Amlodipin ve Furosemidin Ticari Tablet Dozaj Şekillerinden Eş Zamanlı Tayini İçin Valide Edilmiş Bir RP-HPLC Metodun Geliştirilmesi

Year 2012, Issue: 2, 145 - 158, 01.06.2012

Fevziye Ö. Şimşek Mustafa Sinan Kaynak Nurullah Şanlı Selma Şahin

Abstract

Bu çalışmanın amacı, tablet dozaj formlarından amlodipin AML ve furosemidin FSM eş zamanlı tayini için, hızlı, güvenilir ve valide edilmiş yeni bir HPLC metodunun geliştirilmesidir. Bu ilaçların ayrımı ve miktar tayini için, analiz sırasında 45°C’ye ısıtılan bir C18 kolon FortisTM 250x4.60 mm 5 μm kullanılmıştır. Mobil faz, su 15 mM o-fosforik asit, pH 5.0 ve asetonitril 50:50 h/h ’den oluşmaktadır. Analizler 1.0 mL.dk-1 akış hızında, UV dedektör 238 nm’ye ayarlanarak yapılmıştır. Her bir analizin toplam süresi yaklaşık 5 dakika ve injeksiyon hacmi 20 μL’dir. Geliştirilen metod ICH kılavuzuna göre valide edilmiştir. Önerilen metodun uygulaması için, ticari olarak bulunabilen dört farklı AML 1 referans, Norvasc üç jenerik, Dilapin, Norvasc, Penvasc içeren tablet ve iki farklı FSM bir referans Lasix, bir jenerik Desal içeren tablet piyasadan sağlanmış ve ilaç içerikleri tayin edilmiştir. Belirlenen kromatografik koşullar altında, AML ve FSM sırasıyla 4.28. ve 3.68. dakikalarda ayrılmışlardır. Metod AML ve FSM için sırasıyla 1.0-16.0 μg.mL-1 ve 0.1-12.0 μg.mL-1 konsantrasyon aralığında 0.999 korelasyon katsayısı ile doğrusal bulunmuştur. AML için tayin alt sınırı LOD ve ölçülebilir alt sınır LOQ 0.642 μg.mL-1 ve 2.139 μg.mL-1 ve FSM için tayin alt sınırı LOD ve ölçülebilir alt sınır LOQ 0.010 μg.mL-1 ve 0.031 μg.mL-1’dir. Belirlenen koşullar, FDA tarafından kantitatif kromatografik metodlar için önerilen system uygunluk limitlerine tamamen uymaktadır. Metod, bu ilaçların ticari preparatlarından tayini için başarıyla uygulanmıştır.

Keywords

Amlodipin, Furosemid, HPLC, İlaç Analizleri

Determination of Amlodipine and Furosemide with Newly Developed and Validated RP-HPLC Method in Commercially Available Tablet Dosage Forms

Abstract

The aim of this study was to develop a new, fast, reliable and validated RP-HPLC method for the simultaneous determination of amlodipine and furosemide in tablet dosage forms. A C18 column Fortis™ 250x4.60 mm 5 μm which was heated at 45oC during the analysis, was used for the separation and quantification of these drugs. The mobile phase consisted of water 15 mM o-phosphoric acid, pH 5.0 and acetonitrile 50:50 v/v . Analyses were run at a flow rate 1.0 mL.min-1 and UV detector was set at 238 nm. The injection volume was 20 μL and total run time for an assay was approximately 5 min. The developed method was validated according to the ICH guideline. For the application of the proposed RP-HPLC method, commercially available four different AML containing tablets one reference Norvasc and three generic Dilapin, Monovasc, Penvasc tablets and commercially available two different FSM containing tablets one reference Lasix and one generic Desal tablet were obtained from the market and analyzed for their drug content. Under the given chromatographic conditions, AML and FSM were eluted at 4.28 and 3.68 min., respectively. The method was linear in the concentration range of 1.0 to 16.0 μg.mL-1 and 0.1 to 12.0 μg.mL-1 for AML and FSM, respectively with a correlation coefficient >0.999. LOD and LOQ were 0.642 μg.mL-1 and 2.139 μg.mL-1 for AML 0.010 μg.mL-1 and 0.031 μg.mL-1 for FSM, respectively. Under the conditions used, the analysis completely fulfilled the system suitability test limits suggested by FDA for the quantitative chromatographic methods. The method was successfully applied for the analysis of these drugs in commercially available tablets.

