Marwa A.A. Ragab

Non-parametric linear regression of discrete Fourier transform convoluted chromatographic peak responses under non-ideal conditions of internal standard method.

This manuscript discusses the application of chemometrics to the handling of HPLC response data using the internal standard method (ISM). This was performed on a model mixture containing terbutaline sulphate, guaiphenesin, bromhexine HCl, sodium benzoate and propylparaben as an internal standard. Derivative treatment of chromatographic response data of analyte and internal standard was followed by convolution of the resulting derivative curves using 8-points sin x(i) polynomials (discrete Fourier functions). The response of each analyte signal, its corresponding derivative and convoluted derivative data were divided by that of the internal standard to obtain the corresponding ratio data. This was found beneficial in eliminating different types of interferences. It was successfully applied to handle some of the most common chromatographic problems and non-ideal conditions, namely: overlapping chromatographic peaks and very low analyte concentrations. For example, a significant change in the correlation coefficient of sodium benzoate, in case of overlapping peaks, went from 0.9975 to 0.9998 on applying normal conventional peak area and first derivative under Fourier functions methods, respectively. Also a significant improvement in the precision and accuracy for the determination of synthetic mixtures and dosage forms in non-ideal cases was achieved. For example, in the case of overlapping peaks guaiphenesin mean recovery% and RSD% went from 91.57, 9.83 to 100.04, 0.78 on applying normal conventional peak area and first derivative under Fourier functions methods, respectively. This work also compares the application of Theils method, a non-parametric regression method, in handling the response ratio data, with the least squares parametric regression method, which is considered the de facto standard method used for regression. Theils method was found to be superior to the method of least squares as it assumes that errors could occur in both x- and y-directions and they might not be normally distributed. In addition, it could effectively circumvent any outlier data points. For the purpose of comparison, the results obtained using the above described internal standard method were compared with the external standard method for all types of linearity.

High performance liquid chromatography with photo diode array for separation and analysis of naproxen and esomeprazole in presence of their chiral impurities: Enantiomeric purity determination in tablets

A stereoselective high performance liquid chromatographic method with diode array detection (HPLC-DAD) was introduced for S-naproxen and esomeprazole determination in tablets. The separation was achieved on a Kromasil Cellucoat chiral column using a mobile phase consisting of hexane: isopropanol: trifluoroacetic acid (TFA) (90:9.9:0.1 v/v/v). The proposed system was found to be suitable for the enantioseparation of naproxen and omeprazole biologically active isomers. After optimization of the chromatographic conditions, resolution values of 3.84 and 2.17 could be obtained for naproxen and omeprazole isomers, respectively. The method was fully validated for the determination of S-isomers of each drug in their dosage form. Also, the enentiomeric purity was determined in commercial tablet containing S-naproxen and esomeprazole. The enantiomeric purity was calculated for each drug and the chiral impurities (R-isomers) could be determined at 1% level. The method was validated and good results with respect to linearity, precision, accuracy, selectivity and robustness were obtained. The limits of detection (LOD) and quantification (LOQ) were 2.00, 6.50 and 0.10, 0.35μgmL-1 for S-naproxen and esomeprazole, respectively.

Development and Validation of a High-Performance Thin-Layer Chromatographic Method for the Simultaneous Determination of Two Binary Mixtures Containing Ketorolac Tromethamine with Phenylephrine Hydrochloride and with Febuxostat

A validated and highly selective high-performance thin-layer chromatography (HPTLC) method was developed for the determination of ketorolac tromethamine (KTC) with phenylephrine hydrochloride (PHE) (Mixture 1) and with febuxostat (FBX) (Mixture 2) in bulk drug and in combined dosage forms. The proposed method was based on HPTLC separation of the drugs followed by densitometric measurements of their spots at 273 and 320 nm for Mixtures 1 and 2, respectively. The separation was carried out on Merck HPTLC aluminum sheets of silica gel 60 F254 using chloroform-methanol-ammonia (7:3:0.1, v/v) and (7.5:2.5:0.1, v/v) as mobile phase for KTC/PHE and KTC/FBX mixtures, respectively. Linear regression lines were obtained over the concentration ranges 0.20-0.60 and 0.60-1.95 µg band(-1)for KTC and PHE (Mixture 1), respectively, and 0.10-1.00 and 0.25-2.50 µg band(-1) for KTC and FBX (Mixture 2), respectively, with correlation coefficients higher than 0.999. The method was successfully applied to the analysis of the two drugs in their synthetic mixtures and in their dosage forms. The mean percentage recoveries were in the range of 98-102%, and the RSD did not exceed 2%. The method was validated according to ICH guidelines and showed good performances in terms of linearity, sensitivity, precision, accuracy and stability.

