Analytical Quality Research Articles

Stability-indicating green HPLC method for fixed-dose tablets containing remogliflozin etabonate and teneligliptin: an AQbD approach

Background In June 2021, the Central Drug Standards Control Organization approved a fixed-dose combination tablet containing remogliflozin etabonate (100 mg) and teneligliptin (10 mg) to manage type II diabetes. Objective This study aims to develop a stability-indicating RP-HPLC method for quantifying remogliflozin etabonate and teneligliptin in tablet formulations via analytical quality by design (AQbD) principles. Methods Risk assessment, Plackett–Burman design, and central composite design were employed to understand the impact of independent variables on critical analytical attributes. The stationary phase was a HyperClone BDS C18 column, and the mobile phase consisted of acetonitrile and phosphate buffer (20 mM, pH 5) at a 45:55% (v/v) ratio. Results The method, validated per ICH Q2 (R1), resulted in retention times of 3.395 and 12.308 min for teneligliptin and remogliflozin etabonate, respectively. Forced degradation studies confirmed robustness, with clear peak separation and no interference from degradation products. The AGREE score of 0.65 supports its green applicability for tablet analysis in quality control. Conclusion The AQbD-assisted RP-HPLC method developed in this study offers environmental friendliness, efficient separation with well-defined peaks, and simple mobile phase combination.

Analytical quality by design-based stability-indicating high performance liquid chromatography method for the estimation of evogliptin tartrate in bulk and tablet dosage form.

The present study utilized Analytical Quality by Design (AQbD) approach to develop a stability-indicating high-performance liquid chromatography (HPLC) method for estimating evogliptin tartrate using design expert software. The key parameters were methodically optimized, contours were plotted, and stability was evaluated using various forced degradation conditions. Using an Agilent HPLC system with a photo diode array (PDA) detector along with Fortis C18 column (250 × 4.6mm, 5μm) effectively separated the drug from its degradants. The mobile phase used was methanol: water (pH adjusted to 3.0, 76:24; v/v) at 0.8mL/min flow rate. Evogliptin was eluted at 2.98 min, at a detection wavelength of 267 nm. The proposed method was found to be specific, precise, linear and robust. The drug was sensitive to acidic, basic, oxidative, thermal, and photodegradation resolving six degradation products. Thus, the developed AQbD-based stability-indicating HPLC method is applicable in analyzing evogliptin in bulk, tablet dosage form and stability samples.

Anion exchange-HPLC method for evaluating the encapsulation efficiency of mRNA-loaded lipid nanoparticles using analytical quality by design

Lipid nanoparticles (LNPs) are emerging nucleic acid delivery systems in the development of mRNA therapeutics such as the severe acute respiratory syndrome coronavirus 2 vaccines. However, a suitable analytical method for evaluating the encapsulation efficiency (EE) of the LNPs is required to ensure drug efficacy, as current analytical methods exhibit throughput issues and require long analysis times. Hence, we developed and validated an anion-exchange HPLC method using Analytical Quality by Design. Three critical method parameters (CMPs) were identified using risk assessment and Design of Experiments: column temperature, flow rate, and sodium perchlorate concentration. The CMPs were optimized using Face-Centered Central Composite Design. The discriminating power of the optimized HPLC method and RiboGreen assay was comparable. The main advantage of this method is that LNPs can be directly injected into the HPLC system without bursting the LNPs loaded with encapsulated poly(A). The optimized HPLC method was validated as robust, high-throughput, and sufficiently sensitive according to the ICH Q2 guidelines. We believe our findings could promote efficient LNPs-based drug development.

