Nagiba Y. Hassan

Separation and determination of clopidogrel and its impurities by capillary electrophoresis.

Clopidogrel bisulphate, an anti-platelet drug, has been separated from its impurities, namely impurity A, B and C by capillary zone electrophoresis (CZE) using uncoated fused-silica capillary (50.0 microm internal diameter, 31.2cm total length). Four factors affected the separation: buffer concentration, pH of the buffer, concentration of the chiral selector and the applied voltage. Optimization and robustness studies were performed with the aid of reduced central composite experimental design. The buffer used was triethylamine-phosphoric acid and the chosen chiral selector was sulphated beta-cyclodextrin (SCD). The best separation was achieved by using 10mM buffer, pH 2.3, containing 5% (mass/volume (m/v)) SCD. Reversed polarity mode was used with an applied voltage of -12kV and the capillary temperature was maintained at 20 degrees C. The method was validated for quantitative determination of the drug. It offered a limit of detection (LOD) of 0.13 microg/ml, a limit of quantitation (LOQ) of 0.4 microg/ml, and a linearity range of 0.4-300 microg/ml. Commercial bulk samples were analyzed using the developed method.

A comparative study of novel spectrophotometric methods based on isosbestic points; application on a pharmaceutical ternary mixture

This work represents the application of the isosbestic points present in different absorption spectra. Three novel spectrophotometric methods were developed, the first method is the absorption subtraction method (AS) utilizing the isosbestic point in zero-order absorption spectra; the second method is the amplitude modulation method (AM) utilizing the isosbestic point in ratio spectra; and third method is the amplitude summation method (A-Sum) utilizing the isosbestic point in derivative spectra. The three methods were applied for the analysis of the ternary mixture of chloramphenicol (CHL), dexamethasone sodium phosphate (DXM) and tetryzoline hydrochloride (TZH) in eye drops in the presence of benzalkonium chloride as a preservative. The components at the isosbestic point were determined using the corresponding unified regression equation at this point with no need for a complementary method. The obtained results were statistically compared to each other and to that of the developed PLS model. The specificity of the developed methods was investigated by analyzing laboratory prepared mixtures and the combined dosage form. The methods were validated as per ICH guidelines where accuracy, repeatability, inter-day precision and robustness were found to be within the acceptable limits. The results obtained from the proposed methods were statistically compared with official ones where no significant difference was observed.

Stability-indicating determination of meropenem in presence of its degradation product

Stability-indicative determination of meropenem (MERM) in the presence of its open-ring degradation product, the metabolite, is investigated. The degradation product has been isolated, via acid-degradation, characterized and confirmed. Selective quantification of MERM, singly in bulk form, pharmaceutical formulations and/or in the presence of its major degradate is demonstrated. The indication of stability has been undertaken under conditions likely to be expected at normal storage. Among the analytical techniques adopted for quantification are spectrophotometry [first-derivative ((1)D), first-derivative of ratio spectra ((1)DD) and bivariate analysis], as well as chromatography [coupled TLC-densitometry and HPLC].

Univariate versus Multivariate Spectrophotometric Methods for Simultaneous Determination of Complex Binary Mixtures with Overlapped Spectra: A Comparative Study

Two novel simple and accurate univariate spectrophotometric methods were described for the determination of a binary mixture of Sodium cromoglicate (SCG) and Fluorometholone (FLU) namely; ratio subtraction coupled with extended ratio subtraction (RS-EXRSM) and constant center (CCSM) spectrophotometric methods. The methods were able to recover the zero order absorption spectra of both drugs from their binary mixture through simple mathematical calculations. The linearity ranges were found to be (2.5–25 µg/mL) and (4- 25 µg/mL) for SCG and FLU, respectively. A comparative study was conducted between the proposed univariate methods versus multivariate ones, based on using principle component regression (PCR) and partial least squares (PLS-2). Upon applying the methods for the determination of SCG and FLU in laboratory prepared mixtures, the (CCSM and PLS-2) proved to be of higher predictive ability rather than (RS-EXRSM and PCR) in prediction of mixtures containing low concentration of one of the components. The proposed methods were validated in compliance with the ICH guidelines. These methods could be alternative to different HPLC techniques in quality control laboratories lacking the required facilities for those expensive techniques

