European Journal of Chemistry 2013, 4(1), 29-34 | doi: | Get rights and content

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Comparative determination of miconazole, nystatin, hydrocortisone and neomycin by HPTLC/HPLC-DAD

Ismail Salama (1,*) , Mohamed Sayed Gomaa (2)

(1) Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Suez Canal University, Ismailia 41552, Egypt
(2) Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Suez Canal University, Ismailia 41552, Egypt
(*) Corresponding Author

Received: 29 Dec 2012 | Accepted: 20 Jan 2013 | Published: 31 Mar 2013 | Issue Date: March 2013


Two new validated methods for the quantification of miconazole (MIC), nystatin (NYS), hydrocortisone acetate (HCA) and neomycin (NS) by high performance thin layer chromatography (HPTLC)-densitometry and reverse-phase high-performance liquid chromatography procedure coupled with photodiode array detector (RP-HPLC-DAD) were developed and compared. HPTLC separations were performed using a mobile phase, ethylacetate:methanol:glacial acetic acid (60:40:0.4, v:v:v) for MIC, NYS, HCA and ethylacetate for NS. For HPLC-DAD analysis, using an isocratic elution system, separation of all compounds was achieved. Good resolution and quantization were achieved, accuracy and precision, as well as detection and quantitation limits of the two methods, were evaluated and compared. Excellent linearity was observed for all of the standard calibration curves, and the correlation coefficients were above 0.9997. HPTLC limits of quantitation were 15.37×10-2, 9.8×10-2, 13.32×10-2 and 15.19×10-2 µg/mL for MIC, NYS, HCA and NS respectively, whereas HPLC limits were 6.80×10-2, 6.56×10-2, 1×10-1 and 10.16×10-2 µg/band for MIC, NYS, HCA and NS respectively. In comparison with HPLC, HPTLC is less expensive and faster procedure, requiring 2-3 h to analyse 10-12 samples on a single plate. The developed methods were proved to be specific, robust and accurate for the determination of cited drugs in pharmaceutical preparations.



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European Journal of Chemistry


HPTLC; Nystatin; Neomycin; HPLC-DAD; Miconazole nitrate; Hydrocortisone acetate

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DOI: 10.5155/eurjchem.4.1.29-34.721

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Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt



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DOI: 10.1093/jaoacint/qsac074

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[1]. Reynolds, J. E. F. (ed). Martindale: The Extra Pharmacopoeia, 29th ed., Pharmaceutical Press, London, 1989, pp. 430, 666.

[2]. Hardman, J. G.; Limbird, L. E. (eds). Goodman & Gilman's: The Pharmacological Basis of Therapeutics, 9th ed., McGraw- Hill, New York, 1996.

[3]. USP DI, 24th edition, Drug Information for the Health Care Professional, 2004.

[4]. Sweetman, S. C. (ed). Martindale: The complete drug reference, 35th ed., Pharmaceutical Press, London, 2002, p. 392.

[5]. Lemus, G. J. M.; Perez, A. J. Chromatographia 2002, 55, 749-753.

[6]. Waksman, S. A. Neomycin. Nature, Formation, Isolation, and Practical Application, Rutgers University Press, New Brunswick, NJ, 1953.

[7]. Waksman, S. A.; Lechevalier, H. A. Guide to the Classification and Identification of the Actinomycetes and their Antibiotics, The Williams & Wilkins Co., Baltimore, MD, 1953.

[8]. Waksman, S. A. Neomycin. Its Nature and Practical Application, The Williams & Wilkins Co., Baltimore, MD, 1958.

[9]. Rinehart, K. L. The neomycins and related antibiotics, in: Chemistry of Microbial Products E. R. Squibb Lectures on Chemistry of Microbial Products, Institute of Microbiology, John Wiley & Sons, Inc., New York, NY, 1964.

[10]. Kobylinska, M.; Kobylinska, K.; Sobik, B. J. Chromatogr B: Biomed. Appl. 1996, 685, 191-195.

[11]. Goger, N. G.; Gokcen, L. Anal. Lett. 1999, 32, 2595-2602.

[12]. Wrobel, K.; Wrobel, K.; de la Garza Rodriguez, I. M.; Lopezde-Alba, P. L.; Lopez Martinez, L. J. Pharm. Biomed. Anal. 1999, 20, 99-105.

[13]. Khashaba, P. Y.; El-Shabouri, S. R.; Emara, K. M.; Mohamed, A. M. J. Pharm. Biomed. Anal., 2003, 22, 363-376.

[14]. Akay, C.; Ozkan, S. A.; Senturk, Z.; Cevheroglu, S. Il Farmaco 2003, 57, 953-957.

[15]. Lupan, L.; Bandula, R.; Vasilescu, M.; Bercu, C. Fresenius J. Anal. Chem. 1996, 355, 409-411.

[16]. Botsoglou, N. A.; Fletouris, D. J. J. Agr. Food Chem. 1996, 44, 1271-1274.

