SEPARATION AND DETERMINATION OF THE R-ISOMER OF ETODOLAC IN A BULK DRUG SUBSTANCE BY NORMAL-PHASE LIQUID CHROMATOGRAPHY

ABSTRACT Objective:  The objective of the this work is to develop and validate a novel, simple,rapid and reliable analytical method for separation and determination of R-isomer impurity in Etodolac bulk drug material by normal-phase high-performance liquid chromatography as per International Conference on Harmonization guidelines. Methods: The Etodolac R- isomer and S-isomer were separated on a Chiralcel OD-H (150 x 4.0 mm, 5 micron) column by using Ethanol : n-Hexane:Trifluoroacetic acid (50:50:0.1 v/v.) mobile phase with equipped detector at wavelength 225 nm and 25 °C column oven temperature. The resolution between R-isomer and S-isomer were more than two recorded on chromatogram. The specified method was developed and validated for various parameters like reproducibility, limit of detection, limit of quantification, linearity and range, robustness, solution stability and mobile phase stability according to the International Conference on Harmonization (ICH) guidelines.  Results: Linearity were found for Etodolac R-isomer over the concentration range of 600–6000 ng/ml, with the linear regression (Correlation coefficient R = 0.998) and proved to be robust. Limit of detection and limit of quantification of Etodolac R-isomer was found to be 200 and 600 ng/ml. The retention time of R-isomer was considered to be 2.8 min. The percentage recovery of Etodolac R-isomer has been ranged from 97.0 to 102.0 in bulk drug material sample. The proposed analytical method has been found to be suitable, precise,reliable and accurate for the separation and quantitative determination of Etodolac R-isomer in bulk drug sample.                                                                                                                   Conclusion: A novel, speedy, accurate, precise, reliable and rugged analytical method has been developed and validated for normal phase high performance liquid chromatography to determine R-isomer impurity in Etodolac bulk drugs material as per ICH guideline. Keywords: Etodolac, HPLC, Known Impurity. Normal Phase, Validation

SEPARATION AND DETERMINATION OF THE R-ISOMER OF KETOPROFEN IN A BULK DRUG SUBSTANCE BY NORMAL PHASE LIQUID CHROMATOGRAPHY

Objective: The objective is defined to develop and validate simple, rapid, precise, and accurate for separation and determination of R-isomer impurity of ketoprofen bulk drug material by the normal phase high-performance liquid chromatographic method as per the International Conference of Harmonization Guideline (ICH) guidelines.Methods: The R-isomer and S-isomer were baseline resolved on a Chiralcel OJ-H, 150 mm × 4.6 mm, and 5 µm stationary phase column. Mobile phase system containing n-hexane:isopropyl alcohol:glacial acetic acid (50:50:0.1 v/v.). The detector wavelength has been selected 254 nm and column oven temperature 25°C. The chromatographic resolutions between R-isomer and S-isomer were more than two. The developed method was extensively validated according to ICH guidelines.Results: Ketoprofen linearity was found for R-isomer over the concentration range of 600-6000 ng/ml, with the linear regression (Correlation coefficient R=0.999) and proved to be robust. The limit of detection and limit of quantification of ketoprofen R-isomer were found to be 200 and600 ng/ml. The percentage recovery of R-isomer has been ranged from 97.5 to 102.0 in bulk drug material samples of ketoprofen. The proposed method was found to be suitable and accurate for the quantitative determination of R-isomer of ketoprofen in bulk drug sample.Conclusion: A simple, rapid, precise, and accurate normal phase liquid chromatography method has been developed and validated to determineR-isomer impurity of ketoprofen in bulk drugs material as per ICH.Keywords: Ketoprofen, High-performance liquid chromatography, Known impurity, Normal phase, Chiralcel OJ-H, Validation

RAPID SEPARATION AND DETERMINATION OF RIZATRIPTAN N-OXIDE IMPURITY IN RIZATRIPTAN BENZOATE IN A BULK DRUG SUBSTANCE BY REVERSE PHASE LIQUID CHROMATOGRAPHY

Objective: The objective is described to develop and validate a simple, reliable, precise, and specific analytical method for rapid separation and determination of N-oxide impurity in rizatriptan Benzoate active pharmaceutical ingredient bulk drug substances by reverse phase liquid chromatography as per the International Conference on Harmonization (ICH) guidelines.Methods: The methodology utilized as reverse phase liquid chromatography with gradient composition. The mobile phase proportion was compromised mobile A containing 0.25 mm potassium dihydrogen phosphate buffer pH 2.0 and methanol (95:5 v/v) and mobile B containing Acetonitrile. ODS 3V, 250 × 4.6 mm, 5 µm. column. The flow rate is 1.0 ml/minute using an LC system detector at wavelength 280 nm, and the column oven temperature is 40°C. The chromatographic separation performed in reverse phase by the gradient composer over run time was 35 minute. The resolution between N-oxide and rizatriptan was recorded on the chromatogram was more than six. The developed analytical method was validated according to the ICH guidelines.Results: Linearity was found in rizatriptan N-oxide over the concentration range of 450-11000 ng/ml, with the linear regression (Correlation coefficient R = 0.999) and proved to be robust. Limit of detection and limit of quantification of the rizatriptan N-oxide was found 150 and 450 ng/ml. The retention time of rizatriptan and rizatriptan N-oxide was recorded 22.6 and 24.7 minutes, respectively. The percentage recovery of N-oxide has been ranged from 96.0 to 102.0 in the bulk drug material sample. The proposed analytical method has been found suitable, precise, reliable, and accurate for the separation and quantitative determination.Conclusion: A specific, simple, accurate, reliable, and rapid reproducible analytical method has been developed and validated for reverse phase high- performance liquid chromatography to determine N-Oxide impurity in rizatriptan benzoate from bulk drugs material as per ICH guideline.Keywords: Rizatriptan benzoate, N-oxide impurity, High performance liquid chromatography, Reverse phase, ODS column and validation.Â