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  • Title: Application of an experimental design for the optimization and validation of a new HPLC method for the determination of vancomycin in an extemporaneous ophthalmic solution.
    Author: Enrique M, García-Montoya E, Miñarro M, Orriols A, Ticó JR, Suñé-Negre JM, Pérez-Lozano P.
    Journal: J Chromatogr Sci; 2008 Oct; 46(9):828-34. PubMed ID: 19007487.
    Abstract:
    An experimental design has been used to develop and optimize a new high-performance liquid chromatographic (HPLC) method for the determination of Vancomycin in an extemporaneous ophthalmic solution. After the preliminary studies and literature review, the optimized method was carried out on a second generation of a C18 reverse-phase column (Luna 150 x 4.6 mm i.d., 5 microm particle size) and using methanol as organic phase, a less toxic solvent than acetonitrile, described in the extended literature. The experimental design consisted of a Placket-Burman design where six different variables were studied (flow rate, mL/min; temperature, degrees C; pH mobile phase; % buffer solution; wavelength; and injection volume) to obtain the best suitability parameters (Capacity factor-K', tailing factor, resolution, and theoretical plates). After the optimization of the chromatographic conditions and statistical treatment of the obtained results, the final method uses a mixture of a buffer solution of water-phosphoric acid (85%) (99.83:0.17, v/v) adjusted to pH 3.0 using triethylamine and mixed with methanol (87:13, v/v). The separation is achieved using a flow rate of 1.0 mL/min at 35 degrees C. The UV detector was operated at 280 nm. The validation study carried out, demonstrates the viability of the method, obtaining a good selectivity, linearity, precision, accuracy, and sensitivity.
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