Chromatography Methodology Research Articles

Development of a liquid chromatography method for the analysis of oil extract of artemisia cina

Introduction. The problem of development and implementation of own drugs in Kazakhstan today is acute and urgent. The solution to this problem is possible through the use of own resources – domestic medicinal plant raw materials. A promising medicinal raw material is Artemisia cina – an endemic medicinal plant of the of the Turkestan region of Kazakhstan.Aim. Designing a liquid chromatography methodology to analyze the oil extract obtained from Artemisia cina.Materials and methods. Standard sample (SS) of santonin, Artemisia cina’s oil extract. The substance was identified and quantitatively determined using an Agilent Technologies 1200 liquid chromatograph (USA) equipped with ChemStation software.Results and discussion. The main active substance of the oil extract was analyzed using high-performance liquid chromatography (HPLC) method. Choosing the separation conditions for the column involves determining the best composition of the mobile phase and the rate of elution. There were used a reversed-phase system with sorbent C18 and a mobile phase consisting of acetonitrile and a phosphate buffer with a pH of 6.8. Chromatographic studies of the substance were carried out in a gradient mode at an analytical wavelength of 237 nm. The retention time of the standard sample (SS) of santonin matched the retention time (tR) of santonin isolated from the oil extract of Artemisia cina and amounted to 14.3 minutes.Conclusion. A liquid chromatography method has been established for the identification and quantitative determination of the oil extract, focusing on the main active substance, santonin.

Quality by Design Approach Driven Development and Validation of Reverse Phase‐High‐Performance Liquid Chromatography Methodology for Dual Quantification of Aspirin and Rivaroxaban

ABSTRACTClinical trials have shown that combining a modest dose of Rivaroxaban (RIV) with Aspirin (ASP) is more effective than ASP alone in reducing cardiovascular mortality, myocardial infarction, and stroke. A high‐performance liquid chromatography method developed using Quality by Design (QbD) effectively estimates ASP and RIV for routine pharmaceutical analysis. The Plackett‐Burman design assessed key factors including organic phase volume, mobile phase pH, flow rate, temperature, and injection volume using an Ishikawa fish‐bone diagram for risk assessment. These parameters were then optimized via Box‐Behnken design in 15 trials. The method meets the International Council For Harmonization standards with a mobile phase ratio of 45:55 (v/v) acetonitrile: buffer (pH 3, adjusted with orthophosphoric acid), a flow rate of 1 mL/min, a column temperature of 30°C, and an injection volume of 20 µL. A Phenomenex Luna C18 column (250 × 4.6 mm, 5 µm particle size) with detection at 251 nm ultra‐violet wavelength. The retention times of 4.54 min for ASP and 5.81 min for RIV were obtained. Validation confirmed linear ranges: 40–200 µg/mL for ASP and 2–10 µg/mL for RIV, with excellent recovery rates (ASP: 99.30%–101.9%; RIV: 98.52%–101.84%). Developed using QbD principles, the method proved specific, accurate, precise, sensitive, and robust for routine quality control of both compounds in pharmaceutical formulations.

Enhanced enantioselective separation of racemic menthol via reverse-phase high-performance liquid chromatography: Method development and computational insights for pre-screening

A robust and efficient enantioselective separation of racemic menthol was achieved on a standard C18 column with reverse-phase high-performance liquid chromatography (RP-HPLC) and UV detector. (R)-α-hydroxy-4-methylbenzeneacetic acid was utilized as the pre-column derivatization reagent. The impact of mobile phase composition on diastereomer selectivity was thoroughly investigated, resulting in a high resolution of 2.11 under optimized conditions. The method was rigorously validated for linearity, precision, accuracy, limit of detection (LOD) and limit of quantification (LOQ). Notably, a separation pre-screening mechanism (SPM) and a prediction model was developed based on density functional theory (DFT) studies. This model elucidated the relationship between molecular polarity differences (△MPI) and chromatographic behavior, facilitating the interpretation and prediction of racemic menthol resolution with various chiral derivatization reagents. The present work not only presents an efficient and economical approach for menthol enantiomeric separation, but also offers valuable insights for the innovative design and advancement of chromatographic methodologies for a wide array of chiral enantiomers.

