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Coupling of on-Line Pre-Column Oxidative Cleavage and Solid-Phase Enrichment with Liquid Chromatography Using an Eco-Friendly Analytical Procedure to Determine Low Levels of Methotrexate

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DOI: 10.4236/jasmi.2012.24030    3,168 Downloads   5,749 Views   Citations

ABSTRACT

A simple, sensitive and precise green high-performance liquid chromatographic method including on-line pre-column oxidation combined by column switching with a short Hypersil ODS analytical column (100 mm × 4.0 mm i.d.) for enrichment and separation was developed and validated to determine low levels of methotrexate (MTX). The method was based on oxidative cleavage of MTX into highly fluorescence products, 2,4-diaminopteridine-6-carboxaldehyde and the corresponding 2,4-diaminopteridine-6-carboxylic acid, during the flow of phosphate buffer (0.04 M, pH 3.4) containing the analyte through the packed reactor of cerium (IV) trihydroxyhydroperoxide (CTH) at a flow-rate of 0.2 mL/min and 40℃. The fluorescent products were enriched on the head of ODS analytical column for the final separation. The separation was performed at room temperature using an environmentally friendly mobile phase consisting of ethanol and phosphate buffer (0.04 M, pH 3.4) in the ratio of 10:90 (v/v). The eluent was monitored at emission and excitation wavelengths of 463 and 367 nm, respectively. The method was successfully applied, without any interference from the excipients, for the determination of drug in tablets and vials with a detection limit of 0.06 ng/mL from 500 ?L of sample MTX.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

S. Emara, W. Zarad, M. Kamal and R. EL-Bagary, "Coupling of on-Line Pre-Column Oxidative Cleavage and Solid-Phase Enrichment with Liquid Chromatography Using an Eco-Friendly Analytical Procedure to Determine Low Levels of Methotrexate," Journal of Analytical Sciences, Methods and Instrumentation, Vol. 2 No. 4, 2012, pp. 194-202. doi: 10.4236/jasmi.2012.24030.

