – A Retrospective Study" is awarded Best Article of Vol 13 issue 06 Special issue Modern approaches for diagnosis of COVID-19 and current status of awareness

A Study by Gainneos PD et al. entitled "A Comparative Evaluation of the Levels of Salivary IgA in HIV Affected Children and the Children of the General Population within the Age Group of 9 – 12 Years – A Cross-Sectional Study" is awarded Best Article of Vol 13 issue 05 Special issue on Recent Advances in Dentistry for better Oral Health

A Study by Alkhansa Mahmoud et al. entitled "mRNA Expression of Somatostatin Receptors (1-5) in MCF7 and MDA-MB231 Breast Cancer Cells" is awarded Best Article of Vol 13 issue 06

A Study by Chen YY and Ghazali SRB entitled "Lifetime Trauma, posttraumatic stress disorder Symptoms and Early Adolescence Risk Factors for Poor Physical Health Outcome Among Malaysian Adolescents" is awarded Best Article of Vol 13 issue 04 Special issue on Current Updates in Plant Biology to Medicine to Healthcare Awareness in Malaysia

A Study by Kumari PM et al. entitled "Study to Evaluate the Adverse Drug Reactions in a Tertiary Care Teaching Hospital in Tamilnadu - A Cross-Sectional Study" is awarded Best Article for Vol 13 issue 05

A Study by Anu et al. entitled "Effectiveness of Cytological Scoring Systems for Evaluation of Breast Lesion Cytology with its Histopathological Correlation" is awarded Best Article of Vol 13 issue 04

A Study by Sharipov R. Kh. et al. entitled "Interaction of Correction of Lipid Peroxidation Disorders with Oxibral" is awarded Best Article of Vol 13 issue 03

A Study by Tarek Elwakil et al. entitled "Led Light Photobiomodulation Effect on Wound Healing Combined with Phenytoin in Mice Model" is awarded Best Article of Vol 13 issue 02

A Study by Mohita Ray et al. entitled "Accuracy of Intra-Operative Frozen Section Consultation of Gastrointestinal Biopsy Samples in Correlation with the Final Histopathological Diagnosis" is awarded Best Article for Vol 13 issue 01

A Study by Badritdinova MN et al. entitled "Peculiarities of a Pain in Patients with Ischemic Heart Disease in the Presence of Individual Combines of the Metabolic Syndrome" is awarded Best Article for Vol 12 issue 24

A Study by Sindhu Priya E S et al. entitled "Neuroprotective activity of Pyrazolone Derivatives Against Paraquat-induced Oxidative Stress and Locomotor Impairment in Drosophila melanogaster" is awarded Best Article for Vol 12 issue 23

A Study by Habiba Suhail et al. entitled "Effect of Majoon Murmakki in Dysmenorrhoea (Usre Tams): A Standard Controlled Clinical Study" is awarded Best Article for Vol 12 issue 22

A Study by Ghaffar UB et al. entitled "Correlation between Height and Foot Length in Saudi Population in Majmaah, Saudi Arabia" is awarded Best Article for Vol 12 issue 21

A Study by Siti Sarah Binti Maidin entitled "Sleep Well: Mobile Application to Address Sleeping Problems" is awarded Best Article for Vol 12 issue 20

A Study by Avijit Singh"Comparison of Post Operative Clinical Outcomes Between “Made in India” TTK Chitra Mechanical Heart Valve Versus St Jude Mechanical Heart Valve in Valve Replacement Surgery" is awarded Best Article for Vol 12 issue 19

A Study by Sonali Banerjee and Mary Mathews N. entitled "Exploring Quality of Life and Perceived Experiences Among Couples Undergoing Fertility Treatment in Western India: A Mixed Methodology" is awarded Best Article for Vol 12 issue 18

A Study by Jabbar Desai et al. entitled "Prevalence of Obstructive Airway Disease in Patients with Ischemic Heart Disease and Hypertension" is awarded Best Article for Vol 12 issue 17

A Study by Juna Byun et al. entitled "Study on Difference in Coronavirus-19 Related Anxiety between Face-to-face and Non-face-to-face Classes among University Students in South Korea" is awarded Best Article for Vol 12 issue 16

