Sütte Glukokortikoidlerin Analizi İçin Metot Karşılaştırması

Year 2024, Volume: 35 Issue: 1, 91 - 98, 04.07.2024

Gülay Dudaklı , Handan Aydın Vural

https://doi.org/10.35864/evmd.1470307

Abstract

Betametazon, Deksametazon, Prednizolon ve Metilprednizolon büyükbaş hayvanlarda glukokortikoidlere duyarlı çeşitli akut ve kronik hastalıklarda yaygın olarak kullanılan sentetik glukokortikoidlerdir. Glukokortikoidler sütte kalıntı yapmaları sonucu insanlarda ciddi sağlık sorunlarına yol açabilmektedir. Bu çalışmada sütte glukokortikoidlerin belirlenmesine yönelik hassas metodun seçilebilmesi için sıvı-sıvı ekstraksiyon, katı faz ekstraksiyon ve dispersif katı faz ekstraksiyon metotları karşılaştırılmış ve en hassas metot seçilerek valide edilmiştir. Kromatografik ayırım, Agilent XDB-C18 kolonu (150 mm x 4,6 mm, 5µm ) ile gerçekleştirilmiştir. Hedef analitler Diyot Dizi Dedektörü (Diode Array Detector, DAD) ile bütünleşik Agilent 1100 serisi Yüksek Persformanslı Sıvı Kromatografi ( High Performance Liquid Chromatography, HPLC) ile tanımlanmıştır. Betametazon 0.12, 0.3, 0.45 μg/kg seviyesinde analit eklenmiş örneklerin geri alımı %97 ila % 102 arasında, deksametazon 0.12, 0.3, 0.45 μg/kg seviyesinde analit eklenmiş örneklerin geri alımı %97 ila % 102 arasında, metilprednizolon 0.2, 2, 3 μg/kg seviyesinde analit eklenmiş örneklerin geri alımı %94 ila % 100 arasında, prednizolon 0.6, 6, 9 μg/kg seviyesinde analit eklenmiş örneklerin geri alımı %97 ila % 99 arasında değişmiştir.

Keywords

Betametazon, Deksametazon, Prednizolon, Metilprednizolon, HPLC, süt, DAD, metot karşılaştırma, sıvı sıvı ekstraksiyon, katı faz ekstraksiyon, dispersif katı faz ekstraksiyon

Thanks

Katkılarından ötürü Ekrem ERDOĞAN, Dr.T. Zilhan ÇABUK, Dr.Ali ERKURT ve Dr.Selçuk PEKKAYA'ya teşekkür ederiz.

COMPARISON OF METHODS FOR THE ANALYSIS OF GLUCOCORTICOIDS IN MILK

Abstract

Betamethasone, Dexamethasone, Prednisolone and Methylprednisolone are synthetic glucocorticoids that are widely used in various acute and chronic diseases sensitive to glucocorticoids in cattles. Glucocorticoids can cause serious health problems in humans as a result of residues in milk. In this study, in order to choose the sensitive method for the determination of glucocorticoids in milk, liquid-liquid extraction, solid phase extraction and dispersive solid phase extraction methods were compared and the most sensitive method was selected and validated. Chromatographic separation was performed with Agilent XDB-C18 column (150 mm x 4.6 mm, 5µm). Target analytes were identified by Agilent 1100 series High Performance Liquid Chromatography (HPLC) combined with Diode Array Detector (DAD). The recovery of samples spiked with betamethasone 0.12, 0.3, 0.45 μg/kg ranged from 97% to 102%, the recovery of samples spiked with dexamethasone 0.12, 0.3, 0.45 μg/kg ranged from 97% to 102%, the recovery of samples spiked with methylprednisolone analyte at 0.2, 2, 3μg/kg ranged from 94% to 100%, and the recovery of samples spiked with prednisolone at 0.6, 6, 9μg/kg ranged from 97% to 99%.

Keywords

Bethamethasone, dexamethasone, prednisolone, methylprednisolone, HPLC, milk, DAD, method comparison, liquid liquid extraction, solid phase extraction