Validación de un método de cromatografía líquida (HPLC-UV/Vis) para la cuantificación de aldehídos en agua de lluvia y aire

Aldehydes are compounds in the atmosphere formed by the photochemical degradation of other organic compounds in the troposphere and can be emitted by natural or anthropogenic sources. [Objective] An analytical method was implemented to quantify aldehyde samples in matrices such as air and rainwater. [Methodology] Aldehyde sampling and analysis were conducted using method TO-11 from the United States Environmental Protection Agency (EPA). Sampling was carried out by using adsorption cartridges coated with a 2,4-DNPH solution, forming a hydrazone with the aldehydes present in the rainwater and the air, which were tested using a liquid chromatograph coupled to a UV-visible detector (HPLC-UV/Vis). To validate the analysis technique and the analytical quality of the results, the following was determined: linearity, sensitivity, quantification limits, detection limits, repeatability, reproducibility, and recovery percentage. In the case of repeatability, the comparison of the Horwitz coefficient of variation was used with the percentages of relative standard deviation (% RSD) of the samples. [Results] The detection and quantification limits obtained range between 0.18 μg/m3 and 3.20 μg/m3 for acetaldehyde and acrolein, respectively, while quantification limits are between 0.62 μg/m3 for acetaldehyde and 4.70 μg/m3 for heptanal, data that characterizes the method’s analytical quality. [Conclusions] In general, the analysis method for aldehydes showed linearity, with R2 values equal to or greater than 0.9991 for each calibration curve, and relative deviation percentage values less than 2.25 %, indicating good precision in the analysis.

THE ROLE OF DATA ANALYTICS IN MAKING MANAGEMENT DECISIONS BY THE LOGISTICS INTERMEDIARIES

Data analytics plays a crucial role in increasing the effectiveness of management decisions by the logistics of intermediaries. The aim of the article is to identify the extent to which the quality of data analytics affects the effectiveness of decision-making in the logistics intermediaries, in particular, the speed of delivery of the studied companies.The study employed regression and correlation analysis to identify key influencing factors in terms of data analytics on the effectiveness of management decisions of the logistics intermediaries. The significance of investment in the qualification of analysts (with a coefficient of -1.6754), analytical tools (with a coefficient of -1.2575), and integration of analytics in decision-making processes (with a coefficient of -3.2511) directly affect the reduction of delivery time.It is emphasized that each analytical project contributes to the reduction of delivery time by 0.48 hours. Correlation analysis confirmed the relationship between the efficiency of logistics and the level of qualification of analysts (-0.283617), investment in analytical tools (-0.257322), the number of analytical projects (-0.343792), the level of integration of analytics (-0.712058). The strongest correlation was observed for the integration of analytics in management decision-making.It is recommended to focus on the development of analytical competencies, increase of investment in tools, intensification of projects, and integration of analytics in strategic management. Further research is planned on the use of artificial intelligence to optimize management decisions in logistics as part of ensuring the company’s sustainable development.

Various Analytical Methods for Analysis of Sitagliptin – A Review

Sitagliptin is a dipeptidyl- peptidase inhibitor used to treat high blood sugar levels caused by type2 diabetes. Absorption of Sitagliptin is 87% orally bioavailable and taking it with or without food does not affect its pharmacokinetics. Sitagliptin reaches maximum plasma concentration in 2 hours. Now in this present analytical research world quality by design or design by expert technique is used to get improved method for method validation. This concise review work can guide an analyst to choose most appropriate method for a best analytical method development and validation. This assessment encompasses various analytical methods such as UV Spectrophotometry, High performance liquid chromatography [HPLC], Liquid chromatography – Mass spectrometry (LC-MS), High performance thin layer chromatography (HPTLC) and Ultra performance liquid chromatrography (UPLC) for the estimation of sitagliptin in single and/or in combination.

