Linagliptin'in İç Standart Olarak Kullanıldığı Analitik Standart Çözeltilerde Vildagliptin Miktar Tayini: LC-MS/MS Yönteminin Geliştirilmesi ve Validasyonu

Öz

Bu çalışmada, linagliptin’in iç standart olarak kullanıldığı, analitik standart çözeltilerde vildagliptin tayinine yönelik bir LC–MS/MS yöntemi geliştirilmiş ve değerlendirilmiştir. Kalibrasyon ve kalite kontrol (QC) çözeltilerinin hazırlanmasında asetonitril içerisinde çözülmüş referans materyaller kullanılmış ve yardımcı madde kaynaklı girişim açısından bağımsız olarak hazırlanan altı plasebo numunesi incelenmiştir. Kromatografik ayırım, 0,3 mL dk⁻¹ akış hızında ve gradyan elüsyon koşullarında çalıştırılan bir ACE C18 kolon (50 mm × 2,1 mm, 5 µm) kullanılarak gerçekleştirilmiş; sulu ve organik mobil faz bileşenleri sırasıyla 10 mM amonyum asetat ve asetonitril’den oluşmuş ve her iki bileşene de %0,01 formik asit eklenmiştir. Deteksiyon için pozitif iyon elektrosprey ve çoklu reaksiyon izleme (MRM) teknikleri kullanılmıştır. Valide edilen 0,125–4,0 ng mL⁻¹ aralığı boyunca, kalibrasyon modeli R² = 0,9951 değerini vermiştir. Enjekte edilen çözeltilerdeki enstrümantal LOD ve LOQ değerleri sırasıyla 0,028 ve 0,084 ng mL⁻¹ olarak bulunmuştur. En yüksek gün içi veya günler arası kesinlik değeri %2,15 RSD olarak hesaplanmıştır. Üç farklı QC seviyesinde ortalama geri kazanım %99,12–101,7 ve geri kazanım RSD'si %2,00–3,29 (n = 6) olarak belirlenmiştir. Her bir kromatografik döngü 7,5 dakika sürmüştür. İyon baskılanması ve artışı kantitatif olarak ölçülmemiş olmasına rağmen, plasebo preparatlarında vildagliptin MRM piki ile girişime yol açan herhangi bir sinyal saptanmamıştır. Bu validasyon hazırlanan analitik çözeltiler için geçerli olup, dozaj formlarındaki veya biyolojik matrislerdeki performansı teşkil etmemektedir

Anahtar Kelimeler

• Vildagliptin
• LC-MS/MS
• linagliptin
• analitik yöntem validasyonu
• miktar tayini

Development and Validation of an LC–MS/MS Method for the Quantitative Determination of Vildagliptin in Analytical Standard Solutions Using Linagliptin as an Internal Standard

Öz

An LC–MS/MS procedure was developed and evaluated for measuring vildagliptin in analytical standard solutions, with linagliptin serving as the internal standard. Reference materials dissolved in acetonitrile were used to produce the calibration and quality-control (QC) solutions, and six independently prepared placebo samples were examined for excipient-related interference. An ACE C18 column (50 mm × 2.1 mm, 5 µm) was operated under gradient conditions at 0.3 mL min⁻¹ with 10 mM ammonium acetate and acetonitrile as the aqueous and organic components; each contained 0.01% formic acid. Positive-ion electrospray and multiple-reaction monitoring were used for detection. Across the validated interval of 0.125–4.0 ng mL⁻¹, the calibration model gave R² = 0.9951. Instrumental LOD and LOQ values in the injected solutions were 0.028 and 0.084 ng mL⁻¹, respectively. The highest intra-day or inter-day precision value was 2.15% RSD. At the three QC levels, mean recovery was 99.12–101.7%, and recovery RSD was 2.00–3.29% (n = 6). Each chromatographic cycle lasted 7.5 min. No interfering vildagliptin MRM peak was detected in the placebo preparations, although ion suppression and enhancement were not measured quantitatively. The validation applies to prepared analytical solutions and does not establish performance in dosage forms or biological matrices.

Anahtar Kelimeler

• Vildagliptin
• LC-MS/MS
• linagliptin
• analytical method validation
• quantitative determination

Kaynakça

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