İnsan İdrarında Valasiklovirin Tayini için RPLC Yöntemin Geliştirilmesi ve Validasyonu

Sunulan çalışmada, herpes ve megalovirüslerin sebep olduğu enfeksiyonların tedavisinde tercihli olarak kullanılan ilaçlardan valasiklovirin katkılandırılmış idrarda ters faz sıvı kromatografik yöntemle tayini için yöntem geliştirilmiştir. Yöntem geliştirmede, valasiklovirin sistematik olarak pH-kapasite faktörü ilişkisini değerlendirilip, mobil faz optimizasyonu gerçekleştirilmiştir. İnceleme sonunda, optimum ayırma koşulu %4 asetonitril-su (h/h), pH 5,0 ve 37oC kolon sıcaklığı olarak belirlenmiştir. Sonrasında, belirlenen optimum koşulun Uluslararası Harmonizasyon Topluluğu (ICH) yönergelerine göre validasyonu gerçekleştirilmiştir. Geliştirilen yöntem 2-12 μg/mL derişim aralığında muhteşem doğrusallık göstermiştir ve idrar ortamında gerçekleştirilen geri kazanma sonuçları (%) valasiklovirin iki farklı derişimi için 100,147±0,800 ve 100,208±0,604 olarak bulunmuştur. Ayrıca, geliştirilen yönteme sağlamlık testleri de uygulanmıştır ve elde edilen sonuçlar t-testiyle değerlendirilmiştir. Elde edilen validasyon ve sağlamlık testleri sonucunda, geliştirilen yöntemin hassas, tekrar edilebilir, doğru, kesin ve sağlam olduğu bulunmuştur. Bundan dolayı, geliştirilen yöntemin rutin analizler için uygun olduğu belirlenmiştir.

Anahtar Kelimeler:

Antiherpes, valasiklovir, metot geliştirme, pKa, RPLC

Development and Validation of the RPLC Method for the Determination of Valaciclovir in Human Urine

In the present study, a method has been developed for the determination of valaciclovir, one of the preferred drugs for the treatment of infections caused by herpes and megaloviruses, in spiked urine by reversed-phase liquid chromatographic method. In developing the method, the pH-capacity factor relationship of valaciclovir was systematically evaluated and mobile phase optimization was performed. At the end of the examination, the optimum separation condition was determined as 4% acetonitrile-water (v/v), pH 5.0, and 37oC column temperature. Subsequently, the determined optimum condition was validated according to the guidelines of the International Conference on Harmonization (ICH). The developed method showed excellent linearity in the concentration range of 2-12 μg/mL, and the recovery results (%) performed in urine were found to be 100.147±0.800 and 100.208±0.604 for two different concentrations of valaciclovir. In addition, robustness tests were also performed for the developed method and the results were evaluated using the t-test. The results of the validation and robustness tests showed that the developed method is sensitive, reproducible, accurate, precise and robust. Therefore, the developed method was found to be suitable for routine analysis.

Keywords:

Antiherpes, Valaciclovir, method development, pKa, RPLC,

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