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Many therapeutic agents are commercialized under their racemic form. The enantiomers can show differences in the pharmacokinetic and pharmacodynamic profile. The use of a pure enantiomer in pharmaceutical formulations may result in a better therapeutic index and fewer adverse effects. Atropine, an alkaloid of Atropa belladonna, is a racemic mixture of l-hyoscyamine and d-hyoscyamine. It is widely used to dilate the pupil. To quantify these enantiomers in ophthalmic solutions, an HPLC method was developed and validated using a Chiral AGP column at 20C. The mobile phase consisted of a buffered phosphate solution (containing 10 mM 1-octanesulfonic acid sodium salt and 7.5 mM triethylamine, adjusted to pH 7.0 with orthophosphoric acid) and acetonitrile (99 + 1, v/v). The flow rate was 0.6 mL/min, with UV detection at 205 nm. In the concentration range of 14.026.0 g/mL, the method was found to be linear (r &amp;gt; 0.9999), accurate (with recovery of 100.1100.5), and precise (RSD system 0.6; RSD intraday 1.1; RSD interday 0.9). The method was specific, and the standard and sample solutions were stable for up to 72 h. The factorial design assures robustness with a variation of 10 in the mobile phase components and 2C of column temperature. The complete validation, including stress testing and factorial design, was studied and is presented in this research.

Determination of Atropine Enantiomers in Ophthalmic Solutions by Liquid Chromatography Using a Chiral AGP Column