Simultaneous determination of terbinafine HCL and triamcinolone acetonide by UV derivative spectrophotometry and spectrodensitometry

A binary mixture of terbinafine hydrochloride and triamcinolone acetonide was determined by three different methods. The first one concerned with determination of both drugs using first derivative (D(1)) spectrophotometric technique at 297 and 274 nm over concentration ranges of 5-30 and 4-24 microg ml(-1), with mean percentage accuracies 99.90+/-0.67 and 100.25+/-0.49, respectively. The second method depends on ratio-spectra 1st derivative (RSD(1)) spectrophotometry at 298 and 248 nm over the same concentration ranges with mean percentage accuracies 100.22+/-0.51 and 99.93+/-0.56, respectively. The spectrodensitometric analysis provides a rapid and precise method for the separation and quantitation of both terbinafine hydrochloride and triamcinolone acetonide. The method depends on the quantitative densitometric evaluation of thin layer chromatogram of terbinafine hydrochloride and triamcinolone acetonide at 283 and 238 nm over concentration ranges of 5-25 and 2.5-22.5 microg spot(-1), with mean percentage accuracies 100.66+/-0.51 and 100.27+/-0.73, respectively. The suggested procedures were checked using laboratory prepared mixtures and were successfully applied for the analysis of their pharmaceutical preparations. The three methods retained their accuracy and precision when applying the standard addition technique. The results obtained by applying the proposed methods were statistically analysed and compared with those obtained by a reported method.

Selective determination of ertapenem in the presence of its degradation product

Stability-indicative determination of ertapenem (ERTM) in the presence of its beta-lactam open-ring degradation product, which is also the metabolite, is investigated. The degradation product has been isolated, via acid-degradation, characterized and elucidated. Selective quantification of ERTM, singly in bulk form, pharmaceutical formulations and/or in the presence of its major degradant is demonstrated. The indication of stability has been undertaken under conditions likely to be expected at normal storage conditions. Among the spectrophotometric methods adopted for quantification are first derivative ((1)D), first derivative of ratio spectra ((1)DD) and bivariate analysis.

Membrane electrodes for the determination of glutathione

Four glutathione (GSH)-selective electrodes were developed with different techniques and in different polymeric matrices. Precipitation-based technique with bathophenanthroline-ferrous as cationic exchanger in polyvinyl chloride (PVC) matrix was used for sensor 1 fabrication. beta-Cyclodextrin (beta-CD)-based technique with either tetrakis(4-chlorophenyl)borate (TpClPB) or bathophenanthroline-ferrous as fixed anionic and cationic sites in PVC matrix was used for fabrication of sensors 2 and 3, respectively. beta-CD-based technique with TpClPB as fixed anionic site in polyurethane (Tecoflex) matrix was used for sensor 4 fabrication. Linear responses of 1x10(-5) to 1x10(-4)M and 1x10(-6) to 1x10(-3)M with slopes of 37.5 and 32.0mV/decade within pH 7-8 were obtained by using electrodes 1 and 3, respectively. On the other hand, linear responses of 1x10(-5) to 1x10(-2) and 1x10(-5) to 1x10(-3)M with slopes of 47.9 and 54.3mV/decade within pH 5-6 were obtained by using electrodes 2 and 4, respectively. The percentage recoveries for determination of GSH by the four proposed GSH-selective electrodes were 100+/-1, 100.5+/-0.7, 100+/-1 and 99.0+/-0.8% for sensors 1, 2, 3 and 4, respectively. Determination of GSH in capsules by the proposed electrodes revealed their applicability for determination of GSH in its pharmaceutical formulations. Also, they were used to determine GSH selectively in presence of its oxidized form (GSSG). Sensor 4 was successfully applied for determination of glutathione in plasma with average recovery of 100.4+/-1.11%. The proposed method was compared with a reported one. No significant difference for both accuracy and precision was observed.

