Development and Evaluation of Chemometric Models for the Estimation of Sumatriptan in the Presence of Naproxen and a Degradation Product Using UV Spectrophotometry.

BACKGROUND: Chemometrics is a discipline that allows the spectral resolution of drugs in a pharmaceutical formulation along with degradation product and it is as an alternative to chromatographic methods. OBJECTIVE: Sumatriptan (SUM) is co-formulated with naproxen (NAP) and used in acute migraine attacks. SUM which has physiological importance has not been subjected to any stability-indicating chemometric approaches yet so there is a need for accurate and safe method for the assay of the cited drug in their preparation. SUM was determined in pharmaceutical formulation along with NAP and in presence of alkali-induced degradation product with simple and cost-effective multivariate approaches using spectrophotometric data. The greenness and blueness assessment was applied using different ecological metrics, including green analytical procedure index (GAPI), analytical greenness metric (AGREE), analytical eco-scale (AES) & new "blueness" evaluation using BAGI tool, respectively. METHODS: Three chemometric approaches were applied for the stability-indicating determination of SUM in presence of NAP. Classical least squares (CLS), Partial least squares (PLS) and Principal components regression (PCR), three multivariate calibration numerical models that were performed on the UV- spectra of the mixtures, were used to achieve the best resolution. RESULTS: Sumatriptan was analyzed with mean accuracies for PLS (100.29 ± 1.318) and for PCR (100.60 ± 1.564). The presented methods were compared and validated for their quantitative analyses. Moreover, statistical comparison between the results obtained by the proposed models and the official methods showed no significant differences. CONCLUSION: The proposed multivariate calibrations were accurate and specific for quantitative analysis of the studied component. PLS is the best method that has the capacity for qualitative analysis of SUM and it is suitable for routine analysis and stability studies of it in quality control laboratories. Various ecological assessment metrics confirmed the long-standing eco-friendliness of the suggested models.

A high-throughput UPLC-MS-MS Bio-analytical method for the analysis of veterinary pharmaceutical residues in Chicken Tissues, Application of efficient-valid-green (EVG) Framework as a Competence Tool.

Antibacterial medications are receiving the most attention due to hypersensitivity reactions and the emergence of bacterial mutants resistant to antibiotics. Treating Animals with uncontrolled amounts of antibiotics will extend beyond their lives and affect humans. This study aims to determine the concentration of the residues of sulfadimidine, sulfaquinoxaline, diaveridine, and vitamin K3 in the tissues of poultry (muscles and liver) after treatment with the combined veterinary formulation. A UPLC-MS-MS method was developed using Poroshell 120 ECC18 and a mobile phase composed of acetonitrile and distilled water, containing 0.1 % formic acid, in the ratio of (85:15 v/v) at a flow rate of 0.6 mL/min. Sample extraction solvent was optimized using response surface methodology (RSM) to be acetonitrile: methanol in the ratio (49.8: 50.2 v/v), and the method was validated according to the FDA bioanalytical method validation protocol over the range (50-1000 µg/Kg) for sulfaquinoxaline and (50-750 µg/Kg) for the other 3 drugs. The greenness of the sample preparation and analytical method was assessed by applying Analytical Eco-scale (AES) and AGREE coupled with AGREEprep. The Competence of the study was evaluated via the EVG framework known as Efficiency, validation, and greenness, to achieve a balance point represented by a radar chart. The method was applied to decide the time required for poultry products to be safe for human use after administration of the studied drugs. It was found that, after the administration of the last dose, minimally 7 days are required till the levels of the drugs drop to the maximum residue limit determined by the FDA/WHO in animal tissues.

An integrated framework to develop an efficient valid green (EVG) HPLC method for the assessment of antimicrobial pollutants with potential threats to human health in aquatic systems.

