Multivariate Optimization and Validation of HPLC Method for Determination of Spiramycin I in Tablets

Spiramycin (SPY) is an antibiotic belonging to the class of macrolides. Its main structurally related components are SPY I, SPY II and SPY III. The aim of this work was to optimize and validate a simple and fast method of quantification of SPY I in tablets in the presence of SPYs II and III by HPLC based on multivariate approach. Initially, different conditions were tested by means of a complete factorial design 2 3 . Afterward, the chromatographic parameters were optimized using the Doehlert Matrix and response surface methodology, which allowed to obtain an efficient separation for SPY I with relatively short retention time (16 min). The new method was validated using phosphoric acid 0.1% and methanol (67:33, v/v) as mobile phase, flow rate at 1.0 mL min −1 , C8 column (250 mm × 4.6 mm × 5 µm) as stationary phase and UV detection at 232 nm. The method proved to be selective, linear, accurate, with good reproducibility, robust, useful, simple, and efficient for the quantification of SPY I in pharmaceutical tablets. Thus, multivariate approach proved to be an excellent strategy for method optimization with a reduced number of experiments without the need for additional cost with expensive equipment or chromatographic columns.