A Study by Sudha Ramachandra & Vinay Chavan entitled "Enhanced-Hybrid-Age Layered Population Structure (E-Hybrid-ALPS): A Genetic Algorithm with Adaptive Crossover for Molecular Docking Studies of Drug Discovery Process" is awarded Best article for Vol 12 issue 15

A Study by Varsha M. Shindhe et al. entitled "A Study on Effect of Smokeless Tobacco on Pulmonary Function Tests in Class IV Workers of USM-KLE (Universiti Sains Malaysia-Karnataka Lingayat Education Society) International Medical Programme, Belagavi" is awarded Best article of Vol 12 issue 14, July 2020

A study by Amruta Choudhary et al. entitled "Family Planning Knowledge, Attitude and Practice Among Women of Reproductive Age from Rural Area of Central India" is awarded Best Article for special issue "Modern Therapeutics Applications"

A study by Raunak Das entitled "Study of Cardiovascular Dysfunctions in Interstitial Lung Diseas epatients by Correlating the Levels of Serum NT PRO BNP and Microalbuminuria (Biomarkers of Cardiovascular Dysfunction) with Echocardiographic, Bronchoscopic and HighResolution Computed Tomography Findings of These ILD Patients" is awarded Best Article of Vol 12 issue 13

A Study by Kannamani Ramasamy et al. entitled "COVID-19 Situation at Chennai City – Forecasting for the Better Pandemic Management" is awarded best article for Vol 12 issue 12

A Study by Muhammet Lutfi SELCUK and Fatma entitled "Distinction of Gray and White Matter for Some Histological Staining Methods in New Zealand Rabbit's Brain" is awarded best article for Vol 12 issue 11

A Study by Anamul Haq et al. entitled "Etiology of Abnormal Uterine Bleeding in Adolescents – Emphasis Upon Polycystic Ovarian Syndrome" is awarded best article for Vol 12 issue 10

A Study by entitled "Estimation of Reference Interval of Serum Progesterone During Three Trimesters of Normal Pregnancy in a Tertiary Care Hospital of Kolkata" is awarded best article for Vol 12 issue 09

A Study by Ilona Gracie De Souza & Pavan Kumar G. entitled "Effect of Releasing Myofascial Chain in Patients with Patellofemoral Pain Syndrome - A Randomized Clinical Trial" is awarded best article for Vol 12 issue 08

A Study by Virendra Atam et. al. entitled "Clinical Profile and Short - Term Mortality Predictors in Acute Stroke with Emphasis on Stress Hyperglycemia and THRIVE Score : An Observational Study" is awarded best article for Vol 12 issue 07

A Study by K. Krupashree et. al. entitled "Protective Effects of Picrorhizakurroa Against Fumonisin B1 Induced Hepatotoxicity in Mice" is awarded best article for issue Vol 10 issue 20

A study by Mithun K.P. et al "Larvicidal Activity of Crude Solanum Nigrum Leaf and Berries Extract Against Dengue Vector-Aedesaegypti" is awarded Best Article for Vol 10 issue 14 of IJCRR

A study by Asha Menon "Women in Child Care and Early Education: Truly Nontraditional Work" is awarded Best Article for Vol 10 issue 13

A study by Deep J. M. "Prevalence of Molar-Incisor Hypomineralization in 7-13 Years Old Children of Biratnagar, Nepal: A Cross Sectional Study" is awarded Best Article for Vol 10 issue 11 of IJCRR

A review by Chitra et al to analyse relation between Obesity and Type 2 diabetes is awarded 'Best Article' for Vol 10 issue 10 by IJCRR.

A study by Karanpreet et al "Pregnancy Induced Hypertension: A Study on Its Multisystem Involvement" is given Best Paper Award for Vol 10 issue 09

List of Awardees

A Study by Ese Anibor et al. "Evaluation of Temporomandibular Joint Disorders Among Delta State University Students in Abraka, Nigeria" from Vol 13 issue 16 received Emerging Researcher Award

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