Advanced analytical techniques for analysis of Some therapeutic agents used in the treatment of cardiovascular diseases in dosage forms and biological fluids /

Abstract

This thesis consists of three parts. • Part I: General introduction and literature review This part includes a general introduction to the pharmacological and biological effects of cardiovascular drugs of interest. This part presents the drugs of the first combination in this study, felodipine and metoprolol, and the second co administered combination, bisoprolol, rosuvastatin, and clopidogrel, this section includes all the structure-related details and physical characteristics of the compounds of interest, along with their pharmacological action. A complete and updated literature review of the different methods of analysis was summarized for the studied compounds. • Part II: Determination of Cardiovascular Drugs in Bulk and Pharmaceutical Dosage Form by Different Spectrophotometric Methods. This part is divided into four sections. Section A: Ratio Spectra Derivative for Determination of Felodipine and Metoprolol in their Pure and Pharmaceutical Dosage Form. In this section, the determination of felodipine and metoprolol was achieved by calculating first derivative ratio spectra using the numerical differentiation method despite their spectral overlap in zero-order measurements. The proposed method was adopted for the simultaneous estimation of both drugs in the range of 0.30- 15.00 µg/mL for FDP and 0.40- 22.00 µg/mL for MTP in their pure forms, laboratory prepared mixtures, and in their pharmaceutical dosage form. The statistical comparison of the results obtained with those of a reported HPLC method was made. Section B: Mean Centering of Ratio Spectra Method for Determination of Felodipine and Metoprolol in their Pure and Pharmaceutical Dosage form. In this section, the determination of felodipine and metoprolol was achieved by calculating the mean centering of ratio spectra method despite their spectral overlap in zero-order measurements. The proposed method was adopted for the simultaneous estimation of both drugs in the range of 0.30- 15.00 µg/mL for FDP and 0.40- 22.00 µg/mL for MTP in their pure forms, laboratory prepared mixtures, and in their pharmaceutical dosage form. The statistical comparison of the results obtained with those of a reported HPLC method was made. Section C: Ratio Difference Spectrophotometric Method for Determination of Felodipine and Metoprolol in their Pure and Pharmaceutical Dosage Form. In this section, the determination of felodipine and metoprolol was achieved by calculation of ratio difference after obtaining first ratio spectra despite their spectral overlap in zero-order measurements. The proposed method was adopted for the simultaneous estimation of both drugs in the range of 0.30- 15.00 µg/mL for FDP and 0.40- 22.00 µg/mL for MTP in their pure forms, laboratory prepared mixtures, and in their pharmaceutical dosage form. The statistical comparison of the results obtained with those of a reported HPLC method was made. Section D: Dual Wavelength Spectrophotometric Method for Determination of Felodipine and Metoprolol in their Pure and Pharmaceutical Dosage form. In this section, the determination of felodipine and metoprolol was achieved by the dual-wavelength method in the zero-order spectra by calculating the difference between two wavelengths for both drugs despite their spectral overlap in zero-order measurements. The proposed method was adopted for the simultaneous estimation of both drugs in the range of 0.30- 15.00 µg/mL for FDP and 0.40- 22.00 µg/mL for MTP in their pure forms, laboratory prepared mixtures, and in their pharmaceutical dosage form. The statistical comparison of the results obtained with those of a reported HPLC method was made. Part III: Simultaneous Estimation of Cardiovascular Drugs in Bulk, Pharmaceutical Dosage Forms, and spiked human plasma by Different HPLC Methods. This part is divided into three sections. Section A: Eco-friendly, Reversed Phase High Performance Liquid Chromatography (RP-HPLC) Method with UV-detection for Simultaneous Estimation of Felodipine and Metoprolol in their Pure Powder and Pharmaceutical Dosage Form. In this section, simultaneous determination of felodipine and metoprolol was performed using high-performance liquid chromatography coupled with a UV detector using a C18 column and gradient programming adjusted at (70:30, v/v) of potassium dihydrogen phosphate: ethanol then shifted to (20:80, v/v) at 4 minutes till the end of the run at pH 2.5. Olopatadine HCL was used as an internal standard. UV detection was performed at 237 nm for felodipine and 221nm for metoprolol and olopatadine. The proposed method was further applied to analyze both drugs in their pure forms, laboratory prepared mixtures as well as their pharmaceutical dosage form. The results were statistically compared to those obtained from a reported HPLC method. Section B: Eco-friendly, Bioanalytical Reversed Phase High Performance Liquid Chromatography (RP-HPLC) Method Coupled with Fluorescence Detection for Simultaneous Estimation of Felodipine and Metoprolol. In this Section, simultaneous determination of felodipine and metoprolol was performed using high-performance liquid chromatography coupled with a Fluorescence detector using C18 column and using isocratic elution with flow rate of the mobile phase at 1 mL/min adjusted at (40:60, v/v) of potassium dihydrogen phosphate: ethanol at pH of 2.5. Fluorescence detection was programmed to measure felodipine, metoprolol, and tadalafil at excitation wavelengths of 230 nm, 275 nm, and 367 nm and emission wavelengths of 300 nm, 335 nm, and 440 nm, respectively. The proposed method was further applied to analyze both drugs in their pure forms, laboratory-prepared mixtures, pharmaceutical dosage form, and spiked human plasma. Bioanalytical method validation was applied to confirm the method's applicability to human plasma. The results were statistically compared to those obtained from a reported HPLC method. Section C: Reversed Phase High Performance Liquid Chromatography (RP HPLC) Method Coupled with UV Detection for the Simultaneous Estimation of Bisoprolol, Rosuvastatin, and Clopidogrel. In this section, simultaneous determination of the co-administered drugs bisoprolol, rosuvastatin, and clopidogrel along with an internal standard olopatadine HCL using high-performance liquid chromatography coupled with UV detector using C18 column and gradient programming adjusted at (70:30, v/v) of potassium dihydrogen phosphate: acetonitrile then shifted to (20:80, v/v) at 6.5 minutes till the end of the run at pH 3. UV detection was programmed to measure bisoprolol, rosuvastatin, and clopidogrel at wavelengths 225 nm, 240 nm, and 230 nm, respectively. The proposed method was further applied to analyze co-administered drugs in their pure forms, laboratory-prepared mixtures, and pharmaceutical dosage forms. The results were statistically compared to those obtained from a reported HPLC method.