DEVELOPMENT AND VALIDATION OF BIOANALYTICAL HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF CILNIDIPINE AND NEBIVOLOL IN HUMAN PLASMA


Aruna G., Bharathi K, Kvsrg Prasad

Abstract


Objective: To develop and validate a modified isocratic reversed-phase high performance liquid chromatographic (RP-HPLC) method for determination of cilnidipine and nebivolol in human plasma to be used for pharmacokinetic studies.

Methods: The drug was extracted from plasma samples by direct protein precipitation technique using acetonitrile. Amlodipine was used as internal standard (IS). Samples were analyzed on BDS C18 column (250 x 4.6 mm, 5 µm), applying ortho phosphoric acid (0.1%): Acetonitrile, at a ratio of 45:55 v/v in isocratic mode as a mobile phase at a flow rate of 1 ml/min to attain adequate resolution. Separations were performed at room temperature and monitored at a wavelength of 260 nm after injection of 50μl samples into the HPLC system. The analytical method was validated according to FDA bioanalytical method validation guidance. The method was applied for pharmacokinetic study of cilnidipine and nebivolol tablets-10 mg and 5 mg were administered as a single dose to 6 healthy male rabbits under fasting condition. Twelve blood samples were withdrawn from each rabbit over 24 h periods. From the plasma concentration-time data of each individual, the pharmacokinetic parameters; Cmax, Tmax, AUC0-t and AUC0-∞ were calculated.

Results: A peak area was obtained for cilnidipine and nebivolol at 3.943 and 4.719 min retention time respectively. Linearity was established at a concentration range of 0.20-20 μg/ml (r2=0.999, n=8) for cilnidipine and 0.02-2 μg/ml (r2=0.999, n=8) for nebivolol. The lower limit of quantitation (LLOQ) was identifiable and reproducible at 0.2μg/ml for cilnidipine and 0.02 μg/ml for nebivolol. The coefficients of variation (%cv) of the intra-day and inter-day precision of cilnidipine at 600, 1000 and 1600ng/ml levels were found to be 6.90%, 6.19%, 5.22%; and 7.74%, 6.54%, 5.77%, respectively, which are lower than the accepted criteria limits (15-20 %). The mean recovery (%) cilnidipine at 600, 1000, and 1600ng/ml was found to be 101.03%, 99.27% and 104.87%, and for nebivolol 60, 100, and 160 ng/ml was found to be 106.13%, 107.03% and 98.06% respectively. Stability at different conditions and in autosampler was also established. The mean pharmacokinetic parameters; Cmax, Tmax, AUC0-t and AUC0-∞ were 6 ng/ml, 2 hr, 96.76 mg. hr/ml, 63.45 mg. hr/ml for cilnidipine and 5.8ng/ml, 2hr, 74.78 mg. hr/ml, 100.25 mg. hr/ml for nebivolol respectively.

Conclusion: The present analytical method was found to be specific, sensitive, accurate and precise for quantification of cilnidipine and nebivolol in human plasma. It can be successively applied for pharmacokinetics, bioavailability and bioequivalence studies.


Keywords


Cilnidipine, Nebivolol, HPLC, Human plasma, Pharmacokinetics

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References


Sharma T, Rajesh P. Development and validation of UV spectrophotometric method for determination of nebivolol hydrochloride following ICH guidlines and study of its degradation profile. Asian J Pharm Clin Res 2012;5:69-72.

Kokilambiga KS, Lakshmi KS. Analytical methodologies for determination of cilnidipine an overview. Int J Pharm Pharm Sci 2014;6:36-8.

Reema H Rupareliya, Hitendra S Joshi. Stability indicating simultaneous validation of telmisartan and cilnidipine with forced degradation behavior study by RP-HPLC in tablet dosage form. ISRN Chromatography 2013;6:1-7.

Shah DM, Doshi DB. Development and validation of HPTLC method for simultaneous estimation of nebivolol hydrochloride and cilnidipine in combined pharmaceutical tablet dosage form. Int J Pharma Res Rev 2016;5:1-7.

