Fevziye Ö. ŞİMŞEK, Mustafa Sinan KAYNAK, Nurullah ŞANLI, Selma ŞAHİN

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

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.

Anahtar Kelimeler:

Amlodipin, Furosemid, HPLC, İlaç Analizleri

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

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,

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