Eco-Friendly First-Derivative UV-Spectrophotometric Method for Simultaneous Estimation of Dapagliflozin and Eplerenone in Bulk and Synthetic Mixture

Eco-Friendly First-Derivative UV-Spectrophotometric Method for Simultaneous Estimation of Dapagliflozin and Eplerenone in Bulk and Synthetic Mixture | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Eco-Friendly First-Derivative UV-Spectrophotometric Method for Simultaneous Estimation of Dapagliflozin and Eplerenone in Bulk and Synthetic Mixture Tandel prexa Manojbhai, Hiralben Mehta, Ratna Musale This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9264545/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 8 You are reading this latest preprint version Abstract Background: Dapagliflozin and Eplerenone are commonly used together for the management of diabetes-related cardiovascular and renal disorders, necessitating a reliable analytical approach for their combined analysis. Objective: The present work describes the development and validation of an environmentally benign first-order derivative UV-spectrophotometric method for the simultaneous determination of Dapagliflozin and Eplerenone in a synthetic mixture, in accordance with ICH Q2(R2) guidelines. Methods: First-order derivative spectra were employed to resolve spectral overlap between the two drugs. Dapagliflozin exhibited a zero-crossing point at 232 nm, while Eplerenone showed a zero-crossing at 254 nm, enabling selective quantification without mutual interference. Results: The method demonstrated linearity over concentration ranges of 1–6 µg/mL for Dapagliflozin and 2.5–15 µg/mL for Eplerenone, with correlation coefficients approaching unity. Validation studies confirmed acceptable precision, accuracy, specificity, robustness, and sensitivity. No interference from excipients was observed, and the limits of detection and quantification were suitable for routine analysis. Conclusion: Overall, the proposed method is simple, accurate, cost-effective, and eco-friendly, making it suitable for routine quality control of pharmaceutical preparations containing these drugs. Physical sciences/Chemistry Biological sciences/Drug discovery Dapagliflozin Eplerenone Eco-friendly Simultaneous Estimation Synthetic Mixture Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 NOVELTY STATEMENT A routine quality control first order eco spectrophotometric of UV derivative for reliable simultaneous measurement of Dapagliflozin and Eplerenone in synthetic mixtures and no complex chromatographic systems is presented within the work. The method also improves zero crossing analysis of spectral resolution and simultaneous measurement of components. Rapid analysis, low consumption of reagents, and simplicity of the method make the approach alternative for sustainable analysis in laboratory systems for quality control. INTRODUCTION Dapagliflozin (DAPA) is a selective sodium-glucose co-transporter-2 (SGLT2) inhibitor, widely prescribed in the management of type 2 diabetes mellitus and chronic heart failure. By inhibiting renal glucose reabsorption, it effectively lowers blood glucose levels and provides cardiovascular and renal protection. [ 1 ] Eplerenone (EPL) is a selective aldosterone antagonist indicated for hypertension and post-myocardial infarction heart failure. It blocks aldosterone binding at mineralocorticoid receptors, thereby reducing blood pressure and preventing cardiac remodeling. [ 2 ] The therapeutic combination of DAPA and EPL is gaining clinical importance due to their synergistic action in cardio-renal disease management. Consequently, the availability of a reliable analytical method for their simultaneous estimation in combined formulations or synthetic mixtures is of great significance for routine quality control and pharmaceutical analysis. [ 2 ] Chemical structure of Dapagliflozin and Eplerenone is shown in Fig. 1 , 2 . Several analytical techniques, such as HPLC, LC-MS, and HPTLC, have been reported for individual or combined estimation of related drug classes. However, these techniques are expensive, time-consuming, and require complex instrumentation. In contrast, UV-spectrophotometry offers a rapid, simple, and economical alternative. The challenge in simultaneous UV estimation arises from spectral overlap between the two drugs at their absorption maxima (232 nm for DAPA and 254 nm for EPL), making direct quantification difficult. Derivative spectrophotometry is a powerful approach to overcome such limitations. By converting normal spectra to their derivative forms, it enhances selectivity, minimizes background interference, and enables accurate quantification at zero-crossing points. Among the different orders, the first order derivative method is widely used due to its simplicity, reproducibility, and suitability for routine applications. The present study focuses on the Eco-friendly UV-spectrophotometric method Development and validation for the simultaneous estimation of Dapagliflozin and Eplerenone in a synthetic mixture. Validation Studies [ 10 ] The method was validated as per ICH Q2(R2) guidelines, evaluating parameters such as linearity, accuracy, precision, sensitivity, and robustness. This proposed method offers a cost-effective and reliable tool for quality control analysis of pharmaceutical formulations containing these drugs. Green Analytical Method [ 11 , 12 ] Green first-derivative UV analytical method development focuses on creating an environmentally sustainable, highly selective, and reagent-free spectroscopic approach. By eliminating toxic solvents, reducing sample volume, and relying solely on UV absorbance behavior, this technique minimizes chemical waste and energy consumption. The use of first-derivative processing enhances resolution between overlapping spectra, allowing accurate quantification without complex extractions or chromatographic systems. This eco-friendly strategy supports rapid, cost-effective, and precise analysis while aligning with the core principles of Green Analytical Chemistry and sustainable laboratory practices.12 principle of Green Analytical method was shown in Fig. 3 . MATERIALS AND METHODS Materials Apparatus and Instruments Double beam UV-1780 Shimadzu, Japan Analytical Balance: Shimadzu, Japan Model AP225WD Ultrasonicator: FS 4 Frantline, India Volumetric Flasks: 10ml, 50ml, 100ml Pipettes: 1ml, 2ml, 5ml Beaker: 50ml, 100ml, 250ml, 500ml Reagent and Chemicals Dapagliflozin Alembic Research Center. Eplerenone Triveni Chemicals pvt, Ltd. UV Spectroscopic Conditions Solvent: Methanol Mode: Spectrum Scanning range: 200-400nm Baseline correction: Methanol Diluent: Methanol Standard Solution Preparation Dapagliflozin Standard stock solution (100 µg/ml) Preparation After being carefully weighed, 10mg of Dapagliflozin had been transferred to 100ml volumetric flask. Methanol was incorporated into the flasks, the volume was suitably adjusted, and the flask were subjected to a 15-minute sonication. After the sonication make up the volume with methanol. Eplerenone Standard stock solution (100 µg/ml) Preparation Eplerenone 10mg had been carefully weighed as well as then transferred to various 100ml volumetric flask. Following the addition of methanol and appropriate volume adjustment, the flask were sonicated for 15 minutes. After the sonication make up the volume with methanol. Analytical wavelength Selection for Dapagliflozin and Eplerenone From the Standard stock solution of Dapagliflozin and Eplerenone: 10ml from the drug solution are transferred into 100ml volumetric flasks separately, then completed to volume with methanol. From the solution obtained above