Bazı Azol Grubu Antifungal Bileşiklerin Analizi için Ters Faz Sıvı Kromatografi Metot Optimizasyonu
Yıl 2023,
, 347 - 354, 29.12.2023
Dilara Başat Dereli
,
Ebru Çubuk Demiralay
,
İlkay Konçe
Öz
Bu çalışmada azol türevi antifungal ilaç etken maddeleri klotrimazol, lanokonazol ve klimbazolün ters faz sıvı kromatografi (RPLC) yöntemi kullanılarak geliştirilen metot optimizasyonunda, bileşiklerin kapasite faktörü değerleri ve analizlendikleri hareketli fazdaki metanol derişimi ilişkisi esas alınmıştır. İyonlaşan gruba sahip bu bileşiklerin optimizasyonunda hareketli faz pH değeride optimizasyonda bir diğer etkendir. İzokratik bir ayırma için kapasite faktörü değerlerinin 1-5 aralığında olması tercih edilmektedir.
RPLC yöntemiyle gerçekleştirilen klotrimazol, lanokonazol ve klimbazol bileşiklerinin aynı anda tayini için yapılan optimizasyonda %50, %55 ve %60 (v/v) metanol içeren metanol-su ikili karışımları kullanılmıştır. Ayırma için çalışılan ortamlarda iki hareketli faz pH değerindeki (pH 4,0 ve 5,5) alıkonma zamanları belirlenmiştir. Hidrofobik özellikteki bileşiklerin tayini için Zorbax SB-CN (4,6x150mm) kolon tercih edilmiştir. Kolon sıcaklığı 37oC’de ve akış hızı da 1 mL/dakikada sabitlenmiştir. Optimizasyon koşullarının belirlenmesinde logk- % (v/v) metanol ilişkisi ve bileşiklerin ayrılmalarını gösteren seçicilik faktörü (α) parametresinden yararlanılmıştır.
Destekleyen Kurum
Süleyman Demirel Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi
Proje Numarası
TYL-2022-8833
Teşekkür
Bu çalışma Süleyman Demirel Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi (TYL-2022-8833) tarafından desteklenmiştir.
Kaynakça
- [1] Brunton, L. L., Lazo, J. S., Parker, K. L. 2008. 11th ed.; Goodman & Gilman's The pharmacology basis of therapeutics, McGrawHill eBooks, New York.
- [2] Kadavakollu, S., Stailey, C., Kunapareddy, C. S., White, S. 2014. Clotrimazole as a Cancer Drug: A Short Review. Med Chem, 4(11), 722–724.
- [3] Baghi, N., Shokohi, T., Badali, H., Makimura, K., Rezaei-Matehkolaei, A., Abdollahi, M., Didehdar, M., Haghani, I., Abastabar, M. 2016. In vitro activity of new azoles luliconazole and lanoconazole compared with ten other antifungal drugs against clinical dermatophyte isolates. Medical Mycology, 54(7), 757–763.
- [4] Pan, G. C., Peng, F. J., Ying, G. G. 2018. Removal, biotransformation and toxicity variations of climbazole by freshwater algae Scenedesmus obliquus. Environmental Pollution, 240, 534-540.
- [5] Iqbal, D. N., Ashraf, A., Iqbal, M., Nazir, A. 2020. Analytical method development and validation of hydrocortisone and clotrimazole in topical dosage form using RP-HPLC. Future Journal of Pharmaceutical Sciences, 6(49), 1-7.
- [6] Kumaraswamy, G., Kumar, D. S., Joru, P., Emmadi, S. 2017. RP-HPLC Method Development and Validation for Simultaneous Estimation of Lignocaine Hydrochloride and Clotrimazole Hydrochloride in Ear Drops. Asian Journal of Pharmaceutical Analysis, 7(3),163-168.
- [7] Paz-Alvarez, M., Pudney, P. D. A., Hadgraft, J., Lane, M. E. 2018. Topical delivery of climbazole to mammalian skin. International Journal of Pharmaceutics, 549(1-2), 317-324.
- [8] Zhong, Y., Chen, Z. F., Liu, S. S., Dai, X., Zhu, X., Zheng, G., Liu, S., Liu, G., Cai, Z. 2017. Analysis of azole fungicides in fish muscle tissues: Multi-factor optimization and application to environmental samples. Journal of Hazardous Materials, 324, 535–543.
- [9] Üstün, Z., Demiralay, E. Ç. 2017. Simultaneous Quantitative Determination of Imidazole Antimycotics in Human Urine by using RPLC Technique. Eurasian Journal of Analytical Chemistry, 12(6), 953-962.
