Determination of atropine enantiomers in ophthalmic solutions by liquid chromatography using a Chiral AGP column.
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 20 degrees C. 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 14.0-26.0 microg/mL, the method was found to be linear (r > 0.9999), accurate (with recovery of 100.1-100.5%), and precise (RSD system < or = 0.6%; RSD intraday < or = 1.1%; RSD interday < or = 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 2 degrees C of column temperature. The complete validation, including stress testing and factorial design, was studied and is presented in this research.
Soares
Renata
R
University of São Paulo, Department of Pharmacy, Faculty of Pharmaceutical Sciences, CP 66083, CEP 05315-970, São Paulo, Brazil.
Singh
Anil Kumar
AK
Kedor-Hackmann
Erika Rosa Maria
ER
Santoro
Maria Inês Rocha Miritello
MI
D001285:Atropine / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D007202:Indicators and Reagents; D009184:Mydriatics / Q000032:analysis*; D009883:Ophthalmic Solutions / Q000032:analysis; D009961:Orosomucoid / Q000737:chemistry*; D015203:Reproducibility of Results; D013237:Stereoisomerism
D007202:Indicators and Reagents; D009184:Mydriatics; D009883:Ophthalmic Solutions; D009961:Orosomucoid; D001285:Atropine