prepare the working standards of Dapagliflozin (1–6 µg/ml) and Eplerenone (2.5–15 µg/ml) in 10 ml volumetric flask using methanol as solvent. They were scanned from 200 to 400 nm and D 0 spectra is collected using UV spectrophotometer. All D 0 spectra of Dapagliflozin and Eplerenone were converted to the D 1 spectrum using UV probe 2.70 software. Comparison of D 1 spectra of Dapagliflozin and Eplerenone A). D 0 and D 1 spectra were overlaid to confirm the position. D₀ spectrum This is the zero-order spectrum (normal UV spectrum). It’s just the absorbance vs wavelength (A–λ plot). Peaks are broad, as per Beer–Lambert law. D₁ spectrum This is the first derivative spectrum. It represents the slope of the D₀ spectrum (dA/dλ vs λ). Instead of normal peaks, you see positive and negative bands crossing the baseline. The zero-crossing point of one analyte can be used to selectively measure another analyte in mixtures → this is why first-order derivative UV is useful for simultaneous estimation. In summary: D₀ = zero-order = normal absorbance spectrum. D₁ = first-order derivative = slope spectrum (dA/dλ vs λ), useful for resolving overlapping peaks. Dapagliflozin and Eplerenone synthetic Mixture Preparation: To create the final concentration of Dapagliflozin (1µg/ml) along with Eplerenone (2.5µg/ml), aliquots of 1ml from Standard solution of Dapagliflozin (10µg/ml), 2.5ml from the Standard solution of eplerenone (10µg/ml), had been placed into a common volumetric flask, and make up the volume with methanol up to 10ml. Validation of the First order derivative UV Spectroscopic Method Specificity Dapagliflozin and Eplerenone both alone and in combination. No interference from excipients or other formulation components was observed at the selected analytical wavelengths, indicating that the method is specific for the drugs under study. Linearity and Range Calibration studies were conducted by the preparation of various concentrations of solutions. The response of Dapagliflozin was linear in the concentration range of 1–6 µg/mL, whereas Eplerenone was linear over a concentration range of 2.5–15 µg/mL. The calibration curves were found to be well correlated with the correlation coefficients of nearly one, and %RSD values less than 2% indicated that the method is linear in these ranges. Accuracy Accuracy The reliability of the method was determined by performing recovery experiments in three different level: 80, 100 and 120% of target concentration. Synthetic mixtures were compound by adding known amounts of the standards. The percentage recoveries were found between 98 and 102%, thus demonstrating that the method is accuracy and matrix effect free. Precision Intraday precision: Three concentrations of Dapagliflozin (2, 4, and 6 µg/mL) and Eplerenone (5, 10, and 15 µg/mL) were injected in triplet for the same day. Interday precision: Three concentrations were measured for 3 days in a row. In both cases, the %RSD values were lower than 2%; hence this modality delivers replicable results. Detection and Quantitation Limits [ 10 ] The sensitivity of the method relates to the limit of detection (LOD) and limit of quantitation (LOQ) that were derived from three calibration curves created to establish method linearity. These values were determined based on the standard deviations of the intercepts of the calibrations and the slopes of the calibration curves by applying the following equations: LOD = 3.3 × (SD / Slope) LOQ = 10 × (SD / Slope) The values obtained show that the method is sensitive enough to detect and quantify low amounts of both drugs. Robustness To determine robustness, slight changes were made to the experimental conditions, such as minor changes to the detecting wavelength (± 2 nm). As these changes did not lead to significant changes in the attained results, this demonstrated that the method is robust and reliable. Assay The validated method was successfully used to estimate both Dapagliflozin and Eplerenone in mixture. The results of the assay were satisfactory, confirming that the method is applicable for routine quality control analysis. RESULTS Figure 4 indicate Selecting an appropriate wavelength in Eco-friendly first-order derivative UV spectrophotometry, is essential for achieving spectral clarity while maintaining sustainability by avoiding chemical reagents. The derivative spectra of Dapagliflozin and Eplerenone provided distinct zero-crossing points, enabling selective measurement of each drug without mutual spectral overlap which is shown in Fig. 5 and Fig. 6 and synthetic mixture first order derivative spectra shown in Fig. 7 The corresponding calibration plots demonstrated excellent linearity between derivative amplitude and concentration, confirming adherence to the Beer–Lambert law within the optimized green analytical range. Specificity experiments showed that common excipients and matrix components did not interfere at the chosen zero-crossing wavelengths, reflecting the selectivity of this solvent-free approach. The correlation coefficients in Tables 1 in the reports confirmed that linearity was achieved over from 1 to 6 µg/ml for Dapagliflozin and 2.5 to 15 µg/ml for Eplerenone. And their calibration curve is shown in Figs. 8 and 9 respectively Table 1 Calibration Response of Dapagliflozin & Eplerenone Conc. (µg/ml) Dapagliflozin Mean Absorbance ± SD %RSD Conc. (µg/ml) Eplerenone Mean Absorbance ± SD %RSD 1 0.0050 ± 0.0000158 0.316 2.5 0.0130 ± 0.0000380 0.292 2 0.0076 ± 0.0000381 0.501 5 0.0249 ± 0.0000450 0.181 3 0.0112 ± 0.0000277 0.247 7.5 0.0378 ± 0.0000320 0.084 4 0.0149 ± 0.0000500 0.335 10 0.0480 ± 0.0000400 0.083 5 0.0181 ± 0.0000400 0.221 12.5 0.0610 ± 0.0000180 0.029 6 0.0220 ± 0.0000339 0.154 15 0.0727 ± 0.0000550 0.076 Linearity range (µg/ml) 1–6 Linearity range (µg/ml) 2.5–15 Regression equation Y = 0.0034x − 0.0011 Regression equation Y = 0.0048x − 0.0012 Correlation coefficient (R²) 0.9975 Correlation coefficient (R²) 0.9994 Slope 0.0034 Slope 0.0048 Intercept −0.0011 −0.0012 Calibration curves were constructed by plotting first-derivative absorbance against concentration over the ranges of 1–6 µg/ml for dapagliflozin and 2.5–15 µg/ml for eplerenone. Results are expressed as mean ± SD (n = 3), with corresponding %RSD values indicating precision. The regression equations and high correlation coefficients confirm excellent linearity and proportional analytical response for both analytes. Accuracy , evaluated by standard-addition techniques, yielded recoveries of 98.50–99.10% for Dapagliflozin and 99.10–99.70% for Eplerenone showed in Table 2 , indicating strong trueness of the green method. Table 2 Recovery assessment for Dapagliflozin & Eplerenone Drug Recovery level (%) Target conc. (µg/ml) Added (µg/ml) Final conc. (µg/ml) Mean %Recovery ± SD %RSD Dapagliflozin 80 1 0.8 1.8 98.88 ± 0.554 0.56 100 1 1.0 2.0 98.50 ± 1.00 1.02 120 1 1.2 2.2 98.93 ± 0.693 0.70 Eplerenone 80 2.5 2.0 4.5 99.25 ± 0.342 0.34 100 2.5 2.5 5.0 99.33 ± 0.305 0.31 120 2.5 3.0 5.5 99.33 ± 0.695 0.70 Accuracy of the proposed method was evaluated at 80%, 100%, and 120% levels by standard addition. Results are expressed as mean % recovery ± SD (n = 3) with %RSD. Recovery values close to 100% with low variability confirm the accuracy and reliability of the method for both analytes. Table 3 Assessment of method precision within the same day & different day Drug Precision type Conc. (µg/ml) Mean response SD %RSD Dapagliflozin Intra-day 1 0.0069 0.0000079 0.114 2 0.0082 0.0000367 0.447 3 0.0114 0.0000158 0.139 Inter-day 1 0.0065 0.0000790 0.112 2 0.0079 0.0000177 0.224 3 0.0118 0.0000367 0.311 Eplerenone Intra-day 2.5 0.0146 0.0000367 