Mixed Micellar Electrokinetic Chromatographic Analysis of Colistin, Polypeptide Antibiotic, Using Laser-Induced Fluorescence Detection

Abstract

The main goal of this work was to quantify the detection of colistin at low levels in urine samples through the practical application of mixed surfactant micellar electrokinetic chromatography–laser-induced fluorescence (MEKC-LIF) analysis method using its advantage of sensitivity and to examine direct injection of biological samples. Colistin (po- lymyxin E) has neither strong UV chromophore nor fluorophore. So, its assay for metabolism, pharmacokinetics studies for bioavailability and bioequivalence are difficult because of poor detectability. Therefore an enhanced UV or fluores-cence detection by chemical derivatization is required. MEKC-LIF method was proposed for colistin with a 488/520 nm argon-ion laser using a pre-CE derivatization with fluorescein isothiocyanate (FITC). Borate buffer was used as background buffer (BGB). The different parameters affecting the proposed derivatization reaction including concentration of the derivatizing reagent, reaction time and temperature were studied and optimized. The derivative was stable for up to 3 days. Different micelles (TX-100 and SDS) were examined as BGB additives separately but negative-charged mixed micelles (SDS/TX-100) were shown to be the best additive to BGB for the analysis of colistin particularly in human urine as they enhance both selectivity and sensitivity of the proposed method. BGB was used with pH 9.5, 10 kV, 8 s inj time, capillary length 75 cm × 75 μm ID (66 cm effective length), detection was LIF Ex 488 nm; Em 520 nm. The method was applied to colistin analysis in human urine and the recovery was > 98% (n = 5). LOD and LOQ in urine after pre-column derivatization using FITC were 100 and 250 ng/ml, respectively. Urine samples were analysed by direct injection without sample pre-treatment. The mechanism of enhancement of fluorescence of the derivative by surfactant was proposed.

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H. Ahmed, F. Elbarbry and B. Clark, "Mixed Micellar Electrokinetic Chromatographic Analysis of Colistin, Polypeptide Antibiotic, Using Laser-Induced Fluorescence Detection," American Journal of Analytical Chemistry, Vol. 3 No. 3, 2012, pp. 233-241. doi: 10.4236/ajac.2012.33031.

