Reversed phase HPLC method development and validation for the analysis of amlodipine besylate in tablets dosage form and human plasma.
https://doi.org/10.51412/psnnjp.2023.38
Keywords:
Pharmacokinetics, validation, amlodipine besylate, method development, HPLCAbstract
Introduction: Several analytical methods for quantitative determination of amlodipine either in pharmaceuticals or biological fluids have suffered different setbacks. Therefore, we aimed at developing a cost effective, simple, robust, and sensitive reversed phase high performance liquid chromatography (RP-HPLC- UV) method for quantification of amlodipine in dosage form and in human plasma.
Method: The separation was performed on an analytical column 150 mm x 4.6 mm i.d., Agilent Eclipse XDB C18 Column packed with 5 μm particle size and protected by a guard column (30 mm x 4.6 mm i.d.). The mobile phase composition contained acetonitrile: phosphate buffer (25 mM KH2PO4) adjusted to pH 3.1 with orthophosphoric acid in the ratio of 45:55 v/v. The flow rate was 1.2 mL/min and the UV detector was set at a wavelength of 240 nm. The column was maintained at ambient temperature.
Results: The calibration curve gave good linearity in the concentration range studied. The column efficiency was highly evident by high theoretical plates and small height equivalent to theoretical plate (HETP). The percent relative standard deviation (% RSD) was less than 3% in the inter- and intra-day o assay. Amlodipine, 50 ng/mL in plasma stored frozen at -80 C was stable There was no statistical difference in the precision of amlodipine assayed after 12 months when compared to the initial assay (P>0.05). The % recovery of amlodipine in human plasma was 97.79±0.17% to 99.52±0.63%.
Conclusion: The new RP-HPLC method is sensitive, reproducible, precise, accurate and robust. We conclude that the method is suitable for estimation of amlodipine besylate in human plasma for pharmacokinetic studies and assay of pharmaceutical dosage form
References
Mahendra D, Chakraborthy G (2021) A Novel Bioanalytical RP-HPLC Method Development and Validation for Simultaneous Determination of Amlodipine Besylate on Human Plasma. International Journal of Drug Development and Research 13(3): 156-159.
Osman RAM, Elbashir AA (2017) A HighPerformance Liquid Chromatographic (HPLC) Method for the Determination of Amlodipine Drug in Dosage form Using 1,2-Naphthoquine-4-Sulfonate (NQS). Journal of Analytical & Pharmaceutical Research 6(1): 00163. https://doi.org/10.15406/japlr.2017.06.00163
Wankhede SBK, Raka C, Wadkar SB, Chitlange SS (2010) Spectrophotometric and HPLC Methods for Simultaneous Estimation of Amlodipine Besilate, Losar-tan Potassium and Hydrochlorothiazide in Tablets. Indian Journal of Pharmaceutical Sciences 72(1):136-140. https://doi.org/10.4103%2F0250- 474X.62239
Kul D, Dogan-Topal B, Kutucu T, Uslu B, O z k a n S A ( 2 0 1 0 ) H i g h - P e r f o r m a n c e L i q u i d Chromatographic and First Derivative of the Ratio Spectrophotometric Determination of Amlodipine and Valsartan in Their Binary Mixtures. Journal of AOAC International 93(3):882-890.
Yaşin DS, BingülAA, KaraküçükA, Teksin ZS (2021) Development and validation of an HPLC method using an experimental design for analysis of amlodipine besylate and enalapril maleate in a fixed-dose combination. Turkish journal of pharmaceutical sciences 18(3):306-318. https://doi.org/10.4274/tjps.galenos.2020.89725
Argekar AP, Powar SG (2000) Simultaneous determination of atenolol and amlodipine in tablets by high-performance thin-layer chromatography. Journal of pharmaceutical and biomedical analysis 21(6):1137–1142. https://doi.org/10.1016/S0731-7085(99)00210-1.