The persistence of antimicrobial drugs in aquatic environments has raised critical concerns about their possible impact on drinkable water quality and human health. The Nile River is experiencing water pollution owing to increasing discharges of highly contaminated home and industrial effluents and inadequate water management systems. Investigations of the presence of three antimicrobial agents, ciprofloxacin (CIP), sulfamethoxazole (SMZ), and albendazole (ALB), in the Egyptian aquatic system are recommended using a chromatographic method because of their reported existence in the African aquatic environment. In this study, an integrated framework, Efficient Valid Green (EVG), for analytical techniques is proposed and displayed via its radar chart. The EVG framework is achieved through three main pillars: efficiency, validation, and greenness. The proposed EVG-HPLC method was developed and optimized using the AQbD methodology via a face-centered composite (FCC) design by identifying the proper critical method parameters (CMPs) that influence critical quality attributes (CQAs). The method was fully validated according to ICH guidelines, including a factorial robustness study within concentration ranges of 1-100 µg mL-1, 2-100 µg mL-1, and 10-100 µg mL-1 for CIP, SMZ, and ALB, respectively. The proposed method was evaluated in terms of greenness using AGREE (score 0.55) and ComplexGAPI metrics. The optimized chromatographic conditions included a C18 column and a mobile phase of water : acetonitrile : methanol in a ratio of 60 : 19 : 21, v/v/v, respectively, with an aqueous solution of pH 3.5 adjusted with phosphoric acid at a flow rate of 1.57 mL min-1 at 285 nm. The raw water samples collected from Nile River freshwater at different locations were treated using Oasis® PRiME HLB cartridges with satisfactory recoveries for the three analytes (>90%), and the three drugs were detected using the proposed EVG-HPLC method.

Sustainable chromatographic quantitation of multi-antihypertensive medications: application on diverse combinations containing hydrochlorothiazide along with LC-MS/MS profiling of potential impurities: greenness and whiteness evaluation.

Cardiovascular disorders are among the leading causes of death worldwide, especially hypertension, a silent killer syndrome requiring multiple drug therapy for appropriate management. Hydrochlorothiazide is an extensively utilized thiazide diuretic that combines with several antihypertensive drugs for effective treatment of hypertension. In this study, sustainable, innovative and accurate high performance liquid chromatographic methods with diode array and tandem mass detectors (HPLC-DAD and LC-MS/MS) were developed, optimized and validated for the concurrent determination of Hydrochlorothiazide (HCT) along with five antihypertensive drugs, namely; Valsartan (VAL), Amlodipine besylate (AML), Atenolol (ATN), Amiloride hydrochloride (AMI), and Candesartan cilextil (CAN) in their diverse pharmaceutical dosage forms and in the presence of Chlorothiazide (CT) and Salamide (DSA) as HCT officially identified impurities. The HPLC-DAD separation was achieved utilizing Inertsil ODS-3 C18 column (250 × 4.6 mm, 5 µm) attached with photodiode array detection at 225.0 nm. Gradient elution was performed utilizing a mixture of solvent A (20.0 mM potassium dihydrogen phosphate, pH 3.0 ± 0.2, adjusted with phosphoric acid) and solvent B (acetonitrile) at ambient temperature. Linearity ranges were 0.1-100.0 µg/mL for HCT, VAL, AML and CAN, 0.05 -100.0 µg/mL for both ATN and AMI and 0.05-8.0 µg/mL for both CT and DSA. Additionally, this work describes the use of liquid chromatography-electrospray-tandem mass spectrometry for the accurate detection and quantification of the impurities; CT and DSA in the negative mode utilizing triple quadrupole mass spectrometry. The linearity ranges for those impurities were 1.0-200.0 ng/mL and 5.0-200.0 ng/mL for CT and DSA, respectively. Developed methods' validation was achieved in accordance with International Conference on Harmonization (ICH) guidelines. Upon applying liquid chromatographic techniques for the drug analysis, a green and sustainable assessment have to be handled due to the consumption of energy and many solvents. Through the use of the HEXAGON, Analytical Greenness (AGREE) and White Analytical Chemistry (WAC) tools, greenness and sustainability have been statistically assessed. The optimized HPLC-DAD and LC-MS/MS methods were fast, accurate, precise, and sensitive, and consequently could be applied for conventional analysis and quality control of the proposed drugs in their miscellaneous dosage forms for the purpose of reducing laboratory wastes, time of the analysis time, effort, and cost.

Coupling of physical extraction and mathematical filtration in spectrophotometric analysis of natural therapy essential for prophylaxis and treatment of COVID-19 infection - Comparative study along with greenness evaluation.

