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.
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.
Enrique
Montse
M
Unit of Pharmaceutical Technology, Pharmacy and Pharmaceutical Technology Department, School of Pharmacy, University of Barcelona, Avda Joan XXIII s/n 08028. Barcelona, Spain.
García-Montoya
Encarna
E
Miñarro
Montserrat
M
Orriols
Anna
A
Ticó
Joseph Ramon
JR
Suñé-Negre
Joseph Maria
JM
Pérez-Lozano
Pilar
P
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D009883:Ophthalmic Solutions / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014640:Vancomycin / Q000032:analysis*
D009883:Ophthalmic Solutions; D014640:Vancomycin