Publication: Nilotinib’in proses ve bozunma safsızlıklarının belirlenmesi için stabilite göstergeli bir HPLC yönteminin geliştirilmesi ve metot validasyonu
Abstract
Bu çalışmada, nilotinib etken maddesi proses ve bozunma safsızlıklarının bir arada analizi için stabilite göstergeli HPLC yöntemi geliştirildi. Bu yöntemde YMC-Pack ODS-AQ, 150 mm x 4,6 mm, 3,0 µm kolon kullanılmıştır. Potasyum dihidrojen fosfat tamponu ve asetonitril hareketli fazı, gradient elüsyon ile 1,0 mL/ dk akış hızında ayarlanarak maddelerin ayrılması gerçekleştirilmiş ve deteksiyon dalga boyu 265 nm’ye ayarlanmıştır. Çözücü olarak saf su ve etanol 50:50 (h:h) karışımı kullanılmıştır. Yöntemin analitik yöntem validasyonu yapıldı. Seçicilik, doğrusallık, kesinlik, doğruluk, sağlamlık, çözelti stabilitesi, tanıma sınırı ve tayin sınırı parametreleri incelendi. Nilotinib etken maddesi ve bozunma safsızlıkları için yapılan doğrusallık çalışmasında 0,997’den büyük korelasyon katsayısı elde edildi. Yöntem kesinliği için aynı gün içinde yapılan analizlerde 0,63 ile 1,41, farklı günlerde yapılan analizlerde 0,67 ile 1,39 arasında değişen %RSD değerleri elde edildi. Yöntem doğruluğu için Nilotinib için ortalama 100,75 ve bozunma safsızlıkları için ise 87,66 ile 99,26 arasında değişen %geri kazanım elde edildi. Tespit limit sınırı 2,4 – 3,0 ng/ mL arasında; hesaplanabilirlik limiti 8,0 – 10,0 ng/ mL arasında bulundu. Çözeltilerin 15 °C ’de 48 saat stabil olduğu stabil olduğu görüldü. Yöntemin sağlamlığı için bazı çalışmalar yapıldı ve kromatografik koşullardaki değişikliklerin yöntem performansını etkilemediği görüldü. İlaç kapsülü için zorla bozundurma çalışmaları (asidik, bazik, oksidatif, ışık ve ısı) yapıldı. Kütle denklikleri yapılarak %95,43 -%101,40 arasında değişen verim elde edildi.
In this study, the HPLC method with stability indicator was developed for the combined analysis of nilotinib active ingredient, process and degradation impurities. In this method, YMC-Pack ODS-AQ, 150 mm x 4.6 mm, 3.0 µm column was used. The separation of substances was achieved by adjusting the potassium dihydrogen phosphate buffer and acetonitrile mobile phase at a flow rate of 1.0 ml/ min by gradient elution, and the detection wavelength was adjusted to 265 nm. A 50:50 (v:v) mixture of distilled water and ethanol was used as the solvent. Analytical method validation of the method was performed. Selectivity, linearity, precision, accuracy, robustness, solution stability, limit of dedection and limit quantitation parameters were examined. A correlation coefficient greater than 0.997 was obtained in the linearity study for the active ingredient nilotinib and its degradation impurities. For the precision of the method, % RSD values ranging from 0.63 to 1.41 were obtained in the analyzes performed on the same day, and from 0.67 to 1.39 in the analyzes performed on different days. An average recovery rate of 100.75% was obtained for Nilotinib for method accuracy and between 87.66 and 99.26% for degradation impurities. The limit of dedection was found between 2.4 – 3.0 ng/ mL; the limit of quantitation was found between 8.0 – 10.0 ng/ mL. The solutions were found to be stable for 48 hours at 15 °C. Some studies were carried out for the robustness of the method and it was seen that changes in chromatographic conditions did not affect the method performance. Forced degradation studies (acidic, basic, oxidative, light and heat) were performed for the drug capsule. By making mass equivalences, efficiency ranging between 95.43% -101.40% was obtained.
In this study, the HPLC method with stability indicator was developed for the combined analysis of nilotinib active ingredient, process and degradation impurities. In this method, YMC-Pack ODS-AQ, 150 mm x 4.6 mm, 3.0 µm column was used. The separation of substances was achieved by adjusting the potassium dihydrogen phosphate buffer and acetonitrile mobile phase at a flow rate of 1.0 ml/ min by gradient elution, and the detection wavelength was adjusted to 265 nm. A 50:50 (v:v) mixture of distilled water and ethanol was used as the solvent. Analytical method validation of the method was performed. Selectivity, linearity, precision, accuracy, robustness, solution stability, limit of dedection and limit quantitation parameters were examined. A correlation coefficient greater than 0.997 was obtained in the linearity study for the active ingredient nilotinib and its degradation impurities. For the precision of the method, % RSD values ranging from 0.63 to 1.41 were obtained in the analyzes performed on the same day, and from 0.67 to 1.39 in the analyzes performed on different days. An average recovery rate of 100.75% was obtained for Nilotinib for method accuracy and between 87.66 and 99.26% for degradation impurities. The limit of dedection was found between 2.4 – 3.0 ng/ mL; the limit of quantitation was found between 8.0 – 10.0 ng/ mL. The solutions were found to be stable for 48 hours at 15 °C. Some studies were carried out for the robustness of the method and it was seen that changes in chromatographic conditions did not affect the method performance. Forced degradation studies (acidic, basic, oxidative, light and heat) were performed for the drug capsule. By making mass equivalences, efficiency ranging between 95.43% -101.40% was obtained.