Diosmin (DIO) and hesperidin (HSP) are important classes of flavonoid glycoside effectively used to prevent comorbid diseases that are commonly associated with COVID-19. An innovative, green, ccurate, effective, cost, and timeless spectrophotometric strategy was established to analyze such challengeable mixture in a co-formulated tablet namely Diosed C® tablets that comprises DIO, HSP and vitamin C (VIT. C) in the ratio of (450 mg: 50 mg: 100 mg) necessary for prevention and treatment of COVID-19. Vitamin C was resolved through physical extraction using de-ionized water while DIO and HSP were extracted via spectrophotometric methods using two different solvents [0.1 M NaOH or solvent blend consisting of DMSO and methanol (1:1)]. Mathematical filtration technique is successfully applied to recover the parent spectra of both DIO and HSP via three methods which are absorbance resolution (AR), Induced absorbance resolution (IAR) and ratio extraction (RE). VIT. C was successfully analyzed in de-ionized water using its maxima at 266.0 nm in a linearity range 2.0-20.0 µg/mL, DIO was effectively determined in 0.1 M NaOH at 372.0 nm in a linearity range of 7.0-70.0 µg/mL as well as in solvent blend at 344.0 nm in linearity range of 5.0-55.0 µg/mL while HSP was accurately analyzed in 0.1 M NaOH at 240.0 nm in linearity range of 3.5-50.0 µg/mL as well as in solvent blend at 285.0 nm in linearity range of 4.0-50.0 µg/mL. Satisfactory results were accomplished when conducting ICH guidelines for assuring the methods validation. Comparative study was introduced in the analysis of such critical combination and was prosperously devoted for the effective analysis of pharmaceutical dosage form. The proposed extraction pathways undergo the guidelines of green analytical chemistry using Analytical Eco-Scale (AES), AGREE and GAPI greenness assessment tools which confirmed their eco-friendly nature with priority to 0.1 M NaOH. The obtained results of the suggested methods were set side by side with those of official/reported methods statistically and show satisfactory implications. The presented methods were simple, affordable, smoothly applicable and their results were acceptable that enhances their usage and application in the quality control laboratories.

Sustainable spectrophotometric determination of antihypertensive medicines reducing COVID-19 risk via paired wavelength data processing technique - Assessment of purity, greenness and whiteness.

Recent studies have reported that using certain antihypertensive therapies such as angiotensin II receptor blockers (ARBs) is associated with mitigation of fatal outcomes and enhancing clinical features of patients having hypertension during coronavirus pandemic. Thus, in the current work an innovative, effective, white and sustainable spectrophotometric technique called paired wavelength data processing technique (PWDPT) was developed for evaluation of recommended antihypertensive combination therapies incorporating candesartan cilexetil (CAN) and hydrochlorothiazide (HCT). This technique included three methods, namely, absorbance resolution (AR), amplitude resolution (PR) and ratio extraction (RE). Linearity ranges were (5.0 µg/mL - 50.0 µg/mL) and (2.0 µg/mL - 24.0 µg/mL) for CAN and HCT, respectively. Validation and confirmation of all suggested methods were conducted in accordance with ICH guidelines, producing satisfactory results within the accepted limits. Statistical comparison was achieved between the attained results from suggested methods and those attained from official methods, in which insignificant difference was existed. The suggested methods were successfully employed for identification of the studied drugs as well as determination of their spectral recognition and evaluation of the purity in their combined formulations. The proposed methods followed the principles of green analytical chemistry, where their greenness was evaluated and compared with the official potentiometric and HPLC methods via using four tools, namely, National Environmental Methods Index (NEMI), the Analytical Eco-Scale, the Green Analytical Procedure Index (GAPI) and Analytical greenness metric (AGREE) which affirmed the eco-friendly nature of the proposed methods. Moreover, studying the whiteness features was performed using the recently introduced RGB12 model. The acceptable results along with the sustainability, simplicity, affordability and low-cost of the proposed methods encourages their utilization in the quality control laboratories.

Simultaneous determination of hydrochlorothiazide, amlodipine, and telmisartan with spectrophotometric and HPLC green chemistry applications.

For the quantifiable amounts of Telmisartan (TLM) and Hydrochlorothiazide (HYD) in the presence of Amlodipine (AML) in a ternary mixture of synthetic laboratory mixture, a novel, sensitive, quick, and practical reversed-phase high-performance liquid chromatography (RP-HPLC) method was given. In order to separate, a Waters Spherisorb ODS-2 C18 column was used. For HYD, TLM, and AML, these techniques were viable over linearity ranges of 4-12 µg/mL, 4-25 µg/mL, and 5-40 µg/mL, respectively. The mobile phase system was acetonitrile:methanol: phosphate buffer at pH 2.5 (65:5:30 v/v/v), and the flow rate was 1.5 mL/min. Novel spectrophotometric methods were applied for active substances to determine simultaneously. The first method is absorptivity centering using factorized spectrum, and the second method is dual amplitude difference coupled with absorbance subtraction. These approaches have been effectively applied to bulk, laboratory synthetic mixtures to employ active components quantitatively. Correlation coefficients were found to be higher than 0.99 and the limit of detection values lower than 0.49 µg/mL in both spectrophotometric methods. The methodologies were validated following ICH recommendations. In the developed HPLC method, the limit of detection values was found to be 0.01 µg/mL for HYD and 0.02 µg/mL for AML and TLM. The correlation coefficients for the HPLC method were found to be 0.9971 for HYD, 0.9990 for AML, and 0.9983 for TLM. The suggested HPLC technique is a simple, effective, sensitive, environmentally friendly, and time-saving approach for determining TLM and HYD in the presence of AML.