References

[1] S. Shen, T. O’Brien, L. M. Yap, H. M. Prince and C. J. McCormack, “The Use of Methotrexate in Dermatology: A Review,” Australasian Journal of Dermatology, Vol. 53, No. 1, 2102, pp. 1-18.
[2] M. Barca, M. Ilie, D. L. Baconi, A. M. Ciobanu, D. Balalau and G. T. Burcea, “Spectrofluorimetric Methotrexate Assay in Human Plasma,” Farmacia, Vol. 58, No. 1, 2010, pp. 95-101.
[3] S. M. Sabry, M. Abdel-Hady, M. Elsayed, O. T. Fahmy and H. M. Mahr, “Study of Stability of Methotrexate in Acidic Solution. Spectrofluorimetric Determination of Methotrexate in Pharmaceutical Preparations through Acid-Catalyzed Degradation Reaction,” Journal of Pharmaceutical and Biomedical Analysis, Vol. 32, No. 3, 2003, pp. 409-423. Hdoi:10.1016/S0731-7085(03)00239-5
[4] A. Espinosa-Mansilla, I. Duran Meras, A. Zamora Madera, L. Pedano and C. Ferreyra, “Kinetic Fluorimetric Determination of the Antineoplastic Methotrexate (MTX) in Human Serum,” Journal of Pharmaceutical and Biomedical Analysis, Vol. 29, No. 5, 2002, pp. 851-858. Hdoi:10.1016/S0731-7085(02)00212-1
[5] H. Hayashi, C. Fujimaki, S. Tsuboi, T. Matsuyama, T. Daimon and K. Itoh, “Application of Fluorescence Polarization Immunoassay for Determination of Methotrexate-Polyglutamates in Rheumatoid Arthritis Patients,” The Tohoku Journal of Experimental Medicine, Vol. 215, No. 1, 2008, pp. 95-101. Hdoi:10.1620/tjem.215.95
[6] H. L. Cheng, S. S. Chiou, Y. M. Liao, C. Y. Lu, Y. L. Chen and S. M. Wu, “Analysis of Methotrexate and Its Eight Metabolites in Cerebrospinal Fluid by Solid-Phase Extraction and Triple-Stacking Capillary Electrophoresis,” Analytical and Bioanalytical Chemistry, Vol. 398, No. 5, 2010, pp. 2183-2190. Hdoi:10.1007/s00216-010-4152-3
[7] H. L. Cheng, Y. M. Liao, S. S. Chiou and S. M. Wu, “On-Line Stacking Capillary Electrophoresis for Analysis of Methotrexate and Its Eight Metabolites in Whole Blood,” Electrophoresis, Vol. 29, No. 17, 2008, pp. 3665-3673. Hdoi:10.1002/elps.200800029
[8] S. Fang, C. P. Lollo, C. Derunes and M. J. LaBarre, “Development and validation of a Liquid Chromatography Method for Simultaneous Determination of Three Process-Related Impurities: Yeastolates, Triton X-100 and Methotrexate,” Journal of Chromatography B, Vol. 879, No. 30, 2011, pp. 3612-3619. Hdoi:10.1016/j.jchromb.2011.10.003
[9] E. Klapkova, J. Kukacka, K. Kotaska, I. Suchanska, R. Urinovska and R. Prusa, “The Influence of 7-OH Methotrexate Metabolite on Clinical Relevance of Methotrexate Determination,” Clinical Laboratory, Vol. 57, No. 7-8, 2011, pp. 599-606.
[10] K. Michail and M. S. Moneeb, “Determination of Methotrexate and Indomethacin in Urine Using SPE-LC-DAD after Derivatization,” Journal of Pharmaceutical and Biomedical Analysis, Vol. 55, No. 2, 2011, pp. 317-324. Hdoi:10.1016/j.jpba.2011.01.032
[11] L. Hu, Y. Liu and S. Cheng, “Simultaneous Determination of Six Analytes by HPLC-UV for High Throughput Analysis in Permeability Assessment,” Journal of Chromatographic Science, Vol. 49, No. 2, 2011, pp. 124-128. Hdoi:10.1093/chrsci/49.2.124
[12] T. Sartori, F. S.Murakami, A. P. Cruz and A. M. de Campos, “Development and Validation of a Fast RP-HPLC Method for Determination of Methotrexate Entrapment Efficiency in Polymeric Nanocapsules,” Journal of Chromatographic Science, Vol. 46, No. 6, 2008, pp. 505-509.
[13] D. A. el-Hady, N. A. el-Maali, R. Gotti, C. Bertucci, F. Mancini and V. Andrisano, “Methotrexate Determination in Pharmaceuticals by Enantioselective HPLC,” Journal of Pharmaceutical and Biomedical Analysis, Vol. 37, No. 5, 2005, pp. 919-925. Hdoi:10.1016/j.jpba.2004.07.046
[14] M. Uchiyama, T. Matsumoto, T. Matsumoto, S. Jimi, Y. Takamatsu, K. Tamura and S. Hara, “Simple and Sensitive HPLC Method for the Fluorometric Determination of Methotrexate and Its Major Metabolites in Human Plasma by Post-Column Photochemical Reaction,” Biomedical Chromatography, Vol. 26, No. 1, 2012, pp. 76-80. Hdoi:10.1002/bmc.1628
[15] L. van Haandel and J. F. Stobaugh, “Bioanalytical Method Development for a Generation 5 Polyamidoamine Folic Acid Methotrexate Conjugated Nanoparticle,” Analytical and Bioanalytical Chemistry, Vol. 397, No. 5, 2010, pp. 1841-1852. Hdoi:10.1007/s00216-010-3716-6