Quantification of lactic acid in wines using an amperometric biosensor

The objective of this study was to develop an analytical method that allows for the rapid and cost-effective detection and quantification of lactic acid in wines. The analysis of lactic acid is a time-consuming and costly process in the food industry. Therefore, the use of an enzymatic amperometric sensor that employs lactate oxidase (LacOx) immobilized on an oxygen electrode enabled the determination of lactic acid in wine. This study utilizes a voltage of −500mV and recorded the amperometric signal was obtained during oxygen consumption while lactic acid oxidation occurred in the sensor reactor. The detection time was at 10 s. A positive linear relationship was obtained between oxygen consumed as a function of time (mg O2/L x s−1). The lactic acid concentration ranged from 1 to 7 mM with a coefficient R2 = 0.994. The sensor exhibited good specificity KM = 0.675. The immobilized enzyme retained over 85% activity for 60 days, allowing for the reuse of the previously modified membrane up to 22 times. In comparison to other methods documented in the literature, the results demonstrate a favourable analytical quality. Furthermore, other aspects, such as the low residual formation, also exhibit a positive outcome. The analysis of lactic acid showed good correlation when compared to HPLC methodology, which occupies the same linear range. This validates the use of the sensor as an alternative method for quantifying lactic acid in wine. The value of this study lies in its presentation of a technique that is more straightforward to implement than traditional methods for quantifying lactic acid in grape wines.

An automated method for thrombocyte counting in capillary microsamples.

We aimed to develop an automated, low-volume method for thrombocyte counting in capillary blood using the Sysmex predilution (PD) mode. Microsamples were prepared by resuspension of 50 μL blood in 300 μL DCL CellPack. Thrombocyte counting was done in the impedance (PLT-I) and fluorescence (PLT-F) channels. The imprecision and bias was evaluated in >394 microsamples from adult blood. Preanalytical factors (skin-piercing, storage, and transportation in our pneumatic tube system) was assessed, and studies on pediatric microsamples were made for comparison. The improvement in analytical quality and turnaround time was examined. For PLT-F, the imprecision was 1.1%-3.7%, and the bias was 10.1% (95% CI: 8.8-11.3). After skin-piercing, the bias was 8.1% (95% CI: 5.6-10.6) and the imprecision 1.9% (95% CI: 1.3-2.5). Thrombocyte counts kept stable after 4 h at room temperature (94.8% [95% CI: 93.2-96.4]) and after pneumatic tube transportation [6.7% (95% CI: 4.8-8.6)]. The bias of the PD mode for pediatric microsamples was 13.0% (95% CI: -8.4-34.4) in the PLT-F channel. The automated method had a considerably lower imprecision than the existing manual thrombocyte counting method and reduced turnaround times. The automated microsample method offers a low-volume alternative for measurement of thrombocytes. The method appears useful also in pediatric samples.

Integrated Application of Risk Management Techniques in Developing an Analysis Method for Traditional Chinese Medicine: A Case Study of a Percolation Solution for Xiaochaihu Capsules

Risk management should run through the entire process of method development, utilization, and maintenance. Based on the analytical quality by design (AQbD) concept, various integrated risk management techniques were used in this study to develop an analysis method for the percolation solution of Xiaochaihu capsules. During the development of the analysis method, risk assessment was conducted using an Ishikawa diagram and failure mode effects analysis, followed by method optimization using experimental design. The probability of nonconformance calculated via an exhaustive Monte Carlo method quantitatively characterized the risk magnitude of method parameter failures, leading to the establishment of a operable design region method based on risk magnitude. Validation experiments and robustness tests of the data were utilized for model refinement and initial risk review. Methodological validation of the developed method was performed, and control strategies for the analysis method were presented through a decision tree. Stability experiments demonstrated that the samples remained stable at 4 °C for 24 h. The average recovery rate fell between 98.8% and 105%, with relative standard deviations ranging from 2.73% to 4.48%. The results showed that the established analysis method exhibited robustness. This analysis method can simultaneously determine the contents of uridine, adenine, 5-hydroxymethylfurfural, and guanosine. This method can also be employed for process control during percolation. This study integrated various risk management techniques to develop and maintain the analysis method, and this approach can potentially be extended to other analytical methods.

External quality assurance experience with Royal College of Pathologists of Australasia Program at an academic hospital in South Africa.

