A comparative analysis of univariate versus multivariate eco-friendly spectrophotometric manipulations for resolving severely overlapped spectra of vonoprazan and amoxicillin new combination.

Vonoprazan and amoxicillin are pharmacological combinations that demonstrate synergistic effects in treating Helicobacter pylori (H. pylori), a global public health concern associated with peptic ulcer disease and gastric cancer. Four spectrophotometric methods were developed, including two univariate techniques (Fourier self-deconvolution and ratio difference) and two multivariate chemometric approaches (partial least squares and principal component regression). These methods provide innovative solutions for effectively resolving and accurately quantifying the overlapping spectra of vonoprazan and amoxicillin. The concentration ranges covered were 3-60 µg ml-1 for vonoprazan and 5-140 µg ml-1 for amoxicillin. To assess the environmental sustainability of the methodologies, various measures such as the Green Analytical Procedure Index (GAPI), National Environmental Method Index (NEMI), Analytical GREEnness Calculator, and Analytical Eco-scale, as well as RGB12 and hexagon toll were implemented. The validation of the developed techniques was carried out in compliance with ICH standards. The present study is highly significant because it is the first time that the mixture has been determined using the current approaches. The comparative analysis demonstrated no significant difference in terms of accuracy and precision compared to reference HPLC method (p = 0.05). The established spectrophotometric methods offer a straightforward, rapid, and cost-effective alternative to complex analytical techniques for determining the vonoprazan and amoxicillin mixture. They show potential for routine analysis in research laboratories and pharmaceutical industries.

Different Eco-Friendly Spectrophotometric Approaches Including Direct and Manipulations of Zero and Ratio Spectra for Simultaneous Determination of Novel Nasal Spray Combination Used in Seasonal Allergic Rhinitis.

BACKGROUND: The presentation of rhinitis has drawn increasing attention in recent years due to the possibility of overlap or confusion between allergic rhinitis symptoms and those of COVID-19. Azelastine hydrochloride (AZH) and mometasone furoate (MOF) are two of the most efficient combinations for enhancing the symptoms of seasonal allergic rhinitis. OBJECTIVE: This work concerns applying and validating different accurate and simple spectrophotometric approaches for simultaneous quantification of the binary mixture of AZH and MOF in raw material, laboratory-prepared mixtures, and pharmaceutical preparation. Moreover, assessment of the environmental impact of the applied approaches on the environment was also a key goal of this study. METHODS: AZH was determined using the direct spectrophotometric (D0) method, while four reliable spectrophotometric approaches namely, induced dual wavelength (IDW), ratio subtraction (RS), ratio difference (RD), and ratio derivative (1DD) were used for MOF determination. RESULTS: The methods were validated in line with the International Conference of Harmonization standards. In the AZH range of (5-56 µg/mL) and MOF range of (2-20 µg/mL), the linearity of the proposed approaches was investigated with high accuracy findings. There were no significant differences between the obtained results and those of the reported method when compared statistically. Furthermore, the applied spectrophotometric methods were deemed to be eco-friendly according to Green Analytical Procedure Index (GAPI) and Analytical Greenness Calculator (AGREE) assessment metrics. CONCLUSIONS: The applied spectrophotometric methods are simpler, more eco-friendly, and take a shorter time to precisely estimate many measurements compared to the only reported chromatographic analysis. HIGHLIGHTS: Neither publications of novel spectrophotometric methods nor reported green ones have been available for simultaneous determination of the binary mixture of AZH and MOF, so this work has a great significance and novelty in the area of pharmaceutical analysis.

Valuation of environmental influence of recently invented high-performance liquid chromatographic method for hypoglycemic mixtures of gliflozins and metformin in the presence of melamine impurities: Application of molecular modeling simulation approach.

Molecular modeling is the science of representing molecular structures numerically and simulating their behavior with the equations of quantum and classical physics. Coupling molecular modeling and simulation with chromatographic resolution for pharmaceutical products constitutes a new technique in pharmaceutical analysis. An innovative high-performance liquid chromatographic (HPLC) methodology was developed for the quantification of metformin hydrochloride (MET), empagliflozin (EMP), and canagliflozin (CAN) in bulk, laboratory-developed combinations, pharmaceutical tablets, and in the presence of melamine. Chromatographic separation was accomplished using a Symmetry column with 0.03 M potassium dihydrogen phosphate buffer and 0.02 M heptane sulphonic acid: acetonitrile as the mobile phase. Molecular modeling using molecular operating environment software was applied to properly select the stationary phase suitable for the developed HPLC method. Additionally, molecular modeling estimates and validates binding between the studied analytes and the stationary phase to clarify and explain the chromatographic separation and elution order. In accordance with the International Conference of Harmonization recommendations, the method was validated in terms of linearity, accuracy, precision, and selectivity. The linearity ranges (µg/ml) were 200-1500 (MET), 2-15 (EMP), and 20-150 (CAN) and the limit of detection values were in the ranges of 0.17-54.58 µg/ml. Analysis of pharmaceutical tablets using the suggested approach yielded satisfactory outcomes. As a result, it might be used in quality control laboratories to analyze the aforementioned medications.

