Determination Of Active Pharmaceutical Ingredient Research Articles

Review on Analytical Method Validation for Trithioparamethoxy Phenylpropene & Chlorpheniramine Maleate in Pharmaceutical Dosage Form

Analytical method validation is a crucial step in ensuring the reliability and accuracy of analytical procedures employed for the determination of active pharmaceutical ingredients (APIs) in pharmaceutical dosage forms. This study focuses on the validation of analytical methods for the quantification of trithioparamethoxy phenylpropene and chlorpheniramine maleate in pharmaceutical dosage forms. The methods were developed using high- performance liquid chromatography (HPLC) or using ultra performance liquid chromatography (UPLC) with appropriate detection techniques. Parameters such as specificity, linearity, precision, accuracy, robustness, ruggedness, range, stability, LOD, LOQ and system suitability were evaluated according to International Conference on Harmonization (ICH) guidelines. The validated methods demonstrated excellent specificity, linearity over a wide concentration range, precise and accurate results, robustness against variations in method parameters, and suitable system suitability. The validated methods are suitable for routine quality control analysis of pharmaceutical formulations containing trithioparamethoxy phenylpropene and chlorpheniramine maleate, ensuring the reliability and consistency of drug products.

Unveiling Impurities: A Comprehensive RP-HPLC Method for Accurate Atorvastatin Impurity Analysis in Pharmaceuticals Using Accuracy Profile Approach.

Accurate determination of active pharmaceutical ingredients and impurities is essential for ensuring the safety and effectiveness of medications. This study focuses on the validation of a high-performance liquid chromatography (HPLC) method for quantifying Atorvastatin and its impurities, addressing a critical aspect of pharmaceutical analysis. The primary objective is to conduct a comprehensive validation study for the HPLC method, covering specificity assessment, response function establishment, and a detailed analysis of precision, trueness, and tolerance intervals. The emphasis is on demonstrating the method's precision, accuracy, and stability-indicating capabilities across various concentrations and compounds. The HPLC method is validated through rigorous assessments, including specificity, response function establishment, and analyses of precision, trueness, and tolerance intervals. Induced degradation experiments are conducted to explore Atorvastatin's behavior under extreme conditions. Insights into the compound's synthesis and degradation pathways are provided through a proposed mechanism for intramolecular esterification. The results affirm the precision, accuracy, and stability-indicating capabilities of the validated HPLC method. The method effectively differentiates between Atorvastatin and its impurities, showcasing its suitability for pharmaceutical quality control. The validated HPLC method emerges as a robust and reliable tool for Atorvastatin analysis, contributing significantly to pharmaceutical research and quality control. Its application ensures the safety and efficacy of medications, reinforcing its role in pharmaceutical analysis. This study not only validates a crucial HPLC method for Atorvastatin analysis but also provides insights into the compound's behavior under extreme conditions and its synthesis and degradation pathways. The validated method serves as a cornerstone in pharmaceutical research and quality control, ensuring medication safety and efficacy.

Greenness-sustainability metrics for assessment smart-chemometric spectrophotometric strategy for evaluation of the combination of six gastric proton-pump inhibitors with two selected impurities

Green analytical approaches are employed for the determination of active pharmaceutical ingredients, in conjunction with their impurities. Smart chemometric spectrophotometric techniques, including orthogonal partial least square (OPLS), variable selection such as genetic algorithm (GA-OPLS), and interval selection (i-OPLS), were utilized. These chemometric models were implemented for assessing six proton-pump inhibitors Omeprazole, Esomeprazole, Lansoprazole, Pantoprazole, Rabeprazole, and Dexlansoprazole along with two selected official impurities, namely 4-Desmethoxy omeprazole impurity and Rabeprazole-impurity B. Experimental design was implemented to separate impurities, in the process of multivariate calibration, a five-level eight-factor calibration design consisting of 25 samples was selected. This design was deliberately selected to guarantee that the components were mutually orthogonal to assess the model's performance and reliability, a separate validation set of 15 samples was constructed. The best-performing of the proposed techniques were identified by considering the least favorable values of the Correlation Coefficient (R ≥ 0.9995), the Root Mean Square Error of Prediction (RMSEP) values between (0.0102–0.5622), and the Relative Error of Prediction (REP) values between (0.2961–1.1917). The proposed and reported methods' greenness-sustainability was quantitatively evaluated, and a comparative study of the greenness profile was established through a spider chart, the National Environmental Method Index tool, advanced and modified NEMI along with the Hexagon tool, and the whiteness qualities of the presented approaches were assessed by implementing the recently adopted Red-Green-Blue paradigm and White Analytical Chemistry tool. These approaches are well-suited for use in quality control laboratories due to their observed acceptance, long-term sustainability, simplicity, and affordability.