Simultaneous determination of montelukast and fexofenadine using Fourier transform convolution emission data under non- parametric linear regression method.

New hybrid chemometric method has been applied to the emission response data. It deals with convolution of emission data using 8-points sin xi polynomials (discrete Fourier functions) after the derivative treatment of these emission data. This new application was used for the simultaneous determination of Fexofenadine and Montelukast in bulk and pharmaceutical preparation. It was found beneficial in the resolution of partially overlapping emission spectra of this mixture. The application of this chemometric method was found beneficial in eliminating different types of interferences common in spectrofluorimetry such as overlapping emission spectra and self- quenching. Not only this chemometric approache was applied to the emission data but also the obtained data were subjected to non-parametric linear regression analysis (Theil’s method). The presented work compares the application of Theil’s method in handling the response data, with the least-squares parametric regression method, which is considered the de facto standard method used for regression. So this work combines the advantages of derivative and convolution using discrete Fourier function together with the reliability and efficacy of the non-parametric analysis of data. Theil’s method was found to be superior to the method of least squares as it could effectively circumvent any outlier data points.New hybrid chemometric method has been applied to the emission response data. It deals with convolution of emission data using 8-points sin xi polynomials (discrete Fourier functions) after the derivative treatment of these emission data. This new application was used for the simultaneous determination of Fexofenadine and Montelukast in bulk and pharmaceutical preparation. It was found beneficial in the resolution of partially overlapping emission spectra of this mixture. The application of this chemometric method was found beneficial in eliminating different types of interferences common in spectrofluorimetry such as overlapping emission spectra and self- quenching. Not only this chemometric approache was applied to the emission data but also the obtained data were subjected to non-parametric linear regression analysis (Theil’s method). The presented work compares the application of Theil’s method in handling the response data, with the least-squares parametric regression method, which is considered the de facto standard method used for regression. So this work combines the advantages of derivative and convolution using discrete Fourier function together with the reliability and efficacy of the non-parametric analysis of data. Theil’s method was found to be superior to the method of least squares as it could effectively circumvent any outlier data points.

Convolution of Emission Derivative Ratio Curves of Closely Related Fluorescent Reaction Products Using Discrete Fourier Functions and Non-Parametric Linear Regression Method

A spectrofluorimetric method was used for the estimation of closely related fluorescent reaction products, Fluoxetine and Olanzapine, in their mixture after derivatization of both drugs using 4-chloro-7- nitrobenzo - 2 -oxa-1,3 - diazole (NBD-Cl) in borate buffered medium (pH 9.5) to form highly fluorescent products. The method based on the use of first and second derivative ratio of the emission data along with their convolution using 8-points sin xi or cos xi polynomials (discrete Fourier functions). The proposed method facilitates their simultaneous determination despite the presence of a minor component (Olanzapine) and strong overlapped spectra of the two NBD-Cl fluorescent products of fluoxetine and olanzapine. The accurate and precise estimation of the minor component was achieved after the convolution of the derivative ratio curves. Moreover, the obtained data were subjected to non-parametric linear regression analysis (Theil’s method). The work combines the advantages of convolution of derivative ratio curves using discrete Fourier functions together with the reliability and efficacy of the non-parametric analysis of data.

Experimental design and machine learning strategies for parameters screening and optimization of Hantzsch condensation reaction for the assay of sodium alendronate in oral solution

An experimental design was adopted to attain the optimum reaction parameters of chemical derivatization of anhydrous sodium alendronate in an oral solution formula via Hantzsch condensation reaction. All reaction controlling variables, namely, time of reaction, temperature, reagent ratio and volume and buffer type, pH and volume were studied using the Plackett–Burman screening design to determine significant variables. Reaction temperature and pH of the buffer solution were found to be significant variables. Optimization was performed using the central composite design to get the optimum levels of these variables. Moreover, a comparison was made with artificial neural networks and support vector machines. The same results were obtained with low percentage relative error. After carrying out the spectrophotometric analysis, interferences from oral solution excipients were eliminated with a simple extraction procedure before measuring the absorbance at 340 nm. Satisfactory results of sample analysis were obtained and they were in good agreement with the label claim. A linear calibration graph of absorbance versus concentration was obtained with very low value of intercept and high value of correlation coefficient (0.9999) in the range of 2.44–34.10 μg mL−1. The proposed spectrophotometric method was fully validated in accordance with ICH guidelines. Statistical comparison with a reported reference method showed similar results with respect to accuracy and precision.