Laboratories use their performance in external quality assurance (EQA) to establish quality planning strategies and to assess whether testing processes require improvement. The EQA performance of the hematology and coagulation test parameters on the Royal College of Pathologists of Australasia EQA program was evaluated over a 4-year cycle at an academic hospital in Johannesburg, South Africa. The test performance was determined from analytical quality specification (APS) and/or z-scores. Bias and imprecision were used to calculate sigma (σ) metric scores. Specifications from European Federation of Laboratory Medicine and/or biological variation were applied. The laboratory achieved a mean testing score of 98.7±4.0%. There were 103 (10.7%) unacceptable results. On investigation, root causes included: presurvey issues (83%), transcription errors (9%), random errors (6%), and test performance errors (3%). All test parameters evaluated achieved an acceptable median APS during the study period. The mean z-scores, however, were >2 and unacceptable for mean cell hemoglobin concentration and hematocrit. On investigation, this was attributed to significant delay in transport and storage of full blood count samples. White cell count and d-dimer achieved a σ ≥ 6. EQA participation assisted the laboratory in maintaining a quality system. Close monitoring is necessary for international laboratories to avoid sample delays that can affect result quality.

Analytical Procedure Development and Proposed Established Conditions: A Case Study of a Mass Spectrometry Based NDSRI Analytical Procedure

With the finalization of the ICH Q14 Analytical Procedure Development guideline, how to apply enhanced approaches (such as analytical quality by design (AQbD)) to develop an analytical procedure, and to propose Established Conditions (ECs) and corresponding reporting categories, is increasingly being discussed. To gain practical experience in applying an enhanced approach for method development and identifying ECs, we developed, validated, and implemented an analytical procedure for a nitrosamine drug substance-related impurity (NDSRI). Here, as an example of the application of Q12 Lifecycle Management guideline principles in regards to analytical procedures, we briefly elaborate how: 1) the principles documented in the ICH Q14 guideline for analytical procedure development were applied, with the focus on identifying an Analytical Target Profile (ATP), knowledge management and risk assessment; 2) analytical procedure robustness according to the recommendations in ICH Q2(R2) Validation of Analytical Procedure guideline and Q14, were evaluated; and 3) mass spectrometry ECs and associated proposed reporting categories were proposed.

Green analytical chemistry and quality by design: A combined approach towards simultaneous determination of Letrozole with its co-administered Zoledronic Acid for cancer patients

Nowadays, breast cancer is the most affecting and globally diagnosed malignancy among women, yet Letrozole (LTZ) was considered the first-line treatment as hormonal anticancer drug. Unfortunately, LTZ develops osteoporosis as a main side effect which was overcome by using the co-administered; Zoledronic Acid (ZDA). Thus, there was a crucial need for this simultaneous quantification innovation, especially there were no any previously reported methods regarding both drugs together. In this study, an integrated framework was conducted between the experimental analytical quality-by-design (AQbD) approach and green analytical chemistry (GAC), emerging sensitive and robust RP-HPLC method. Box-Behnken Design was the developed model for optimizing an isocratic chromatographic separation on C18 Equisil® ODS (4.6 × 250 mm, 5.0 μm) column at ambient temperature, using the mobile phase of 0.1 % aqueous trifluroacetic acid (pH 2.8): acetonitrile (54.5:45.5, v/v), at 1.0 mL/min flow rate with PDA detection at 254.0 nm and 210.0 nm for LTZ and ZDA, respectively. Model statistical and residual plots analysis was significant and normally distributed. Method was fully validated as per ICH guidelines, where good linearity was 0.20–10.00 µg/mL for both drugs in presence of Tadalafil (TDF) as an internal standard, obtaining adequate correlation coefficients (r) values. Calculated LOD results were 0.058 and 0.040 µg/mL while calculated LOQ results were 0.175 and 0.122 µg/mL for LTZ and ZDA, respectively. The proposed method was effectively applied on bulk, pharmaceutical dosage forms, and spiked human plasma. Statistical comparison of the anticipated results with the reported ones was performed. Greenness assessment was evaluated using Green Analytical Procedure Index (GAPI) and Analytical Greenness (AGREE) tools; where superiority results were achieved relative to other reported methods. Finally, an EVG method evaluation tool was assessed, and the attained results were represented through its radar chart.