PH-responsive subcutaneous implants prepared via hot-melt extrusion and fluidised-bed spray coating for targeted invasive predator control

Globally, invasive mammalian predators are responsible for the depredation and extinction of threatened species. The development of innovative control methods that target the most impactful of these predators is crucial to ensure the successful conservation of threatened species. Feral cats (Felis catus) are one of the most damaging invasive mammalian predators, particularly in Australia, where they are estimated to kill more than 1.5 billion mammals, birds, and reptiles annually. Mammals are disproportionally depredated by feral cats, and efforts to reintroduce or conserve remnant populations of threatened mammal species are hampered by shortfalls in current control methods. To address this challenge, we have prototyped the population-protecting implant (PPI), a subcutaneous implant that is designed to target the most dangerous and efficient predators, so called ‘problem individuals’. Conceptually, the implant has a reverse enteric coating that is stable in the subcutaneous environment of a prey animal but dissolves rapidly in the gastric environment of a predator, resulting in the release of a toxic payload and death of the predator. In this work we report a novel approach for the manufacture of PPIs, that combines hot-melt extrusion and fluidised-bed spray coating, to respectively afford implants with accurate drug loading and a uniform reverse-enteric coating. Most of the implants bearing a 300 μm coating were found to effectively contain their payload for more than 280 d in vitro, in a simulated subcutaneous environment (pH 7.4, 37 °C) but release it within 2.5 h in vitro under simulated gastric conditions (pH 1.5, 37 °C). However, two failures were observed at 64 d and 200 d, likely due to the increased osmotic pressure caused by the soluble core. This work highlights a revised implant design that may improve implant stability in vivo and facilitate the translation of the PPI to field use, for the conservation of threatened species. We also introduce a novel and facile approach to determine the loading of non-chromophoric payloads in hot-melt extruded systems that is highly relevant for the determination of active pharmaceutical ingredients in drug-delivery formulations.

Green Chromatographic Method for Determination of Active Pharmaceutical Ingredient, Preservative, and Antioxidant in an Injectable Formulation: Robustness by Design Expert.

We report an efficient HPLC method for simultaneous qualitative and quantitative analysis of lincosamide antibiotic injectable formulations containing Clindamycin phosphate (CMN), benzyl alcohol (BA), and ethylenediaminetetraacetic acid (EDTA) as major ingredients. The three components were separated by Phenomenex prodigy C8 (250 mm × 4.6 mm, 5 μm) HPLC column, flow rate 1.1 mL/min, injection volume 30 μL, and column temperature 35 °C, using 0.05 M sodium acetate buffer (pH 4.5) with acetonitrile (ACN) in the ratio of 80:20 (v/v). The detection wavelength was set as 240 nm. The method was validated as per International Conference on Harmonization (ICH) guidelines and was confirmed to be specific, precise, accurate, and linear. Method robustness was executed by utilizing quality in the design of the experiment. Accuracy results were found to be 99.3–100.5% for CMN, 99.3–100.8% for BA, and 99.1–100.3% for EDTA. Precision results were obtained as % relative standard deviation (RSD): 0.6% for CMN, 0.4% for BA, and 0.4% for EDTA. Correlation coefficient (r2) values were obtained as >0.999 for the three components. Analytical solutions are stable for 48 h at benchtop and refrigerator conditions. The greenness of the analytical method was evaluated by the Green Analytical Procedure Index (GAPI), National Environmental Method Index (NEMI), analytical eco-scale, and Analytical Greenness (AGREE) tools to confirm that the method is eco-friendly.

Structure of the active pharmaceutical ingredient bismuth subsalicylate

Structure determination of pharmaceutical compounds is invaluable for drug development but remains challenging for those that form as small crystals with defects. Bismuth subsalicylate, among the most commercially significant bismuth compounds, is an active ingredient in over-the-counter medications such as Pepto-Bismol, used to treat dyspepsia and H. pylori infections. Despite its century-long history, the structure of bismuth subsalicylate is still under debate. Here we show that advanced electron microscopy techniques, namely three-dimensional electron diffraction and scanning transmission electron microscopy, can give insight into the structure of active pharmaceutical ingredients that are difficult to characterize using conventional methods due to their small size or intricate structural features. Hierarchical clustering analysis of three-dimensional electron diffraction data from ordered crystals of bismuth subsalicylate revealed a layered structure. A detailed investigation using high-resolution scanning transmission electron microscopy showed variations in the stacking of layers, the presence of which has likely hindered structure solution by other means. Together, these modern electron crystallography techniques provide a toolbox for structure determination of active pharmaceutical ingredients and drug discovery, demonstrated by this study of bismuth subsalicylate.