A novel application of hydrophilic interaction liquid chromatography for the identification of compounds with intramolecular hydrogen bonds

The incorporation of intramolecular hydrogen bonds (IMHB) into small molecules constitutes an interesting optimization strategy to afford potential drug candidates with enhanced solubility as well as permeability and consequently improved bioavailability (if metabolic stability is high). Common methods to assess IMHB rely on spectroscopic or diffraction techniques, which, however, have limited throughput when screening for hit compounds in early phases of drug discovery. Inspired by literature findings using supercritical fluid chromatography (SFC) as an indirect method for IMHB identification in a screening context, we aimed at developing a secondary chromatographic methodology taking advantage of commonly used HPLC-MS instrumentation. In this work, we explored hydrophilic interaction liquid chromatography (HILIC) and developed a method for discriminating compounds based on their hydrogen bonding features. By quantifying retention of different matched molecular pairs (MMP) and using information about their low energy conformations from quantum-mechanical calculations, we defined a hydrogen bonding-driven adsorption (kads) chromatographic parameter to assess a compound’s propensity to forming IMHB. In addition to the MMP analysis, we found that the kads parameter allows for the differentiation of analytes forming IMHB regardless of the comparison with control compounds.

Plasmodium SEY1 is a novel druggable target that contributes to imidazolopiperazine mechanism of action.

The precise mode of action of ganaplacide (KAF156), a phase III antimalarial candidate, remains elusive. Here we employ omics-based methods with the closely related chemical analog, GNF179, to search for potential Plasmodium targets. Ranking potential targets derived from chemical genetics and proteomic affinity chromatography methodologies identifies SEY1, or Synthetic Enhancement of YOP1, which is predicted to encode an essential dynamin-like GTPase implicated in homotypic fusion of endoplasmic reticulum (ER) membranes. We demonstrate that GNF179 decreases Plasmodium SEY1 melting temperature. We further show that GNF179 binds to recombinant Plasmodium SEY1 and subsequently inhibits its GTPase activity, which is required for maintaining ER architecture. Using ultrastructure expansion microscopy, we find GNF179 treatment changes parasite ER and Golgi morphology. We also confirm that SEY1 is an essential gene in P. falciparum. These data suggest that SEY1 may contribute to the mechanism of action of imidazolopiperazines and is a new and attractive druggable target.

Optimized reversed phase liquid chromatography methodology for the determination of vonoprazan fumarate impurities: Towards Six Sigma quality standards and sustainability assessment

Vonoprazan fumarate (VPZ), a potent potassium-competitive acid blocker, holds great promise as a therapeutic option for addressing acid-related disorders. This study introduces a refined reversed phase liquid chromatography methodology tailored for the comprehensive analysis of eight related substances, including starting materials, byproducts, and degradants within VPZ. Our approach integrates response surface methodology and tolerance analysis to achieve six sigma quality standards in chromatographic performance. By embedding specifications into the optimization process, we ensure robustness during method development. Chromatographic separation was executed using an XSelect CSH Phenyl-Hexyl column under stepped gradient conditions, employing a mobile phase comprising 0.1 % trifluoroacetic acid aqueous solution and acetonitrile. The flow rate was maintained at 1.3 mL/min, with UV absorbance at 252 nm, and a column temperature set at 25 °C. To evaluate the stability indicating ability of the method, forced degradation studies were conducted. Importantly, identified degradants did not interfere with the accurate quantification of VPZ and its associated impurities. Validation of the method was achieved through accuracy profiles. A greenness assessment was conducted using National Environmental Methods Index (NEMI), carbon footprint analysis, Analytical Greenness Calculator (AGREE), and Complementary Green Analytical Procedure Index (Complex GAPI). Additionally, blueness and whiteness assessments were conducted using the Blue Applicability Grade Index (BAGI) and Red-Green-Blue 12 (RGB 12) algorithms, respectively. The proposed method exhibited a green profile in NEMI and Complex GAPI. The carbon footprint was calculated at 0.055 kg CO2 equivalent per sample. The AGREE score was 0.67, BAGI was 80.0, and the whiteness score from the RGB12 algorithm was 83.5.This methodological framework holds promise for utilization in process development and quality assurance of VPZ in bulk drug manufacturing, particularly in the absence of official monographs within recognized compendia.

