Development of Validated Analytical Methods for Selected Cardiovascular Drugs in Formulations and Its Application in In-vitro Interaction studies

Shivashankar, V (2015) Development of Validated Analytical Methods for Selected Cardiovascular Drugs in Formulations and Its Application in In-vitro Interaction studies. Doctoral thesis, The Tamilnadu Dr. M.G.R. Medical University, Chennai.


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The development and validation of different analytical techniques for the selected cardiovascular drugs (dronedarone hydrochloride, levosimendan, rivaroxaban and ticagrelor) were successfully carried out. All the developed methods have been validated as per ICH guideline for validation of analytical method. The analytical methods developed are having its own advantages for using them in routine quality control process. The in-vitro drug interaction of selected CVD drugs with certain coadministered drugs was carried out by applying the developed RP-HPLC method. The second order derivative UV spectroscopic method for determination of dronedarone hydrochloride is a simple and accurate method. The second order spectrum were in good resolution when compare to zero order spectrum of dronedarone hydrochloride tablets. The spectrofluroimetric method developed utilized simple oxidation method with acidic CAS solution. The developed spectrofluorimetric method is highly sensitive for the determination of dronedarone hydrochloride. The HPTLC method developed was less time consuming method and resulted excellent response during detection of dronedarone hydrochloride. The HPLC method developed utilized simple mobile phase and the retention time was less than ten minutes demonstrate faster separations of dronedarone hydrochloride. The developed HPLC method applied in determination of in-vitro protein binding interaction studies demonstrate the method is capable of separating both the study and interacting drug (aspirin/ atorvastatin). The retention times of the drugs were showing good resolution between the drug peaks and the amount of unbound dronedarone hydrochloride was determined accurately. Thus the effect of interacting drugs on protein binding of dronedarone hydrochloride was studied by the developed HPLC method. The UV spectrophtotometric method and spectrofluorimetric method developed for determination of levosimendan are very simple. The validation results of these methods demonstrate they are highly reliable. The developed HPTLC method resulted lower LOD and LOQ values of levosimendan described the greater sensitivity of the method. The HPLC method developed used simple mobile phase composition and the method is not laborious as reported methods. The stability tests carried out for levosimendan revealed the HPLC method is stability indicating one and the degradants formed did not interfere with the levosimendan retention time. The application of HPLC method for in-vitro interaction studies has proven it is appropriate for the determination of percentage protein binding of levosimendan. The interaction study with aspirin, clopidogrel and atorvastatin explored significant variation of percentage in protein binding of levosimendan. The results of spectrophotometric method for the determination of rivaroxaban revealed it is accurate, stable and precise. The spectroflurimetric method results described the method is extremely sensitive for the determination of rivaroxaban in very low concentrations and it is a good alternative for high cost methods. The HPTLC method for determination of rivaroxaban provides highly selective quantitative stability indicating method in presence of it degraded products. The HPLC method developed for rivaroxaban shown highly acceptable resolution and recovery results revealed excellent accuracy of the method. The HPLC method explored application of it in in-vitro interaction studies of rivaroxaban with aspirin and clopidogrel. The displacement of rivaroxaban from protein caused by the interacting drug was accurately measured by this method. The UV spectrophotometric method for determination of ticagrelor was having greater sensitivity than the HPLC method. The developed HPTLC method for ticagrelor revealed the method is simple and time saving for the analysis of multiple samples in single run. The recoveries of ticagrelor were also found to be excellent. The stability indicating HPLC method developed revealed degraded products formed by stress were not interfering with the separation of ticagrelor. The method has shown good resolution and has high specificity for ticagrelor determination. All the developed analytical methods were validated as per ICH recommendations. The validation process described the analytical methods has good repeatability, specificity, precision and robustness. The regression analysis revealed acceptable correlation coefficient values that demonstrated excellent linearity response. The results of slight variations in experimental and instrumental parameters proved the robustness of the method. The low values of %RSD reflected usefulness of the method in assay of the formulation. The assays of the formulations of the selected drugs were successfully performed using the developed analytical methods. The mean assay concentrations found described the agreeable label claim of the formulation. The assay results also revealed the additives present in the formulations has not interfered in the assay and hence the methods can be applied in routine quality control process. The statistical analysis of the difference in percentage protein binding shown there is extreme significance of variations in percentage protein binding of the selected drugs due to interaction. The significant difference in protein binding necessitates the monitoring of selected drugs levels when co-administered with interacting drugs. The alteration in the free drug concentration might affect the desired outcome of the therapy and might result in serious adverse events due to the DDI. The developed HPLC method can be extended for the therapeutic monitoring of the selected CVD drugs which will help in adjusting the dosed of these drugs during co-administration. The application of developed analytical methods those are highly sensitive can be extended to study in the biological fluids.

Item Type: Thesis (Doctoral)
Uncontrolled Keywords: Development, validated analytical methods, Cardiovascular Drugs, formulations, application, in-vitro interaction studies.
Subjects: PHARMACY > Pharmaceutical Chemistry
Depositing User: Subramani R
Date Deposited: 20 Aug 2017 05:23
Last Modified: 27 Oct 2022 15:19

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