Simultaneous Determination of Active Pharmaceutical Ingredients and Water-Soluble Polymers: Analysis of Dissolution Profiles from Sustained-Release Formulations and Mechanisms Involved.

The dissolution behaviors of base excipients from sustained-release formulations have been investigated using various methodologies. However, the dissolution of polymers has not been fully evaluated because differences between formulations are still verified only by the release of active pharmaceutical ingredients (APIs). In our previous study, we proposed a quick and simultaneous analysis of dissolved APIs and water-soluble polymers by ultra HPLC using charged aerosol and photodiode array detectors. The purpose of this study was to verify whether the analysis system could be adapted to other water-soluble polymers. Dissolution tests were conducted using matrix model tablets prepared from three polymers and three APIs (propranolol, ranitidine, and cilostazol) with different solubilities. The dissolution profiles of the polymers and APIs were determined using the proposed analysis system and compared. The results clarified differences in the dissolution behaviors of the APIs and polymers. The polymers, especially hydroxypropyl cellulose, exhibited the dissolution properties characteristic of each model formulation. Propranolol and ranitidine showed the diffusion type, while cilostazol showed the erosion type release mechanism due to their different solubilities. The release of cilostazol was delayed in all models compared to the polymer, which may be due to the aggregation of cilostazol in the gel layer. This analytical method can be used to study the dissolution behavior (diffusion or erosion) of APIs from matrix tablets containing various polymers. This method will provide useful information on release control, which will make it easier and more efficient to design appropriate formulations and analyze the release mechanisms.

Modified aqueous mobile phases: A way to improve retention behavior of active pharmaceutical compounds and their impurities in liquid chromatography

Most commonly used analytical technique for determination of active pharmaceutical ingredients and their impurities in quality control throughout all phases of drug research, development and manufacture is definitely reversed-phase high performance liquid chromatography (RP-HPLC). However, pharmaceutical industry professionals are often faced with various challenges in RP mode, which cannot be resolved with common variations in the composition of the mobile phase. These challenges often occur when analyzing compounds that contain basic ionizable groups, possess large differences in polarities and require consumption of high amounts of toxic organic solvents. Among available strategies for addressing the aforementioned issues, the most convenient one includes RP-HPLC mobile phase modifications by an addition of the proper chemical compounds. In that respect, RP-HPLC method can be easily adapted to the needs of the analysis without time-consuming and expensive equipment procurement. In this review the chaotropic chromatography, micellar liquid chromatography, and cyclodextrin modified RP-HPLC systems are presented and discussed in details. Special attention is devoted to the theoretical background, the possibility of retention modeling and applications in various fields of pharmacy, as well as their prospective in further research.

A novel electronic tongue using electropolymerized molecularly imprinted polymers for the simultaneous determination of active pharmaceutical ingredients

The combination of chemometrics and electrochemical sensors modified with molecularly imprinted polymers (MIPs) towards the development of MIP-based electronic tongues (ETs) was explored herein. To demonstrate the potential of such an approach, the simultaneous determination of paracetamol, ascorbic acid and uric acid mixtures in pharmaceutical samples was evaluated. To this aim, MIP-based sensors for the different compounds were prepared by in situ electropolymerization of pyrrole in the presence of p-toluenesulfonate anion (pTS−), which acted as functional doping ion of the polypyrrole (PPy) MIP backbone. Morphological characterization of the MIPs was done by scanning electron microscopy (SEM), while functionalization of the electrodes was monitored electrochemically. Under the optimized measuring conditions, the developed sensors showed a good performance, with good linearity at the μM level (R2 > 0.992, limits of detection between 1 and 24 μM) as well as good repeatability (intra- and inter-day RSD values between 3 and 6% over 30 consecutive measurements). Finally, the quantification of the individual substances in different pharmaceutical samples was achieved by an artificial neural networks (ANNs) model, showing satisfactory agreement between expected and obtained values (R2 > 0.987).

