KİSTİK FİBROZİS İLAÇLARI TEZACAFTOR VE IVACAFTOR'UN EŞ ZAMANLI KANTİFİKASYONU İÇİN RPHPLC-PDA YÖNTEMİNDE STRATEJİK QBD UYGULAMASI

Öz

Amaç: Kistik fibrozis tedavisinde kullanılan Tezacaftor ve Ivacaftor ilaçlarının eş zamanlı ölçümü için Tasarıma Göre Kalite (QbD) yaklaşımı kullanılarak yeni bir RP-HPLC PDA yöntemi geliştirildi. Gereç ve Yöntem: Optimizasyon, metanolün mobil faz oranı, tamponun pH'ı ve akış hızının faktör olarak seçilmesi ve tepkilerin, yani alıkonma süresi ve kuyruk faktörünün değerlendirilmesi yoluyla Merkezi Bileşik Tasarım ile gerçekleştirildi. Bu teknik, bir foto diyot dizisi dedektörü ile donatılmış bir Waters modülü ile birlikte bir Inertial ODS C18 kolonundan (250 x 4,6 mm, 5 μm parçacık boyutunda) yararlanır. 1.0 ml/dak'lık bir akış hızı, 45:55 oranında metanol ve tampondan oluşan bir mobil faz ve 210 nm'lik bir tespit dalga boyunu içeren kromatografik koşullar, Tezacaftor ve Ivacaftor'u etkili bir şekilde ayırmak için iyi bir şekilde tasarlanmıştır. Sonuç ve Tartışma: Yöntem, ivacaftor için %99.69 ve tezacaftor için %100.06'lık ortalama geri kazanımlarla dikkate değer bir doğruluk gösterdi. Sistem uygunluğu, yöntem kesinliği ve ara kesinlik için % miktar tayini sonuçları sürekli olarak %99.91 ile %100.37 aralığında tespit edildi. Doğrusallık verileri, hem Tezacaftor hem de Ivacaftor için bir korelasyon katsayısı değeri gösterdi. Tezacaftor ve Ivacaftor için LOD ve LOQ değerleri sırasıyla 0.56, 0.57, 1.69 ve 1.74 olarak belirlendi. Doğrulama parametrelerinden elde edilen sonuçlar, QbD yaklaşımı kullanılarak geliştirilen bu RP-HPLC yönteminin sağlam ve güvenilir olduğunu göstermektedir. Rutin analizler için bu teknik değerli bir araç olarak hizmet eder ve kistik fibroz tedavisi alanında biyoanalitik ve biyoeşdeğerlik araştırmalarında önemli bir rol oynar. Bu yöntem, kombinasyon tablet formülasyonlarında Tezacaftor ve Ivacaftor'un kesin ve doğru miktarının belirlenmesini sağlar.

Anahtar Kelimeler

• Merkezi kompozit tasarım
• ivacaftor
• RP-HPLC
• tezacaftor
• yöntem geliştirme
• yöntem validasyonu

STRATEGIC QBD IMPLEMENTATION IN RPHPLC-PDA METHOD FOR SIMULTANEOUS QUANTIFICATION OF CYSTIC FIBROSIS DRUGS TEZACAFTOR AND IVACAFTOR

Öz

Objective: A novel RP-HPLC PDA method was developed using a Quality by Design (QbD) approach for the simultaneous quantification of Tezacaftor and Ivacaftor medications employed in the management of cystic fibrosis. Material and Method: Optimization was performed by Central Composite Design by selecting mobile phase ratio of methanol, pH of buffer and flow rate as factors and evaluating responses namely retention time and tailing factor. This technique makes use of an Inertial ODS C18 column (250 x 4.6 mm, 5 μm particle size) in conjunction with a Waters module fitted with a photo diode array detector. The chromatographic conditions including a flow rate of 1.0 ml/min, a mobile phase composed of methanol and buffer in a 45:55 ratio, and a detection wavelength of 210 nm, were thoughtfully designed to effectively separate Tezacaftor and Ivacaftor. Result and Discussion: The method demonstrated remarkable accuracy, with average recoveries of 99.69% for ivacaftor and 100.06% for tezacaftor. The % assay results for system suitability, method precision, and intermediate precision consistently fell within the range of 99.91% to 100.37%. Linearity data exhibited correlation coefficient values of one for both Tezacaftor and Ivacaftor. The LOD and LOQ values for Tezacaftor and Ivacaftor were determined to be 0.56, 0.57, 1.69, and 1.74, respectively. The results obtained from the validation parameters demonstrate that this RP-HPLC method, developed using the QbD approach, is robust and dependable. It serves as a valuable tool for routine analysis and plays a pivotal role in bioanalytical and bioequivalence research within the realm of cystic fibrosis treatment. This method ensures precise and accurate quantification of Tezacaftor and Ivacaftor in combination tablet formulations.

Anahtar Kelimeler

• Central composite design
• ivacaftor
• method development
• method validation
• RP-HPLC
• tezacaftor

Kaynakça

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