İnsan İdrarında Bazı Tetrasiklinlerin Analizine Yönelik Yeşil RPLC Yaklaşımı

Abstract

Gerçekleştirilen çalışmada geniş etki spektrumuna sahip tetrasiklin grubu bileşiklerden 4-epitetrasiklin, 4-epioksitetrasiklin ve tetrasiklinin katkılandırılmış insan idrarında tayini için organik çözücü olarak asetonitril, metanol gibi toksik organik çözücülerin yerine toksik olmayan etanolün kullanıldığı yeşil bir RPLC yöntem geliştirilmiştir. Sıvı kromatografik yöntem geliştirmede pH 2,5-7.0 aralığında bileşiklerin sıvı kromatografik davranışları sistematik olarak incelenmiş ve %10 (h/h) etanol içeren pH'sı 3,0 olan ve 37 oC kolon sıcaklığına sahip koşul optimum koşul olarak belirlenmiştir. Sonrasında geliştirilen yöntemin ICH yönergelerine göre validasyonu ve sağlamlık testleri gerçekleştirilmiştir. Validasyon ve sağlamlık testleri sonucunda geliştirilen yöntemin rutin analizlerinde kullanım için uygun olduğu belirlenmiştir. Geliştirilen yöntem 0,5-3,0 µg/mL aralığında doğrusaldır ve çalışılan bileşiklerin katkılandırılmış insan idrarından geri kazanım sonuçları %100'e yakın olarak bulunmuştur. Ayrıca bu çalışmada geliştirilen yöntemin yeşilliği Analitik Yeşillik Hesaplayıcı (AGREE) ve Yeşil Analitik Prosedür İndeksi (GAPI) programları kullanılarak değerlendirilmiştir.

Keywords

• Tetrasiklinler
• yeşil RPLC
• metot geliştirme
• pKa

A Green RPLC Approach to Some Tetracyclines Analysis in Human Urine

Abstract

In the study conducted, an environmentally friendly RPLC method using non-toxic ethanol instead of toxic organic solvents such as acetonitrile and methanol organic solvents was developed for the determination of 4-epitetracycline, 4-epioxytetracycline, and tetracycline, which are compounds of the tetracycline group with a broad spectrum of activity, in spiked human urine. In developing the liquid chromatographic method for the compounds studied, their liquid chromatographic behavior in the pH range of 2.5-7.0 was systematically investigated and the condition with 10% (v/v) ethanol in its structure, pH 3.0, and a column temperature of 37 oC was determined to be the optimum condition. Validation and robustness tests of the developed method were then carried out by the ICH guidelines. As a result of the validation and robustness tests, it was determined that the developed method is suitable for use in routine analyses. The developed method is linear in the range of 0.5-3.0 µg.mL-1 and the recovery results of the investigated compounds from spiked human urine were close to 100%. In addition, the greenness of the method developed in this study was evaluated using the Analytical Greenness Metric Approach (AGREE) and Green Analytical Procedures Index (GAPI) programs.

Keywords

• Tetracyclines
• green RPLC
• method development
• pKa

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