Development of a Validated HPLC Method for the Estimation of Metformin HCl and Propranolol HCl
Venkata Srikanth Meka *
School of Pharmacy, International Medical University, Kuala Lumpur, Malaysi and Department of Pharmacy, A.U. College of Pharmaceutical Sciences, Andhra University, Visakhapatnam-530003, India.
Janaki Ram Battu
Department of Pharmacy, Govt. Polytechnic, Visakhapatnam, India.
Senthil Rajan Dharmalingam
School of Pharmacy, International Medical University, Kuala Lumpur, Malaysia.
Adinarayana Gorajana
School of Pharmacy, International Medical University, Kuala Lumpur, Malaysia.
Venkata Ramana Murthy Kolapalli
Department of Pharmacy, A.U. College of Pharmaceutical Sciences, Andhra University, Visakhapatnam-530003, India.
*Author to whom correspondence should be addressed.
Abstract
The present investigation targets to develop a simple, specific, sensitive and accurate reverse phase high performance liquid chromatographic (RP-HPLC) method in human plasma for the estimation of metformin HCl and propranolol HCl from bulk drug and also from the marketed products. Human plasma samples were subjected to correct procedure for protein precipitation by methanol and protein free plasma samples were directly injected into HPLC C18 column. Chromatographic determination was performed on a reversed phase C18 column (3.9 mm X 300 mm, particle size 5 µm) using a mixture of acetonitrile and 0.1M pH 4.5 potassium dihydrogenortho phosphate buffer (mL) (40:60) at a flow rate of 0.8 mL/min and maintained at a pressure of 140 to 150 Kg/cm2. The retention time for metformin HCl and propranolol HCl was found to be 9.084 min and 6.132 min respectively at 232 nm without any interference of endogenous compounds in the plasma. The method was linear in the range between 50-2000 ng/mL. The peak areas were reproducible as indicated by low coefficient of variation. It was found that the excipients in the tablet dosage form do not interfere in the quantification of active drug by proposed method.
Keywords: Metformin HCl, propranolol HCl, validation, HPLC method, human plasma, accuracy, precision.