Application of Analytical Quality by Design to the development and validation of reduced and non-reduced capillary electrophoresis analytical procedures for mAb purity determination

Capillary electrophoresis-sodium dodecyl sulfate (CE-SDS) is a common analytical procedure used to quantitate critical quality attributes relating to the purity and glycosylation of monoclonal antibodies (mAbs). In this study, the application of an Analytical Quality by Design framework incorporating Design of Experiments was used to develop and validate both non-reduced (CE-NR) and reduced (CE-R) versions of this analytical procedure. Formal risk assessments were used to identify critical method attributes for optimization based on their potential impacts to performance criteria outlined in an analytical target profile. The resulting response surfaces connecting these critical factors to method performance were then utilized to generate a harmonized procedure to reduce execution risk across CE-R and CE-NR applications. Validation of these procedures according to regulatory guidelines support that they meet their required performance criteria, and a multivariate assessment of procedure robustness indicates that method parameters are in a sufficient state of control to ensure appropriate quantitation of mAb quality. Overall, this study demonstrates the utility of adopting an Analytical Quality by Design framework to leverage multidimensional knowledge from multiple critical method parameters to ensure an analytical procedure is fit-for-purpose.

An assessment of analytical performance using the six sigma scale in second-trimester maternal prenatal screening practices in China

ObjectivesWe aimed to evaluate the analytical performance of second-trimester maternal serum screening in China, and to compare if there are differences in sigma levels across different methods and months. MethodsA retrospective study was conducted to assess the analytical quality levels of laboratories by calculating the Sigma metrics with prenatal screening biomarkers: AFP, Total β-hCG, free β-hCG, uE3. Data from 591 laboratories were selected. Sigma metrics were computed using the formula: Sigma metrics(σ) = (%TEa - |%Bias|)/%CV. The Friedman test and Mann-Whitney test were used to compare differences across various methods and different months. The Hodges–Lehmann was used for determining 95 % confidence intervals of pseudo-medians. ResultsOnly uE3 showed significant monthly variations in sigma calculations. However, around 8 % of laboratories across all four analytes demonstrated sigma levels both above 6 and below 3 in different months. Laboratories utilizing time-resolved fluorescence methods significantly outperformed those using chemiluminescence in sigma level. For AFP, the pseudo-median difference between these methods lies within a 95 % confidence interval of (−3.22, −1.93), while for uE3, it is at (−2.30, −1.40). Notably, the median sigma levels for all analytes reached the 4-sigma threshold, with free β-hCG even attaining the 6-sigma level. ConclusionWith current standards, China's second-trimester maternal serum screening is of relatively high analytical quality, and variations in sigma levels exist across different months and methods.

The performance of quantitative D-dimer assays in Chinese clinical laboratories by analyzing data from National External quality Assessment Scheme

Background and aimsTo investigate performance of D-dimer assays in China and address analytical quality issues. Materials and methodsD-dimer assays data were collected from China National External Quality Assessment Scheme (China NEQAS) from 2014 to 2022. We analyzed reagents, assay results, reporting unit and cutoffs in 2022 China NEQAS. Interlaboratory coefficient variations (CVs) and influence of modified/unmodified test systems on CVs were investigated over 9 years. ResultsThere were 82 reagent brands in China NEQAS, but 55 reagent instructions did not indicate expression unit (DDU or FEU). Up to 7-fold of the ratio of max-to-min mean results was shown among different assays with same unit on the same sample. A prevalence of FEU (63.4%) over DDU (17.1%) was observed. Although 669 laboratories (37.9%) among 1766 laboratories used reagents without VTE exclusion claim, they also reported cutoffs. The CVs of only two assays were decreasing over years. CVs of modified test systems were higher than those of unmodified systems before improvement. ConclusionsExpression unit should be required to label in package inserts by regulatory authority. Laboratory professionals should follow instructions for use and prefer unmodified test systems for clinical safely application. Harmonization of reporting units through collaborative efforts is the promising step.