Gas evolution analyses of Ni-rich Li-ion and Li-metal batteries at cylindrical jelly-roll configurations

Understanding gas evolution during battery operation is pivotal for elucidating the underlying mechanisms of battery behavior, a cornerstone for fostering innovation in energy storage technologies. Despite the wealth of quantitative analyses available, the realm of large-scale, practical, full-cell investigations remains relatively unexplored. This study bridges this gap by unveiling a novel, robust gas chromatography methodology designed to meticulously quantify gas generation within 18650 cylindrical cells Li-ion and anode-free Li-metal Ni-rich batteries across a spectrum of operational conditions. Our approach not only reaffirms the capability for precise, reliable quantitative measurements but also illuminates new findings on the dynamics of gas evolution. This includes the significant impact of temperature and cathode material composition on gas generation, alongside a pioneering exploration into the behavior of anode-free Li-metal batteries. These insights not only advance our understanding of battery performance and safety at a practical cell level but also pave the way for the development of more efficient, durable, and safer energy storage solutions.

The role of the cation and anion in aqueous liquid chromatography with sodium dodecyl sulphate and imidazolium-based ionic liquids as mobile phase reagents

BackgroundIn reversed-phase liquid chromatography, solute retention is primarily influenced by interactions between a nonpolar stationary phase and a moderately polar hydro-organic mobile phase, based on the solute lipophilicity. However, challenges regarding retention and peak tailing can arise due to ionic interactions between positively charged analytes and free silanols present on silica-based stationary phases. To address these challenges, incorporating surfactants and ionic liquids (ILs) into the mobile phase offers an effective solution. These additives synergistically enhance chromatographic performance through electrostatic and lipophilic interactions, which enable fine-tuning of selectivity and improved separation efficiency. ResultsThis study explores the chromatographic behaviour of several basic compounds in aqueous mixtures containing the anionic surfactant sodium dodecyl sulphate (SDS), above its critical micellar concentration, combined with various 1-alkyl-3-methylimidazolium-based ionic liquids (ILs) featuring chloride, tetrafluoroborate, and hexafluorophosphate anions, all without the addition of organic solvents. Specifically, this research investigates the influence of different anion types within the ILs and considers the impact of the IL cations. Analysis of solute peak profiles reveals narrow and symmetrical peaks. By introducing tetrafluoroborate and hexafluorophosphate IL anions into a mobile phase that contains an anionic surfactant, the study sheds light on the interactions occurring within the chromatographic column. This enhanced understanding of the combined effects of surfactants and ILs contributes to refining chromatographic methodologies. SignificanceThis research highlights the importance of carefully selecting the appropriate IL when incorporating it into a micellar mobile phase alongside SDS. This combination results in practical retention times that surpass the performance achieved with either the surfactant or IL alone in the mobile phase. The study particularly emphasises the impact of the IL anion, especially in the absence of SDS and organic solvents. This unveils interactions that are otherwise obscured in micellar and hydro-organic media, providing new insights into chromatographic dynamics.

