title: Bioanalytical method development and validation of natamycin in rabbit tears and its application to ocular pharmacokinetic studies.
abstract: A new selective and sensitive high-performance liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method was developed for the quantification of natamycin in rabbit tears using amphotericin B as internal standard (IS). Chromatographic separation was achieved on a Luna Cyano column (100 mm × 2 mm, 3 μm) using ammonium acetate buffer (pH 4; 3.5mM): methanol (10:90, v/v) as the mobile phase. The run time was 5 min. Detection was performed by negative ion electrospray ionization in multiple reaction monitoring (MRM) mode. The calibration curve was linear over the concentration range from 25 to 800 ng/ml, and lower limit of detection of 12.5 ng/ml. The accuracy and precision of the method were within the acceptable limit of ± 20% at the lower limit of quantitation and ± 15% at other concentrations. Natamycin was stable during the battery of stability studies viz., bench-top, auto-sampler, freeze/thaw cycles and 30 days storage in a freezer at -70 ± 10 °C. The method was successfully applied to the ocular pharmacokinetic studies of natamycin eye drops in New Zealand rabbit tears.
authors.affiliation: Pharmacokinetics and Metabolism Division, Central drug Research Institute, CSIR, Lucknow 226001, India. rabi.cdri@gmail.com
authors.forename: R S
authors.initials: RS
authors.lastname: Bhatta
authors.forename: H
authors.initials: H
authors.lastname: Chandasana
authors.forename: C
authors.initials: C
authors.lastname: Rathi
authors.forename: D
authors.initials: D
authors.lastname: Kumar
authors.forename: Y S
authors.initials: YS
authors.lastname: Chhonker
authors.forename: G K
authors.initials: GK
authors.lastname: Jain
mesh_terms: D000666:Amphotericin B; D000818:Animals; D000935:Antifungal Agents / Q000008:administration & dosage / Q000032:analysis* / Q000493:pharmacokinetics*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004305:Dose-Response Relationship, Drug; D016903:Drug Monitoring / Q000379:methods*; D004355:Drug Stability; D015394:Molecular Structure; D010866:Natamycin / Q000008:administration & dosage / Q000032:analysis* / Q000493:pharmacokinetics*; D009883:Ophthalmic Solutions; D011817:Rabbits; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods; D013666:Tears / Q000737:chemistry*; D013997:Time Factors
chemical_list: D000935:Antifungal Agents; D009883:Ophthalmic Solutions; D000666:Amphotericin B; D010866:Natamycin