Jaldeepsinh V, Rathod, Dilip G, Maheshwari. Development and validation of second order derivative spectrophotometric method for simultaneous estimation of cilnidipine and valsartan in synthetic mixture. Am J PharmTech Res 2015;5:314-24.

Lakshmi Aswini G, Dachinamoorthy D, Seshagiri Rao J. Stability indicating RP-HPLC-PDA method for simultaneous estimation of olmesartan, cilnidipine and chlorthalidone with forced degradation behavior study in bulk and in its tablet dosage form. Am J PharmTech Res 2015;5:379-92.

Mohammad Yunoos, Gowri Sankar D. Validated stability indicating RP-HPLC method for simultaneous quantitative estimation of hydrochlorothiazide and nebivolol hydrochloride in bulk and combined tablet dosage form. Am J PharmTech Res 2015;5:507-19.

Manzoor A, Rashmi DR, Satishkumar Shetty A, Anil Kumar SM, Ravi MC, Kuppast IJ. RP-HPLC method development and validation for simultaneous estimation of cilnidipine and olmesartan medoxomil in combined tablet dosage form. World J Pharm Pharm Sci 2015;4:785-95.

Zeel T Doshi, Jignesh S, Shah, Dilip G, Maheshwari. A review on analytical method for determination of calcium channel blocker in different dosage forms. J Global Trends Pharm Sci 2015;6:2829–39.

Ravisankar P, Devala Rao G. Rapid separation and quantification of certain anti-hypertensive agents in their dosage forms by reversed phase-high performance liquid chromatography. Indo Am J Pharm Res 2015;5:2335-58.

Prasad S, Virkar, Satish G, Pingale, Kiran V, Mangaonkar. Development and validation of a high performance liquid chromatography method for determination of telmisartan in rabbit plasma and its application to a pharmacokinetic study. Anal Bioanal Techniques 2012;3:1-5.

Jaafar JI, Tamimi A. Development and validation of HPLC/UV method for determination of meloxicam in human plasma and application in pharmacokinetic studies. Int J Pharm Pharm Sci 2014;7:370-8.

Pharne AB, Santhakumari BA, Ghemud S, Jain HK, Kulkarni MJ. Bioanalytical method development and validation of vildagliptin a novel dipeptidyl peptidase IV inhibitor by RP-HPLC method. Int J Pharm Pharm Sci 2012;4:119-23.

Swathimutyam P, Prabhakar B. Bio analytical method development and validation of temozolomide in rat plasma using RP-HPLC method. Int J Pharma Sci Res 2016;7:1298-301.

Hendriks G. Review theoretical models in LC based bioanalytical method development. J Pharm Biomed Anal 2009;46:1-10.

International Conference on Harmonization (ICH), Validation of analytical methods definitions and terminology. ICH Q2 A; 1994.

International Conference on Harmonization (ICH), Validation of analytical methods. Methodology, ICH Q2B; 1996.




About this article

Title

DEVELOPMENT AND VALIDATION OF BIOANALYTICAL HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF CILNIDIPINE AND NEBIVOLOL IN HUMAN PLASMA

Keywords

Cilnidipine, Nebivolol, HPLC, Human plasma, Pharmacokinetics

DOI

10.22159/ijpps.2017v9i10.20237

Date

02-10-2017

Additional Links

Manuscript Submission

Journal

International Journal of Pharmacy and Pharmaceutical Sciences
Vol. 9, Issue 10, 2017 Page: 253-259

Online ISSN

0975-1491

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Authors & Affiliations

Aruna G.
Department of Quality Assurance, Krishna Teja Pharmacy College, Tirupati

Bharathi K
Institute of Pharmaceutical Technology, Sri Padmavati Mahila Visvavidyalayam (Women’s University), Tirupati, Andhra Pradesh, India
India

Kvsrg Prasad
Institute of Pharmaceutical Technology, Sri Padmavati Mahila Visvavidyalayam (Women’s University), Tirupati, Andhra Pradesh, India
India


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