Sütteki Sülfonamidlerin Tayini için Yeni bir Yüksek Performanslı Sıvı Kromatografisi Yönteminin Geliştirilmesi ve Valide Edilmesi

Yıl 2024, Cilt: 14 Sayı: 4, 1615 - 1625, 01.12.2024

Tulay Oymak

https://doi.org/10.21597/jist.1523734

Öz

Bu çalışmada sülfonamidler (SAs) grubunda bulunan sülfadiazin (SDZ), sülfamerazin (SMR) ve sülfometaksazol’ün (SMX) süt numunelerinde analizi için hızlı, hassas ve basit bir yüksek performanslı sıvı kromatografi (HPLC) yöntemi geliştirilmiş ve valide edilmiştir. SDZ, SMR ve SMX’in sütten ekstraksiyonunda protein çöktürme yöntemi kullanılmıştır. Protein çöktürme yönteminde çöktürücü reaktif türü ve miktarı optimize edilmiştir. Çalışma matriks etkisinin analizlere olan etkisinin gözlemlenmesi amacıyla tüm validasyon parametreleri süt numuneleri kullanılarak çalışılmıştır. Yöntemin validasyonunda ICH klavuzu uyarınca doğrusallık, sağlamlık, doğruluk ve kesinlik testleri gerçekleştirilmiştir. Süt numunelerinde sülfonamidlerin tayini için geliştirilen yöntemin gözlenebilme sınırları (LOD) SDZ, SMR ve SMX için sırasıyla 0.11, 0.10 ve 0.14 µg/mLve tayin sınırları (LOQ) 0.34, 0.30 ve 0.43 µg/mL’dir. SAs’lerin sütteki ortalama geri kazanımları ≥ %93.0±4.4’tür. Yöntemin gün içi ve günler arası tekrarlanabilirliğini % bağıl standart sapma cinsinden sırasıyla ≤ %6.0 ve ≤% 5.4’dir. Geliştirilen yöntem doğru ve güvenilir olup gıda kontrol ve toksikolojik analiz laboratuvarlarında süt numunelerinin içeriğindeki SAs'lerin rutin analizi için kullanılabilir.

Anahtar Kelimeler

Sülfonamidler, HPLC, Yöntem validasyonu

Development and Validation of a New High Performance Liquid Chromatography Method for the Determination of Sulfonamides in Milk

Öz

In this study, a rapid, sensitive and simple high performance liquid chromatographic (HPLC) method was developed and validated for the analysis of sulfadiazine (SDZ), sulfamerazine (SMR) and sulfamethoxol (SMX) in milk samples which are in the group of sulfonamides (SAs). Protein precipitation method was used in the extraction of SDZ, SMR and SMX from milk. The precipitant reagent type and its amount were optimized in the protein precipitation method. All validation parameters were studied using milk samples in order to observe the effect of the study matrix effect on the analyses. Validation of the method was carried out with linearity, robustness, accuracy and precision in accordance with the ICH guideline. The limits of detection (LOD) of the method developed for the determination of sulfonamides in milk samples were 0.11, 0.10 and 0.14 µg/mL and the limits of quantification (LOQ) were 0.34, 0.30 and 0.43 µg/mL for SDZ, SMR and SMX, respectively. The average recoveries of SAs in milk are ≥ 93.0 ± 4.4%. The intra-day and inter-day reproducibility of the method in terms of % relative standard deviation are ≤ 6 % and ≤5.4%, respectively. The developed method is accurate and reliable and can be used in routine analysis of SAs in milk samples in food control and toxicological analysis laboratories.