[16] S. Chen and Z. Zhang, “Molecularly Imprinted Solid-Phase Extraction Combined with Electrochemical Oxidation Fluorimetry for the Determination of Methotrexate in Human Serum and Urine,” Spectrochimica Acta Part A, Vol. 70, No. 1, 2008, pp. 36-41. Hdoi:10.1016/j.saa.2007.07.009
[17] H. Li, W. Luo, Q. Zeng, Z. Lin, H. Luo and Y. Zhang, “Method for the Determination of Blood Methotrexate by High Performance Liquid Chromatography with Online Post-Column Electrochemical Oxidation and Fluorescence Detection,” Journal of Chromatography B, Vol. 845, No. 1, 2007, pp. 164-168. Hdoi:10.1016/j.jchromb.2006.07.026
[18] I. Duran Meras, A. Espinosa Mansilla and M. J. Rodríguez Gomez, “Determination of Methotrexate, Several Pteridines, and Creatinine in Human Urine, Previous Oxidation with Potassium Permanganate, Using HPLC with Photometric and Fluorimetric Serial Detection,” Analytical Biochemistry, Vol. 346, No. 2, 2005, pp. 201-209. Hdoi:10.1016/j.ab.2005.07.038H
[19] E. A. McCrudden and S. E. Tett, “Improved High-Performance Liquid Chromatography Determination of Methotrexate and Its Major Metabolite in Plasma Using a Poly (Styrene-Divinylbenzene) Column,” Journal of Chromatography B, Vol. 721, No. 1, 1999, pp. 87-92. Hdoi:10.1016/S0378-4347(98)00439-3H
[20] Z. Yu, D. Westerlund and K. S. Boose, “Determination of Methotrexate and Its Metabolite 7-Hydroxymethotrexate by Direct Injection of Human Plasma into a Column-Switching Liquid Chromatographic System Using Post-Column Photochemical Reaction with Fluorimetric Detection,” Journal of Chromatography B, Vol. 689, No. 2, 1997, pp. 379-386. Hdoi:10.1016/S0378-4347(96)00357-X
[21] T. Suzuki, H. Hashimoto and N. Ichinose, “Microanalysis of Methotrexate by High-Performance Liquid Chromatography Using a Fluorescence Detector,” Fresenius Journal of Analytical Chemistry, Vol. 351, No. 8, 1995, pp. 806-807. Hdoi:10.1007/BF00323644
[22] H. Kubo, Y. Umiguchi, M. Fukumoto and T. Kinoshita, “Fluorimetric Determination of Methotrexate in Serum by High-Performance Liquid Chromatography Using in-Line Oxidation with Hydrogen Peroxide,” Analytical Sciences, Vol. 8, No. 6, 1992, pp. 789-792. Hdoi:10.2116/analsci.8.789
[23] J. Salamoun, M. Smrz, F. Kiss and A. Salamounova, “Column Liquid Chromatography of Methotrexate and Its Metabolites Using a Post-Column Photochemical Reactor and Fluorescence Detection,” Journal of Chromatography B, Vol. 419, 1987, pp. 213-223. Hdoi:10.1016/0378-4347(87)80279-7
[24] J. Salamoun and J. Frantisek, “Determination of Methotrexate and Its Metabolites 7-Hydroxymethotrexate and 2,4-Diamino-N10-methylpteroic Acid in Biological Fluids by Liquid Chromatography with Fluorimetric Detection,” Journal of Chromatography B, Vol. 378, No. 1, 1986, pp. 173-181.
[25] J. A. Nelson, B. A. Harris, W. J. Decker and D. Farquhar, “Analysis of Methotrexate in Human Plasma by High-Performance Liquid Chromatography with Fluorescence Detection,” Cancer Research, Vol. 37, No. 11, 1977, pp. 3970-3973.
[26] S. Chen, Z. Zhang, D. He, Y. Hu, H. Zheng and C. He, “Flow-Injection-Electrochemical Oxidation Fluorimetry for Determination of Methotrexate,” Luminescence, Vol. 22, No. 4, 2007, pp. 338-342. Hdoi:10.1002/bio.968
[27] S. Emara, “Determination of Methotrexate in Pharmaceutical Formulations by Flow Injection Analysis Exploiting the Reaction with Potassium Permanganate,” II Farmaco, Vol. 59, No. 10, 2004, pp. 827-833. Hdoi:10.1016/j.farmac.2004.06.005
[28] R. I. Stefan, R. G. Bokretsion, J. F. van Staden and H. Y. Aboul-Enein, “Simultaneous Determination of L- and D-Methotrexate Using a Sequential Injection Analysis/ Amperometric Biosensors System,” Biosensors and Bioelectronics, Vol. 19, No. 3, 2003, pp. 261-267. Hdoi:10.1016/S0956-5663(03)00210-0
[29] S. Emara, S. Razee, A-N. El-Shorbagy and T. Masujima, “Flow Injection Method for the Determination of Methotrexate with a Column-Packed Oxidizing Agent,” Analyst, Vol. 121, No. 2, 1996, pp. 183-188. Hdoi:10.1039/an9962100183
[30] United States Pharmacopeial Convention, “The United States Pharmacopeia and National Formulary (USP 28-NF 23),” The Official Compendia of Standards, Asian Edition, The United States Pharmacopeial Convention Inc., Rockville, 2005.

  
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