Eco-friendly spectrophotometric evaluation of triple-combination therapies in the treatment strategy of patients suffering from hypertension during coronavirus pandemic - Spectralprint recognition study.

Recent studies have reported that using certain antihypertensive therapies such as angiotensin II receptor blockers (ARBs) and calcium channel blocker (CCBs) is associated with reduction of fatal outcomes and improving clinical characteristics of patients suffering from hypertension during coronavirus pandemic. Thus, in the current work an effective, innovative and eco-friendly spectrophotometric manner namely, parent spectrum extraction (PSE)was established for evaluation of recommended triple antihypertensive combination therapies incorporate valsartan (VAL) as ARBs, amlodipine besylate as CCBs (AML) and hydrochlorothiazide (HCT)as diuretic into single-pill in challengeable ratio. PSE manner composed of two complementary steps, auxiliary resolution coupled with data analysis resolution(DAR)and it is characterized by resolving the spectral bands of the drugs and extraction of their discrete parent spectra (D0); accordingly, enabling determination of each analyte at its λmax. Auxiliary resolution of AML in triple mixture was applied to decrease complexity of overlapped spectra via constant multiplication (CM) followed by spectrum subtraction (SS) to obtain resolved mixture of VAL and HCT while data analysis resolution (DAR) of this binary mixture was applied via one of three novel methods namely, absorbance extraction (AE), peak-amplitude extraction (PE) and ratio extraction (RE) along with SS method. The proposed methods had analyzed VAL, AML and HCT in the range of 4.0-44.0 µg/mL, 4.0-40.0 µg/mL and 2.0-24.0 µg/mL, respectively with an excellent correlation coefficient (r ≥ 0.9999). Further, the proposed methods in PSR manner were validated as stated by ICH guidelines and it was found that accuracy and precision results are within the acceptable limit. The suggested procedures were effectively utilized for the concurrent quantification of VAL, AML and HCT in synthetic mixtures and tablets. The greenness of the proposed spectrophotometric methods was evaluated by National Environmental Methods Index (NEMI), the Analytical Eco-Scale, the Green Analytical Procedure Index (GAPI) and Analytical greenness metric (AGREE) where the four tools affirmed the eco-friendly nature of the proposed methods. A comparison between the outcomes of the studied methods with the official and reported ones was performed and no statistical difference was arisen between the methods regarding to accuracy and precision.The achieved results along with the simplicity, affordability and low-cost of the proposed methods recommended their appropriateness for the regular quality control examination and analysis of pure materials and pharmaceutical formulations as well as their applicability for the spectralprint recognition of the studied drugs.

Sticking - pulling strategy for assessment of combined medicine for management of tough symptoms in COVID-19 Pandemic using different windows of spectrophotometric Platform-Counterfeit products' detection.

This study presents a comprehensive comparative study of different green spectrophotometric approaches without any physical separation on processing a ternary mixture of Aceclofenac (ACE), Paracetamol (PAR) and Rabeprazole (RAB) in combined medicine for managing tough symptoms in the COVID-19 Pandemic. The different univariate complementary resolutions according to the response used for the assay of the cited drugs after applying the processing steps were implemented using successive in-silico sample enrichment for resolving the ternary mixture via different windows of spectrophotometric platform using sticking - pulling strategy (SPS). Window I; based on manipulation of the data of zero order absorption spectrum of the mixture using novel Extended absorbance difference (EAD) and Absorbance difference (AD) methods coupled with corresponding spectrum subtraction method (SS). Window III; based on manipulation of the data of ratio spectra via Constant value coupled with constant subtraction (CV-CS) and novel Induced dual amplitude difference (IDAD) method coupled with corresponding spectrum subtraction method (SS). Finally, window IV; based on manipulation of the data of derivative of the ratio spectrum of the mixture via novel Factorized derivative ratio null contribution (FDD-NC) and Factorized unlimited derivative ratio (FUDD) methods coupled with corresponding spectrum subtraction method (SS). Synthetic mixtures and commercial medicine were constructively analyzed using the proposed methods while maintaining calibration graphs to be linear over ranges; 4.0-40.0 µg/mL for ACE, 2.0-14.0 µg/mL for PAR and 4.0-30.0 µg/mL for RAB. Moreover, methods' validation was confirmed via performing exhaustive statistical treatment of the experimental findings. The proposed methodologies can be used for the routine analysis of the cited drugs in quality control laboratories. Additionally, Spectral Similarity Index (SSI) was calculated to detect counterfeit products and methods' greenness profile was finally guaranteed through analytical greenness (AGREE) metric assessment tool.