A comparative study of progressive versus successive spectrophotometric resolution techniques applied for pharmaceutical ternary mixtures

This work represents a comparative study of a novel progressive spectrophotometric resolution technique namely, amplitude center method (ACM), versus the well-established successive spectrophotometric resolution techniques namely; successive derivative subtraction (SDS); successive derivative of ratio spectra (SDR) and mean centering of ratio spectra (MCR). All the proposed spectrophotometric techniques consist of several consecutive steps utilizing ratio and/or derivative spectra. The novel amplitude center method (ACM) can be used for the determination of ternary mixtures using single divisor where the concentrations of the components are determined through progressive manipulation performed on the same ratio spectrum. Those methods were applied for the analysis of the ternary mixture of chloramphenicol (CHL), dexamethasone sodium phosphate (DXM) and tetryzoline hydrochloride (TZH) in eye drops in the presence of benzalkonium chloride as a preservative. The proposed methods were checked using laboratory-prepared mixtures and were successfully applied for the analysis of pharmaceutical formulation containing the cited drugs. The proposed methods were validated according to the ICH guidelines. A comparative study was conducted between those methods regarding simplicity, limitation and sensitivity. The obtained results were statistically compared with those obtained from the official BP methods, showing no significant difference with respect to accuracy and precision.

COMPARATIVE STUDY OF NEW SPECTROPHOTOMETRIC METHODS; AN APPLICATION ON PHARMACEUTICAL BINARY MIXTURE OF CIPROFLOXACIN HYDROCHLORIDE AND HYDROCORTISONE

ABSTRACT Simple, specific, accurate and precise spectrophotometric methods were developed and validated for simultaneous estimation of Ciprofloxacin hydrochloride (CIP) and Hydrocortisone (HYD) in their pure form and pharmaceutical dosage form. Two new spectrophotometric methods were applied: extended ratio subtraction (EXRSM) and ratio difference (RDSM) methods. The results were compared to three well-established methods: Mean centering of ratio spectra method (MCR), Isoabsorpative point spectrophotometric method (IsoM) and Absorbance ratio method (ARM). The linearity range for the spectrophotometric methods was found to be (2-14 µg/mL) and (1-14 µg/mL) for CIP and HYD respectively. The selectivity of the developed methods was investigated by analyzing laboratory prepared mixtures of the two drugs and their combined dosage form. The results obtained from the proposed methods were statistically compared using one-way analysis of variance (ANOVA) where no significant difference was observed between the new (EXRSM and RDSM) and the well-established methods which prove their validity for the analysis of this binary mixture. The methods were validated as per ICH guidelines regarding accuracy, precision, repeatability and robustness; which were found to be within the acceptable limits. The new methods were simple, sensitive and don’t need a special program, so they could be easily applied as alternative methods to LC methods in quality control laboratories lacking the required facilities for those techniques.

A comparative study of different aspects of manipulating ratio spectra applied for ternary mixtures: Derivative spectrophotometry versus wavelet transform

This work represents a comparative study of different aspects of manipulating ratio spectra, which are: double divisor ratio spectra derivative (DR-DD), area under curve of derivative ratio (DR-AUC) and its novel approach, namely area under the curve correction method (AUCCM) applied for overlapped spectra; successive derivative of ratio spectra (SDR) and continuous wavelet transform (CWT) methods. The proposed methods represent different aspects of manipulating ratio spectra of the ternary mixture of Ofloxacin (OFX), Prednisolone acetate (PA) and Tetryzoline HCl (TZH) combined in eye drops in the presence of benzalkonium chloride as a preservative. The proposed methods were checked using laboratory-prepared mixtures and were successfully applied for the analysis of pharmaceutical formulation containing the cited drugs. The proposed methods were validated according to the ICH guidelines. A comparative study was conducted between those methods regarding simplicity, limitation and sensitivity. The obtained results were statistically compared with those obtained from the reported HPLC method, showing no significant difference with respect to accuracy and precision.