Keywords

Amlodipine, Furosemide, HPLC, Drug Analysis

References

• Martindale “The Extrapharmacopoeia”, 35th Ed., Sweetman, S.C., (Eds), London, U.K., Pharmaceutical Press, (2007).
• Physician Desk Reference (PDR), 61st Ed., Sanborn, K.D., (Eds), New Jersey, USA, Med- ical Economics Co., Montvale, (2007).
• Goodman and Gilman’s, “The Pharmacological Basis of Therapeutics”, 11th Ed., Brun- ton, L.L., (Eds), New York, USA, McGraw-Hill Medical Publications Division, (2006).
• Levine, C.B., Fahrbach, K.R., Frame, D., Connelly, J.E., Estok, R.P., Stone, L.R., Luden- sky, V.: Effect of amlodipine on systolic blood pressure. Clin Ther, 25(1), 35 (2003)
• SevillanoCabeza, A., CampinsFalco, P., SerradorGarcia, M.C.: Extractive-spectrophoto- metric determination of furosemide with sodium 1,2-naphthoquinone-4-sulphonate in pharmaceutical formulations. Anal Lett, 30(1), 91 (1997)
• Okuda, T., Yamashita, K., Motohashi, M.: High-performance liquid chromatography using on-line solid-phase extraction: determination of furosemide in human serum. J Chromatogr B Biomed Appl, 682(2), 343 (1996)
• Abou-Auda, H.S., Al-Yamani, M.J., Morad, A.M., Bawazir, S.A., Khan, S.Z., Al-Khamis, K.I.: High-performance liquid chromatographic determination of furosemide in plasma and urine and its use in bioavailability studies. J Chromatogr B Biomed Sci Appl, 710(1-2), 121 (1998)
• Radeck, W., Heller, M.: Improved method for the determination of furosemide in plasma by high-performance liquid chromatography. J Chromatogr, 497, 367 (1989)
• Uchino, K., Isozaki, S., Saitoh, Y., Nakagawa, F., Tamura, Z., Tanaka, N.: Quantitative determination of furosemide in plasma, plasma water, urine and ascites fluid by high- performance liquid chromatography. J Chromatogr, 308, 241 (1984)
• Abdel-Hamid, M.E.: High-performance liquid chromatography-mass spectrometric analysis of furosemide in plasma and its use in pharmacokinetic studies. Farmaco, 55(6-7), 448 (2000)
• Fakhari, A.R., Nojavan, S., Haghgoo, S., Mohammadi, A.: Development of a stability- indicating CE assay for the determination of amlodipine enantiomers in commercial tablets. Electrophoresis, 29(22), 4583 (2008)
• Almani, F., Rind, F.M.A., Memon, A.H., Mughal, U.R., Laghari, M.G.H., Memon, N., Maheshwari, M.L., Khuhawar, M.Y.: Spectrophotometric Determination of Amlodipine Besylate Using 2-Hydroxynaphthaldehyde as a Derivatizing Reagent. Asian J Chem, 22(2), 1205 (2010)
• Kul, D., Dogan-Topal, B., Kutucu, T., Uslu, B., Ozkan, S.A.: High-Performance Liquid Chromatographic and First Derivative of the Ratio Spectrophotometric Determination of Amlodipine and Valsartan in Their Binary Mixtures. J AOAC Int, 93(3), 882 (2010)
• Çelebier, M., Kaynak, M.S., Altınöz, S., Sahin, S.: Validated HPLC method development: The simultaneous analysis of amlodipine and valsartan in samples for liver perfusion studies. Hacettepe Univ J Fac Pharm, 28(1), 15 (2008)
• Alsarra, I.A.: High-performance liquid chromatographic method for quantitative de- termination of amlodipine in human plasma and pharmaceutical dosage form and its application to pharmacokinetic studies. J Chromatogr Sci, 47(10), 863 (2009)
• Kasawar, G.B., Farooqui, M.N.: Simultaneous determination of amlodipine besylate and benazepril hydrochloride in pharmaceutical dosage form by LC. Anal Sci, 25(12), 1495 (2009)
• Mohammadi, A., Rezanour, N., Ansari Dogaheh, M., Ghorbani Bidkorbeh, F., Hash- em, M., Walker, R.B.: A stability-indicating high performance liquid chromatographic (HPLC) assay for the simultaneous determination of atorvastatin and amlodipine in commercial tablets. J Chromatogr B, 846(1-2), 215 (2007)
• Naidu, K.R., Kale, U.N., Shingare, M.S.: Stability indicating RP-HPLC method for simul- taneous determination of amlodipine and benazepril hydrochloride from their combi- nation drug product. J Pharm Biomed Anal, 39(1-2), 147 (2005)
• Bahrami, G., Mirzaeei, S.: Simple and rapid HPLC method for determination of amlo- dipine in human serum with fluorescence detection and its use in pharmacokinetic studies. J Pharm Biomed Anal, 36(1), 163 (2004)
• Tatar, S., Atmaca, S.: Determination of amlodipine in human plasma by high-perfor- mance liquid chromatography with fluorescence detection. J Chromatogr B, 758(2), 305 (2001)
• Demiralay, E.C., Alsancak, G., Ozkan, S.A.: Determination of pKa values of nonsteroi- dal antiinflammatory drug-oxicams by RP-HPLC and their analysis in pharmaceutical dosage forms. J Sep Sci, 32(17), 2928 (2009)
• Gumustas, M., Sanli, S., Sanli, N., Ozkan, S.A.: Determination of pK(a) values of some antihypertensive drugs by liquid chromatography and simultaneous assay of lercani- dipine and enalapril in their binary mixtures. Talanta, 82(4), 1528 (2010)
• Sanli, N., Sanli, S., Sizir, U., Gumustas, M., Ozkan, S.A.: Determination of pK (a) Val- ues of Cefdinir and Cefixime by LC and Spectrophotometric Methods and Their Analy- sis in Pharmaceutical Dosage Forms. Chromatographia, 73(11-12), 1171 (2011)
• http://www.wedgewoodpetrx.com/items/amlodipine-furosemide-capsule.html. 23.04.2013).
• US Pharmacopeial Convention, USP31 NF26. 2008, Portcity Press: Baltimore, USA.
• Method Validation in Pharmaceutical Analysis: A Guide to Best Practice, 1st Ed., Ermer, J. and Miller, J.H., (Eds), Germany, Wiley-VCH Verlag GmbH & Co, (2005).
• Analytical Method Development and Validation, Swartz, M.E. and Krull, I.S., (Eds), New York, USA, Marcel Dekker, (1997).
• ICH, Q2(R1) Validation of Analytical Procedures: Text and Methodology, International Conference on Harmonisation of Technical Requirements for Registration of Pharma- ceuticals for Human Use (ICH), C.f.m.P.f.H., Editor. 2005, ICH Secretariat: Geneva, Switzerland.
• Development and Validation of Analytical Methods, Riley, C.M. and Rosanske, T.W., (Eds), Amsterdam, The Netherlands, Elsevier, (1996).