Moving Toward Economic and Digital Sustainability in Marketing Analytics

Despite analytical advancements, firms have yet to realize the full potential of big data marketing analytics (BDMA) because the poor quality data restricts customer predictions and insightful decisions. Technology and market uncertainty create challenges in understanding business needs, choosing analytical tools, determining customer insights, and market trends. This study aims to assess the quality of marketing analytics, including technology and information quality. Data were collected from 236 North American respondents working in firms with at least limited experience in the deployment of BDMA. The analysis tool was PLS-SEM. The findings supported the hypothesis that technology and market uncertainty negatively influence the quality of analytical outcomes. This study makes a significant theoretical and methodological contribution to BDMA literature by assessing the quality of analytics as an integrated formative construct.

Analytical quality by design (AQbD) based optimization of RP-UPLC method for determination of nivolumab and relatlimab in bulk and pharmaceutical dosage forms

BackgroundThe Analytical Quality by Design (AQbD) methodology extends the application of Quality by Design (QbD) principles to the management of the analytical procedure life cycle, encompassing method creation, optimization, validation, and continuous improvement. AQbD assists in creating analytical procedures that are robust, reliable, precise, and cost-efficient. Opdualag™ is a combination of Nivolumab and Relatlimab, which are antibodies that block programmed death receptor-1 (PD-1) and lymphocyte-activation gene 3 (LAG-3) receptors, used to treat advanced melanoma. This work aims to develop and validate a reversed-phase ultra-performance liquid chromatography (RP-UPLC) method using AQbD principles to determine NLB and RTB in pharmaceutical products.ResultsA central composite design (CCD) comprising three factors arranged in five distinct levels was implemented via Design-expert® software to optimize the chromatographic conditions. A mathematical model was constructed and the effects of three independent factors namely flow rate (X1), percentage of methanol in the mobile phase (X2), and temperature (X3) on responses including retention time (Y1–2), resolution factor (Y3), theoretical plates (Y4–5), and tailing factor (Y6–7) were investigated. The software determined the optimal chromatographic conditions for the separation of NLB and RTB, which were as follows: 32.80% methanol in the mobile phase, 0.272 mL/min flow rate, 29.42 °C column temperature, and 260 nm UV detection. The retention time for NLB and RTB were 1.46 and 1.88 min, respectively. The method exhibited linearity across the concentration ranges of 4–24 µg/mL for RTB and 12–72 µg/mL for NLB. The limits of detection (LOD) and limit of quantification (LOQ) for NLB and RTB, respectively, were 0.89 µg/mL, 2.69 µg/mL and 0.15 µg/mL and 0.46 µg/mL. The percentage relative standard deviation (%RSD) of intraday and interday precision for NLB and RTB was below 2. The recovery percentages for NLB and RTB were determined to be 99.57–100.43% and 99.59–100.61%, respectively. Both drugs were found to be susceptible to oxidative and photolytic degradation in forced degradation studies.ConclusionsEmploying the AQbD-based methodology, a straightforward, fast, accurate, precise, specific, and stability-indicating RP-UPLC method has been established for the quantitative analysis of NLB and its RTB in pharmaceutical formulations.

