THE COMBINED EFFECTS OF PH AND ACETONITRILE COMPOSITION ON THE SEPARATION OF TWO LINCOSAMIDE ANTIBIOTICS
Reversed- phase chromatography has become the method of choice to separate compounds which are similar in structure to each other. Selectivity, resolution and retention time can be improved by controlling different variants. The choice of the mobile phase for the analysis is important to achieve the best chromatographic results. The retention time, shape and width of chromatographic peaks are dependent on the buffer pH and also on the amount of organic modifier in the mobile phase. The aim of this study is to separate two lincosamide antibiotics- clindamycin phosphate and lincomycin hydrochloride by using different mobile phases. Each mobile phase was made up of a phosphate buffer and acetonitrile. The mobile phases had different buffer pHs and different percentage compositions of acetonitrile. Analysis was carried out using nine different mobile phases to observe which of them gave the best results in terms of resolution, retention time and peak shape. The best results were given when using a mobile phase having 30% acetonitrile with a buffer pH of 3.0. This work shows the importance of adjusting the amount of the organic modifier in the mobile phase together with the buffer pH. The amount of organic modifier should not be too large as to cause loss of resolution between two neighbouring peaks and not too small so as to result in too long of a retention time. The pH should be chosen to have the greatest percentage of ionised or unionised species of the analyte of interest present.
Rafferty JL, Siepmann JI, Schure MR. Mobile phase effects in reversed-phase liquid chromatography: A comparison of acetonitrile/water and methanol/water solvents as studied by molecular simulation. J Chromatogr A 2011;1218(6):2203-13.
Schoenmakers PJ, van Molle S, Hayes CM, Uunk LG. Effects of pH in reversed- phase liquid chromatography. Anal Chim Acta 1991;250:1 19.
Mendham J, Denney RC, Barnes JD, Thomas MJ. Vogelâ€™s Textbook of Quantitative Chemical Analysis. 6th ed. Essex: Pearson Education Ltd.; 2000.
McMaster MC. HPLC. A practical Userâ€™s Guide. New York: VCH Publishers; 1994.
Hendriks G. Theoretical models in LC based bioanalytical method development. J Pharm Biomed Anal 2009;49(1):1-10.
Lopes Marques RM, Schoenmakers PJ. Modelling retention in reversed- phase liquid chromatography as a function of pH and solvent composition. J Chromatogr A 1992;592(1-2):157-82.
Galaon T, David V. The influence of mobile phase pH on the retention and selectivity of related basic compounds in reversed - Phase liquid chromatography. Rev Roum Chim 2012;57:131-40.
Zambrano D. Clindamycin in the Treatment of Human Infections. Michigan: Pharmacia and Upjohn; 1997.
Verdier L, Bertho G, Gharbi-Benarous J, Girault JP. Lincomycin and clindamycin conformations. A fragment shared by macrolides, ketolides and lincosamides determined from TRNOE ribosome-bound conformations. Bioorg Med Chem 2000;8(6):1225-43.
Du Bok C, Seong Ryeol R, On You C, Pemin Y, Kyu - Hyuk K. Recovery and purification of lincomycin from the culture broth of Streptomyces lincolnensis. J Ind Eng Chem 2005;11(6):932-7.
Williams DN, Crossley K, Hoffman C, Sabath LD. Parenteral clindamycin phosphate: Pharmacology with normal and abnormal liver function and effect on nasal staphylococci. Antimicrob Agents Chemother 1975;7(2):153-8.
Silverman RB. The Organic Chemistry of Drug Design and Drug Action. 2nd ed. Amsterdam: Elsevier Academic Press; 2004.
Stella VJ, Charman WN, Naringrekar VH. Prodrugs. Do they have advantages in clinical practice. Drugs 1985;29:455-73.
Herr RR, Slomp G. Lincomycin. II. Characterization and gross structure. J Am Chem Soc 1967;89(10):2444-7.