Pandya KK, Satia M, Gandhi TP, Modi IA, Modi RI, Chakravarthy BK (1995) Detection and determination of total amlodipine by high-performance thin-layer chromatography: a useful technique for pharmacokinetic studies. Journal of chromatography B, B i o m e d i c a l a p p l i c a t i o n s 6 6 7 ( 2 ) : 3 1 5 – 3 2 0 . https://doi.org/10.1016/0378-4347(95)00016-c
Sarkar A, Das D, Selvan P, Gowda K, Mandal U, et al. (2008) Simultaneous determination of Metoprolol succinate and Amlodipine besylate in human plasma by liquid chromatography–tandem mass spectrometry method and its application in bioequivalence study. Journal of chromatography B, Analytical technologies in the biomedical and life sciences 873(1): 77-85. https://doi.org/10.1016/j.jchromb.2008.07.040
Marzo A, Bo LD, Mazzucchelli P, Monti NC, Crivelli F, Ismaili S, Uhr MR, Commare PL (2000) Amlodipine bioequivalence achieved with a very sensitive liquid chromatography tandem mass spectrometric bioassay. Arzneimittelforschung 50(8): 688–694. https://doi.org/10.1055/s-0031-1300274
Narapusetti A, Bethanabhatla S, Sockalingam A, Repaka N, Saritha V (2015) Simultaneous determination of rosuvastatin and amlodipine in human plasma using tandem mass spectrometry: Application to disposition kinetics. Journal of advanced research 6(6): 931-940. https://doi.org/10.1016/j.jare.2014.08.010
Bahrami GH, Mirzaeei SH (2004) Simple and rapid HPLC method for determination of amlodipine in human serum with fluorescence detection and its use in pharmacokinetic studies. Journal of pharmaceutical and b i o m e d i c a l a n a l y s i s 3 6 ( 1 ) : 1 6 3 - 8 . https://doi.org/10.1016/j.jpba.2004.05.016
K a d i o g l u Y, O z t u r k M ( 2 0 1 2 ) Spectrofluorimetric determination of amlodipine in human plasma without derivatization. Brazilian Journal of Pharmaceutical Sciences 48(4):719-725.
Korany MA, Mahgoub H, Haggag RS, Ragab MAA, Elmallah OA (2017) Green Chemistry: Analytical and Chromatography. Journal of Liquid Chromatography & R e l a t e d Te c h n o l o g i e s 4 0 ( 1 6 ) : 8 3 9 - 8 5 2 . https://doi.org/10.1080/10826076.2017.1373672
De Brito WA, Dantas MG, Nogueira FHA, Da Silva-Junior EF, De Arauja- Junior JX, De Aquino TM, EAN Riberio, LG da Silva Solon, CFSAragao,APB Gomes (2017) Development and Validation of HPLC-DAD and UHPLC-DAD Methods for the Simultaneous Determination of Guanylhydrazone Derivatives Employing a Factorial Design. Molecules 22(9): 1-15. https://doi.org/10.3390/molecules22091394
Alsarra IA (2009) High-Performance Liquid Chromatographic Method for Quantitative Determination of Amlodipine in Human Plasma and Pharmaceutical Dosage Form and its Application to Pharmacokinetic Studies. Journal of Chromatographic Science 47(10): 863 – 867. https://doi.org/10.1093/chromsci/47.10.863
Kumar S, Ram B (2019) Analytical method development and validation of Amlodipine besylate in tablet dosage form. Journal of Drug Delivery & T h e r a p e u t i c s 9 ( 4 - A ) : 4 6 3 - 4 6 6 http://dx.doi.org/10.22270/jddt.v9i4-A.3459
International Conference on Harmonization (ICH) (1996) Technical Requirements for the Registration of Pharmaceuticals for Human Use, Validation of Analytical Procedures; Methodology, Q2B, Geneva, 3-9.
United States Pharmacopeia 30, National Formulary 25 (2007) Validation of compendial methods. The United States Pharmacopeial Convention, Inc. Rockville, MD., USA, pp. 1225.
Doran Pauline M. (2013) Unit Operations in Bioprocess Engineering Principles (Second Edition), Academic Press, 445-595. https://doi.org/10.1016/B978-0- 12-220851-5.00011-3
El-Gizawy SM, Abdelmageed OH, Omar MA, Deryea SM, Abdel-Megied AM (2012). Development and Validation of HPLC Method for Simultaneous D e t e r m i n a t i o n o f A m l o d i p i n e , Va l s a r t a n , Hydrochlorothiazide in Dosage Form and Spiked Human Plasma. American Journal of Analytical Chemistry 3(6): 422-430 http://dx.doi.org/10.4236/ajac.2012.36055
United States Pharmacopeia and National Formulary (2014). Assay of amlodipine besylate in USP and NF official monographs, The United States Pharmacopeial convention, Rockville, MD, USP 37, NF 32, 2:1761-1762.
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