Analytical Study on Some Amide Containing Drugs / Sara Abd El-Rasheed El- Hanboushy, Supervised By Mohamed AbdelKawy, Hayam Mahmoud Lotfy , Yasmin Mohamed Fayez.

Thesis (M.Sc.) – Cairo University. Faculty of Pharmacy. Department of Analytical Chemistry.

Includes bibliographical references.

(Analytical Study on Some Amide- Containing Drugs)
This thesis consisted of a medical introduction and two parts:
Part I: Simultaneous determination of pseudoephedrine, loratadine in their binary mixture and with paracetamol in their ternary mixture in Trimed Flu® and Clarinase® tablets:
This part was divided into five sections:
Section A: General introduction and literature review:
This section provided a general introduction about the chemistry, mode of action and literature review of the reported analytical methods of pseudoephedrine sulphate (PSE), loratadine (LOR) and paracetamol (PAR).
Section B: Spectrophotometric methods for determination of co-formulated drugs of pseudoephedrine, loratadine and paracetamol in presence of para-aminophenol:
This section consists of two parts:
I.B.1.Simultaneous determination of pseudoephedrine and loratadine in their binary mixture and with paracetamol in their ternary mixture in presence of para-aminophenol by spectrophotometric methods via different manipulation pathways
Different manipulating pathways were applied for simultaneous determination of mixtures of pseudoephedrine (PSE), loratadine (LOR) and paracetamol (PAR) in presence of PAP (the degradation product of paracetamol) in their pure powdered form and pharmaceutical dosage form. PAP could be eliminated first by constant multiplication method to give resolved ternary mixture of PSE, LOR and PAR .Then for analysis of the resolved ternary mixture there are two pathways, the first Pathway where [without LOR enrichment], In this pathway PAR could be determined at its λmax 248 nm without any contribution of both co-formulated drugs while PSE could be determined by derivative ratio method or direct measure at its first derivative after elimination of PAR by constant multiplication method (CM). The second pathway [with LOR enrichment], in this pathway spectral overlapping could be solved by successive, progressive resolution technique and without preliminary resolution technique. In successive resolution where LOR could be obtained using CC followed by its elimination to get resolved binary mixture of PSE and PAR. This resolved binary mixture could be determined by direct determination of PAR at its λmax 248 nm while PSE could be determined either by amplitude correction method (PC) or direct measure at its first derivative after elimination of PAR by constant multiplication method (CM).In progressive resolution LOR could be determined by amplitude correction method (PC) while PAR could be determined directly at its first derivative without any contribution of LOR. In without preliminary resolution techniques, LOR could be determined by dual wavelength and PAR could be determined by constant multiplication method (CM).
Spectrophotometric methods could effectively separate the PAR in presence of up to 90% of its degradation product para-aminophenol (PAP) and it can be employed as a stability indicating method for PAR
I.B.2.Simultaneous determination of pseudoephedrine sulphate and loratadine in their binary mixture by spectrophotometric methods via different manipulation pathways
Zero order absorption spectra, constant multiplication (CM) and induced amplitude modulation method (IAM) could be used for determination of LOR. On the other hand, zero order absorption spectrum at 256.8 nm after spectrum subtraction of LOR (which had been obtained by CM), dual wavelength method (DW), induced dual wavelength method (IDW), ratio difference method (RD) , absorbance correction method (AC) induced amplitude modulation method (IAM) could be used for determination of PSE.
The specificity of the developed methods was investigated by analyzing laboratory prepared mixtures and pharmaceutical dosage form containing the cited drugs with no interference from additives. The proposed methods were validated according to ICH guidelines. The obtained results were statistically compared with those of the official and reported methods; using student’s t-test, F-test, and one-way ANOVA, showing no significant difference with respect to accuracy and precision.
Section C: Simultaneous determination of the ternary mixture of loratadine, pseudoephedrine and paracetamol by chromatographic methods:
HPLC-DAD and TLC-densitometric methods were successfully applied for simultaneous determination of PSE, LOR and PAR in their pure powdered form, laboratory-prepared mixtures and pharmaceutical dosage form.
The HPLC-DAD method was carried out on carried out on Scharlau column KromaPhase 100 C8, (4.6 mm x 150 mm, 5-µm) using step gradient elution. A mobile phase system consisting of 0.05M sodium phosphate monobasic dihydrate solution
(pH2.9), methanol and acetonitrile was used. The separation was achieved with the linear gradient program. The flow rate was 1.0 mL/min. the pH was adjusted with orthophosphoric acid. The mobile phase was filtered using 0.45 μmmillipore membrane filter (Billerica, MA, USA) and delivered at flow rate 1.0 mL/min. The injection volume was 20-µL. The multiple wavelength detector was set at 210 nm for measurement of PSE and PAR and 254 nm for LOR. All the analyses were done at ambient temperature 25°C.
