Application of a New Strategy of Validation Based on "β,γ-Content Tolerance Interval" for Checking the Chiral Chromatography Method for Quantification of the Chiral Impurity of Levofloxacin

A simple and accurate chiral high performance liquid chromatography (HPLC) method was developed for determination of Levofloxacin and its chiral impurity (R)-enantiomer. The chiral impurity Levofloxacin separation method was optimized by Box-Behnken design and validated by an innovative analytical approach recently developed in our laboratory. This last approach as well as uncertainty estimation consists of building a graphical decision-making tool, based on the β,γ-content tolerance interval, called the uncertainty profile. The proposal approach applied to the optimized chiral separation method of Levofloxacin shows its fitness.