A robust ultra performance liquid chromatography method for dacarbazine quantification in nanosponge formulation: A quality by design approach

A new, user-friendly Ultra Performance Liquid Chromatography (UPLC) method was developed using Analytical Quality by Design (QbD) for precise analysis of Dacarbazine in both bulk powder and Dacarbazine-loaded Nanosponges. This method follows ICH guidelines and uses a statistically designed approach to optimize the separation conditions for Dacarbazine. A central composite design was implemented to identify the ideal combination of organic solvent (mobile phase composition) and flow rate. The UPLC method was successfully optimized for rapid separation (1.3 minutes) for Dacarbazine using a mobile phase consisting of 70% Methanol: 30% Orthophosphoric acid (0.1%) at a flow rate of 1.0 mL/min, using a Hypersil C18 column. The method exhibited excellent linearity with a correlation coefficient value of 0.997 across a concentration range of 4-20 μg/mL, accuracy (drug recovery of 96-100%) and sensitivity (LOD 1.15 μg/mL, LOQ 3.50 μg/mL) and it successfully quantified the drug in the formulated Dacarbazine-loaded Nanosponges. This QbD-driven UPLC method offers a valuable tool for reliable Dacarbazine quality control in pharmaceuticals providing rapid analysis, high accuracy, and low detection limits.

AQbD based approach for UPLC procedure development for the concurrent quantification of Metformin, Vildagliptin, Dapagliflozin and Sitagliptin in bulk and tablets: Response surface methodology paradigm

Analytical quality by design and Response Surface Methodology based UPLC procedure has been developed for the concurrent quantification of four oral anti-diabetic drugs Metformin, Vildagliptin, Dapagliflozin and Sitagliptin in bulk and tablets. ANOVA was applied for responses statistical analysis. Retention times and peak resolutions were selected as critical quality attributes. The Critical method parameters (CMPs) considered for the Response surface methodology were the organic composition of the mobile phase (%), pH of the buffer, and flow rate. CMPs of the UPLC procedure were optimized employing a 16-runs 53 Central-composite design. The optimized procedure suggested by the desirability functions approach utilizing 49.3 volumes ethanol and 50.7 volumes pH 3.73, 0.1% Triethylamine buffer with 0.26 mL/min flow rate as mobile phase, Shield RP-18 (100 × 2.1 mm, 1.7 μm) column as a stationary phase with 4 min run time resulted in maximum desirability, 1.000. At 0.737 min, 1.425 min, 1.929 min and 2.645 min Metformin, Vildagliptin, Dapagliflozin and Sitagliptin retention times were observed respectively with acceptable resolution. The optimized procedure was validated and stress studies were executed following the ICHQ2(R1) and ICHQ1A, ICHQ1B guidelines respectively. The proposed technique was evaluated employing greenness assessment tool and 0.86 was the observed greenness score. In accordance with the literature, it is evident that this is the initially reported green UPLC procedure for Metformin, Vildagliptin, Dapagliflozin and Sitagliptin concurrent estimation.

Advanced stability-indicating gradient elution reverse phase-high performance liquid chromatography method development and validation for simultaneous estimation of rutin and curcumin using an analytical quality by design approach

This research delves into a systematic Analytical Quality by Design (AQbD) approach to develop and validate a stability-indicating robust RP-HPLC method for the simultaneous determination of Rutin and Curcumin, compounds known for their anti-inflammatory, antioxidant, and wound-healing properties. Utilizing QbD principles, critical method parameters (CMPs) were identified and screened using design of experiments (DoE) and optimized via response surface methodology (RSM) with Central Composite Design (CCD). The optimized chromatographic separation used a Phenomenex Luna C18 column with gradient elution, using methanol and 0.1% formic acid, a flow rate of 1 ml/min, and detection at 345 nm (isosbestic point). The retention times were 5.63 mins for Rutin and 17.01 mins for Curcumin. The stability indication of the developed HPLC method was performed, and validation parameters were meticulously conducted following the ICH Q2(R1) guidelines, demonstrating linearity over 2-10 μg/ml for Rutin and 5-25 μg/ml for Curcumin, with correlation coefficients of 0.9999 and 0.9998, respectively. Precision studies showed %RSD within acceptable limits, and accuracies ranged between 97-100%. This validated HPLC method offers a reliable, sensitive approach for simultaneous quantifying Rutin and Curcumin, suitable for routine analysis in pharmaceutical laboratories and quality control.