- [10] Roses, M., Bosch, E. 2002. Influence of hareketlie phase acid–base equilibria on the chromatographic behaviour of protolytic compounds. Journal of Chromatography A, 982, 1–30.
- [11] Demiralay, E. Ç., Cubuk, B., Ozkan, S. A., Alsancak, G. 2010. Combined effect of polarity and pH on the chromatographic behaviour of some angiotensin II receptor antagonists and optimization of their determination in pharmaceutical dosage forms. J Pharm Biomed Anal, 53, 475–482.
- [12] Erdemgil, F. Z., Sanli, S., Sanli, N., Ozkan, G., Barbosa, J., Guiteras, J., Beltrán, J.L. 2007. Determination of pKa values of some hydroxylated benzoic acids in methanol–water binary mixtures by LC methodology and potentiometry. Talanta, 72, 489–496.
- [13] Horváth, C., Melander, W., Molnár, I. 1977. Liquid chromatography of ionogenic substances with nonpolar stationary phases. Anal Chem, 49, 142–154.
- [14] Subirats, X., Bosch, E., Rose´s, M. 2006. Retention of ionisable compounds on high-performance liquid chromatography XV. Estimation of the pH variation of aqueous buffers with the change of the acetonitrile fraction of the hareketlie phase. J Chromatogr A, 1121, 170–177.
- [15] Secilmis, H. C., Demiralay, E. C., Alsancak, G., Ozkan, S. A. 2012. The Combined Effect of the Organic Modifier Content and pH of the Hareketlie Phase on the Chromatographic Behavior of Some Arylpropionic and Arylacetic Acids to Optimize Their Liquid Chromatographic Determinations. Chromatographia, 75, 711–720.
- [16] Poole, C. F., Poole, S. K. 1991. Chromatography Today. First Ed., Elsevier Science B.V. Amsterdam, 715s.
- [17] Kazakevich, Y., Lobrutto, Y. 2007. HPLC for Pharmaceutical Scientists, First ed., Wiley-Interscience, USA, 1140s.
- [18] Meyer, V. R. 2010. Practical High-Performance Liquid Chromatography, Fifth ed., John Wiley and Sons, United Kingdom, 428s.
- [19] Swiss ADME program, 2023. http://www.swissadme.ch /index.php (Erişim Tarihi: 26.07.2023).
- [20] Chemicalize program, Kimyasal Hesaplama. https://chemicalize.com/#/calculation (Erişim Tarihi: 10.03.2016).
- [21] Mussini, T., Covington, A. K., Longhi, P., Rondinini, S. 1985. Criteria for Standardization of pH Measurements in Organic Solvents and Water + Organic Solvent Mixtures of Moderate to High Permittivities. Pure and Applied Chemistry, 57(6), 865- 876.
Reverse Phase Liquid Chromatography Method Optimization for Analysis of Some Azole Group Antifungal Compounds
Yıl 2023,
, 347 - 354, 29.12.2023
Dilara Başat Dereli
,
Ebru Çubuk Demiralay
,
İlkay Konçe
Öz
In this study, the capacity factor values of the compounds and the methanol concentration in the mobile phase in which they were analyzed were based on the method optimization developed by using the reverse phase liquid chromatography (RPLC) method of the azole antifungal drug active ingredients clotrimazole, lanoconazole and climbazole. In the optimization of these compounds with ionizing groups, the pH value of the mobile phase is another factor in the optimization. For an isocratic separation, it is preferred that the capacity factor values are in the range of 1-5.
Methanol-water binary mixtures containing 50%, 55% and 60% (v/v) methanol were used in the optimization for the simultaneous determination of clotrimazole, lanoconazole and climbazol compounds by RPLC method. Retention times at pH values of two mobile phases (pH 4.0 and 5.5) were determined in the working media for separation. Zorbax SB-CN (4.6x150mm) column was preferred for the determination of hydrophobic compounds. The column temperature was fixed at 37 °C and the flow rate was fixed at 1 mL/min. The logk-% (v/v) methanol relationship and the selectivity factor (α) parameter, which shows the separation of the compounds, were used to determine the optimization conditions.
Proje Numarası
TYL-2022-8833
Kaynakça
- [1] Brunton, L. L., Lazo, J. S., Parker, K. L. 2008. 11th ed.; Goodman & Gilman's The pharmacology basis of therapeutics, McGrawHill eBooks, New York.
- [2] Kadavakollu, S., Stailey, C., Kunapareddy, C. S., White, S. 2014. Clotrimazole as a Cancer Drug: A Short Review. Med Chem, 4(11), 722–724.
