The method development and validation was done by using waters Alliance 2895 seperation model with a PDA detector 996model.the data was acquired using Enpower 2 software and the column used was Hypersil C18, 150 x4.6mm,5
Geftinib is not official in any Pharmacopoeia. A literature survey on Geftinib revealed that, until now no analytical method was reported for its determination in bulk drug and its capsule dosage form.Since literature did not cite any method for determination of this drug from bulk drug as well as its formulation it was proposed to develop and validate a method by using RP-HPLC method(2).
Instrumentation and analytical conditions:
The method development and validation was done by using Waters Alliance 2895 seperation model with a PDA detector 996model.The data was acquired using Enpower 2 software and the column used was Hypersil C18, 150 x4.6mm,5µ. Labman Digital Ultra sonicator was used for enhancing dissolution of the drug. A Lab india make pH meter was used for pH adjustment.
Chemicals and Reagents:
Pure Geftinib citrate gift sample was obtained from Sura Pharma Reaseach Lab ,Hyderbaded. HPLC grade water, Methanol-HPLC,Acetonitrile-HPLC from Merck ,Mumbai.. Labman Digital Ultra sonicator was used for enhancing dissolution of the drug. A Lab india make pH meter was used for pH adjustment.
Preparation of Mobile phase ,Standard and Sample preparations:
Preparation of Mobile phase:
Accurately measure 150 ml (15%) of HPLC Methanol and 850 ml of HPLC Water (85%) were mixed and degassed with a digital ultrasonicater for 10 minutes and then filtered through 0.45 µ filter under vacuum filtration.The Mobile phase was used as the diluent.
Preparation of Standard solution:
Weigh accurately 10 mg of Geftinib citrate and transfer into a clean, dry 10ml volumetric flask and add 7ml of Methanol, sonicate to dissolve and removal of air completely and make the volume upto the mark with the diluent.Further pipette out 0.75ml of the above Geftinib stock solution into a 10ml volumetric flask and dilute up to the mark with diluent and filtered through 0.45µm membrane filter.
Preparation of Sample solution:
Each capsule was weighed and powered by using mortor and pestle and 10mg equivalent weight of Geftinibcitrate sample was weighed and transferred into 10ml of clean ,dry volumetric flask. Then add 7ml of diluent and sonicate to dissolve for 20min. The volume was made upto 10ml with diluent.
Pipette out 0.75ml of the above solution into a 10ml volumetric flask and dilute up to the mark with diluent and filtered through 0.45µm membrane filter.
Procedure:
Inject the samples by changing the chromatographic conditions and record the chromatograms, note the conditions of proper peak elution for performing validation parameters as per ICH guidelines.
Mobile Phase Optimization:
Initially the mobile phase tried was Water, Methanol: Water and Acetonitrile: Water with varying proportions. Finally, the mobile phase was optimized to Methanol: Water in proportion 15:85 v/v respectively.
Optimization of Column:
The method was carried out with various C18 columns like Phenomenex column, Xterra, and Symmetry C18 column. Hypersil C18 (4.6 x 150mm, 5µm) was found to be ideal as it gave good peak shape and resolution at 1.0ml/min flow The sample was injected using 10 µl fixed loop and PDA detector was set at a wavelength of 284nm and the run time is 6min.