The power of High Impact Amplitude Manipulation (HIAM) technique for extracting the basic spectra of two Fixed-dose combinations (FDC) -Spectrophotometric purity analysis via spectral contrast angle.

HIAM technique allows the extraction of the original constant signal of each single component out of interference signals of a mixture and further transformed into basic spectrum (D0). It includes the methods: ratio subtraction coupled with unified constant subtraction (RS-UCS), constant center (CC) and constant extraction (CE). The technique was introduced for the analysis of two pharmaceutical formulations used to treat cardiovascular diseases. The formulations are binary combinations of Amlodipine (AML) with either Atorvastatin (ATR) or Candesartan (CND) which shows interefernce absorbance signals. The technique was valid over the linearity range of (5.0-35.0 µg/ml) for AML, ATR and CND with recovery percentage 100.40 ± 1.88 , 100.00 ± 0.86 and 99.83 ± 1.07, respectively . The extracted signals were tested for its purity by spectral contrast angle (cos θ) to illustrate the efficency of the HIAM technique where cos θ values ranges from (0.9902 to 0.9986). The presented technique was fully validated regarding ICH guidelines and were statistically compared using one-way ANOVA at 95% confidence.

Smart Spectrophotometric Methods for Concurrent Determination of Furosemide and Spironolactone Mixture in Their Pharmaceutical Dosage Forms

Abstract Simple, precise, accurate and specific spectrophotometric methods are progressed and validated for concurrent analysis of Furosemide (FUR) and Spironolactone (SPR) in their combined dosage form depend on spectral analysis procedures. Furosemide (FUR) in the binary mixture could be analyzed at its λmax 274 nm using its recovered zero order absorption spectrum using constant multiplication method (CM). Spironolactone (SPR) in the mixture could be analyzed at its λmax 238 nm by ratio subtraction method (RS). Concurrent determination for FUR and SPR in their mixture could be applied by amplitude modulation method (AM), absorbance subtraction method (AS) and ratio difference (RD). Linearity ranges of FUR and SPR were (2.0µg/mL-22.0 µg/mL) and (3.0µg/mL-30.0 µg/mL), respectively. Specificity of the proposed spectrophotometric methods was examined by analyzing the prepared mixtures in laboratory and was applied successfully for pharmaceutical dosage form analysis which have the cited drugs without additives contribution. The proposed spectrophotometric methods were also validated as per as the guidelines of ICH. Statistical comparison was performed between the obtained results with those from the official methods of the cited drugs, using one-way ANOVA, F-test and student t-test. The results are exhibiting insignificant difference concerning precision and accuracy

Evaluation of in silico and in lab sample enrichment techniques for the assessment of challengeable quaternary combination in critical ratio.

A comparative study of successive spectrophotometric resolution technique for the simultaneous determination of a challengeable quaternary mixture of Chlorpheniramine maleate (CPM), Pseudoephedrine hydrochloride (PSE), Ibuprofen (IBU) and Caffeine (CAF) is presented, without preliminary physical separation steps. Several successive steps were applied on built-in spectrophotometer software utilizing zero and/or derivative and/or ratio spectra of the studied components. These methods, namely, Dual amplitude difference (DAD) as a novel method, Constant multiplication coupled with spectrum subtraction method (CM-SS), Factorized first derivative coupled with derivative transformation method (FD1 -DT) and Derivative ratio method (DD1). The calibration graphs are linear over the concentration range of 10.0-80.0 µg/mL,150.0-900.0 µg/mL, 200.0-1400.0 µg/mL and 3.0-30.0 µg/mL for CPM, PSE, IBU and CAF, respectively. The specificity of suggested methods was studied via laboratory prepared (diverse ratios) mixtures and were successfully applied for Antiflu® capsules' analysis. Moreover, sample enrichment via In Silico (via software of spectrophotometer) and In Lab (via spiking with pure sample) techniques was elected for a pharmaceutical dosage form analysis comprising CPM and PSE as minor components. Accuracy, precision and specificity were between the valid limits. Validation steps were done in accordance with the ICH guidelines. Moreover, statistical comparison was carried out between the obtained and reported results for pure powder form and no significant difference appeared.