Green-assisted spectrophotometric techniques utilizing mathematical and ratio spectra manipulations to resolve severely overlapped spectra of a cardiovascular pharmaceutical mixture.

Cardiovascular diseases, in particular hypertension and hypercholesterolemia, are two of the main causes of death worldwide. These conditions are silent killer syndromes that need a variety of pharmacological treatments to be effectively controlled. This study introduces novel, environmentally friendly spectrophotometric techniques for the simultaneous determination of telmisartan (TMS) and rosuvastatin calcium (RVS) in their pharmaceutical dosage forms. For the simultaneous determination of the binary mixture, the suggested methods included the dual wavelength method (DWM) which utilizes mainly the absorbance difference at 233 nm and 253 for TMS determination and, the absorbance difference at 274 nm and 310 for RVS determination as the selected wavelengths for each drug is directly proportional to the drug of interest independent on the other interfering component. The Fourier-self deconvolutions method (FSDM) depends on compressing their bandwidth to resolve the overlap. Ratio difference spectrophotometric method (RDSM) that utilizes TMS 35 µg.mL-1 and RVS 20 µg.mL-1, respectively as divisors to produce the ratio spectra for each drug. Further manipulation of the produced ratio spectra was applied for the determination of the two drugs. Mean centering method (MCM) where a suitable wavelength range was chosen to exclusively use the informative portions and prevent experimental spectrum noises. The investigated methods showed good levels of detection and quantification together with excellent linearity. The suggested methods' greenness was evaluated using two different greenness evaluation tools, which showed that the methods were green in terms of several factors, including the safety of the chemicals, instruments, and waste. The validity of the methodologieswas investigated by resolving prepared laboratory mixtureswith varying TMS and RVS ratios. The standard addition method also assured the newly added methods. Finally, statistical analysis using the reported method did not reveal any appreciable differences in terms of accuracy and precision. The developed methods can be employed in quality control laboratories to ascertain the binary mixture due to their high precision and affordability.

A green spectrophotometric method for the simultaneous determination of nasal binary mixture used in respiratory diseases: Applying isosbestic point and chemometric approaches as a resolving tool, greenness evaluation.

Nasal drug combination is a very useful therapy for elevating the symptoms of various respiratory diseases as seasonal allergic rhinitis and infectious respiratory illness as pandemic COVID-19. One of best combination is Fluticasone propionate (FLU) and Azelastine (AZE). In this study, different UV spectrophotometric and chemometric methods have been applied for quantitative analysis of FLU and AZE without previous separation in their pure form, laboratory prepared mixture and pharmaceutical dosage form. Absorbance subtraction (AS) and Amplitude modulation (AM) spectrophotometric methods have been applied for the simultaneous determination of the cited drugs. Besides, three well-known chemometric techniques; namely, classical least squares (CLS), partial least square (PLS), and principal component regression (PCR) have been applied for the simultaneous analysis of both drugs by using spectrophotometric data. To be friendly to the environment, the greenness of the proposed methods was taken into consideration and evaluation of the analytical methods' greenness was done using two green analytical chemistry metrics known as, Analytical Greenness Calculator and an eco-scale scoring method. They indicated that the methods were environmentally friendly in relation to numerous approaches like instrument, reagents, and safety of waste. Analyzing laboratory prepared mixtures including different quantities of FLU and AZE, as well as their marketed dose form, was used to assess the selectivity of the applied methods. The validity of the developed methods was investigated by applying the standard addition technique. The resulting data were statistically compared to those obtained by the official or reported HPLC methods for FLU and AZE, which revealed no significant difference in accuracy and precision at p = 0.05.

Different approaches for the assessment of greenness of spectrophotometric methodologies utilized for resolving the spectral overlap of newly approved binary hypoglycemic pharmaceutical mixture.

