Development of Spectrophotometric Determination Method for Some Pharmaceutical Preparation Using Cloud Point Extraction and Study Biological Application

Abstract

The assessment of medicaments in the biological and ecological samples, not just in drug formulations, has become important issues for the present. Although most of the analysis of the medicaments at trace levels in these matrices implements by utilizing highly instruments at the time being, these instruments may not be accessible in some laboratories. In reality, the detachment and extraction of trace amounts of medications in environmental and biological samples did not get widespread attention utilizing the so-called cloud point extraction (CPE). In such a manner, we accept that the present work will add to open new skylines in the design of analytical techniques relating to this subject and expand this system application of extraction in one of the significant fields of clinical and ecological sciences. The current study deals with three main chapters. They are as follows: Chapter One: includes general introduction and division of derivatization reactions utilized in analytical chemistry with an emphasis on only two significant types as well as a short prologue to the medication under study (Methyldopa, Sulfacetamide.Na and Bromohexin.HCl) and the chemical literature review of analytical techniques for their assessment. This part also deals in detail with how to gain proficiency with the significance to utilizing the surface-active substances, kinds, basic principles, and uses of the cloud point extraction in analytical chemistry. finally, the chapter examined the aims of this study. Chapter Two: incorporates an outline of the instrumental analytical procedure, general contraption and chemicals utilized in this investigation. Full subtleties analytical techniques for CPE which have been intended for the evaluation of the drugs under examination are made in this thesis. This section contained five fundamental parts: - Part One: Complexation reaction between the medication (methyldopa) with metal as reagent (FeII) . III Part Two: Diazotization reaction coupled between the medications (Sulfacetamide.Na and Bromhexin.HCl) with reagents (p- cresol or β-naphthol) respectively. Part Three: Cloud point extraction for all medications utilizing in the study with the same reagents. Part Four: We have synthesized methyldopa-Fe(II) complex, through treating methyldopa with Fe ( II) in basic media. As well as, we have synthesized new azo compounds of SAC & BRH drugs with p-cresol & β - naphthol in presence nitrous acid, respectively. Part Five: Includes a method of studying the biological activity of the prepared compounds are screened in vitro for antibacterial activity against (gram –ve) bacteria, such as (Escherichia Coli and Klebsiellasp.) and (gram +ve) bacteria, like ( Staphylococcus Epidermidis and Staphylococcus Aureus ); and antifungal activity against (Candida albicans). Chapter Three: features the significant part of this study, which manages with the results and discussion of two significant issues of the analytical applications, specifically, separation enrichment by utilizing complexing reactions, azo-coupling reactions and cloud point extraction procedure with the adoption of some reactions to form colored compounds with medications under investigation and their evaluation spectrophotometrically. This section included three main parts; The first part contains an established procedure for the complexation and evaluation of methyldopa( MDP) drug and cloud point extraction with visible spectrophotometry. In the first, the technique depends on the complexing reaction of methyldopa with Fe(II) as a reagent in an alkaline medium to form the colour complex water-soluble compound and the absorbance measured at λmax of 555 nm. Beer’s law is obeyed in the concentration range of (5-50)μg mL-1 (r2 = 0.9994) for MDP. It got the detection and quantitation limits of 0.164 and 0.541 μg mL-1 respectively, and a superb sensitivity in terms of the molar absorptivity of 1562.22 L.mol- 1 .cm -1 . The proposed technique was approved and applied for the IV estimation of methyldopa drug in different samples of the pharmaceutical formulations. In the second, the procedure depends on the CPE of the medication (MDP) with Fe (II) as a reagent in basic medium to give a complex coloured compound which can be easily separated by a nonionic surfactant TX-100 and methyldopa assessed under the established ideal conditions, accomplishing the detection of limits 0.0486 μg mL-1 , the linear range of (1-50) μg mL-1 (r2 = 0.9991) with sensitivity in terms of the molar absorptivity of 6080 L.mol-1 .cm-1 for MDP. The proposed technique is compared with reported methods published in the chemical literature and utilized for the estimation of (MDP) in different samples. The second part includes two developed methods for the isolation, enrichment and assessment of (SAC & BRH) drugs utilizing azo-coupling reaction and cloud point extraction combined with visible spectrophotometry. In the first, the technique depends on the reaction of the diazonium salts of (SAC & BRH) with (p- cresol &β - naphthol), respectively to form an intense Orange and Red-orange water-soluble compounds and the absorbance measured spectrophotometrically at the absorption maximum of 450 and 495 nm. For SAC drug, the beer’s law is obeyed in the concentration range of (5-100)μg mL-1 (r2 = 0.9996). It was obtained the detection limit and quantitation limits of 0.2142 and 0.707 μg mL-1, and a superb sensitivity in terms of the molar absorptivity of 1488.249 L.mol-1 .cm-1 and for BRH drug, the beer’s law is obeyed in the concentration range of (5-60)μg mL-1 (r2 = 0.9993). It was obtained the detection limit and quantitation limits of 0.163 and 0.5379 μg mL-1 , and a superb sensitivity in terms of the molar absorptivity of 2104.209 L.mol-1 .cm-1 . The proposed technique is validated and applied for the evaluation of (SAC & BRH) in different samples of the pharmaceutical formulations. In the second, the technique includes the reaction of diazotized SAC & BRH with p- cresol &β -naphthol as reagents to form an azo dye orange and red-orange coloured product which can be separated into micelles of TX-100 & TW80 , respectively and the drugs SAC & BRH achieving determined in the visible region of the spectrum at λmax of 450 and 495 nm. For both drugs SAC & BRH, the detection and quantitation limits of (0.070 and 0.231),( 0.0215 and 0.0710 ) μg V mL-1 , with linear range of (1.0 - 60),( 2.5-30 ) μg mL-1 (r2 = 0.9995, 0.9992 ) and superb sensitivity in terms of the molar absorptivity of (7417.622, 16503.6) L.mol-1 .cm-1 . The developed technique is combined with reported methods published in the chemical literature and utilized for the assessment of SAC and BRH, respectively in various samples. The third part, all the synthesized compounds (MDP-Fe) and azo compounds of SAC & BRH were characterized by their physical properties and spectral data, such as FTIR spectrophotometer . The fourth part includes: studying the biological activity of synthesized compounds, show that methyldopa-complex give the biological activity against Staphylococcus aureus , Klebsiella sp. and Candida albicans. Also, the SAC-azo compound show the boilogical activity against Staphylococcus aureus , Klebsiella sp. and Candida albicans . Finally, the BRH-azo compound give the boilogical activity against Staphylococcus aureus , Staphylococcus epidermidis, Escherichia coli , Klebsiella sp. and Candida albicans .