0.251 5 0.0258 0.0000750 0.291 7.5 0.0445 0.000224 0.503 Inter-day 2.5 0.0158 0.0000316 0.200 5 0.0235 0.0000381 0.162 7.5 0.0437 0.0000791 0.181 Precision of the proposed method was evaluated at multiple concentration levels under intra-day (repeatability) and inter-day (intermediate precision) conditions. Results are expressed as mean absorbance ± SD (n = 3) with corresponding %RSD values. The low %RSD observed across all concentration levels indicates excellent precision and reproducibility of the method for both analytes. Table 4 Wavelength variation effect on Dapagliflozin & Eplerenone Drug Sr. No. Nominal wavelength response + 2 nm response −2 nm response Dapagliflozin (232 nm) 1 0.00667 0.00663 0.00669 2 0.00668 0.00665 0.00670 3 0.00667 0.00664 0.00668 Mean 0.006673 0.00664 0.00669 SD 0.00001 0.00001 0.00001 %RSD 0.1498 0.1506 0.1495 Eplerenone (254 nm) 1 0.0138 0.0135 0.0139 2 0.0135 0.0133 0.0137 3 0.0137 0.0136 0.0140 Mean 0.0137 0.0135 0.0139 SD 0.000158 0.000158 0.000158 %RSD 1.15 1.17 1.14 Robustness of the proposed method was evaluated by measuring derivative response at the selected wavelengths (232 nm for dapagliflozin and 254 nm for eplerenone) and at ± 2 nm variations. Results are expressed as mean ± SD (n = 3) with %RSD values. Minimal variation in analytical response indicates that the method is robust and unaffected by small changes in wavelength. Table 5 Detection and Quantitation limits of the proposed method Name of Drug Limit of Detection(µg/ml) Limit of quantitation(µg/ml) Dapagliflozin 0.0153 0.0465 Eplerenone 0.0380 0.0792 Limits of detection (LOD) and quantitation (LOQ) for dapagliflozin and eplerenone were determined based on the standard deviation of the response and the slope of the calibration curve, in accordance with ICH guidelines. The low LOD and LOQ values indicate high sensitivity of the proposed method for the quantification of both analytes. Table 6 Assay results for Synthetic Mixture analysis Theoretical Amount (mg) Measured Amount (mg) Assay value (%) Dapagliflozin Eplerenone Dapagliflozin Eplerenone Dapagliflozin Eplerenone 1 2.5 0.99 2.49 98.67 ± 0.0057 99.06 ± 0.611 1 2.5 0.98 2.46 1 2.5 0.99 2.48 Assay of dapagliflozin and eplerenone in synthetic mixture was performed to evaluate the accuracy of the proposed method. Results are expressed as mean assay values ± SD (n = 3). The assay values close to 100% indicate the suitability of the method for accurate quantification of both analytes in combined formulations. Overlay UV spectra showing the selection of analytical wavelength based on optimal absorbance and suitability for simultaneous estimation. Precision studies, including both intraday and interday precision Tabulated in Table no 3, produced low %RSD values, confirming the method's reproducibility without requiring hazardous chemicals. Minor, intentional variations in analytical conditions (± 2 nm wavelength changes) had minimal impact on results, demonstrating robustness and operational stability showed in Table no 4. The sensitivity parameters detection limit (DL) and quantitation limit (QL) showed in Table no 5 which indicate that the method can efficiently detect and quantify low analyte levels while maintaining environmental friendliness. Table no 6 showed Assay results of the synthetic mixture further validated the approach, providing purity values of 98.67 ± 0.0057% for Dapagliflozin and 99.06 ± 0.611% for Eplerenone. Overall, the findings confirm that the developed first-order derivative UV method is precise, accurate, selective, and inherently green, supporting its suitability for routine eco-friendly analytical applications. Figure 10 shows the overall green analytical assessment of the UV spectrophotometric method evaluated using the AGREE software.” DISCUSSION The Successful development and validation of a green first-order derivative UV spectroscopic method are crucial for ensuring accurate, sustainable, and reliable quantification of drug combinations with overlapping spectral features. This technique inherently supports green analytical principles by eliminating the need for hazardous solvents and reducing overall resource consumption. The use of zero-crossing points significantly enhanced selectivity, allowing clear distinction between Dapagliflozin and Eplerenone without chemical separation steps. Carefully selected wavelengths based on derivative maxima and zero-crossing behavior provided optimal sensitivity for both analytes, contributing to strong analytical performance. The calibration data exhibited excellent linearity across the validated concentration ranges, fulfilling the expectations outlined in ICH guidelines. The high correlation coefficients confirm that the derivative approach maintains a proportional response, a key requirement for dependable quantitative analysis. Accuracy was further validated through recovery studies, which yielded values close to 100%, demonstrating that the method can accurately determine each analyte even in the presence of formulation excipients. These findings align with literature emphasizing accuracy as a cornerstone of pharmaceutical quality control. Precision, assessed through repeatability and intermediate studies, produced consistently low %RSD values, reinforcing the method's reproducibility. Robustness testing indicated that small, deliberate variations in analytical conditions—such as minor wavelength shifts—did not significantly impact results, demonstrating that the method is stable, practical, and suitable for routine use. The sensitivity parameters (DL and QL) reflected the method’s capability to detect and quantify low analyte levels, making it advantageous for impurity monitoring and low-dose formulations. Assay values consistent with pharmacopeial specifications further validate the applicability of this green derivative method for active pharmaceutical ingredient determination in synthetic mixtures. Overall, the method meets all essential ICH validation criteria, including specificity, linearity, accuracy, precision, robustness, and sensitivity. Its simplicity, minimal environmental impact, and reliable analytical performance make it a strong candidate for eco-friendly routine quality assessment. Future work may incorporate chemometric techniques to enhance spectral resolution and further broaden the scope of green UV derivative applications. CONCLUSION An Eco-friendly reliable and straightforward simplified spectrum derivative for the first order of the UV for the method of spectrophotometry was developed and validated for the first time for the simultaneous measurement of Dapagliflozin and Eplerenone in a synthetic mixture. The method attained the ICH Q2 (R2) validation levels for the analysis of pharmaceuticals in the fields of accuracy, precision, specificity, and robustness. The method was reproducible, not interfering at the predetermined wavelengths. The method is suitable for routine pharmaceutical quality control due to the low sensitivity and the straightforward nature of the method to use. Declarations ACKNOWLEDGEMENTS The authors sincerely thank to Alembic Research Center and Triveni chemicals for providing the pure drug samples and necessary resources that made this study possible. We also express our gratitude to Parul University, Parul Institute of Pharmacy and Research, for offering laboratory facilities, encouragement, and continuous support throughout the research process. Finally, we appreciate all individuals who contributed to this work for their cooperation and valuable assistance. Author’s contributions Prexa Tandel performed the experimental work and drafted the manuscript. Hiralben Mehta & Ratna Musale supervised the study and critically revised the manuscript. All authors read and approved the final manuscript. Competing interests The authors declare no competing interests. Data Availability All data generated or analyzed during this study are included in this published article. Ethics declaration and consent to participate Not applicable. Consent for publication Not applicable. Funding Open access funding provided by Parul University. None received. References An, J., Niu, F. & Sim, J. J. Cardiovascular and kidney outcomes of spironolactone or eplerenone in combination with ACEI/ARBs in patients with diabetic kidney disease. Pharmacotherapy 41 , 998–1008 (2021). Abdel-Mooty, M. Albuminuria-lowering effect of dapagliflozin, eplerenone, and their combination in patients with chronic kidney disease: a randomized crossover clinical trial. J. Am. Soc. Nephrol. 