Synchronized Assessment of Lobeglitazone Sulfate and Metformin Hydrochloride in Tablet by Robust, High-performance Thin-layer Chromatographic Method

Background: A combination of fixed-doses containing 0.5 mg lobeglitazone sulfate and 500 mg metformin hydrochloride has demonstrated efficacy in enhancing glycemic control in diabetes. Aims: The projected work aimed to establish and validate a high-performance thin-layer chromatographic methodology for the quantification of both drugs in tablet formulations. Objectives: The task involves creating and validating a method in accordance with ICH guidelines to quantify two particular drugs in tablet formulations accurately. Methods: The high-performance thin-layer chromatographic analysis utilized aluminum plates layered with silica gel 60F254, and the solvent system consisted of acetonitrile, 1 M ammonium acetate (methanol), toluene, and triethyl amine (1.5:2.5:4:0.2 v/v/v/v), followed by densitometric scanning at 237 nm. Results: The methodology exhibited linearity in the range of 100-1500 ng/band for lobeglitazone sulfate and 1000-15000 ng/band for metformin hydrochloride, with correlation coefficients of 0.9991 and 0.9992, correspondingly. Exceptional sensitivity was observed, with detection limits of 8.17 ng/band for lobeglitazone sulfate and 271.34 ng/band for metformin hydrochloride, along with quantification limits of 24.75 ng/band for lobeglitazone sulfate and 822.24 ng/band for metformin hydrochloride. The method demonstrated precision (% relative standard deviation of peak area <2) and accuracy (recovery between 96 and 103%). Conclusion: The suggested methodology is fit for the concurrent quantification of both drugs in tablet formulations, making it applicable for routine quality control assessments in laboratories.

A UPLC Method for Simultaneous Estimation of Trastuzumab and Hyaluronidase in Bulk and Pharmaceutical Dosage Form

Objective: The proposed research aimed at establishing a quick, accurate, and exact ultra-performance liquid chromatography (UPLC) methodology for contemporaneously analyzing trastuzumab and hyaluronidas in bulk and pharmaceutical formulation, with a focus on the stability-indicating properties of the assay. Method: Using a Water X-Bridge C18 column (50 mm x 4.6 mm x 2.5 μ) and a mobile phase of ACN: Buffer (pH 2.5) [50% v/v], pushed at 0.5 mL/min, compounds were separated chromatographically. An ultraviolet (UV) detector fixed at 260 nm was employed to identify the isolated compounds. Results: The findings showed that 1.3529 and 2.404 minutes were optimal for separating Trastuzumab and Hyaluronidase, correspondingly. The current procedure has been verified in accordance with ICH standards Q2 R1, and stability-indicating tests have been performed in accordance with ICH standards Q1A R2. Both the intra- and inter-day precisions were determined to be satisfactory. The suggested approach showed linearity between 30 to 180 μg/mL of trastuzumab and between 12.50-75μg/ mL of hyaluronidase. It was determined that the limit of detection (LoD) and limit of quantitation (LoQ) for trastuzumab were 0.36 and 1.2 μg/mL, correspondingly, whereas those for hyaluronidase were 0.15 and 0.5 μg/mL. The approach was shown to have a recovery rate around 98.6 to 99.7%. Conclusion: The suggested approach successfully separated the chemical compounds from their by-products. Therefore, trastuzumab and hyaluronidase routine evaluation and stability-indicating tests were both effectively implemented using the approach.

Quantitative determination of lobeglitazone sulfate and glimepiride in combined tablet by robust high‐performance thin layer chromatographic method

AbstractA combination of fixed dose tablet containing 0.5 mg lobeglitazone sulfate (LBZ) and 1 mg glimepiride (GLM) has demonstrated efficacy in enhancing glycemic control in diabetes. The projected work aimed to develop and validate a high‐performance thin‐layer chromatographic (HPTLC) methodology for the precise quantification of both the drugs in tablet formulations. The HPTLC analysis utilized aluminium plates layered with silica gel 60F254, and the solvent system consisted of ethyl acetate, benzene, and hexane (4:3:1 v/v/v) followed by densitometric scanning at 238 nm. The Rf value was found to be 0.68 ± 0.001 for LBZ and 0.48 ± 0.002 for GLM. The methodology exhibited linearity in the range of 100–2000 ng/band for LBZ and 200–4000 ng/band for GLM, with correlation coefficients of 0.9988 and 0.9981, respectively. Exceptional sensitivity was observed, with detection limits of 23.86 ng/band for LBZ and 58.26 ng/band for GLM, along with quantification limits of 72.32 ng/band for LBZ and 176.55 ng/band for GLM. The method demonstrated precision (% relative standard deviation of peak area < 2) and accuracy (recovery between 97% and 102%). The suggested method is suitable for quantifying both the drugs in tablets, making it useful for routine quality control in laboratories.