References

• Arioli, F., Gamberini, M.C., Pavlovic, R., Di Cesare, F., Draghi, S., Bussei, G., Mungiguerra, F., Casati, A. ve Fidani, M. (2022). Quantification of cortisol and its metabolites in human urine by LC-MSn: applications in clinical diagnosis and anti-doping control. Anal Bioanal Chem, 414, 6841-6853. doi: 10.1007/s00216-022-04249-3
• Buchwald, P. ve Bodor, N. (2004). Soft glucocorticoid design: structural elements and physicochemical parameters determining receptor-binding affinity. Die Pharmazie, 59, 396–404.
• Caretti, F., Gentili, A., Ambrosi, A., Rocca,M.L., Delfini, M., Di Cocco, M.E. ve D’Ascenzo, G. (2010). Residue analysis of glucocorticoids in bovine milk by liquid chromatography–tandem mass spectrometry. Analytical and Bioanalytical Chemistry, 397, 2477-2490.
• Cherlet, M., De Baere,S. ve De Backer, P.(2004) Quantitative determination of dexamethasone in bovine milk by liquid chromatography–atmospheric pressure chemical ionization–tandem mass spectrometry. Journal of Chromatography B, 805, 57-65. doi:https://doi.org/10.1016/j.jchromb.2004.02.015
• Courtheyn D., Le Bizec, B., Brambilla, G., De Brabander, H.F., Cobbaert, E., Van De Wiele, M., Vercammen, J. ve De Wasch K.(2002). Recent developments in the use and abuse of growth promoters. Analytica Chimica Acta, 473, 71-82.
• Comparison Between Reversed Phase C18 and C8 SPE Cartridge. Hawach Scientific. https://www.specartridge.com/comparison-between-reversed-phase-c18-and-c8-spe-cartridge/
• Cun, L., Yinliang, W., Ting, Y. ve Yan, Z. (2010). Rapid simultaneous determination of dexamethasone and betamethasone in milk by liquid chromatography tandem mass spectrometry with isotope dilution. Journal of Chromatography A,1217, 411-414. doi: https://doi.org/10.1016/j.chroma.2009.12.015
• Dirikolu, L. (2013). Glucocorticoids, Mineralocorticoids and Adrenolytic Drugs. Riviere, J. Ve Papich, M.(Ed.), Veterinary Pharmacology And Therapeutics, 9.
• Dolan, J.W. (2017). Back to Basics: The Role of pH in Retention and Selectivity. LCGC International, 35, 22-28.
• Guo,C., Wu, C., Zhang,Z., Tan, S., Chen, S. ve Chen, G. (2024). Simultaneous determination of 58 glucocorticoid residues in milk by ultra-high performance liquid chromatography-tandem mass spectrometry. Journal of Chromatography A, 1719. doi: https://doi.org/10.1016/j.chroma.2024.464734
• Islas, G., Ibarra, I.S., Hernandez, P., Miranda, J.M. ve Cepeda, A.(2017). Dispersive Solid Phase Extraction for the Analysis of Veterinary Drugs Applied to Food Samples: A Review. Hindawi International Journal of Analytical Chemistry, 2017. doi: https://doi.org/10.1155/2017/8215271
• Kyle, P.B. (2017). Toxicology: GCMS. Mass Spectrometry for the Clinical Laboratory, 131-163. doi: https://doi.org/10.1016/B978-0-12-800871-3.00007-9
• Luo, Y., Uboh, C.E., Soma, L.R., Guan, F.Y., Rudy, J.A., ve Tsang, D.S. (2005). Simultaneous analysis of twenty-one glucocorticoids inequine plasma by liquid chromatography/tandem massspectrometry. Rapid Communications in Mass Spectrometry, 19,1245-1256.
• Malone, E.M., Elliott, C., Kennedy, D.G. ve Regan, L. (2010). Screening and Quantitative Confirmatory Method for the Analysis of Glucocorticoids in Bovine Milk Using Liquid Chromatography-Tandem Mass Spectrometry. Journal Of Aoac Internatıonal, 93, 1656-1665. doi: 10.1093/jaoac/93.5.1656
• Mcdonald, M., Granelli, K. ve Sjöberg, P. (2007). Rapid multi-residue method for the quantitative determination and confirmation of glucocorticosteroids in bovine milk using liquid chromatography–electrospray ionization–tandem mass spectrometry. Analytica Chimica Acta, 588, 20-25. doi:https://doi.org/10.1016/j.aca.2007.01.075
• Ötles, S. ve Kartal, C. (2016). Solid-Phase Extraction (Spe): Principles and Applications in Food Samples. Acta Sci. Pol. Technol. Aliment, 15(1), 5-15. doi: 10.17306/J.AFS.2016.1.1
• Schacke, H., Döcke, W.D. ve Asadullah,K. (2002). Mechanisms involved in the side effects of glucocorticoids. Pharmacology & Therapeutics, 96, 23-43.
• Schimmer, B.P. ve Funder, J.W. (2022). Adrenocorticotropic Hormone, Adrenal Steroids, and the Adrenal Cortex. Goodman & Gilman's: The Pharmacological Basis of Therapeutics, 13.
• TC Tarım ve Orman Bakanlığı. (2017). Türk Gıda Kodeksi Hayvansal Gıdalarda Bulunabilecek Farmakolojik Aktif Maddelerin Sınıflandırılması ve Maksimum Kalıntı Limitleri Yönetmeliği. (30000). The European Commission.(2021). Commission Implementing Regulation (EU) 2021/808. https://eur-lex.europa.eu/eli/reg_impl/2021/808/oj ‘den erişilmiştir.
• Tölgyesi, A., Tölgyesi, L., Sharma, V.K., Sohn, M. ve Fekete, J.(2010). Quantitative determination of corticosteroids in bovine milk using mixed-mode polymeric strong cation exchange solid-phase extraction and liquid chromatography–tandem mass spectrometry. Journal Of Pharmaceutical And Biomedical Analysis, 53(4), 919-928. doi: https://doi.org/10.1016/j.jpba.2010.06.026
• Turhan, E., Kabil, E., Dudaklı, G. ve Dirikolu, L. (2018, Mart). A New Approach For The Determination Of Betamethasone And Dexamethasone By Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS). (Poster). International Conference Of Racing Analysts And Veterinarians, Dubai.
• UHPLC Separation of Nine Corticosteroids in Under Four Minutes. Thermo Scientific. https://lcms.cz/labrulez-bucket-strapi-h3hsga3/AB_123_HPLC_Corticosteroids_UHPLC_28_Jan2011_LPN_2729_8e265e33bf/AB123-HPLC-Corticosteroids-UHPLC-28Jan2011-LPN2729.pdf ‘ den erişilmiştir.