Development and validation of stability indicating RP-HPLC method for Simultaneous Estimation of Imipramine Hydrochloride and Diazepam in combined pharmaceutical dosage form
Keywords:Diazepam Imipramine HCl, RP HPLC, anxiolytic, Antidepressant
A simple, precise, and stability indicating high performance liquid chromatography (HPLC) method was developed and validated for the simultaneous determination of Imipramine hydrochloride and Diazepam in pharmaceutical dosage form.
Materials and Methods: The method involves the use of easily available inexpensive laboratory reagents. The separation was achieved on Phenomenex Prodigy C-18 column (150*4.6 mm, i.d., 5 ?m particle size) with isocratic flow with UV detector. The mobile phase at a flow rate of 1.0 mL/min consisted of methanol and 0.1%v/v ortho phosphoric acid (pH 3) in the ratio of 70:30 v/v.
Results: A linear response was observed over the concentration range 5.00-50.00 ?g/mL of diazepam and the concentration range 5.00-50.00 ?g/mL of Imipramine HCl. Limit of detection and limit of quantitation for Imipramine were 2.73 ?g/mL and 1.27 ?g/mL, and for Diazepam were 1.80 ?g/mL and 1.98 ?g/mL, respectively. The method was successfully validated in accordance to ICH guidelines acceptance criteria for linearity, accuracy, precision, specificity, robustness.
Conclusion: The analysis concluded that the method was selective for simultaneous estimation of Imipramine HCl and Diazepam can be potentially used for the estimation of these drugs in combined dosage form.
Maj J, Miroslawa M, Ewa M, Wladyslawa D. Different pharmacokinetic and pharmacological effects following acute and chronic treatment with imipramine. J Neu Trans 1982; 54(3-4): 219-228.
Klotz U, Antonin KH, Bieck PR. Pharmacokinetics and plasma binding of diazepam in man, dog, rabbit, guinea pig and rat. J Pharmacol Exp Ther 1976; 199: 67-73.
Choudhury T, Ghosh J, Bagchi D. Reverse phase high performance liquid chromatographic method and method validation of imipramine by using single mobile phase. Int J Pharma Sci Tech 2010; 4(1): 54-62.
Rouini MR, Ardakani YH. An improved HPLC method for rapid quantitation of diazepam and its major metabolites in human plasma. Talanta 2008; 75: 671–676.
Moghaddam KA, SolataniUddin MNF, Samanidou VF, Papadoyannis IN. Development and validation of an HPLC method for the determination of benzodiazepines and tricyclic antidepressants in biological fluids after sequential SPE. J Sep Sci 2008, 31(13): 2358-70.
Samanidou VF, Nika MK, Papadoyannis IN. Development of an HPLC method for the monitoring of tricyclic antidepressants in biofluids. J Sep Sci 2007; 30(15): 2391-400.
Surya Prakash G, Neeraj U, Gopal G. Development and validation of spectrophotometric, HPTLC and HPLC methods for the determination of Imipramine and Chlordiazepoxide in pharmaceutical dosage forms. Der Pharmacia Sinica 2012; 3(2): 185-192.
Patel S. Development and Validation of HPTLC Method for Simultaneous Determination of Diazepam and Propranolol Hydrochloride in Tablet Dosage Form. Int J Pharma Front Res 2011; 1(3): 29-37.
Lavinia G. Hinescu, Cristina M. Ranetti, Mihaela I, Elena I, Constantin D, Constantin M, Cristiana C, Victor AV. Hplc method for the simultaneous determination of the components of an aqueous antidote solution. Farmacia 2011; 59(1): 97-105.
Tania VM, Maria JR, Quezia BC, Maria ET. Development and Optimization of a HPLC–DAD Method for the Determination of Diverse Pharmaceuticals in Estuarine Surface Waters. J Chromato Sci 2010, 48: 176-182.
Tulja Rani G, Gowri shankar D, Kadgapathi P and Satyanarayana B. Development of an RP-HPLC Method for the Simultaneous Estimation of Propranolol Hydrochloride and Diazepam in Combined Dosage form. Ind J Pharm Edu Res 2011; 45(4): 296-300.
Mohammad NU, Victoria FS, Ioannis NP. Simultaneous determination of 1,4-benzodiazepines and tricyclic antidepressants in saliva after sequential spe elution by the same hplc conditions. J Chinese Chem Soc 2011, 58: 142-154.
Wael AD, Ahmad AH, Kamal S, Khalid M, Eyad AN. Simultaneous high performance liquid chromatographic analysis of oxicams in pharmaceutical formulations. Int J Pharm 2012; 2(4): 687-695.
Ashraful ISM, Shamima S, Muhammad SBS, Irin D. Uv-spectrophotometric and rp-hplc methods for the simultaneous estimation of acetaminophen and caffeine: validation, comparison and application for marketed tablet analysis. Int J Pharm 2012; 2(1): 39-45.
Palleshwar Rao G, Rao JVLNS, Lanka ARP, Srinivasu P. Development and validation of a new stability indicating hplc method for quantification of process related and degradation
impurities of bicalutamide in tablet dosage forms. Int J Pharm 2012; 2(1): 218-223.
ICH, Q2 (R1) Validation of Analytical Procedure: Test and Methodology, International Conference on Harmonization, Geneva, 2005.
ICH, Q2A Validation of Analytical Procedures: Consensus Guidelines; ICH Harmonized Tripartite Guidelines, 1994.
ICH, Q2B Validation of Analytical Procedures: Methodology, Consensus Guidelines ICH Harmonized Tripartite Guidelines, 1996
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