- [3] Baghi, N., Shokohi, T., Badali, H., Makimura, K., Rezaei-Matehkolaei, A., Abdollahi, M., Didehdar, M., Haghani, I., Abastabar, M. 2016. In vitro activity of new azoles luliconazole and lanoconazole compared with ten other antifungal drugs against clinical dermatophyte isolates. Medical Mycology, 54(7), 757–763.
- [4] Pan, G. C., Peng, F. J., Ying, G. G. 2018. Removal, biotransformation and toxicity variations of climbazole by freshwater algae Scenedesmus obliquus. Environmental Pollution, 240, 534-540.
- [5] Iqbal, D. N., Ashraf, A., Iqbal, M., Nazir, A. 2020. Analytical method development and validation of hydrocortisone and clotrimazole in topical dosage form using RP-HPLC. Future Journal of Pharmaceutical Sciences, 6(49), 1-7.
- [6] Kumaraswamy, G., Kumar, D. S., Joru, P., Emmadi, S. 2017. RP-HPLC Method Development and Validation for Simultaneous Estimation of Lignocaine Hydrochloride and Clotrimazole Hydrochloride in Ear Drops. Asian Journal of Pharmaceutical Analysis, 7(3),163-168.
- [7] Paz-Alvarez, M., Pudney, P. D. A., Hadgraft, J., Lane, M. E. 2018. Topical delivery of climbazole to mammalian skin. International Journal of Pharmaceutics, 549(1-2), 317-324.
- [8] Zhong, Y., Chen, Z. F., Liu, S. S., Dai, X., Zhu, X., Zheng, G., Liu, S., Liu, G., Cai, Z. 2017. Analysis of azole fungicides in fish muscle tissues: Multi-factor optimization and application to environmental samples. Journal of Hazardous Materials, 324, 535–543.
- [9] Üstün, Z., Demiralay, E. Ç. 2017. Simultaneous Quantitative Determination of Imidazole Antimycotics in Human Urine by using RPLC Technique. Eurasian Journal of Analytical Chemistry, 12(6), 953-962.
- [10] Roses, M., Bosch, E. 2002. Influence of hareketlie phase acid–base equilibria on the chromatographic behaviour of protolytic compounds. Journal of Chromatography A, 982, 1–30.
- [11] Demiralay, E. Ç., Cubuk, B., Ozkan, S. A., Alsancak, G. 2010. Combined effect of polarity and pH on the chromatographic behaviour of some angiotensin II receptor antagonists and optimization of their determination in pharmaceutical dosage forms. J Pharm Biomed Anal, 53, 475–482.
- [12] Erdemgil, F. Z., Sanli, S., Sanli, N., Ozkan, G., Barbosa, J., Guiteras, J., Beltrán, J.L. 2007. Determination of pKa values of some hydroxylated benzoic acids in methanol–water binary mixtures by LC methodology and potentiometry. Talanta, 72, 489–496.
- [13] Horváth, C., Melander, W., Molnár, I. 1977. Liquid chromatography of ionogenic substances with nonpolar stationary phases. Anal Chem, 49, 142–154.
- [14] Subirats, X., Bosch, E., Rose´s, M. 2006. Retention of ionisable compounds on high-performance liquid chromatography XV. Estimation of the pH variation of aqueous buffers with the change of the acetonitrile fraction of the hareketlie phase. J Chromatogr A, 1121, 170–177.
- [15] Secilmis, H. C., Demiralay, E. C., Alsancak, G., Ozkan, S. A. 2012. The Combined Effect of the Organic Modifier Content and pH of the Hareketlie Phase on the Chromatographic Behavior of Some Arylpropionic and Arylacetic Acids to Optimize Their Liquid Chromatographic Determinations. Chromatographia, 75, 711–720.
- [16] Poole, C. F., Poole, S. K. 1991. Chromatography Today. First Ed., Elsevier Science B.V. Amsterdam, 715s.
- [17] Kazakevich, Y., Lobrutto, Y. 2007. HPLC for Pharmaceutical Scientists, First ed., Wiley-Interscience, USA, 1140s.
- [18] Meyer, V. R. 2010. Practical High-Performance Liquid Chromatography, Fifth ed., John Wiley and Sons, United Kingdom, 428s.
- [19] Swiss ADME program, 2023. http://www.swissadme.ch /index.php (Erişim Tarihi: 26.07.2023).
- [20] Chemicalize program, Kimyasal Hesaplama. https://chemicalize.com/#/calculation (Erişim Tarihi: 10.03.2016).
- [21] Mussini, T., Covington, A. K., Longhi, P., Rondinini, S. 1985. Criteria for Standardization of pH Measurements in Organic Solvents and Water + Organic Solvent Mixtures of Moderate to High Permittivities. Pure and Applied Chemistry, 57(6), 865- 876.