33 , 1569–1580 (2022). National Center for Biotechnology Information. PubChem compound summary for dapagliflozin (CID: 9887712). Available at: https://pubchem.ncbi.nlm.nih.gov/ (2025). National Center for Biotechnology Information. PubChem compound summary for eplerenone (CID: 443872). Available at: https://pubchem.ncbi.nlm.nih.gov/ (2025). Jani, B. R., Shah, K. V. & Kapupara, P. P. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-9264545","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":619455106,"identity":"4d832617-cf3a-418d-83e4-825fa3660ee7","order_by":0,"name":"Tandel prexa Manojbhai","email":"","orcid":"","institution":"Parul Institute of Pharmacy and Research","correspondingAuthor":false,"prefix":"","firstName":"Tandel","middleName":"prexa","lastName":"Manojbhai","suffix":""},{"id":619455107,"identity":"e19fd7de-e2a0-4c31-9c4a-e9883f74df82","order_by":1,"name":"Hiralben 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Musale","email":"","orcid":"","institution":"Parul Institute of Pharmacy and Research","correspondingAuthor":false,"prefix":"","firstName":"Ratna","middleName":"","lastName":"Musale","suffix":""}],"badges":[],"createdAt":"2026-03-30 08:56:21","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-9264545/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9264545/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":106578501,"identity":"ca6d99b3-9354-43b6-928c-762223aebb47","added_by":"auto","created_at":"2026-04-10 06:12:28","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":42235,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eChemical structure of Dapagliflozin\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-9264545/v1/f301eacb98bd18542f1c18ab.png"},{"id":106725817,"identity":"71df1907-0b69-41c6-b424-a53fce63030b","added_by":"auto","created_at":"2026-04-12 18:33:58","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":45001,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eChemical structure of Eplerenone\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-9264545/v1/545c1d748d41c0224233752f.png"},{"id":106578510,"identity":"7a6f1a53-6da7-4145-9ff8-d426e73b3bbc","added_by":"auto","created_at":"2026-04-10 06:12:29","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":146474,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e12 Principle of Green Analytical Method\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIllustration of the key principles of green analytical chemistry, highlighting strategies such as reduction of solvent consumption, energy efficiency, waste minimization, and safer analytical practices applied during method development.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-9264545/v1/8fd9ad13168a6414a54aadb0.png"},{"id":106725274,"identity":"5e589d94-dfb5-4dc2-a46e-e8c8ef832f05","added_by":"auto","created_at":"2026-04-12 18:32:11","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":113244,"visible":true,"origin":"","legend":"\u003cp\u003eSelection of Wavelength\u003c/p\u003e\n\u003cp\u003eOverlay UV spectra showing the selection of analytical wavelength based on optimal absorbance and suitability for simultaneous estimation.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-9264545/v1/4b5b9f99bb4013c8a98a7b5d.png"},{"id":106578504,"identity":"f55648ee-3260-4df7-85dc-0b4287d7fdf0","added_by":"auto","created_at":"2026-04-10 06:12:28","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":139403,"visible":true,"origin":"","legend":"\u003cp\u003eFirst order spectra of Dapagliflozin.\u003c/p\u003e\n\u003cp\u003eFirst-order derivative UV spectrum of dapagliflozin showing characteristic peaks and zero-crossing points used for its selective quantification in the proposed method.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-9264545/v1/c82398fd259124f60763c8ee.png"},{"id":106728463,"identity":"2d656e5d-9f7d-4f7f-8120-a5fe72d9b96c","added_by":"auto","created_at":"2026-04-12 18:42:52","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":198969,"visible":true,"origin":"","legend":"\u003cp\u003eFirst order spectra of Eplerenone.\u003c/p\u003e\n\u003cp\u003eFirst-order derivative UV spectrum of eplerenone illustrating distinct spectral features \u0026nbsp;enabling its accurate determination without interference.\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-9264545/v1/9df91939c5a18e158fbbe71c.png"},{"id":106725875,"identity":"83fd4cbc-7639-4f2a-b92e-3ec25cb85c04","added_by":"auto","created_at":"2026-04-12 18:34:14","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":118862,"visible":true,"origin":"","legend":"\u003cp\u003eFirst order spectra of Mixture.\u003c/p\u003e\n\u003cp\u003eOverlay first-order derivative spectra of dapagliflozin and eplerenone in combined form, demonstrating the absence of spectral interference and suitability for simultaneous estimation.\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-9264545/v1/931443764a3e826220fb31a5.png"},{"id":106578506,"identity":"dd7dea17-b4da-46b6-b761-09ca81693fa7","added_by":"auto","created_at":"2026-04-10 06:12:28","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":34173,"visible":true,"origin":"","legend":"\u003cp\u003eCalibration Curve of Dapagliflozin\u003c/p\u003e\n\u003cp\u003eCalibration curve obtained by plotting first-order derivative absorbance against concentration for dapagliflozin, showing a linear relationship within the studied range with high correlation.\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-9264545/v1/c74ce25d3f4052edba559e98.png"},{"id":106725887,"identity":"db46a6a6-e81a-426a-b13f-bd984bea4b23","added_by":"auto","created_at":"2026-04-12 18:34:18","extension":"png","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":36977,"visible":true,"origin":"","legend":"\u003cp\u003eCalibration Curve of Eplerenone\u003c/p\u003e\n\u003cp\u003eCalibration curve of eplerenone representing the linear relationship between derivative \u0026nbsp;absorbance and concentration, confirming the method’s reliability for quantitative analysis.\u003c/p\u003e","description":"","filename":"9.png","url":"https://assets-eu.researchsquare.com/files/rs-9264545/v1/73570450f9136bf764b12be4.png"},{"id":106578508,"identity":"8da1f063-6efa-4c78-b7fe-d671156ad805","added_by":"auto","created_at":"2026-04-10 06:12:28","extension":"png","order_by":10,"title":"Figure 10","display":"","copyAsset":false,"role":"figure","size":217545,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eResult of Green Analysis of the Method (\u003c/strong\u003eAnalysis was perform using \u003cstrong\u003eAGREE \u003c/strong\u003eSoftware)\u003cstrong\u003e \u003c/strong\u003eGraphical representation of the greenness assessment of the proposed analytical method evaluated using AGREE (Analytical GREEnness) software. The circular diagram summarizes compliance with the 12 principles of green analytical chemistry, with the overall score indicating the environmental sustainability of the method. The high AGREE score reflects minimal solvent usage, reduced waste generation, and energy-efficient analytical conditions, confirming the eco-friendly nature of the developed method.