Derivative emission spectrofluorimetry: Application to the analysis of newly approved FDA combination of ibuprofen and famotidine in tablets

A new combination of ibuprofen (NSAID) and famotidine (H2 receptor antagonist) was recently approved by the FDA. It was formulated to relief pain while decreasing the risk of ulceration, which is a common problem for patients receiving NSAID. A rapid and simple derivative emission spectrofluorimetric method is proposed for the simultaneous analysis of this combination in their pharmaceutical preparation. The method is based upon measurement of the native fluorescence intensity of the two drugs at λex = 233 nm in acetonitrile. The emission data were differentiated using the first (D1) derivative technique. The plots of derivative fluorescence intensity versus concentration were rectilinear over a range of 2-35 and 0.4-8 µg/mL for both ibuprofen (IBU) and famotidine (FAM), respectively. The method was sensitive as the limits of detection were 0.51 and 0.12 µg/mL and limits of quantitation were 1.70 and 0.39 µg/mL, for IBU and FAM respectively. The proposed derivative emission spectrofluorimetric method was successfully applied for the determination of the two drugs in their synthetic mixtures and tablets with good accuracy and precision. The proposed method was validated as per ICH guidelines.

Green chemistry: Analytical and chromatography

ABSTRACT Nowadays, the environment protection and the personal health and safety are given more consideration in the field of chemistry, thus resulting in an increased number of published researches about how to work according to green instructions, to follow up the recommendations of environmental agencies and to obtain better clean handling of chemistry. In this review, green chemistry definition, importance, principles, and some recent applications in the field of green chemistry were discussed. In addition, the review summarizes the evolution of green analytical chemistry (GAC) with its specific principles and how to make the analytical process more environmentally benign with special emphasis on recent applications of GAC. Moreover, the green chromatography, its methods, and some of its applications were outlined. Finally, different techniques available up till now for the assessment of greening of the methods were also presented. GRAPHICAL ABSTRACT

KINETIC INVESTIGATION OF PENTOXIFYLLINE BASED ON NONPARAMETRIC LINEAR REGRESSION OF DERIVATIVE AND CONVOLUTED DERIVATIVE CHROMATOGRAPHIC AND SPECTROPHOTOMETRIC RESPONSES

A novel application of chemometrics to the chromatographic peak responses in the kinetic investigation of Pentoxiphylline was introduced using nonparametric linear regression method as the statistical method of analysis. The kinetic study of Pentoxiphylline was conducted under two different stress conditions using DAD-HPLC. The chemometric methods were applied to the HPLC and spectrophotometric data of the kinetic study of Pentoxiphylline. First and second derivative treatment of chromatographic and spectrophotometric response data were followed by convolution of the resulting derivative curves using 8-points sin xi polynomials (discrete Fourier functions). The study also presents a comparison between parametric and non-parametric regression methods of analysis. Moreover, application of Arrhenius equation to the determination of the half life of Pentoxiphylline in alkaline condition was studied before and after the chemometric treatment of the data. The results obtained indicated that chemometric treatment of data with the application of non-parametric method enhances the linearity parameters obtained during the kinetic investigation. These linearity parameters were further used to estimate the degradation rate constant (K) and half life of the drug at room temperature which is very important for the estimation of the stability of the drug on shelf with accuracy and with minimum experimental work and errors.

Comparative study of some robust statistical methods: weighted, parametric, and nonparametric linear regression of HPLC convoluted peak responses using internal standard method in drug bioavailability studies

AbstractThis manuscript discusses the application and the comparison between three statistical regression methods for handling data: parametric, nonparametric, and weighted regression (WR). These data were obtained from different chemometric methods applied to the high-performance liquid chromatography response data using the internal standard method. This was performed on a model drug Acyclovir which was analyzed in human plasma with the use of ganciclovir as internal standard. In vivo study was also performed. Derivative treatment of chromatographic response ratio data was followed by convolution of the resulting derivative curves using 8-points sin xi polynomials (discrete Fourier functions). This work studies and also compares the application of WR method and Theils method, a nonparametric regression (NPR) method with the least squares parametric regression (LSPR) method, which is considered the de facto standard method used for regression. When the assumption of homoscedasticity is not met for analytical data, a simple and effective way to counteract the great influence of the high concentrations on the fitted regression line is to use WR method. WR was found to be superior to the method of LSPR as the former assumes that the y-direction error in the calibration curve will increase as x increases. Theils NPR method was also found to be superior to the method of LSPR as the former assumes that errors could occur in both x- and y-directions and that might not be normally distributed. Most of the results showed a significant improvement in the precision and accuracy on applying WR and NPR methods relative to LSPR. FigureComparison between RSD (percent) and Er (percent) calculated for Acyclovir, for area, D1, and D1/FF in the internal standard methods, using the three types of regression models: parametric, nonparametric, and weighted