Photogenerated Iodine for Determination of Active Pharmaceutical Ingredients in Tetralgin Drug

Photogenerated Iodine for Determination of Active Pharmaceutical Ingredients in Tetralgin Drug

DEVELOPMENT AND VALIDATION OF METHODS FOR QUANTITATIVE DETERMINATION OF ACTIVE PHARMACEUTICAL SUBSTANCES IN NASAL SPRAY

Intranasal administration of H1-histamine receptor blockers may be a promising approach to the treatment of allergic rhinitis. Earlier, an original composition of a nasal spray containing fexofenadine hydrochloride and ammonium glycyrrhizinate and demonstrating a high level of therapeutic efficacy, was developed.The aim of the study was to develop and validate a method of the quantitative determination of active pharmaceutical ingredients fexofenadine hydrochloride and ammonium glycyrrhizinate in a spray for intranasal administration.Materials and methods. During the development and validation of the method of the fexofenadine hydrochloride and ammonium glycyrrhizinate quantitative determination in a nasal spray, the method of high performance liquid chromatography was used: a Dionex Ultimate 3000 UV chromatograph with a Luna C18 column (2) containing octadecylsilicagel with a 5 μm grain size as a sorbent. The analysis and validation procedures were performed in accordance with the requirements of the State Pharmacopoeia of the Russian Federation, the XIVth edition.Results. The study showed that for the simultaneous quantitative determination of fexofenadine hydrochloride and ammonium glycyrrhizinate, the optimal elution regime is a gradient mode with a mobile phase containing 50 mmol/L potassium dihydrogen phosphate solution with methanol (45:55), which ensured the separation of the components in the 20 minutes interval. The validation procedures showed that the developed methodology correspond to all the criteria of validity in terms of the following indicators: correctness, precision, specificity and linearity in the analytical area.Conclusion. The obtained results indicate the possibility of using the method of high-performance liquid chromatography in a gradient elution mode with a mobile phase of the composition of a 50 mmol/L solution of potassium dihydrogen phosphate with methanol (45:55) for the simultaneous quantitative determination of active pharmaceutical ingredients – fexofenadine hydrochloride and ammonium glycyrrhizinate as parts of a promising nasal spray for the allergic rhinitis treatment.

Semiautomatic and fully functional electrochemical microanalyzer BO-05 suitable for scientific, didactic, and analytical applications: The use in the potentiometric analysis of drugs

This article presents the potentiometric method of determination of chlorides using the original BO-05 electrochemical microanalyzer. The quantification of chlorides is one of the frequently performed methods, both in the indirect determination of active pharmaceutical ingredients (API) and impurities in pharmaceutical raw materials, pharmacopoeial substances or pharmaceutical formulations as well. Successfully validated method was used to the analysis of chlorides in the preparations containing verapamil hydrochloride in form of tablets Staveran® and Verapamil®. The mean content of the studied API calculated to one tablet was close to the declared and equal to 123.6±1.5 mg and 122.6±1.1 mg, respectively. The presence of excipients have no significant impact on the final results. Thus shown that the electrochemical microanalyzer BO-05 is suitable for scientific, didactic and analytical applications.

Development of a Comprehensive HPLC Method for Determination of Product-related Impurities and Assay of Active Ingredients in Papaverine Hydrochloride Products

Abstract. Papaverine hydrochloride products are used as anticonvulsants in routine medical practice. Most of the approved product specification files include thin-layer chromatography for assessment of product-related impurities and UV spectrophotometry for determination of active pharmaceutical ingredients. An HPLC assay is not used for determination of papaverine hydrochloride in drug dosage forms.The aim of the study was to develop an HPLC test method for determination of product-related impurities and for quantification of papaverine hydrochloride in solutions for injection, tablets, and rectal suppositories.Materials and methods: samples of the following Russian-made papaverine products were used in the study: Papaverine, solution for injection, 20 mg/mL; Papaverine, rectal suppositories, 20 mg; Papaverine, tablets, 40 mg. The Agilent 1260 Infinity II DAD System was used for the HPLC assay, and the Agilent 8453Е UV-Vis System was used for recording UV spectra. The determination of product-related impurities and the assay of active ingredients were performed simultaneously by HPLC using a reversed-phase column Kromasil 100-5-C18, 250×4.6 mm, 5 μm, the gradient elution mode, and detection at 238 nm. Papaverine Hydrochloride USP RS, 99% purity, and Noscapine EP CRS were used as reference standards.Results: the study demonstrated that determination of product-related impurities and assay of active ingredients in papaverine products can be performed simultaneously using HPLC.Conclusions: the authors proposed an HPLC test method for determination of active ingredients in papaverine products, which is aligned with the “consistent standardisation” principle and can be recommended for inclusion into draft monographs for papaverine products.