Stella V, Borchardt R, Hageman M, Oliyai R, Maag H, Tilley J. Prodrugs: Challenges and Rewards. Part 2. New York: Springer; 2007.
Espinosa S, Bosch E, RosÃ©s M, ValkÃ³ K. Change of mobile phase pH during gradient reversed-phase chromatography with 2,2,2-trifluoroethanol-water as mobile phase and its effect on the chromatographic hydrophobicity index determination. J Chromatogr A 2002;954(1-2):77-87.
Barbosa J, Toro I, BergÃ©s R, Sanz-Nebot V. Retention behaviour of peptides, quinolones, diuretics and peptide hormones in liquid chromatography. Influence of ionic strength and pH on chromatographic retention. J Chromatogr A 2001;915(1-2):85-96.
Canals I, Oumada FZ, RosÃ©s M, Bosch E. Retention of ionizable compounds on HPLC 6. pH measurements with the glass electrode in methanol-water mixtures. J Chromatogr A 2001;911(2):191-202.
RosÃ©s M, Oumada FZ, Bosch E. Retention of ionizable compounds on high-performance liquid chromatography. VII. Characterization of the retention of ionic solutes in a C18 column by mass spectrometry with electrospray ionization. J Chromatogr A 2001;910(2):187-94.
Ahuja S, Rasmussen H. HPLC Method Development for Pharmaceuticals. Amsterdam: Elsevier Academic Press; 2007.
Poole CF, Poole SK. Chromatography Today. Amsterdam: Elsevier Science; 1991.
Snyder LR, Kirkland JJ, Glajch JL. Practical HPLC Method Development. New York: John Wiley and Sons; 1997.
Sanli S, Sanli N, Alsancak G. Spectrophotometric determination of acidity constants of some macrolides in acetonitrile-water binary mixtures. Acta Chim Slov 2010;57(4):980-7.
Christian GD. Analytical Chemistry. 5th ed. New York: John Wiley and Sons; 1994.
Pragst F, Herzler M, Herre S, Erxleben BT, Rothe M. UV Spectra of TOXIC Compounds. London: Pharmaceutical Press; 2003.
Goeringer KE, McIntyre lM, Drummer OH. LC-MS analysis of serotonergic drugs. J Anal Toxicol 2003;27(1):30-5.
Kirkland JJ, van Straten MA, Claessens HA. High pH mobile phase effects on silica- based reversed- phase high- performance liquid chromatographic columns. J Chromatogr A 1995;691:3-19.
Amiri AA, Hemmateenejad B, Safavi A, Sharghi H, Beni AR, Shamsipur M. Structure-retention and mobile phase-retention relationships for reversed-phase high-performance liquid chromatography of several hydroxythioxanthone derivatives in binary acetonitrile-water mixtures. Anal Chim Acta 2007;605(1):11-9.
BergÃ©s R, Sanz-Nebot V, Barbosa J. Modelling retention in liquid chromatography as a function of solvent composition and pH of the mobile phase. J Chromatogr A 2000;869(1-2):27-39.
Espinosa S, Bosch E, RosÃ©s M. Retention of ionizable compounds in high-performance liquid chromatography. IX. Modelling retention in reversed-phase liquid chromatography as a function of pH and solvent composition with acetonitrile-water mobile phases. J Chromatogr A 2002;947(1):47-58.
Espinosa S, Bosch E, RosÃ©s M. Retention of ionizable compounds in high-performance liquid chromatography 14. Acid-base pK values in acetonitrile-water mobile phases. J Chromatogr A 2002;964(1-2):55 66.
Siddartha B, Sudheer Babu I, Ravichandra Gupta CH, Parthiban C. Analytical method development and validation for simultaneous estimation of tamsulosin and tolterodine in bulk and pharmaceutical dosage form by RP-HPLC method. Asian J Pharm Clin Res 2014;7(2):156-60.
Kabra P, Nargung LV, Srinivasa Murthy M. RP- HPLC method for estimation of an antipsychotic drug - Pimozide. Asian J Pharm Clin Res 2014;7(4):49-51.
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