Jelatin bazlı hidrojellerde timolün tayini için stabilite göstergeli yüksek performanslı sıvı kromatografisi yönteminin doğrulanması

Öz

Bu çalışmanın amacı, bal arılarındaki varroosis istilasıyla mücadele için geliştirilen bir jelatin hidrojelin yapısına yüklenen ve bir model salım sistemine salınan timol miktarını belirlemek için analitik bir yöntem geliştirmektir. Kromatografik ayırma, yüksek performanslı sıvı kromatografı, C18 kolon ve asetonitril:su (75:25) hareketli faz kullanılarak, 1 mL/dakika izokratik akış hızıyla gerçekleştirildi. Yöntemi doğrulamak için özgüllük, hassasiyet, doğrusallık, tespit ve ölçüm limiti, doğruluk, sağlamlık, çözelti stabilitesi ve sistem uygunluk parametreleri incelendi. Stres testi için hidrolitik, termal, oksidatif ve fotolitik bozunma çalışmaları yapıldı. Yöntem, 0,25-15 μg/mL (R2 = 0,999) aralığında doğrusaldır. LOD, 6,2261 ng/mL ve LOQ, 18,866 ng/mL olarak hesaplandı. Gün içi ve günler arası hassasiyet çalışması %RSD değerleri ≤ %2 idi. Hidrojel için ortalama geri kazanım %100,3 (±2,12) ve hava örneği için %99,9 (±3,14) idi. Analitik yöntemin özgül, doğrusal, kesin ve sağlam olduğu kanıtlandı. Bu çalışma, jelatin bazlı hidrojel yapısında ve model salım atmosferinde timolün tespiti için hassas, kullanışlı ve pratik bir yöntem sunmaktadır.

Anahtar Kelimeler

• Kontrollü salım
• HPLC
• hidrojel
• timol
• validasyon
• varroosis.

Destekleyen Kurum

T.C. Tarım ve Orman Bakanlığı, Tarımsal Araştırmalar ve Politikalar Genel Müdürlüğü

Proje Numarası

TAGEM/HSGYAD/A/20/A5/P6/1943

Etik Beyan

Bu çalışma Pendik Veteriner Kontrol Enstitüsü Yerel Etik Kurulu tarafından onaylandıktan sonra gerçekleştirilmiştir (karar numarası: 202-17/2018).

Teşekkür

Bu çalışma birinci yazarın “Varroosis Mücadelesi İçin Kontrollü Timol Salım Sisteminin Geliştirilmesi ve Etkinliğinin Araştırılması” (Doktora Tezi, Ankara Üniversitesi, Ankara, Türkiye, 2022) başlıklı doktora tezinden türetilmiştir. T.C. Tarım ve Orman Bakanlığı, Tarımsal Araştırmalar ve Politikalar Genel Müdürlüğü'ne (TAGEM) (proje no: TAGEM/HSGYAD/A/20/A5/P6/1943, Proje Başlangıç-Bitiş tarihi: 01.01.2020-01.01.2022) bu çalışmanın finansal desteğine katkılarından dolayı teşekkür ederiz.

Validation of stability-indicating high-performance liquid chromatography method for the determination of thymol in gelatin-based hydrogels

Öz

The aim of this study was to develop an analytical method to determine the amount of thymol loaded into the structure of a gelatin hydrogel developed to struggle varroosis infestation of honey bees and released into a model release system. Chromatographic separation was achieved using a high-performance liquid chromatograph, C18 column and acetonitrile:water (75:25) mobile phase with an isocratic flow rate of 1 mL/min. To validate the method, specificity, precision, linearity, detection and measurement limit, accuracy, robustness, solution stability and system suitability parameters were studied. Hydrolytic, thermal, oxidative and photolytic degradation studies were performed for stress testing. The method is linear in the range of 0.25-15 μg/mL (R2 = 0.999). LOD was calculated as 6.2261 ng/mL and LOQ as 18.866 ng/mL. Intra-day and inter-day precision study %RSD values were ≤ 2%. The average recovery for the hydrogel was 100.3% (±2.12) and for the air sample was 99.9% (±3.14). The analytical method proved to be specific, linear, precise and robust. This study presents a sensitive, convenient and practical method for the detection of thymol in gelatin-based hydrogel structure and model release atmosphere.

Anahtar Kelimeler

• Controlled release
• HPLC
• hydrogel
• thymol
• validation
• varroosis.

Destekleyen Kurum

T.C. Ministry of Agricultural and Forestry, General Directorate of Agricultural Research and Policies

Proje Numarası

TAGEM/HSGYAD/A/20/A5/P6/1943

Etik Beyan

This study was carried out after were approved by the Local Ethics Committee of Pendik Veterinary Control Institute (decision number: 202-17/2018).

Teşekkür

This study is derived from the first author's PhD thesis titled "Development of a Controlled Timol Release System for Varroosis Struggle and Investigation of its Effectiveness" (PhD Thesis, Ankara University, Ankara, Turkey, 2022). We would like to thank the T.C. Ministry of Agricultural and Forestry, General Directorate of Agricultural Research and Policies (TAGEM) (project no: TAGEM/HSGYAD/A/20/A5/P6/1943, Project Start-End date: 01.01.2020-01.01.2022)) a for their contribution to the financial support for this study.

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