The TLC-densitometric method was developed using developing system consisted of a mixture of ethyl acetate: methanol: ammonia (13:2:0.3, by volume) and UV detection at 208 nm. The densitometric method could effectively separate the PAR in presence of up to 90% of its degradation product para-aminophenol (PAP) and it can be employed as a stability indicating method for PAR
The proposed methods were validated as per ICH guidelines for linearity, range, accuracy, precision and specificity. The results were found to be within the acceptable limits.
Section D: Electrochemical determination of LOR and PSE by ion selective electrode:
The electrochemical method was developed for selective determination of PSE, LOR separately via construction of ion selective electrode for PSE and LOR constructed using a polyvinyl chloride (PVC) based membrane as a polymer, tetraphenyl borate (TPB) and potassium tetrakis (4-chlorohenyl) borate (KTCPB) as a cationic exchanger, Bis-sebacate and nitrophenyloctyl ether (NPOE), were used as a plasticizer for determination of PSE and LOR, respectively.
Section E: Statistical comparison and conclusion:
Statistical comparison of the results was obtained by applying the proposed methods to pure samples of PSE, LOR and PAR versus the official methods where the calculated t and F values were less than the theoretical ones indicating that there was no significant difference between the proposed and the official methods with respect to accuracy and precision. One-way ANOVA was applied for the purpose of comparison of the developed methods with those of the official and reported methods, which showed that there was no significant difference between them for the determination of PSE, LOR and PAR regarding accuracy and precision.
Part II: Simultaneous determination of furosemide and spironolactone Lasilactone® Tablets:
This chapter was divided into four sections:
Section A: General introduction and literature review:
This section provided a general introduction about the chemistry, mode of action and literature review of the reported analytical methods of furosemide (FUR) and spironolactone (SP).
Section B: Simultaneous spectrophotometric determination of furosemide and spironolactone in their binary mixture:
Different manipulating pathways were applied for simultaneous determination of FUR and SP in their pure powdered form and pharmaceutical dosage form without prior separation steps. These pathways were applied either on scanned zero order absorption spectra namely, absorbance subtraction (AS) or on the amplitudes of the ratio spectra of zero order absorption spectra are namely, amplitude modulation (AM) and derivative ratio (DD1), then using the absorbance of the recovered zero order absorption spectra namely, ratio subtraction coupled with constant multiplication (RS-CM).Finally, pathway depend on using the absorbance of the recovered zero order absorption spectra namely, ratio subtraction (RS) coupled with constant multiplication (RS-CM)
The proposed methods were validated according to ICH guidelines. The obtained results were statistically compared with those of the official or reported methods; using student’s t-test, F-test, and one-way ANOVA, showing no significant difference with respect to high accuracy and precision.
Section C: Simultaneous chromatographic determination of furosemide and spironolactone in their binary mixture:
UPLC-DAD and TLC-densitometric methods were successfully applied for simultaneous determination of FUR and SP in their pure powdered form, laboratory-prepared mixtures and pharmaceutical dosage form.
The UPLC-DAD method carried out on Scharlau column C18 (2.1 mm x50 mm, 1.8 µm) analytical column. The mobile phase consisted of a mixture of ammonium acetate buffer: acetonitrile: methanol (32: 59: 9, by volume, pH 5.5) in an isocratic mode. The mobile phase was filtered using 0.45 pm Millipore membrane filter (Billerica, MA) and delivered at a flow rate of 1.0 mL/min. The injection volume wa
10-µL and the detection was done at 232 nm. All the analysis was done at ambient temperature 250C.
The TLC-densitometric method was developed using developing system consisted of a mixture of ethyl acetate: hexane: methanol: acetic acid (7: 7: 0.2: 0.2, by volume) and scanned at 238 nm.
The proposed methods were checked using laboratory-prepared mixtures and were successfully applied for the analysis of pharmaceutical dosage form containing the studied drugs and were validated according to ICH guidelines. The proposed methods could be used for the routine analysis of the studied drugs in their pharmaceutical dosage form in quality control laboratories.
Section D: Statistical comparison and conclusion:
Statistical comparison of the results obtained by applying the proposed methods to pure samples of FUR and SP versus the official and reported methods where the calculated t and F values were less than the theoretical ones indicating that there was no significant difference between them with respect to accuracy and precision. One-way ANOVA was applied for the purpose of comparison of the developed methods with those of the official and reported methods, which showed that there was no significant difference between them for the determination of FUR and SP regarding accuracy and precision.
A general introduction in addition to appendices describing the validation parameters, common materials, apparatus and reagents were given.
The thesis comprises 74 Figures, 63 tables and 315 references from 1973-2018 and ends with summary in Arabic.

Text in English, summaries in English and Arabic.