Trade-off efficacy and data processing strategy in the power of spectral resolution of co-formulated antihypertensive pharmaceuticals.

Versatile, extraction-free univariate spectrophotometric methods have been modified to get effective spectral resolution of mixtures in accordance with their feature challenges. The proposed methods have been applied and validated for analyzing some antihypertensive medicines in their co-formulated medicinal products. Two mixtures have been used: the first one [Mix I (OLM/ADB)] is composed of Olmesartan medoxomil (OLM) and Amlodipine besylate (ADB) with partly-overlapped spectra while the second [Mix II (TEL/HCT)] is made up Telmisartan (TEL) and Hydrochlorothiazide (HCT) with total-overlapped spectra. Induced dual wavelength, absorbance correction and ratio subtraction coupled with constant multiplication methods were applied to Mix I (OLM/ADB), while dual wavelength, advanced absorbance subtraction and constant center coupled with spectrum subtraction methods were applied to Mix II (TEL/HCT). Calibration graphs were established with good correlation coefficients. The methods exhibit significant advantages as simplicity, sensitivity, minimal data manipulation besides optimum robustness. Selectivity was inspected using lab-mixtures with diverse ratios. Accuracy, precision and repeatability were found to be within the acceptable limits. The proposed methods are good enough to be used for co-assay of analytes in combined forms without any interfering from excipients. Moreover, all results were estimated in accordance with ICH criteria and successfully compared with those of the reported methods applying t-test and F-test at 95% confidence level. Generally, these methods can be used efficiently for routine quality control testing.

Impact Study of Mathematical Manipulation on the Resolution Efficiency of the Spectrophotometric Technique-An Application on Veterinary Binary Mixture with Overlapping Absorption Bands.

BACKGROUND: The impact of mathematical manipulation on the efficiency of smart spectrophotometric approaches was considered for analyzing a veterinary binary mixture of tylosin tartrate (TYT) and doxycycline hydrochloride (DOX). OBJECTIVE: Based on subtraction which can be response subtraction via numerical factor calculation between two wavelengths. METHOD: Absorbance subtraction (AS) and amplitude summation (A-Sum), or subtraction of constant value or spectrum, was used to eliminate the contribution of the interfering component, as is done in spectrum subtraction (SS), extended ratio subtraction (EXRS), ratio subtraction (RS), and derivative subtraction (DS), or multiplication of a constant value by the divisor to get the spectrum of one component, or division by the normalized spectrum to modulate the original absorption spectrum to concentration as in concentration value and also detected in amplitude modulation (AM). The derivative transformation method (DT) is a form of mathematical manipulation that transforms a derivative to its original absorption form. RESULTS: The consumption of these methods will become more significant in quality control departments for the repetitive quantitative analysis of different veterinary products in both research and industry laboratories. Some manipulations may be used for simultaneous analysis of DOX and TYT, such as absorbance subtraction, amplitude modulation, ratio subtraction and derivative subtraction coupled with spectrum subtraction, concentration value, and amplitude summation, while others may be used for DOX only, such as derivative transformation and constant value. CONCLUSIONS: The recovery percentages confirmed that the accuracy and the reproducibility were approved by the following ICH guidelines. HIGHLIGHTS: 1. Partitioning methods: which perfectly divide the drugs response at the iso-point such as amplitude modulation, absorbance subtraction and amplitude summation. 2. Extracting methods: which easily calculate zero-order of the two cited drugs in such as ratio subtraction were coupled with extended ratio subtraction method (RS-EXRS), spectrum subtraction (SS), constant multiplication method (CM) or extract their derivative spectra such as derivative subtraction. 3. Transforming methods: which able to change the derivatized spectrum to its original zero spectra such as derivative transformation. 4. Graphical methods: which directly measure the concentration of the drugs from the chart.

Induced mathematical filtration as an innovative strategy for discrimination and estimation of glycemic control drugs in fixed dose combination.

