Analytical study of some anticoccidial drugs

Abstract

This thesis consists of six parts: Part I: General introduction This part involves an introduction about coccidiosis, causative parasites, disease symptoms, negative impact on poultry, classification and mode of action of anticoccidial drugs Part II: Literature review This part includes a review of the physical and chemical properties of the studied anticoccidial drugs, Amprolium hydrochloride and Ethopabate. Also, it includes different analytical methods reported in the literature that aimed for the determination of the selected drugs in pure form, mixtures, pharmaceutical formulation, and biological media. Part III: Spectrophotometric determinations of some anticoccidial drugs This part is divided into six sections: Section A: Simultaneous determination of Amprolium hydrochloride and Ethopabate by derivative ratio spectrophotometric method The first derivative of the ratio spectra for each of Amprolium hydrochloride and Ethopabate is obtained. Amprolium hydrochloride is measured at 234.7 nm and Ethopabate is measured at 306.8 nm with mean recovery percentages of 99.76 ± 0.907 and 100.29 ± 0.847, respectively.

Section B: Simultaneous determination of Amprolium hydrochloride and Ethopabate by mean centering spectrophotometric method The obtained ratio spectra are mean centered and the peak amplitude is measured at 238.8 nm for Amprolium hydrochloride and 313 nm for Ethopabate with mean recovery percentages of 100.26 ± 1.018 and 99.93 ± 1.281, respectively. Section C: Simultaneous determination of Amprolium hydrochloride and Ethopabate by ratio difference spectrophotometric method For the determination of Amprolium hydrochloride in presence of Ethopabate, the amplitude difference of the ratio spectra at 239 and 310 nm is measured with mean recovery percentage of 99.27 ± 0.892. For the determination of Ethopabate in presence of Amprolium hydrochloride, the amplitude difference of the ratio spectra at 239 and 313 nm is measured with mean recovery percentage of 100.40 ± 1.814. Section D: Simultaneous determination of Amprolium hydrochloride and Ethopabate by dual wavelength spectrophotometric method Different sets of wavelengths are tried. Absorbance values of ETH are the same at 235.3 and 308 nm therefore these two wavelengths are selected for determination of AMP with mean recovery percentage of 99.30 ± 1.097. The same for the two wavelengths 244 and 268.4 nm at which the absorbance values of AMP are the same, hence these two wavelengths are selected for determination of ETH with mean recovery percentage of 100.02 ± 1.061. Section E: Simultaneous determination of Amprolium hydrochloride and Ethopabate by area under the curve spectrophotometric method Area under curve of the absorption spectra in the wavelength ranges 235.6-243 nm (λ1 - λ2) and 268.3–275 nm (λ3 – λ4) for each of AMP and ETH is calculated. The concentrations of AMP and ETH can be obtained by applying Cramer’s rule with mean recovery percentages of 100.38 ± 1.006 and 100.39 ± 0.955, respectively. Section F: General discussion In this section, results obtained by the proposed methods are compared with those obtained by a reported first derivative spectrophotometric method. Part IV: Synchronous spectrofluorimetric determination of some anticoccidial drugs This part is divided into three sections: Section A: Introduction to synchronous spectrofluorimetry

This section explains the theory of synchronous spectrofluorimetry and its advantages over conventional spectrofluorimetry

Section B: Direct synchronous spectrofluorimetric determination of Amprolium hydrochloride in presence of Ethopabate using Experimental design A reaction is carried out to convert Amprolium hydrochloride to a fluorescent derivative which is determined by synchronous spectrofluorimetry at 362 nm using constant wavelength difference Δλ = 80 nm. This method is used to determine Amprolium hydrochloride in pure form with mean recovery percentage of 99.08 ± 0.801, synthetic mixtures, pharmaceutical formulation, chicken plasma and food samples.

Section C: First derivative synchronous spectrofluorimetric determination of Ethopabate in presence of Amprolium hydrochloride

The first derivative synchronous fluorescence signal is measured at 288 nm for determination of Ethopabate in presence of Amprolium hydrochloride using constant wavelength difference Δλ = 80 nm. This method is used to determine Ethopabate in pure form with mean recovery percentage of 100.40 ± 0.720, synthetic mixtures, pharmaceutical formulation, chicken plasma and food samples.

Part V: Simultaneous determination of Amprolium hydrochloride, Ethopabate and Levamisole hydrochloride by TLC-densitometric method TLC-densitometric method is proposed for simultaneous determination of Amprolium hydrochloride and Ethopabate in presence of Levamisole hydrochloride, an anthelmintic commonly co-administered with anticoccidial drugs to treat helminthiasis and coccidiosis in chickens with mean recovery percentages of 100.72 ± 1.189, 99.93 ± 1.258 and 100.53 ± 0.654, respectively . Experimental conditions such as developing system and wavelength of detection are optimized and the plates are scanned at 213 nm

Part VI: Potentiometric determination of Amprolium hydrochloride This part is divided into two sections: Section A: Introduction to Ion-selective electrodes (ISEs) This section describes the theory and the different types of ion selective electrodes. Also, it includes their assembly and their use with other fields. Section B: Potentiometric determination of Amprolium hydrochloride by Precipitation and Ionophore based techniques This section describes the determination of Amprolium hydrochloride using potassium tetrakis 4-chloro phenyl borate as anionic exchanger (electrode 1) and using inclusion of the drug into HP β-CD cavity (electrodes 2 and 3) and into Calix-4-arene cavity (electrode 4). Dioctyl phthalate is used as a plasticizer for electrodes 1, 2 and 3, while 2-nitro phenyl octyl ether is used as a plasticizer for electrode 4. All electrodes are fabricated in a polymeric matrix of polyvinyl chloride. The accuracy results for potentiometric determination of Amprolium hydrochloride using electrode 1, electrode 2, electrode 3 and electrode 4 are 100.59 ± 1.721, 100.01 ± 1.241, 100.48 ± 0.672 and 99.91 ± 1.427, respectively. The performance of the four electrodes is assessed according to the IUPAC recommendations and shows that they have fast, stable and nernstian response. Response time, effect of pH and temperature on the response, selectivity and stability of the electrodes are studied to determine the optimum conditions for the determination of Amprolium hydrochloride in pure form, pharmaceutical formulation and food samples. This thesis contains 135 references, 78 figures, 55 tables and ends with Arabic summary.