A voltammetric method coupled with chemometrics for determination of a ternary antiparkinson mixture in its dosage form: greenness assessment

An electroanalytical methodology was developed by direct differential pulse voltammetric (DPV) measurement of Levodopa (LD), Carbidopa (CD) and Entacapone (ENT) mixture using bare glassy carbon electrode (GCE) in Britton Robinson (BR) buffer (pH = 2.0). A multivariate calibration model was then applied to the exported preprocessed voltammetric data using partial least square (PLS) as a chemometric tool. Additionally, the model was cross-validated and the number of latent variables (LVs) were determined to produce a reliable model for simultaneous quantitation of the three drugs either in their synthetic mixtures or in their marketed pharmaceutical formulation with high accuracy and precision. Data preprocessing was used to tackle the problem of lacking bi-linearity which is commonly found in electrochemical data. The proposed chemometric model was able to provide fast and reliable technique for quantitative determination of antiparkinson drugs in their dosage forms. This was successfully achieved by utilizing sixteen mixtures as calibration set and nine mixtures as validation set. The percent recoveries for LD, CD and ENT were found to be 100.05% ± 1.28%, 100.04% ± 0.53% and 99.99% ± 1.25%, respectively. The obtained results of the proposed method were statistically compared to those of a previously reported High Performance Liquid Chromatography (HPLC) methodology. Finally, the presented analytical method strongly supports green analytical chemistry regarding the minimization of potentially dangerous chemicals and solvents, as well as reducing energy utilization and waste generation.

Simultaneous Identification and Quantification of Phenylalanine, Glycine and Lysine amino acids and conjugated heterocyclic amines in Maillard chemical model system using RP-HPLC-DAD

In this study, we developed a simple reversed-phase high-performance liquid chromatography (RP-HPLC) methodology for simultaneous identification and quantification of three types of mix: (1) phenylalanine and PHIP; (2) glycine and MeIQx; (3) lysine and 4,8 DiMeIQx in Maillard chemical model systems. In addition, we investigated the effects of different conditions on decreasing amino acids and heterocyclic amines (HCA) formation in chemical model systems. The results show that the developed methods used to separate the proposed mixes gave a relative standard deviation (RSD)% of less than 2% of peak area and retention time, thus proving the robustness and high reproducibility of the proposed method. The relationship between concentration and peak areas is linear with high statistical significance (p<0.001). The residual and slope of linear regression give a limit of detection LoD) and (LoQ) of phenylalanine and PHIP were (2.92- 0.029 ppm),(9.73- 0.098 ppm),respectively, which indicate the high sensitivity of the developed method. Moreover, the findings illustrate that a water ratio of 20% leads to a decrease in phenylalanine versus the largest amount of PHIP, and phenylalanine is the most active amino acid with a decreasing percentage up to 99%.

Examining the Artificial Sweeteners in Commonly Consumed Beverages, Chewing Gums, Chocolates, and Mouthwashes using HPLC and TLC Methodology

Background: Artificial sweeteners (AS) are synthetic compounds extensively used in food and beverages industriesdue to its techno-functional role as alternatives to table sugar. Many studies report the adverse health effect on useof such compounds beyond its permitted limit. Hence, the aim of present study is to analyzed the AS contents inselected commonly consumed beverages and food products.Methods: High-Performance Liquid Chromatography and Thin Layer Chromatography methodology was usedto carry out the analysis.Results: Concentration of Ace-K ranged from 7 to 14 mg/100 ml, sucralose from 83 to 93 mg/100 ml, and aspartame11mg/100ml in beverages. In mouthwashes, saccharin ranged from 102 to 140 mg/100g. Among chocolates,saccharine was observed from 9 to 13 mg/100g. Chewing gums contained 9 to 193 mg/100 g of aspartame,118mg/100g of Ace-K, and 82 to 155 mg/100g of sucralose.Conclusion: The data could help in public awareness as well as regulatory bodies in monitoring the levels of ASfound in food products and beverages with officially permitted limits.