\u003c/p\u003e","description":"","filename":"10.png","url":"https://assets-eu.researchsquare.com/files/rs-9264545/v1/82ca489577c8f20b0f067e89.png"},{"id":106994043,"identity":"5dfea9b9-0068-457b-b8db-72f8fdd24103","added_by":"auto","created_at":"2026-04-15 15:03:10","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2238156,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9264545/v1/05cdabb6-358d-4ae0-ac2d-739f741a6623.pdf"},{"id":106728170,"identity":"25bacaff-3fa1-44c4-9601-52d9c2ea8db9","added_by":"auto","created_at":"2026-04-12 18:42:03","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":344256,"visible":true,"origin":"","legend":"","description":"","filename":"GraphicalAbstract.docx","url":"https://assets-eu.researchsquare.com/files/rs-9264545/v1/9513e8e4cca2d3ad434b25b9.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Eco-Friendly First-Derivative UV-Spectrophotometric Method for Simultaneous Estimation of Dapagliflozin and Eplerenone in Bulk and Synthetic Mixture","fulltext":[{"header":"NOVELTY STATEMENT","content":"\u003cp\u003eA routine quality control first order eco spectrophotometric of UV derivative for reliable simultaneous measurement of Dapagliflozin and Eplerenone in synthetic mixtures and no complex chromatographic systems is presented within the work. The method also improves zero crossing analysis of spectral resolution and simultaneous measurement of components. Rapid analysis, low consumption of reagents, and simplicity of the method make the approach alternative for sustainable analysis in laboratory systems for quality control.\u003c/p\u003e"},{"header":"INTRODUCTION","content":"\u003cp\u003eDapagliflozin (DAPA) is a selective sodium-glucose co-transporter-2 (SGLT2) inhibitor, widely prescribed in the management of type 2 diabetes mellitus and chronic heart failure. By inhibiting renal glucose reabsorption, it effectively lowers blood glucose levels and provides cardiovascular and renal protection.\u003csup\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]\u003c/sup\u003e Eplerenone (EPL) is a selective aldosterone antagonist indicated for hypertension and post-myocardial infarction heart failure. It blocks aldosterone binding at mineralocorticoid receptors, thereby reducing blood pressure and preventing cardiac remodeling.\u003csup\u003e[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eThe therapeutic combination of DAPA and EPL is gaining clinical importance due to their synergistic action in cardio-renal disease management. Consequently, the availability of a reliable analytical method for their simultaneous estimation in combined formulations or synthetic mixtures is of great significance for routine quality control and pharmaceutical analysis.\u003csup\u003e[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]\u003c/sup\u003e Chemical structure of Dapagliflozin and Eplerenone is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, \u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eSeveral analytical techniques, such as HPLC, LC-MS, and HPTLC, have been reported for individual or combined estimation of related drug classes. However, these techniques are expensive, time-consuming, and require complex instrumentation. In contrast, UV-spectrophotometry offers a rapid, simple, and economical alternative. The challenge in simultaneous UV estimation arises from spectral overlap between the two drugs at their absorption maxima (232 nm for DAPA and 254 nm for EPL), making direct quantification difficult. Derivative spectrophotometry is a powerful approach to overcome such limitations. By converting normal spectra to their derivative forms, it enhances selectivity, minimizes background interference, and enables accurate quantification at zero-crossing points. Among the different orders, the first order derivative method is widely used due to its simplicity, reproducibility, and suitability for routine applications. The present study focuses on the Eco-friendly UV-spectrophotometric method Development and validation for the simultaneous estimation of Dapagliflozin and Eplerenone in a synthetic mixture.\u003c/p\u003e \u003cp\u003e \u003cb\u003eValidation Studies\u003c/b\u003e \u003csup\u003e[\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eThe method was validated as per ICH Q2(R2) guidelines, evaluating parameters such as linearity, accuracy, precision, sensitivity, and robustness. This proposed method offers a cost-effective and reliable tool for quality control analysis of pharmaceutical formulations containing these drugs.\u003c/p\u003e \u003cp\u003e \u003cb\u003eGreen Analytical Method\u003c/b\u003e \u003csup\u003e[\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eGreen first-derivative UV analytical method development focuses on creating an environmentally sustainable, highly selective, and reagent-free spectroscopic approach. By eliminating toxic solvents, reducing sample volume, and relying solely on UV absorbance behavior, this technique minimizes chemical waste and energy consumption. The use of first-derivative processing enhances resolution between overlapping spectra, allowing accurate quantification without complex extractions or chromatographic systems. This eco-friendly strategy supports rapid, cost-effective, and precise analysis while aligning with the core principles of Green Analytical Chemistry and sustainable laboratory practices.12 principle of Green Analytical method was shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eMaterials\u003c/h2\u003e \u003cdiv id=\"Sec4\" class=\"Section3\"\u003e \u003ch2\u003eApparatus and Instruments\u003c/h2\u003e \u003cp\u003eDouble beam UV-1780 Shimadzu, Japan\u003c/p\u003e \u003cp\u003eAnalytical Balance: Shimadzu, Japan Model AP225WD\u003c/p\u003e \u003cp\u003eUltrasonicator: FS 4 Frantline, India\u003c/p\u003e \u003cp\u003eVolumetric Flasks: 10ml, 50ml, 100ml\u003c/p\u003e \u003cp\u003ePipettes: 1ml, 2ml, 5ml\u003c/p\u003e \u003cp\u003eBeaker: 50ml, 100ml, 250ml, 500ml\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e\n\u003ch3\u003eReagent and Chemicals\u003c/h3\u003e\n\u003cp\u003e \u003cstrong\u003eDapagliflozin\u003c/strong\u003e \u003cp\u003eAlembic Research Center.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eEplerenone\u003c/strong\u003e \u003cp\u003eTriveni Chemicals pvt, Ltd.\u003c/p\u003e \u003c/p\u003e\n\u003ch3\u003eUV Spectroscopic Conditions\u003c/h3\u003e\n\u003cp\u003eSolvent: Methanol\u003c/p\u003e \u003cp\u003eMode: Spectrum\u003c/p\u003e \u003cp\u003eScanning range: 200-400nm\u003c/p\u003e \u003cp\u003eBaseline correction: Methanol\u003c/p\u003e \u003cp\u003eDiluent: Methanol\u003c/p\u003e\n\u003ch3\u003eStandard Solution Preparation\u003c/h3\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eDapagliflozin Standard stock solution (100 \u0026micro;g/ml) Preparation\u003c/h2\u003e \u003cp\u003eAfter being carefully weighed, 10mg of Dapagliflozin had been transferred to 100ml volumetric flask. Methanol was incorporated into the flasks, the volume was suitably adjusted, and the flask were subjected to a 15-minute sonication. After the sonication make up the volume with methanol.