Farmasötik Ürünlerde Empagliflozin Tahmini İçin Yeşil bir HPLC Yönteminin Geliştirilmesi ve Validasyonu

Öz

Bu çalışmada, empagliflozinin farmasötik ürünlerdeki miktar tayini için iki farklı kromatografik yöntem geliştirilmiştir. Bunlardan ilki mobil fazda organik değiştirici olarak asetonitril kullanılan klasik yöntem, ikincisi ise mobil fazda organik değiştirici olarak etanol kullanılan yeşil yöntemdir. Klasik yöntemde Extend C18 kolon (250 x 4,6 mm, 5 µm) kullanılmış ve sıcaklık 30 °C'de sabit tutulmuştur. Mobil faz formik asit çözeltisi (ultra saf suda %0,1) ve asetonitril (55/45, v/v) idi ve izokratik elüsyon uygulandı. Mobil fazın akış hızı 1,0 mL dk-1 ve enjeksiyon hacmi 10 µL idi. Tespit 223 nm'de bir UV dedektörü kullanılarak gerçekleştirilmiştir. Yeşil yöntemde organik değiştirici olarak etanol kullanılmıştır. Bu yöntemler arasındaki tek fark organik değiştiriciydi. Yöntemlerin diğer tüm koşulları aynıydı. Her iki kromatografik yöntem de seçicilik, doğrusallık, doğruluk, kesinlik, saptama ve miktar belirleme limiti ve sağlamlık gibi çeşitli parametreler açısından ICH kılavuzlarına göre doğrulanmıştır. Kromatografik yöntemlerin tayin katsayıları 5-30 mg mL-1 gliklazid konsantrasyon aralığında 0.999'dan büyüktü. Geliştirilen kromatografik yöntemler farmasötik formülasyonlara uygulanmıştır. Elde edilen sonuçların ortalamalar açısından karşılaştırmaları Student (t) testi, standart sapmalar açısından karşılaştırmaları ise Fischer (F) testi kullanılarak yapılmıştır. Bu yöntemler arasında anlamlı bir fark bulunmamıştır. Her iki yöntemin çevresel etkisi AGREE ve GAPI yazılımları kullanılarak değerlendirilmiş ve sürdürülebilirlikleri teyit edilmiştir. Bu doğrulanmış metotlar, tablet formülasyonlarında EPG'nin kantitatif analizi için güvenilir ve çevre dostu yaklaşımlar sunarak farmasötik analizlerde daha güvenli ve sürdürülebilir laboratuvar uygulamalarını desteklemektedir. Farmasötik ürünlerdeki EPG'nin kantitatif analizi için yeni yaklaşım, şu anda kullanılan yöntemlere ekonomik ve ekolojik açıdan sorumlu bir alternatif olarak görülebilir.

Anahtar Kelimeler

• Yeşil analitik metod
• empagliflozin
• kromatografik metod

Development and Validation of A Green HPLC Method for Estimation of Empagliflozin in Pharmaceutical Products

Abstract

In this study, two different chromatographic methods were developed for the quantification of empagliflozin in pharmaceutical products. The first was the classical method using acetonitrile as an organic modifier in the mobile phase and the second was the green method using ethanol as an organic modifier in the mobile phase. In the classical method, an Extend C18 column (250 x 4.6 mm, 5 µm) was used and the temperature was kept constant at 30 °C. The mobile phase was the formic acid solution (0.1% in ultrapure water) and acetonitrile (55/45, v/v), and isocratic elution was applied. The flow rate of the mobile phase was 1.0 mL min-1 and the injection volume was 10 µL. Detection was performed using a UV detector at 223 nm. In the green method, ethanol was used as an organic modifier. The only difference between these methods was the organic modifier. All other conditions of the methods were identical. Both chromatographic methods were validated according to ICH guidelines for various parameters such as selectivity, linearity, accuracy, precision, limit of detection and quantification, and robustness. The coefficients of determination of the chromatographic methods were greater than 0.999 in the concentration range of 5-30 mg mL-1 glyclazide. The developed chromatographic methods were applied to pharmaceutical formulations. Comparisons of the results obtained in terms of means were made using the Student (t) test and in terms of standard deviations using the Fischer (F) test. There was no significant difference between these methods. The environmental impact of both methods was evaluated using AGREE and GAPI software, confirming their sustainability. These validated methods provide reliable and environmentally friendly approaches for the quantitative analysis of EPG in tablet formulations, supporting safer and more sustainable laboratory practices in pharmaceutical analysis. For the quantitative analysis of EPG in pharmaceutical products, the new approach can be viewed as an affordable and ecologically responsible alternative to the methods now in use.

Keywords

• Green analytical method
• empagliflozin
• chromatographic method

Kaynakça

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