FÁRMACOS NA PANDEMIA DA COVID-19: UMA TEMÁTICA PARA A EDUCAÇÃO AMBIENTAL CRÍTICA NO ENSINO DE QUÍMICA ANALÍTICA

PHARMACEUTICALS IN THE COVID-19 PANDEMIC: A THEME TO CRITICAL ENVIRONMENTAL EDUCATION ON ANALYTICAL CHEMISTRY TEACHING. During the COVID-19 pandemic, the pursuit of a cure for the disease resulted in an excessive consumption of various pharmaceuticals, even after their inefficacy had been scientifically demonstrated. This scenario highlights the urgency of educational initiatives that improve students’ critical thinking and promote awareness of indiscriminate use of pharmaceuticals and its environmental and social implications. This research described the educational initiative “Pharmaceuticals in the COVID-19 pandemic” based on the study case method, implemented at an analytical chemistry class, and analyzed its potential on promoting Critical Environmental Education through a content analysis guided by the indicators proposed by Luz and Tonso. Students were divided into groups and studied different cases about pharmaceuticals featured in the media during the pandemic. They were expected to gather relevant data regarding the case scenario; to take a position on a socio-environmental dilemma; propose an analytical chromatographic methodology for the pharmaceutical in the case and present their results in a final report. The educational initiative was considered aligned with Critical Environmental Education principals due to the presence of six out of the seven indicators. As a future perspective, we aim to optimize the educational initiative in order to include all the indicators and students’ suggestions.

An analytical framework combining online high-performance liquid chromatography methodologies and biological properties of different extracts of Leonurus cardiaca.

Little or no information is available concerning online high-performance liquid chromatography (HPLC) antioxidants and the antibiofilm effect of Leonurus cardiaca. Five distinct extractions of methanolic, ethyl acetate, dichloromethane, hexane, and water were obtained from L. cardiaca. In the online-HPLC-antioxidant analysis of all examined samples, rosmarinic acid emerged as the primary antioxidant, registering concentrations ranging from 6 to 15ppm at wavelengths of 517 and 734nm. Notably, the water extract exhibited robust antioxidant activity In vitro. Regarding acetylcholinesterase and butrylcholinesterase inhibition, the n-hexane extract exhibited superior inhibition with values of 3.08 and 5.83 galanthamine equivalent, respectively. Except for the water extract, all tested extracts (at a concentration of 20μg/mL) exhibited substantial inhibitory activity against biofilm formation, in many cases superior to 80%, and reached even 94.52% againstEscherichia coli. Although less vigorous, the extracts also acted against the mature biofilm (inhibition up 76.50% againstStaphylococcus aureus). They could work against the metabolism inside an immature and mature biofilm, with inhibition percentages up to 93.18% (vs.Pseudomonas aeruginosa) and 76.50% (vs.Acinetobacter baumannii), respectively. Considering its significant antioxidants, enzyme inhibition, and antimicrobial activity, L. cardiaca emerges as a promising candidate for therapeutic potential.

Statistical analysis-based green planar chromatographic methodology for the quality assessment of food supplements: a case study on Origanum vulgare L. commercial products

Statistical analysis-based green planar chromatographic methodology for the quality assessment of food supplements: a case study on Origanum vulgare L. commercial products

Incidences of aflatoxin contaminations in ingredients, feed and products of poultry from two regions in Ghana