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1] M. E. Evans, D. J. Feola and R. P. Rapp, “Polymyxin B Sulfate and Colistin: Old Antibiotics for Emerging Mul- tiresistant Gram-Negative Bacteria,” Annals of Pharma-cotherapy, Vol. 33, No. 9, 1999, pp. 960-967. doi:10.1345/aph.18426
[2] T. Jensen, S. S. Pedersen, S. Garne, C. Heilmann, N. Hoiby and C. Koch, “Colistin Inhala-tion-Therapy in Cystic-Fibrosis Patients with Chronic Pseudo-monas-Aerugi-nosa Lung Infection,” Journal of Antimicrobial Chemo- therapy, Vol. 19, No. 6, 1987, pp. 831-838. doi:10.1093/jac/19.6.831
[3] D. Decolin, P. Leroy, A. Nicolas and P. Archimbault, “Hyphenated Liquid Chromatographic Method for the Determination of Colistin Residues in Bovine Tissues,” Journal of Chromatographic Science, Vol. 35, No. 12, 1997, pp. 557-564.
[4] A. H. Thomas and I. Holloway, “Thin-Layer Chromatographic Method for Identification of Polymyxins,” Journal of Chromatography, Vol. 161, 1978, pp. 417-420.
[5] T. Suzuki, H. Inouye, K. Fujikawa and Y. Suketa, “Studies on the Chemical Structure of Colistin: I. Fractionation, Molecular Weight Determination, Amino Acid and Fatty Acid Composition,” Journal of Biochemistry, Vol. 54, No. 1, 1963, pp. 25-30.
[6] J. A. Orwa, C. Govaerts, K. Gevers, E. Roets, A. Van Schepdael and J. Hoogmartens, “Study of the Stability of Polymyxins B-1, E-1 and E-2 in Aqueous Solution Using Liquid Chromatography and Mass Spectrometry,” Jour- nal of Pharmaceutical and Biomedical Analysis, Vol. 29, No. 1-2, 2002, pp. 203-212. doi:10.1016/S0731-7085(02)00016-X
[7] B. C. Grande, M. S. G. Falcon, C. Perez-Lamela, M. R. Comesana and J. S. Gandara, “Quantitative Analysis of Colistin and Tiamulin in Liquid and Solid Medicated Premixes by Hplc with Diode-Array Detection,” Chromatographia, Vol. 53, No. 1, 2001, pp. S460-S463. doi:10.1007/BF02490378
[8] P. Perez-Lozano, E. Gar-cia-Montoya, A. Orriols, M. Minarro, J. R. Tico and J. M. Sune-Negre, “Application of a Validated Method in the Stability Study of Colistin Sulfate and Methylparaben in a Veterinary Suspension Formulation by High-Performance Liquid Chromatography with a Diode Array Detector,” Journal of Aoac In- ternational, Vol. 90, No. 3, 2007, pp. 706-714.
[9] G. Morovjan, P. P. Csokan and L. Nemeth-Konda, “Hplc Deter-mination of Colistin and Aminoglycoside Antibiotics in Feeds by Post-Column Derivatization and Fluorescence Detection,” Chromatographia, Vol. 48, No. 1-2, 1998, pp. 32-36. doi:10.1007/BF02467512
[10] B. Cancho-Grande, M. Rodri-guez-Comesana and J. Simal-Gandara, “Simple Hplc Determination of Colistin in Medicated Feeds by Pre-Column Derivatization and Fluorescence Detection,” Chromatographia, Vol. 54, No. 7-8, 2001, pp. 481-484. doi:10.1007/BF02491203
[11] S. Morales-Munoz and M. D. L. De Castro, “Dynamic Ultrasound-Assisted Extraction of Colistin from Feeds with On-Line Pre-Column Derivatization and Liquid Chro- matography-Fluorimetric Detection,” Journal of Chro- matography A, Vol. 1066, No. 1-2, 2005, pp. 1-7. doi:10.1016/j.chroma.2005.01.010
[12] M. Mashat, H. Chrystyn, B. J. Clark and K. H. Assi, “Development and Vali-dation of HPLC Method for the Determination of Tobramycin in Urine Samples Post-In- halation Using Pre-Column Deriva-tisation with Fluo- rescein Isothiocyanate,” Journal of Chro-matography B, Vol. 869, No. 1-2, 2008, pp. 59-66. doi:10.1016/j.jchromb.2008.05.012
[13] D. J. Gmur, C. R. Bredl, S. J. Steele, S. P. Cai, D. R. Vandevanter and P. A. Nardella, “Determination of Po- lymyxin E-1 in Rat Plasma by High-Performance Liquid Chromatography,” Journal of Chro-matography B, Vol. 789, No. 2, 2003, pp. 365-372. doi:10.1016/S1570-0232(03)00162-4
[14] Y. Dotsikas, C. K. Markopoulou, J. E. Koundourellis and Y. L. Loukas, “Validation of a Novel LC-MS/MS Method for the Quantitation of Colistin A and B in Human Plasma,” Journal of Separation Science, Vol. 34, No. 1, 2011, pp. 37-45. doi:10.1002/jssc.201000680
[15] J. W. Kang, T. Vankeirsbilck, A. Van Schepdael, J. Orwa, E. Roets and J. Hoogmartens, “Analysis of Colistin Sulfate by Capillary Zone Electrophoresis with Cyclodextrins as Additive,” Electrophoresis, Vol. 21, No. 15, 2000, pp. 3199-3204. doi:10.1002/1522-2683(20000901)21:15<3199::AID-ELPS3199>3.0.CO;2-8
[16] H. K. Kristensen, and S. H. Hansen, “Separation of Polymyxins by Micellar Electrokinetic Capillary Chromatography,” Journal of Chromatography, Vol. 628, 1993, pp. 309-315. doi:10.1016/0021-9673(93)80013-X
[17] S. Ahuja and N. Jespersen, “Modern Instrumental Analysis,” Elsevier Pub., Amsterdam, 2006.
[18] N. Simeon, E. Chatelut, P. Canal, M. Nertz and F. Couderc, “Anthracycline Analysis by Capillary Electrophoresis: Application to the Analysis of Daunorubicine in Kaposi Sarcoma Tumor,” Journal of Chromatography A, Vol. 853, No. 1-2, 1999, pp. 449-454.
[19] M. Brinkley, “A Brief Survey of Methods for Preparing Protein Conjugates with Dyes, Haptens and Crosslinking Reagents,” Bioconjugate Chemistry, Vol. 3, No. 1, 1992, pp. 2-13. doi:10.1021/bc00013a001
[20] Y. Huang, J. P. Duan, Q. R. Chen and G. N. Chen, “Micellar Electrokinetic Chromatography of Enkephalin-Related Peptides with Laser-Induced Fluorescence Detection,” Electrophoresis, Vol. 25, No. 7, 2004, pp. 1051- 1057. doi:10.1002/elps.200405810
[21] P. P. H. Le Brun, A. I. De Graaf and A. Vinks, “High-Performance Liquid Chromatographic Method for the Determination of Colistin in Serum,” Therapeutic Drug Monitoring, Vol. 22, No. 5, 2000, pp. 589-593. doi:10.1097/00007691-200010000-00014
[22] Y. M. Liu, M. Schneider, C. M. Sticha, T. Toyooka and J. V. Sweedler, “Separation of Amino Acid and Peptide Stereoisomers by Nonionic Micelle-Mediated Capillary Electrophoresis after Chiral Derivatization,” Journal of Chromatography A, Vol. 800, No. 2, 1998, pp. 345-354. doi:10.1016/S0021-9673(97)01137-0
[23] A. B. Wey and W. Thormann, “Capillary Electrophoresis-Electrospray Ionization Ion Trap Mass Spectrometry for Analysis and Confirmation Testing of Morphine and Related Compounds in Urine,” Journal of Chromatography A, Vol. 916, No. 1-2, 2001, pp. 225-238. doi:10.1016/S0021-9673(00)01096-7
[24] F. A. Elbarbry and A. S. Shoker, “Liquid Chromatographic Determination of My-cophenolic Acid and Its Metabolites in Human Kidney Trans-plant Plasma: Pharmacokinetic Application,” Journal of Chro-matography B-Analytical Technologies in the Biomedical and Life Sciences, Vol. 859, No. 1, 2007, pp. 276-281. doi:10.1016/j.jchromb.2007.09.036
[25] F. Elbarbry, K. Wilby and J. Alcorn, “Validation of a Hplc Method for the Determination of P-Nitrophenol Hy-droxylase Activity in Rat Hepatic Microsomes,” Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, Vol. 834, No. 1-2, 2006, pp. 199-203. doi:10.1016/j.jchromb.2006.02.006

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