Simultaneous measurement of saxagliptin hydrochloride (SAG) and dapagliflozin propanediol monohydrate (DAG) in bulk powder, laboratory-prepared mixtures, and pharmaceutical dosage form were applied by utilizing three precise and sensitive spectrophotometric techniques which were developed and validated. The first method was the induced dual-wavelength approach (IDW), which relied primarily on the use of alternative equality factors (F) to abolish the effect of DAG when determining SAG and vice versa. The ratio difference method (RDM) was the second method, which used 25 µg/ml of DAG and 20 µg/ml of SAG as divisors to determine the amplitude difference on the ratio spectrum of SAG and DAG, respectively. SAG was determined at λmax 221 nm after plateau subtraction followed by multiplication by the divisor of DAG 25 µg/ml using the third method, ratio subtraction coupled with extended ratio subtraction method (RSER). Subsequently, using an extension ratio subtraction of the spectra, DAG was determined at λmax 225 nm was determined. The developed methods were effectively used to estimate SAG and DAG in laboratory-prepared mixtures and pharmaceutical dosage forms, with satisfactory recoveries. The methodologies were assessed for their environmental friendliness using the analytical eco-scale, analytical GREEnness calculator, and green analytical procedureindex (GAPI). These methodologies were validated following the International Conference on Harmonisation (ICH) requirements. A statistical comparison of the obtained findings to those of the published method revealed no significant differences in precision and accuracy. Because of their high precision and cost-effectiveness, the developed methods can be used in quality control laboratories to determine the binary mixture.

Application of experimental design in HPLC method optimization and robustness for the simultaneous determination of canagliflozin, empagliflozin, linagliptin, and metformin in tablet.

Gliflozins and gliptins represent two different pharmacological drug classes that exert different and potentially complementary glucose-lowering effect in patients with type II diabetes mellitus. A novel, selective, and sensitive HPLC method was developed for the determination of canagliflozin, empaglifozin, linagliptin, and metformin in pure form, in laboratory prepared mixtures, and in pharmaceutical dosage form. Experimental design optimization was applied by using Plackett-Burman and face-centered composite designs to achieve the best resolution with minimum experimental trials. Three significant variables affecting optimization, namely buffer pH, percentage of methanol, and percentage of acetonitrile, were studied. Chromatographic separation was achieved using an Agilent Eclipse C8 column, and column temperature was kept at 45°C. The mobile phase was composed of dipotassium hydrogen phosphate buffer (0.05 M, adjusted to pH 6 using o-phosphoric acid):acetonitrile:methanol (50:25:25, v/v/v) at a flow rate of 1.5 mL/min. Sharp and well-resolved peaks of the cited drugs were obtained. The method was fully validated in terms of linearity, accuracy, precision, selectivity and robustness in agreement with the International Council of Harmonization (ICH) guidelines Q2 (R1). Satisfactory results were obtained by the analysis of tablets through applying the developed method. Therefore, it could be performed for the analysis of the cited drugs in quality control laboratories.

Smart spectrophotometric methods for the simultaneous determination of newly co-formulated hypoglycemic drugs in binary mixtures.

Achieving good glycemic control in patients with type II diabetes mellitus is essential for preventing both microvascular and macrovascular complications. Combination therapy represents the principle strategy for successful long term control of type II diabetes mellitus with minimal complications. Two sensitive, precise and non-destructive spectroscopic methods were developed for the simultaneous estimation of two new co-formulated hypoglycemic drugs; canagliflozin/metformin (CAG/MEF) and empagliflozin/linagliptin (EMG/LIG) in tablets with no need of previous separation. The first method was amplitude modulation (a normalized spectra-based UV spectrophotometric method) for the analysis of (CAG/MEF) binary mixture. The amplitude of the constant at the plateau region at (264-310 nm) on the ratio spectrum was measured and used for the determination of CAG concentration in the mixture. On the other hand, MEF was estimated by subtracting the previously obtained amplitude from the total amplitude of CAG and MEF at the isosbestic point (λiso) at 250 nm. The second method was chemometric-assisted FTIR spectrophotometric method for the determination of (EMG/LIG) binary mixture. (EMG/LIG) mixture in chloroform was analyzed using FTIR in the region 4000-400 cm-1. The spectral region 3900-2900 cm-1 was selected for (EMG/LIG) determination using principal component regression and partial least squares chemometric methods. The methods were validated according to ICH guidelines. The studied drugs were successfully determined in tablets applying the developed methods. Validation parameters were in agreement with acceptance limits, ensuring methods accuracy and selectivity. Besides, no significant difference was obtained by statistically comparing the obtained results with the reported one.

