STABILITY INDICATING ANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF HPLC FOR PREGABALIN QUANTIFICATION IN BULK AND PHARMACEUTICAL DOSAGE FORMULATIONS

Öz

Objective: To develop and validate a rapid, sensitive, and cost-effective HPLC-UV method for the quantification of pregabalin (PRG) in both bulk drug substance and pharmaceutical dosage formulations, addressing the growing need for robust analytical techniques in pharmaceutical quality control and forensic toxicology. Material and Method: An HPLC-UV method was developed using an Agilent 1100/1200 system equipped with a UV detector and a Merck monolithic silica column (100 mm x 4.6 mm). The mobile phase consisted of acetonitrile and water (5:95, v/v) adjusted to pH 7.0 with orthophosphoric acid and sodium hydroxide, at a flow rate of 2.0 ml/min and a column temperature of 45°C. UV detection was set at 200 nm. The method was validated according to ICH Q2(R1) guidelines, evaluating linearity, sensitivity, accuracy, precision, recovery, and stability-indicating properties through forced degradation studies. Result and Discussion: The developed HPLC-UV method demonstrated rapid analysis with a pregabalin retention time of 1.44 minutes and a total run time under 2 minutes. System suitability parameters met pharmacopeial requirements. The method exhibited excellent linearity in the range of 1 to 150 µg/ml (R² > 0.999), with LOD and LOQ values of 0.25 µg/ml and 0.76 µg/ml, respectively. Intra-day and inter-day accuracy ranged from 96.09% to 99.68% and 95.07% to 98.78%, respectively, with RSD values for precision consistently below 2.22%. Recovery from pharmaceutical formulations was between 99.0% and 101.75%. Forced degradation studies revealed that pregabalin is stable under acidic, oxidative, and thermal stress conditions, as well as during freeze-thaw and autosampler storage, but is susceptible to base hydrolysis. The validated HPLC-UV method is demonstrated to be rapid, accurate, precise, and stability-indicating, making it suitable for routine quality control of pregabalin in pharmaceutical products and for forensic applications.

Anahtar Kelimeler

• Forensic pharmacy
• forensic toxicology
• HPLC-UV
• method development and validation
• pregabalin

PREGABALİN’İN HAM MADDE VE FARMASÖTİK DOZAJ FORMÜLASYONLARINDA MİKTAR TAYİNİ İÇİN STABİLİTE GÖSTERGELİ HPLC YÖNTEMİ GELİŞTİRME VE VALİDASYONU

Öz

Amaç: Farmasötik kalite kontrol ve adli toksikoloji alanlarında güvenilir analitik tekniklere yönelik artan ihtiyacı karşılamak amacıyla, pregabalinin (PRG) hem ham madde hem de farmasötik dozaj formülasyonlarında kantitatif tayini için hızlı, hassas ve uygun maliyetli bir HPLC-UV yöntemi geliştirmek ve valide etmektir. Gereç ve Yöntem: UV dedektörlü Agilent 1100/1200 sistemi ve Merck monolitik silika kolon (100 mm x 4.6 mm) kullanılarak bir HPLC-UV metodu geliştirilmiştir. Mobil faz, asetonitril ve suyun (5:95, v/v) ortofosforik asit ve sodyum hidroksit ile pH 7.0'a ayarlanmış karışımından oluşmaktadır. 2.0 ml/dak. akış hızı ve 45°C kolon sıcaklığı tercih edilmiştir. UV dalgaboyu 200 nm'ye ayarlanmıştır. Metot, ICH Q2(R1) kılavuzlarına göre doğrusallık, hassasiyet, doğruluk, kesinlik, geri kazanım ve zorlu koşullarda bozunma çalışmaları yoluyla stabilite göstergesi özellikler açısından valide edilmiştir. Sonuç ve Tartışma: Geliştirilen HPLC-UV metodu, 1.44 dakikalık pregabalin alıkonma süresi ve 2 dakikanın altında toplam analiz süresi ile hızlı analiz sağlanmıştır. Sistem uygunluk parametreleri farmakope gerekliliklerini karşılamaktadır. Metot, 1 ila 150 µg/ml konsantrasyon aralığında doğrusallık (R² > 0.999) göstermekte ve sırasıyla 0.25 µg/ml ve 0.76 µg/ml LOD ve LOQ değerlerine erişmektedir. Gün içi ve günler arası doğruluk sırasıyla %96.09 ila %99.68 ve %95.07 ila %98.78 arasında değişirken, kesinlik için RSD değerleri sürekli olarak %2.22'nin altında kalmıştır. Farmasötik formülasyonlardan geri kazanım %99.0 ile %101.75 arasında olmuştur. Zorlu koşullarda bozunma çalışmaları, pregabalinin asidik, oksidatif ve termal stres koşullarının yanı sıra dondurma-çözdürme ve otomatik örnekleyici saklama sırasında stabil olduğunu, ancak baz hidrolizine karşı hassas olduğunu ortaya koymuştur. Geliştirilen ve valide edilen HPLC-UV metodu, farmasötik ürünlerdeki pregabalinin rutin kalite kontrolü ve çeşitli adli uygulamalar için uygun olduğu gösterilmiş hızlı, doğru ve kesin bir metottur.

Anahtar Kelimeler

• Adli eczacılık
• adli toksikoloji
• HPLC-UV
• metot geliştirme ve geçerli kılma
• pregabalin

Kaynakça

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