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eEplerenone Standard stock solution (100 µg/ml) Preparation\u003c/h3\u003e\n\u003cp\u003eEplerenone 10mg had been carefully weighed as well as then transferred to various 100ml volumetric flask. Following the addition of methanol and appropriate volume adjustment, the flask were sonicated for 15 minutes. After the sonication make up the volume with methanol.\u003c/p\u003e\n\u003ch3\u003eAnalytical wavelength Selection for Dapagliflozin and Eplerenone\u003c/h3\u003e\n\u003cp\u003eFrom the Standard stock solution of Dapagliflozin and Eplerenone: 10ml from the drug solution are transferred into 100ml volumetric flasks separately,\u0026ensp;then completed to volume with methanol. From the solution obtained above prepare the working\u0026ensp;standards of Dapagliflozin (1\u0026ndash;6 \u0026micro;g/ml) and Eplerenone (2.5\u0026ndash;15 \u0026micro;g/ml) in 10 ml volumetric flask using methanol as solvent. They were scanned from 200 to 400 nm and D\u003csub\u003e0\u003c/sub\u003e\u0026ensp;spectra is collected using UV spectrophotometer. All D\u003csub\u003e0\u003c/sub\u003e spectra of Dapagliflozin and Eplerenone were converted to the D\u003csub\u003e1\u003c/sub\u003e spectrum using UV\u0026ensp;probe 2.70 software. Comparison of D\u003csub\u003e1\u003c/sub\u003e\u0026ensp;spectra of Dapagliflozin and Eplerenone A). D\u003csub\u003e0\u003c/sub\u003e and D\u003csub\u003e1\u003c/sub\u003e spectra were overlaid to confirm the position.\u003c/p\u003e \u003cp\u003eD₀ spectrum\u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eThis is the zero-order spectrum (normal UV spectrum).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eIt\u0026rsquo;s just the absorbance vs wavelength (A\u0026ndash;λ plot).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003ePeaks are broad, as per Beer\u0026ndash;Lambert law.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cp\u003eD₁ spectrum\u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eThis is the first derivative spectrum.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eIt represents the slope of the D₀ spectrum (dA/dλ vs λ).\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eInstead of normal peaks, you see positive and negative bands crossing the baseline.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eThe zero-crossing point of one analyte can be used to selectively measure another analyte in mixtures \u0026rarr; this is why first-order derivative UV is useful for simultaneous estimation.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cp\u003eIn summary:\u003c/p\u003e \u003cp\u003e \u003cul\u003e \u003cli\u003e \u003cp\u003eD₀ = zero-order\u0026thinsp;=\u0026thinsp;normal absorbance spectrum.\u003c/p\u003e \u003c/li\u003e \u003cli\u003e \u003cp\u003eD₁ = first-order derivative\u0026thinsp;=\u0026thinsp;slope spectrum (dA/dλ vs λ), useful for resolving overlapping peaks.\u003c/p\u003e \u003c/li\u003e \u003c/ul\u003e \u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eDapagliflozin and Eplerenone synthetic Mixture Preparation:\u003c/h2\u003e \u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eTo create the final concentration of Dapagliflozin (1\u0026micro;g/ml) along with Eplerenone (2.5\u0026micro;g/ml), aliquots of 1ml from Standard solution of Dapagliflozin (10\u0026micro;g/ml), 2.5ml from the Standard solution of eplerenone (10\u0026micro;g/ml), had been placed into a common volumetric flask, and make up the volume with methanol up to 10ml.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eValidation of the First order derivative UV Spectroscopic Method\u003c/h2\u003e \u003cdiv id=\"Sec13\" class=\"Section3\"\u003e \u003ch2\u003eSpecificity\u003c/h2\u003e \u003cp\u003eDapagliflozin and Eplerenone both alone and in combination. No interference from excipients or other formulation components was observed at the selected analytical wavelengths, indicating that the method is specific for the drugs under study.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eLinearity and Range\u003c/h2\u003e \u003cp\u003eCalibration studies were conducted by the preparation of\u0026ensp;various concentrations of solutions. The response of Dapagliflozin was linear in the concentration range of 1\u0026ndash;6 \u0026micro;g/mL, whereas Eplerenone was linear over a concentration range\u0026ensp;of 2.5\u0026ndash;15 \u0026micro;g/mL. The calibration curves were found to be well correlated\u0026ensp;with the correlation coefficients of nearly one, and %RSD values less than 2% indicated that the method is linear in these ranges.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eAccuracy\u003c/h2\u003e \u003cp\u003eAccuracy\u003c/p\u003e \u003cp\u003eThe reliability of the method was determined by performing recovery experiments in three different level: 80, 100\u0026ensp;and 120% of target concentration. Synthetic mixtures were compound by adding known amounts of the\u0026ensp;standards. The percentage recoveries were found between 98 and\u0026ensp;102%, thus demonstrating that the method is accuracy and matrix effect free.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003ePrecision\u003c/h2\u003e \u003cp\u003eIntraday precision: Three concentrations of Dapagliflozin (2, 4, and 6 \u0026micro;g/mL) and Eplerenone\u0026ensp;(5, 10, and 15 \u0026micro;g/mL) were injected in triplet for the same day.\u003c/p\u003e \u003cp\u003eInterday precision: Three concentrations were measured for 3 days\u0026ensp;in a row.\u003c/p\u003e \u003cp\u003eIn both cases, the %RSD values were lower\u0026ensp;than 2%; hence this modality delivers replicable results.\u003c/p\u003e \u003cp\u003e \u003cb\u003eDetection and Quantitation Limits\u003c/b\u003e \u003csup\u003e[\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eThe sensitivity of the method relates to the limit of detection (LOD) and limit of quantitation (LOQ) that were derived from three calibration curves created to establish method linearity. These values were determined based on the standard deviations of the intercepts of the calibrations and the slopes of the calibration curves by applying the following equations:\u003c/p\u003e \u003cp\u003eLOD\u0026thinsp;=\u0026thinsp;3.3 \u0026times; (SD / Slope)\u003c/p\u003e \u003cp\u003eLOQ\u0026thinsp;=\u0026thinsp;10 \u0026times; (SD / Slope)\u003c/p\u003e \u003cp\u003eThe values obtained show that the method is sensitive enough to detect and quantify low amounts of both drugs.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eRobustness\u003c/h2\u003e \u003cp\u003eTo determine robustness, slight changes were made to the experimental conditions, such as minor changes to the detecting wavelength (\u0026plusmn;\u0026thinsp;2 nm). As these changes did not lead to significant changes in the attained results, this demonstrated that the method is robust and reliable.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eAssay\u003c/h2\u003e \u003cp\u003eThe validated method was successfully used to estimate both Dapagliflozin and Eplerenone in mixture. The results of the assay were satisfactory, confirming that the method is applicable for routine quality control analysis.