Presentation Title

DEVELOPMENT AND VALIDATION OF HPLC METHOD WITH PDA DETECTOR FOR SIMULTANEOUS QUANTIFICATION OF CO-ELUTED COMPOUNDS IN PHARMACEUTICAL SAMPLES

Location

Atrium

Format

Event

Start Date

14-2-2014 12:00 AM

Abstract

Objective. To develop and validate an HPLC method for the separation and quantification of co-eluted dipivefrine, a prodrug, and its active form, epinephrine, in a standardized pharmaceutical sample. Background. The current USP methods for determination of individual compounds are not suitable for their detection when co-eluted. To our knowledge, there is no published analytical method using HPLC with Photo-Diode-Array (PDA) detector for the simultaneous detection of the co-eluent which is required for the development of novel dipivefrine dosage form. Methods. Different strengths of mobile phases were evaluated using methanol and acetonitrile. Several reversedphase C18 and Hilic columns at various pressures and flow rates using Waters Alliance System with PDA detector were evaluated. A 2.0 mg/mL standard stock solution of each compound was prepared in 0.0015N HCL solvent. Standard calibration curves ranging from 20 - 200¼g/mL were injected and plotted for both compounds. The linearity of the injections, the reproducibility of quantification, the minimum detection limit, and the intra and inter assay variation for each compound were tested and validated. Results. Separation was achieved using C18 column 43 (3.9mm X 300mm, 10¼m) with a flow rate of 2.0 ml/min at wavelength of 254 nm at ambient temperature. (More data are being collected and will be presented in the poster). Conclusion. A reliable and sensitive reversed-phase HPLC method was developed and validated for the quantification of dipivefrine and epinephrine from one sample. (Project in progress, more data will be provided to confirm conclusion). Grants. President's Faculty Research & Development Grant FY2014.

This document is currently not available here.

Share

COinS
 
Feb 14th, 12:00 AM

DEVELOPMENT AND VALIDATION OF HPLC METHOD WITH PDA DETECTOR FOR SIMULTANEOUS QUANTIFICATION OF CO-ELUTED COMPOUNDS IN PHARMACEUTICAL SAMPLES

Atrium

Objective. To develop and validate an HPLC method for the separation and quantification of co-eluted dipivefrine, a prodrug, and its active form, epinephrine, in a standardized pharmaceutical sample. Background. The current USP methods for determination of individual compounds are not suitable for their detection when co-eluted. To our knowledge, there is no published analytical method using HPLC with Photo-Diode-Array (PDA) detector for the simultaneous detection of the co-eluent which is required for the development of novel dipivefrine dosage form. Methods. Different strengths of mobile phases were evaluated using methanol and acetonitrile. Several reversedphase C18 and Hilic columns at various pressures and flow rates using Waters Alliance System with PDA detector were evaluated. A 2.0 mg/mL standard stock solution of each compound was prepared in 0.0015N HCL solvent. Standard calibration curves ranging from 20 - 200¼g/mL were injected and plotted for both compounds. The linearity of the injections, the reproducibility of quantification, the minimum detection limit, and the intra and inter assay variation for each compound were tested and validated. Results. Separation was achieved using C18 column 43 (3.9mm X 300mm, 10¼m) with a flow rate of 2.0 ml/min at wavelength of 254 nm at ambient temperature. (More data are being collected and will be presented in the poster). Conclusion. A reliable and sensitive reversed-phase HPLC method was developed and validated for the quantification of dipivefrine and epinephrine from one sample. (Project in progress, more data will be provided to confirm conclusion). Grants. President's Faculty Research & Development Grant FY2014.