A validated LC-MS/MS method for simultaneous determination of linagliptin and metformin in spiked human plasma coupled with solid phase extraction: Application to a pharmacokinetic study in healthy volunteers.

Combination therapy has a pivotal role in type II diabetes mellitus management in patients unable to maintain normal glycemic level using metformin alone. Addition of linagliptin, dipeptidyl peptidase-IV inhibitor, to metformin improves glycemic control. This study is concerned with the development of an HPLC-MS/MS method for simultaneous quantification of linagliptin and metformin in spiked human plasma. The method was applied to evaluate the potential pharmacokinetic interactions between the cited drugs in healthy volunteers. Solid phase extraction was applied using Strata™ X cartridge. Separation was carried out on Symmetry® C18 column using methanol: 10 mM ammonium formate buffer (containing 0.2% formic acid) in a ratio of (95: 5, v/v) as mobile phase at flow rate 0.25 mL min-1. Quantification was performed with multiple reaction monitoring in positive ionization mode. The monitored transitions were set at m/z 473.24 → 419.94, 130.14 → 60.18 and 340.27 → 116.07 for linagliptin, metformin and alogliptin (internal standard), respectively. The method was validated according to FDA guidelines. The method showed excellent linearity over concentration ranges 0.25-10 and 25-2000 ng mL-1 for linagliptin and metformin, respectively. The validated HPLC-MS/MS method was successfully applied to pharmacokinetic study of linagliptin and metformin in healthy volunteers after oral administration of Jentadueto® tablets.

Different resolution techniques for management of overlapped spectra: Application for the determination of novel co-formulated hypoglycemic drugs in their combined pharmaceutical dosage form.

To maintain intensive glucose control early in the type II diabetes mellitus process, novel combinations of canagliflozin/metformin (CAG/MEF) and empagliflozin/linagliptin (EMG/LIG) offer particular treatment benefits. In this study, sensitive and precise spectrophotometric methods were developed for the determination of such hypoglycemic drug combinations in bulk powder and in pharmaceutical dosage form without prior separation. The first method was ratio difference coupled with modified isosbestic point technique where the amplitude difference between 239 and 291 nm on the ratio spectrum of CAG obtained using 4 µg/ml of MEF as divisor was used for determination of CAG. On the other hand, MEF was estimated using a modified isosbestic spectrophotometric method, where the total concentration of CAG and MEF in mixture could be calculated at 250 nm (isosbestic point) after multiplication by a correction factor. Then concentration of MEF could be calculated by subtraction. The second method was ratio subtraction coupled with extended ratio subtraction, where EMG was determined at 225 nm by subtraction of plateau values from the ratio spectrum followed by multiplication with the spectrum of 6.5 µg/ml of LIG (divisor). Then, an extension of the normal ratio subtraction method was performed in order to determine LIG at 226 nm. Linearity was obtained over 5-30, 2.5-16, 2.5-16 and 1.25-8 µg/ml for CAG, MEF, EMG and LIG, respectively. The developed methods were successfully applied for the determination of studied drugs in tablets. Validation parameters were found to be within acceptance limits, thus confirming methods accuracy and selectivity. The obtained results were statistically compared with the reported one showing no significant difference in terms of accuracy and precision. The methods could be applied for routine analysis of the cited drugs in quality control laboratories.

Spectrofluorimetric determination of gemifloxacin mesylate and linezolid in pharmaceutical formulations: Application of quinone-based fluorophores and enhanced native fluorescence.

Quinone-based fluorophores and enhanced native fluorescence techniques were applied for a fast quantitative analysis of gemifloxacin mesylate (GEM) and linezolid (LIN) in pharmaceutical formulations. For this purpose, three sensitive, accurate and precise spectrofluorimetric methods were developed. GEM, as an n-electron donor, reacts with 7,7,8,8-tetracyanoquinodimethane (method A) and 2,5-dichloro-3,6-dihydroxy-p-benzoquinone (method B) as п-electron acceptors, forming charge transfer complexes that exhibit high fluorescence intensity at 441 and 390 nm upon excitation at 260 and 339 nm, respectively. Method C depends on measurement of enhanced native fluorescence of LIN in phosphate buffer (pH 5) at 380 nm upon excitation at 260 nm. Experimental factors affecting fluorescence intensity were optimized. Linearity was obtained over concentration ranges 50-500, 10-60 and 20-400 ng mL-1 for methods A, B and C, respectively. The developed methods were validated and successfully applied for determination of the cited drugs in tablets.