\u003c/p\u003e \u003c/div\u003e"},{"header":"RESULTS","content":"\u003cp\u003eFigure 4 indicate Selecting an appropriate wavelength in Eco-friendly first-order derivative UV spectrophotometry, is essential for achieving spectral clarity while maintaining sustainability by avoiding chemical reagents. The derivative spectra of Dapagliflozin and Eplerenone provided distinct zero-crossing points, enabling selective measurement of each drug without mutual spectral overlap which is shown in Fig. \u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e5\u003c/span\u003e and Fig. \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e6\u003c/span\u003e and synthetic mixture first order derivative spectra shown in Fig. \u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e7\u003c/span\u003e The corresponding calibration plots demonstrated excellent linearity between derivative amplitude and concentration, confirming adherence to the Beer\u0026ndash;Lambert law within the optimized green analytical range. Specificity experiments showed that common excipients and matrix components did not interfere at the chosen zero-crossing wavelengths, reflecting the selectivity of this solvent-free approach.\u003c/p\u003e\n\u003cp\u003eThe correlation coefficients in Tables \u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e in the reports confirmed that \u003cstrong\u003elinearity\u003c/strong\u003e was achieved over from 1 to 6 \u0026micro;g/ml for Dapagliflozin and 2.5 to 15 \u0026micro;g/ml for Eplerenone. And their calibration curve is shown in Figs. \u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e8\u003c/span\u003e and \u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e9\u003c/span\u003e respectively\u0026nbsp;\u003c/p\u003e\n\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eCalibration Response of Dapagliflozin \u0026amp; Eplerenone\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eConc. (\u0026micro;g/ml)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eDapagliflozin Mean Absorbance\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e%RSD\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003eConc. (\u0026micro;g/ml)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003eEplerenone Mean Absorbance\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e%RSD\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.0050\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0000158\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.316\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e2.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0130\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0000380\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.292\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.0076\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0000381\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.501\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0249\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0000450\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.181\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.0112\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0000277\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.247\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e7.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0378\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0000320\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.084\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.0149\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0000500\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.335\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0480\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0000400\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.083\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.0181\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0000400\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.221\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e12.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0610\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0000180\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.029\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.0220\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0000339\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.154\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0727\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0000550\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.076\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eLinearity range (\u0026micro;g/ml)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e\n \u003cp\u003e1\u0026ndash;6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003eLinearity range (\u0026micro;g/ml)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e\n \u003cp\u003e2.5\u0026ndash;15\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eRegression equation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e\n \u003cp\u003eY\u0026thinsp;=\u0026thinsp;0.0034x\u0026thinsp;\u0026minus;\u0026thinsp;0.0011\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003eRegression equation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e\n \u003cp\u003eY\u0026thinsp;=\u0026thinsp;0.0048x\u0026thinsp;\u0026minus;\u0026thinsp;0.0012\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eCorrelation coefficient (R\u0026sup2;)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e\n \u003cp\u003e0.9975\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003eCorrelation coefficient (R\u0026sup2;)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e\n \u003cp\u003e0.9994\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eSlope\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e\n \u003cp\u003e0.0034\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003eSlope\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e\n \u003cp\u003e0.0048\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eIntercept\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e\n \u003cp\u003e\u0026minus;0.0011\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e\n \u003cp\u003e\u0026minus;0.0012\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003eCalibration curves were constructed by plotting first-derivative absorbance against concentration over the ranges of 1\u0026ndash;6 \u0026micro;g/ml for dapagliflozin and 2.5\u0026ndash;15 \u0026micro;g/ml for eplerenone. Results are expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD (n\u0026thinsp;=\u0026thinsp;3), with corresponding %RSD values indicating precision. The regression equations and high correlation coefficients confirm excellent linearity and proportional analytical response for both analytes.\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAccuracy\u003c/strong\u003e, evaluated by standard-addition techniques, yielded recoveries of 98.50\u0026ndash;99.10% for Dapagliflozin and 99.10\u0026ndash;99.70% for Eplerenone showed in Table \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, indicating strong trueness of the green method.\u0026nbsp;\u003c/p\u003e\n\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eRecovery assessment for Dapagliflozin \u0026amp; Eplerenone\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eDrug\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eRecovery level (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003eTarget conc. (\u0026micro;g/ml)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003eAdded (\u0026micro;g/ml)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003eFinal conc. (\u0026micro;g/ml)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003eMean %Recovery\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e%RSD\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e\u003cstrong\u003eDapagliflozin\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e1.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e98.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.554\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e0.56\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e1.