A QbD and Green Analytical Chromatography Technique for the Determination of Active Pharmaceutical Ingredient, Preservative and Antioxidant Contents in Lincosamide Antibiotic Injectable Formulation

A QbD and Green Analytical Chromatography Technique for the Determination of Active Pharmaceutical Ingredient, Preservative and Antioxidant Contents in Lincosamide Antibiotic Injectable Formulation

DETERMINATION OF FAVIPIRAVIR RESIDUES ON THE SURFACES OF PHARMACEUTICAL MANUFACTURING EQUIPMENT BY HPLC, LUMINESCENCE AND SPECTROPHOTOMETRY

Determination of active pharmaceutical ingredients (APIs) in washes from the surface of pharmaceutical equipment is necessary to prevent cross-contamination of the next batch of products. Highly sensitive methods of chromatographic (HPLC), luminescent (Lum) and spectrophotometric (SF) determination of antiviral drug favipivir (FAV) residues in washes during cleaning of pharmaceutical equipment have been developed. Chromatographic and luminescence recording conditions of favipiravir were optimized. The content of FAV in the washes (μg/wash) is determined according to the calibration curve. The proposed methods are validated by the following parameters: specificity, linearity, accuracy, limit of detection and limit of quantification. The coefficients of linear dependences in the studied concentration ranges correspond to the permissible values for the methods of determination of substances whose concentration in the sample is above the limit of quantitative determination. Calibration graphs are presented in concentration ranges 0.02-2.0 μg ml (HPLC), 0.3-3.5 μg/ ml (Lum) and 1.0-20.0 μg/ml (SP), the detection limits are 0.023 μg/ml, 0.12 μg/ml and 0.11 μg/ml, respectively. The degree of extraction of favipivir from applicators and surfaces of pharmaceutical equipment is more than 85 %. Luminescent and SP techniques have a number of advantages over HPLC: less complexity, expressiveness and lower consumption of reagents, while the HPLC method has a greater sensitivity. The developed methods can be recommended for determination of residual amounts of favipivir at quality control of cleaning of the pharmaceutical equipment.

The Challenge for Engineering Pharmaceutical Crystalline Solids: Scientific and Regulatory Affairs Perspectives for Crystal Structure Design and Prediction

The Breakthrough into solid-state research has become emerging approach for structure determination of Active Pharmaceutical Ingredients (APIs) and excipeents that consequently influence their physic-chemical properties, biopharmaceutical and pharmacokinetic profiles. The concept of conventional pharmaceutical salts has been extended to multicomponent crystals which diversity in nature of the non-covalent intermolecular interactions determine the crystal packing patterns within the structures, and thus modulate the native properties of APIs. Therefore, the aim of this review is to highlight how accomplishments in crystallographic research on molecular crystal have influenced their classification and how these new solid phases have been recognized by the regulatory bodies. The advantage to explore the pharmaceutical crystalline solids of one API implies the selection of the form with favorable properties for the development of formulations for pharmaceutical dosage forms.

Near-infrared spectroscopy-based methods for quantitative determination of active pharmaceutical ingredient in transdermal gel formulations

Semi-solid pharmaceutical forms, like topical and transdermal products, are widely used for medical purposes since they provide convenient and pain-free treatment for patients. In this study, two transdermal gel formulations with different active pharmaceutical ingredients were analyzed with near-infrared spectrophotometers with three distinct optical arrangements (dispersive, Fourier transform, and diode-array spectrophotometers) for the measurement of active pharmaceutical ingredient concentration. The reference concentrations were measured with high-performance liquid chromatography methods.The developing of calibration for reference concentrations based on near-infrared spectra was performed using partial least squares regression. The spectra from near-infrared spectrophotometer with diode-array optical arrangement presented the best root mean square error of cross-validation results with 0.045% and 0.144% for the two active pharmaceutical ingredients. The root mean square error of cross-validation using dispersive spectra were 0.125% and 0.193% and using Fourier transform spectra were 0.130% and 0.344% for the active pharmaceutical ingredients. The results of partial least squares regression were confirmed using simulated spectra based on the measured datasets.Near-infrared spectroscopy can be used successfully for the quantitative measurement of active pharmaceutical ingredient concentration in transdermal gel formulations. The type of optical arrangements of spectrophotometers can influence the performance of calibrations and have to be optimized case by case.

