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Enantioseparation of Tedizolid phosphate by RP-HPLC, using 𝛽-Cyclodextrin as a Chiral Mobile Phase Additive

Ajit Anerao, Vikram Dighe, Satish John, Nitin Pradhan.


Enantiomeric separation of Tedizolid phosphate was developed using a reversed phase HPLC (RP-HPLC) with 𝛽-cyclodextrin (𝛽-CD) as a chiral mobile phase additive. Effect of the pH value of aqueous buffer, concentration of chiral additive, composition of mobile phase, and column temperature on the enantioseparation of Tedizolid phosphate was investigated on the Phenomenex Luna, Phenyl-Hexyl, 250 x 4.6mm, 5μm HPLC column. A satisfactory resolution was achieved at column temperature 20⁰C using a mobile phase consisting of a mixture of aqueous buffer of pH 7.0 of disodium hydrogen phosphate with additive 𝛽-cyclodextrin, triethylamine and acetonitrile. This analytical method was evaluated by performing method validation as per ICH guideline. The calibration curve was plotted within the concentration range between 0.30 and 2.25 µg mL−1 and the recoveries between 96.9 and 105.3% were obtained, with regression coefficient R2 is 0.998. The limit of detection (LOD) and limit of quantitation (LOQ) of S- enantiomer of Tedizolid phosphate is 0.10 and 0.30 μg mL−1 The developed method was demonstrated to be accurate, robust and sensitive for the determination of enantiomeric purity of Tedizolid phosphate drug substance.

Key words: Tedizolid phosphate; enantiomer; cyclodextrin; method development

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