Commercial poultry feed required for proper growth of birds and egg production contains essential nutrients with maize as key component. Inadequately dried maize is prone to aflatoxin contamination and therefore when used in feed formulation for poultry may compromise the safety of the feeds and poultry products. This study investigated the levels of aflatoxins in feed ingredients, feed and poultry products sampled from Eastern and Greater -Accra regions of Ghana. The aflatoxin levels of B1, B2, G1, and G2 were determined using a High-Performance Liquid Chromatography (HPLC) methodology. Main feed ingredients used were fishmeal, cotton seed cake, soya meal, rice, wheat and maize bran as well as maize grains. There was correlation between the level of aflatoxins and moisture content in poultry feed ingredients. All the poultry feeds (100 %) analysed showed the presence of aflatoxins with total aflatoxins recorded ranging from 5.32 to 29.88 μg/kg. Maize samples of the poultry feeds, from all two regions, revealed maize to be a major contributor to the overall total aflatoxin contents found in the feed. Five (5) out of ten (10) communities investigated in the two (2) regions where the poultry feeds were examined recorded total aflatoxin levels in maize above the Ghana Standard Authority (GSA) specification of 20 μg/kg. Aflatoxins G1 and G2 were not detected in all samples of chicken meat and eggs. Total aflatoxin levels recorded for all chicken meat and eggs were below GSA specification of 5 μg/kg; implying that these products were safe for consumption.

Densitometric simultaneous assessment of teneligliptin hydrobromide and pioglitazone hydrochloride in combined tablet

AbstractA highly effective high‐performance thin‐layer chromatographic methodology has been developed and validated for the concurrent measurement of teneligliptin hydrobromide hydrate and pioglitazone hydrochloride in tablet offering excellent sensitivity, precision, accuracy, and robustness. The methodology involved using aluminum plates layered with silica gel 60F254 were used, and the solvent system consisted of a mixture of chloroform, methanol, and ammonia (8:1:0.5 v/v/v). The analysis involved scanning of the plate at a wavelength of 229 nm and analyzing the resulting densitograms. The linear correlation was established over a range of concentrations 250–3000 ng/band for teneligliptin hydrobromide hydrate and 187.5–2250 ng/band for pioglitazone hydrochloride. The method displayed detection limits of 21.29 ng/band for teneligliptin hydrobromide hydrate and 25.97 ng/band for pioglitazone hydrochloride. The quantification limits were 64.52 ng/band for teneligliptin hydrobromide hydrate and 78.71 ng/band for pioglitazone hydrochloride. The results of precision parameters showed that the % relative standard deviation of peak area was consistently below 2%, indicating high precision. The accuracy of the method was demonstrated to be between 96% and 103%. Proposed method was found to be superior and capable of overcoming the shortcomings of previously reported methods for the assessment of both the drugs.

Development and validation of high‐performance thin layer chromatographic method for concurrent estimation of dapagliflozin and vildagliptin in combined tablet

AbstractEnhanced glycemic regulation in individuals with diabetes mellitus can be achieved using a fixed‐dose combination of dapagliflozin (10 mg) and vildagliptin (100 mg) in tablet. The primary objective of this research was to develop and validate a high‐performance thin layer chromatographic methodology for accurately measuring the quantities of dapagliflozin and vildagliptin in combined tablet formulation. The methodology involved using aluminum plates layered with silica gel 60F254, and the solvent system comprised of acetonitrile, benzene, and glacial acetic acid (9:1:2 v/v/v). Densitograms were scanned at a wavelength of 210 nm. The linearity of the procedure was established in the series of 200–2500 ng/band for dapagliflozin and 2000–25000 ng/band for vildagliptin, with correlation coefficients (r2) of 0.9931 and 0.9954, correspondingly. The method demonstrated good sensitivity, with detection limits of 21.07 ng/band for dapagliflozin and 154.97 ng/band for vildagliptin; quantification limits of 63.84 ng/band for dapagliflozin and 469.60 ng/band for vildagliptin. The methodology was found to be precise (% relative standard deviation of peak area <2) and accurate (recovery between 97% and 103%). Proposed method was found to be superior and capable of overcoming the shortcomings of previously reported methods for the assessment of dapagliflozin and vildagliptin in combined formulation.