Application of HPLC for Evaluation and Standardisation of Hawthorn Flower Herbal Substance

The process of harmonisation of Russian and foreign approaches and requirements in the field of medicines quality assurance calls for revision of quality control procedures included in various regulations and guidelines. The monograph FS.2.5.0062.18 “Hawthorn flowers” of the State Pharmacopoeia of the Russian Federation XIV edition includes a test procedure for determination of flavonoids by a chromatospectrophotometric method. This procedure does not take into account current scientific capabilities and has a number of shortcomings, therefore it was necessary to revise the existing test procedure and develop a new approach to the standardisation of the hawthorn flower herbal substance.The objectiveof the studywas to develop an assay method for standardisation and evaluation of hawthorn flower using high performance liquid chromatography (HPLC).Materials and methods:the study was performed using samples of hawthorn flowers by Russian manufacturers. Quercetin (USP RS) and Hyperoside (HWI, primary standard) were used as the reference standards. The HPLC analysis was performed using an Infinity II 1260 DAD LC system (Agilent), and the UV spectra were recorded on a Cary 100 Varian spectrophotometer. A TLC Visualizer (CAMAG) was used to obtain digital images of thin layer chromatography plates.Results:the authors developed an HPLC test procedure for quantitative determination of total flavonoids, expressed as hyperoside, in hawthorn flowers. The developed procedure gives reliable and reproducible results and is characterised by high sensitivity and selectivity. The results of quantitative determination of the total flavonoid content in hawthorn flowers were used to propose the standard for the total content of flavonoids, expressed as hyperoside, of “not less than 0.5%”.Conclusions:the developed assay method for determination of active pharmaceutical ingredients in hawthorn flower products by HPLC can be recommended for inclusion into the Assay part of the “Hawthorn Flowers” monograph of the State Pharmacopoeia of the Russian Federation.

Spectroelectrochemical Detection of Pharmaceutical Compounds Using Carbon Screen Printed Electrodes

Several analytical methods have been published concerning the determination of active pharmaceutical ingredients (APIs) in drug formulations or in environmental samples. The most commonly used are chromatographic techniques but these methods are time consuming, expensive, require complicated preconcentration processes and need sophisticated instruments. On the other hand, electrochemical methods present the advantage of high simplicity, high sensitivity, good stability and low cost. These instruments can also be portable, which is particularly suitable for on-site monitoring of pharmaceuticals in field analysis. In the present work, Carbamazepine, Ibuprofen, Clarithromycin, Naratriptan, Dopamine and Acetaminophen have been determined using different electrochemical techniques at screen printed electrodes (SPE). The spectroelectrochemical behavior of the voltammetric response of some of these APIs allows the selective detection of these compounds despite their similar oxidation potentials on SPEs. The analysis was carried out by simultaneously performing an electrochemical and an optical experiment, the corresponding voltabsorptograms obtained are presented, showing the correlation between the derivative voltabsorptograms at the maximum of absorbances and the electrochemical curve. This work is supported by Innovec`EAU (SOE1/P1/F0173) project from the Interreg Sudoe 2014-2020 program funding by European Regional Development Funds

Novel Approach for the Simultaneous Determination of Carbinoxamine Maleate, Pholcodine, and Ephedrine Hydrochloride Without Interference from Coloring Matter in an Antitussive Preparation Using Smart Spectrophotometric Methods.

The presence of coloring matters in syrups usually interferes with the spectrophotometric determination of active pharmaceutical ingredients. A novel approach was introduced to eliminate the interference of sunset yellow (coloring matter) in Cyrinol syrup. Smart, simple, accurate, and selective spectrophotometric methods were developed and validated for the simultaneous determination of a ternary mixture of carbinoxamine maleate, pholcodine, and ephedrine hydrochloride in syrup. Four of the applied methods used ratio spectra: successive derivative subtraction coupled with constant multiplication, successive derivative of ratio spectra, ratio subtraction coupled with ratio difference, and ratio spectra continuous wavelet transforms zero-crossing. In addition, a method that was based on the presence of an isosbestic point, the amplitude summation method, was also established. A major advantage of the proposed methods is the simultaneous determination of the mentioned drugs without prior separation steps. These methods were successfully applied for the determination of laboratory-prepared mixtures and a commercial pharmaceutical preparation without interference from additives, thus proving the selectivity of the methods. No significant difference regarding both accuracy and precision was observed upon statistical comparison of the results obtained by the proposed methods with each other and with those of official or reported ones.