An innovative strategy was developed for the estimation of a fixed dose combination containing Alogliptin (ALO) and pioglitazone (PIO) using induced concept for resolving the overlapped spectra, lacking isoabsorptive point. This strategy is based on coupling factors as numerical values or ratios as spectrum form with the recorded signals leading to induced mathematical filtration of the drug of interest and complete elimination of the interfering one in the combination without prior physical separation. The calculated factors were factor of equality in induced dual wavelength (IDW) or absorptivity factor in induced concentration subtraction method (ICS) while absorptivity ratio spectrum for induced amplitude modulation method (IAM). The calibration curves displayed linearity within 1.0-16.0 µg/mL for ALO and 2.0-22.0 µg/mL for PIO with good correlation coefficients. The induced methods specificity was also assured through the assaying different synthetic mixtures prepared to contain the two drugs in ratios approaching the ratio actually found in the marketed dosage form. The methods were applicable and suitable for estimating ALO and PIO in both bulk form and their fixed dose combination. Induced methods have been extensively validated in accordance with ICH guidelines and results demonstrated the accuracy and reproducibility in comparison to the reported method.

Comprehensive comparative study of eco-friendly Univariate and multivariate methodological approaches on processing multi-component formulation quality.

This study presents comprehensive comparative study of different eco-friendly spectrophotometric approaches without any sample treatment on processing quaternary mixture of sulphadimidine sodium (SDS), sulphaquinoxaline sodium (SQS), diaveridine (DVD) and vitamin K3 (VTK3). The different univariate complementary resolutions according to the response used for the assay of the cited drugs after applying the processing steps were implemented using successive ratio subtraction coupled with constant multiplication (SRS-CM), absorbance subtraction (AS) and amplitude modulation (AM). On the other hand, multivariate spectrophotometric models were developed and validated for simultaneous determination of the cited mixture. Resolution was accomplished by using two multivariate calibration greener models, including principal component regression (PCR) and partial least-squares (PLS). The proposed approaches are considered environmentally friendly since they use only water as reagent, which is cheap and safe for the operator. The calibration graphs are linear over the range of (4.0-13.0) µg/mL for (SDS), (1.0-10.0) µg/mL for (SQS), (1.0-11.0) µg/mL for (DVD) and (1.0-8.0) µg/mL for (VTK3). Specificity of the applied procedures was assessed by analyzing the laboratory-prepared mixtures and their combined dosage form. The outcomes of the developed methods were statistically compared with those of the official and reported methods; using Student's t-test and F-test, showing no significant difference. The proposed methodologies can be used for the routine analysis of the cited drugs in quality control laboratories.

Synchronous UPLC Resolution of Aceclofenac and Diacerin in Their Powdered Forms and Matrix Formulation: Stability Study.

Accurate, rapid and selective reversed phase ultra-performance liquid chromatography method with UV detection has been established and validated for the synchronous determination of aceclofenac (ACE) and diacerin (DIA) in the occurrence of diclofenac sodium and rhein, their main degradation products, respectively. Chromatographic separation was accomplished using Inertsil C-18 column (50 × 2.1 mm i.d., 1.7 µm particle size) in isocratic mode, with mobile phase consisting of 20 mM ammonium acetate buffer:acetonitrile in the ratio of 42:58 (v/v), pH adjusted to 3.00 by using 10% acetic acid, the flow rate of 0.25 mL/min and UV detection was performed at 265 nm. The retention times were 2.00 +/- 0.24, 2.69 +/- 0.19, 4.00 +/- 0.23 and 5.24 +/- 0.25 min for DIA, rhein, ACE and diclofenac sodium, respectively. Excellent linearity was shown over a range of 1.0-150.0 µg/mL and 0.5-87.5 µg/mL with mean percentage recoveries of 98.87 ± 1.19 and 98.84 ± 1.08 for ACE and DIA, respectively. Parameters of precision and accuracy of the method meet the established criteria. The obtained RSD values were quite low and indicate good reproducibility of the method. Thus, the developed method can be used for the combined dosage form analysis and its chemical stability studies.

The concept of Relative Absorptivity Distribution for enhancing disbanding power of spectrophotometric technique to resolve co-formulated tablets: A tool for purity index and uniformity assessment.