Anahtar Kelimeler

Sulfonamides, HPLC, Method validation

Kaynakça

• Arroyo-Manzanares, N., Gámiz-Gracia, L., & García-Campaña, A. M. (2014). Alternative sample treatments for the determination of sulfonamides in milk by HPLC with fluorescence detection. Food Chemistry, 143, 459–464. https://doi.org/10.1016/j.foodchem.2013.08.008
• Chen, Z., Yu, C., Xi, J., Tang, S., Bao, T., & Zhang, J. (2019). A hybrid material prepared by controlled growth of a covalent organic framework on amino-modified MIL-68 for pipette tip solid-phase extraction of sulfonamides prior to their determination by HPLC. Microchimica Acta, 186(6), 393. https://doi.org/10.1007/s00604-019-3513-7
• Chromatography, A. (2014). International Conference on Harmonization, Photostability testing of new drug substances and products, QIB, November 1996. 1(2), 1–4.
• Fu, L., Zhou, H., Miao, E., Lu, S., Jing, S., Hu, Y., … Wu, M. (2019). Functionalization of amino terminated carbon nanotubes with isocyanates for magnetic solid phase extraction of sulfonamides from milk and their subsequent determination by liquid chromatography-high resolution mass spectrometry. Food Chemistry, 289(October 2018), 701–707. https://doi.org/10.1016/j.foodchem.2019.03.097
• Gao, Q., Luo, D., Ding, J., & Feng, Y. Q. (2010). Rapid magnetic solid-phase extraction based on magnetite/silica/poly(methacrylic acid-co-ethylene glycol dimethacrylate) composite microspheres for the determination of sulfonamide in milk samples. Journal of Chromatography A, 1217(35), 5602–5609. https://doi.org/10.1016/j.chroma.2010.06.067
• ICH. (2005). International Conference on Harmonisation of Technical Requirements for Registiration of Pharmacuticals for Human Use. ICH Harmonised Tripartite Guideline. Validation of Analytical Procedures: Text and Methodology ICH Q2(R1). In International Conference on Harmonization Quality Guidelines. Geneva, Switzerland.
• Kim, H. J., Jeong, M. H., Park, H. J., Kim, W. C., & Kim, J. E. (2016). Development of an immunoaffinity chromatography and HPLC-UV method for determination of 16 sulfonamides in feed. Food Chemistry, 196, 1144–1149. https://doi.org/10.1016/j.foodchem.2015.10.014
• Korashy, M. A. R., Gawad, S. A. A., Hassan, N. Y., & Abdelkawy, M. (2022). Solid Phase Extraction and Simultaneous Chromatographic Quantification of some Non-steroidaAnti-inflammatory Drug Residues; an Application in Pharmaceutical Industrial Wastewater Effluent. Brazilian Journal of Pharmaceutical Sciences, 58, 1–14. https://doi.org/10.1590/s2175-97902022e18691
• Li, Y., Wu, X., Li, Z., Zhong, S., Wang, W., Wang, A., & Chen, J. (2015). Fabrication of CoFe2O4-graphene nanocomposite and its application in the magnetic solid phase extraction of sulfonamides from milk samples. Talanta, 144, 1279–1286. https://doi.org/10.1016/j.talanta.2015.08.006
• Mthiyane, Z. L., Makhubela, N., Nyoni, H., Madikizela, L. M., Maseko, B. R., & Ncube, S. (2023). Determination of antibiotics during treatment of hospital wastewater using automated solid-phase extraction followed by UHPLC-MS: occurrence, removal and environmental risks. Environmental Technology (United Kingdom), (May), 1–11. https://doi.org/10.1080/09593330.2023.2209741
• Nasir, A. N. M., Yahaya, N., Zain, N. N. M., Lim, V., Kamaruzaman, S., Saad, B., … Hirota, Y. (2019). Thiol-functionalized magnetic carbon nanotubes for magnetic micro-solid phase extraction of sulfonamide antibiotics from milks and commercial chicken meat products. Food Chemistry, 276(June 2018), 458–466. https://doi.org/10.1016/j.foodchem.2018.10.044
• Pochivalov, A., Cherkashina, K., Shishov, A., & Bulatov, A. (2021). Microextraction of sulfonamides from milk samples based on hydrophobic deep eutectic solvent formation by pH adjusting. Journal of Molecular Liquids, 339, 116827. https://doi.org/10.1016/j.molliq.2021.116827
• Sabir, A. M., Moloy, M., & Bhasin, P. S. (2016). Hplc Method Development and Validation: a Review. International Research Journal of Pharmacy, 4(4), 39–46. https://doi.org/10.7897/2230-8407.04407
• Sarakatsanou, C., Karastogianni, S., & Girousi, S. (2023). Promising Electrode Surfaces, Modified with Nanoparticles, in the Sensitive and Selective Electroanalytical Determination of Antibiotics: A Review. Applied Sciences (Switzerland), 13(9). https://doi.org/10.3390/app13095391
• Shaaban, H., Mostafa, A., Alqarni, A. M., Alsultan, R., shehab, Z. Al, Aljarrash, Z., … Amir, M. (2023). Dispersive liquid-liquid microextraction utilizing menthol-based deep eutectic solvent for simultaneous determination of sulfonamides residues in powdered milk-based infant formulas. Journal of Food Composition and Analysis, 117(December 2022), 105137. https://doi.org/10.1016/j.jfca.2023.105137
• Watkinson, A. J., Murby, E. J., & Costanzo, S. D. (2007). Removal of antibiotics in conventional and advanced wastewater treatment: Implications for environmental discharge and wastewater recycling. Water Research, 41(18), 4164–4176. https://doi.org/10.1016/j.watres.2007.04.005
• Zhao, L., Ji, Y., Sun, P., Deng, J., Wang, H., & Yang, Y. (2019). Effects of individual and combined zinc oxide nanoparticle, norfloxacin, and sulfamethazine contamination on sludge anaerobic digestion. Bioresource Technology, 273(September 2018), 454–461. https://doi.org/10.1016/j.biortech.2018.11.049