P.Palanisamy, B.Jayakar, M.Kumar, R.Margret Chandra, B.S.Venkateshwralu, A.Pasupathi, M.Vkumudhavalli.
Objective: A simple, efficient and reproducible Reverse Phase-High Performance Liquid Chromatography (RP-HPLC) method for determination of Diclofenac sodium,Vitamin B1,Vitamin B6, and Vitamin B12 in soft gelatin capsule pharmaceutical formulation has been developed.
Method & Result: The Diclofenac Sodium, the separation was carried out in Phenomenex ODS C18 column (150 x 4.6mm; 5μm) using mobile phase consisting of mixture of 400mL potassium hydrogen phosphate (pH 3.0) and 600mL of Acetonitrile. The flow rate was 1.0mL/min and effluent was detected at 254nm. The retention time of Diclofenac sodium was 5.4 min. The percentage recovery was within the range between 100.00% and 101.86% for Diclofenac Sodium. The linearity range was found to be 20-400μg/mL (r2 =1) for Diclofenac Sodium. The Vitamin B1 & Vitamin B6, the separation was carried out in Inertsil ODS C18 column (250 x 4.6mm; 5μm) using mobile phase consisting of mixture of 1000mL of Sodium 1-hexane sulfonic acid solution , 730mL of water , 270mL glacial acetic acid and 10ml of methanol The flow rate was 1.5mL/min and effluent was detected at 280nm. The retention time of Vitamin B1 & Vitamin B6 were 3.70 min & 7.74 min respectively. The percentage recovery was within the range between 99.46% and 101.03% & 99.35 and 101.78 respectively for Vitamin B1 & Vitamin B6. The linearity range was found to be 10-200μg/mL (r2 =1) for Vitamin B1 and for Vitamin B6 the linearity range was found to be 10.5-210μg/mL. For Vitamin B12, the separation was carried out in Phenomenex ODS C18 column (150 x 4.6mm; 5μm) using mobile phase consisting of mixture of .25mL trifluroacetic acid, 870mL of water and 130mL of Acetonitrile. The flow rate was 1.2mL/min and effluent was detected at 361nm.The retention time of Vitamin B12 was 6.75 min. The percentage recovery was within the range between 98.86% and 101.94% for Vitamin B12 .The linearity range was found to be 1-20μg/mL (r2 =0.9999) for Vitamin B12. The obtained chromatograms showed good resolutions between the peaks which helps to quantify the drugs in well-defined manner.
Conclusion: The Proposed study describes new and simple RP-HPLC method for the estimation of Diclofenac Sodium,Vitamin B1,Vitamin B6 and Vitamin B12 in soft gelatin capsule dosage form. The method was validated as per ICH guidelines and found to be simple, sensitive, accurate and precise. Therefore the proposed method can be successfully used for the routine lab analysis of estimation of Diclofenac Sodium,Vitamin B1,Vitamin B6, and Vitamin B12 in soft gelatin capsule dosage form.