[17]. Groll, A. H.; Mickiene, D.; Werner, K.; Pistelli, S. C.; Walsh, T. J. J. Chrom. B 1999, 735, 51-57.

[18]. Yeo, S. K.; Lee, H. K.; Li, S. F. Y. J. Chrom. A 1991, 585, 133-137.

[19]. Raith, K.; Althoff, E.; Banse, J.; Neidhardt, H.; Neubert, R. H. H. Electrophoresis 1998, 19, 2907-2911.

[20]. Heneedak, H.; Salama, I.; Mostafa, S.; El-Sadek, M. J. Chrom. Sci. 2012, 50, 855-861.

[21]. Bonazzi, D.; Andrisano, V.; Gatt, R.; Cavrini, V. J. Pharm. Biomed. Anal. 1995, 13, 1321-1329.

[22]. Amin, A. S. Anal. Lett. 1996, 29, 1527-1537.

[23]. Medenica, M.; Ivanovic, D.; Markovic, S.; Malenovic, A.; Jancic, B. Pharmind. 2004, 66, 330-333.

[24]. Gagliardi, L.; De Orsi, D.; Giudice, M. R. D.; Gatta, F.; Porra, R.; Chimenti, P.; Tonelli, D. Anal. Chim. Acta. 2002, 457, 187-198.

[25]. Chauhan, V.; Conway, B. Chromatographia 2005, 61, 555-559.

[26]. Hajkova, R.; Solich, P.; Dvocak, J.; Cicha, J. J. Pharm. Biomed. Anal. 2003, 32, 921-927.

[27]. Jancic-stojanovic, B.; Malenovic, A. J. AOAC Int. 2010, 93, 102-107.

[28]. Hay, M.; Mormed, P. J. Chrom. B. 1997, 702, 33-39.

[29]. Grippa, E.; Santini, L.; Castellano, G.; Gatto, M. T.; Leone, M. G.; Saso, L. J. Chrom. B 2000, 738, 17-25.

[30]. Majid, O.; Akhlaghi, F.; Lee, T.; Holt, D. W.; Trull, A. Ther. Drug Monit. 2001, 23, 163-168.

[31]. Shaikh, B.; Jackson, J.; Guyer, G.; Ravis, W. R. J. Chrom. B: Biomed. Appl. 1991, 571, 189-198.

[32]. Medina, M. B.; Unruh, J. J. J. Chrom. B 1995, 663, 127-135.

[33]. Oertel, R.; Renner, U.; Kirch, W. J. Pharm. Biomed. Anal. 2004, 35, 633-638.

[34]. Oertel, R.; Neumeister, V.; Kirch, W. J. Chrom. A 2004, 1058, 197-201.

[35]. Byoung-Hyoun, K.; Suk Chin, L.; Hye Jin, L.; Jong Hoa, O. Biomed. Chrom. 2003, 17, 396- 403.

[36]. International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use, ICH Harmonised Tripartite Guideline-Validation of Analytical Procedures: Text and Methodology Q2(R1), Current Step 4 version, London, 2005.

[37]. The British Pharmacopoeia, Her Majesty's Stationery Office. London, UK, 2008, pp 404-405, 485-486, 1080-1081, 1382, 2519-2520, 2559-2560, 2592-2593, 2766-2767, 2861.

[38]. Zeaiter, M.; Roger, J. M.; Maurel, V. B.; Rutledge, D. N. Trends. Anal. Chem. 2004, 23, 157-170.

[39]. Mulholland, M. Trends Anal. Chem. 1988, 7, 383-389.

[40]. Heyden, Y. V.; Nijhuis, A.; Verbeke, J. S.; Vandeginste, B. G. M.; Massaret, D. L. J. Pharm. Biomed. Anal. 2001, 24, 723-753.

How to cite

Salama, I.; Gomaa, M. Eur. J. Chem. 2013, 4(1), 29-34. doi:10.5155/eurjchem.4.1.29-34.721
Salama, I.; Gomaa, M. Comparative determination of miconazole, nystatin, hydrocortisone and neomycin by HPTLC/HPLC-DAD. Eur. J. Chem. 2013, 4(1), 29-34. doi:10.5155/eurjchem.4.1.29-34.721
Salama, I., & Gomaa, M. (2013). Comparative determination of miconazole, nystatin, hydrocortisone and neomycin by HPTLC/HPLC-DAD. European Journal of Chemistry, 4(1), 29-34. doi:10.5155/eurjchem.4.1.29-34.721
Salama, Ismail, & Mohamed Sayed Gomaa. "Comparative determination of miconazole, nystatin, hydrocortisone and neomycin by HPTLC/HPLC-DAD." European Journal of Chemistry [Online], 4.1 (2013): 29-34. Web. 4 Jun. 2023
Salama, Ismail, AND Gomaa, Mohamed. "Comparative determination of miconazole, nystatin, hydrocortisone and neomycin by HPTLC/HPLC-DAD" European Journal of Chemistry [Online], Volume 4 Number 1 (31 March 2013)

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