Resolving the spectral bands of the drugs into their discrete constituents is critical for an effective mathematical spectrophotometric method. The spectral resolution is ordinarily affected by the required application. In the current work an innovative, simple and green Relative Absorptivity Distribution (RAD) concept was established for the successful assay and assessing of content uniformity and purity index of tablet co-formulation which is recommended for lowering the blood glucose level and is comprised of Cangliflozin (CGF) and Metformin CGF and MTF. From the spectrophotometric perspective, the investigation of this combination is challenging as the composition of the tablets is CGF:MTF (1:17), where CGF is not only the minor analyte but also the one of lower absorptivity. The RAD concept is based on the production of master Resolving Spectra (RS). Within the RAD concept and according to the manipulation used for the generation of the RS, three different univariate mathematic methods via spectrophotometer software were developed. The proposed methods are characterized by the ability of extracting the raw spectra of each investigated analyte separately, consequently, enabling each analyte to be determined at its λmax without the contribution of the other. The proposed methods had analyzed CGF in the range 1.0-30.0 µg/mL and MTF within the range 1.0-20.0 µg/mL. The guidelines of the ICH were performed for the complete validation of all methods and the results confirmed satisfactory accuracy, precision and selectivity. The accomplished results together with the simplicity and low-cost of all methods suggested their suitability for the routine quality control analysis of bulk materials, assay of the pharmaceutical formulations and evaluating the content uniformity. Statistical comparison of the results with those of reported HPLC method showed good agreement. In addition, the environmental impact of the proposed methods was assessed by utilizing the National Environmental Methods Index (NEMI), the Analytical Eco-Scale and the Green Analytical Procedure Index (GAPI) where the three tools confirmed the environmentally friendly nature of all proposed methods.

Coupling of liquid-liquid extraction and mathematical filtration techniques for the separation and quantification of five components in semisolid dosage form with severely overlapped spectra.

Quadriderm cream was a combination of four components; Clioquinol (CLIO), Betamethasone (BETA), Tolnaftate (TOL), Gentamicin (GEN) in addition to the preservative Chlorocresol (CC). Four components CLIO, TOL, BETA, and CC were extracted in methanol and determined by mathematic filtration spectrophotometric techniques. The partially overlapped spectrum of CLIO was determined by constant value, constant multiplication, and concentration value methods then eliminated via spectrum subtraction (SS) to get the resolved ternary mixture of TOL, BETA, and CC with severely overlapping spectra. TOL was determined by derivative ratio at zero crossing point of BETA using CC as a divisor. While, BETA could be determined using TOL as a divisor at zero crossing of CC. BETA and CC were obtained using novel (DD1FS) followed by SS. By applying these novel procedures, the DD1 spectrum of each component alone was recovered where Pmax-min was directly proportional to its concentration. Liquid-liquid extraction technique was used for the semisolid dosage form where GEN was extracted with a mixture of chloroform: water (50:50, v/v); and the induced fluorescence obtained by derivatization with o-phthalaldehyde was measured at 419 nm after excitation at 359 nm. Accuracy and precision testing of the developed methods showed good results. Specificity of the methods was ensured and was successfully applied for the analysis of pharmaceutical formulation of the five components in combination. ICH guidelines were used for validation of the proposed methods. Statistical data were calculated, and the results were satisfactory revealing no significant difference regarding accuracy and precision.

Novel feature extraction approach for achieving potential spectral resolution: Green analytical application on zofenopril calcium and hydrochlorothiazide in their spectrally overlapping binary mixture.

A novel and green spectrophotometric method, namely constant extraction, based on extracting a spectral feature value (constant ratio) has been developed and validated for simultaneous estimation of a binary mixture of zofenopril calcium (ZOF) and hydrochlorothiazide (HCT) in their bulk and marketable formulation. A comparative study between this newly developed method and four long-established ones; ratio difference, ratio subtraction coupled with constant multiplication, advanced amplitude modulation and absorbance subtraction has been carried out giving very promising results. Various analytical performance parameters like linearity, accuracy, precision, and robustness were investigated in accordance with ICH (Q2B). Satisfying optimized instrumental parameters; absorbance of ZOF and HCT were linearly increased for the previously mentioned methods in a concentration range 5.0-35.0 and 3.0-20.0 µg/mL, respectively showing correlation coefficients ≥0.9990. Moreover, specificity has been checked through analyzing synthetic mixtures of the studied analytes. Feasibility has been successfully assessed by simultaneous quantification of both analytes in the commercially available formulation. As well, validity was examined and no interference from common excipients was noticed. Observed data has been statistically compared with those of the published one concluding that no significant variations between both results have been indicated. Furthermore, greenness profile of these methods was assessed by analytical Eco-Scale and found superior to that of the reported HPLC method as an even greener approach for the simultaneous analysis of ZOF and HCT.