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e2.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e98.50\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e1.02\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e120\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e1.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e2.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e98.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.693\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e0.70\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e\u003cstrong\u003eEplerenone\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e2.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e2.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e4.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e99.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.342\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e0.34\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e2.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e2.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e5.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e99.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.305\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e0.31\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e120\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e2.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e3.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e5.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e99.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.695\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c7\"\u003e\n \u003cp\u003e0.70\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\"\u003eAccuracy of the proposed method was evaluated at 80%, 100%, and 120% levels by standard addition. Results are expressed as mean % recovery\u0026thinsp;\u0026plusmn;\u0026thinsp;SD (n\u0026thinsp;=\u0026thinsp;3) with %RSD. Recovery values close to 100% with low variability confirm the accuracy and reliability of the method for both analytes.\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eAssessment of method precision within the same day \u0026amp; different day\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eDrug\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003ePrecision type\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003eConc. (\u0026micro;g/ml)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003eMean response\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003eSD\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e%RSD\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eDapagliflozin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eIntra-day\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.0069\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0000079\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.114\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.0082\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0000367\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.447\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.0114\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0000158\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.139\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eInter-day\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.0065\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0000790\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.112\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.0079\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0000177\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.224\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.0118\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0000367\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.311\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eEplerenone\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eIntra-day\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e2.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.0146\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0000367\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.251\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.0258\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0000750\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.291\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e7.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.0445\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.000224\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.503\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eInter-day\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e2.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.0158\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0000316\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.200\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.0235\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0000381\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.162\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e7.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.0437\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0000791\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003e0.181\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003ePrecision of the proposed method was evaluated at multiple concentration levels under intra-day (repeatability) and inter-day (intermediate precision) conditions. Results are expressed as mean absorbance\u0026thinsp;\u0026plusmn;\u0026thinsp;SD (n\u0026thinsp;=\u0026thinsp;3) with corresponding %RSD values. The low %RSD observed across all concentration levels indicates excellent precision and reproducibility of the method for both analytes.\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eWavelength variation effect on Dapagliflozin \u0026amp; Eplerenone\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eDrug\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eSr. No.\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003eNominal wavelength response\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e+\u0026thinsp;2 nm response\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e\u0026minus;2 nm response\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eDapagliflozin (232 nm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.00667\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.00663\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.00669\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.00668\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.00665\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.00670\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.00667\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.00664\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.00668\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eMean\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.006673\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.00664\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.00669\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eSD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.00001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.00001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.00001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e%RSD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.1498\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.1506\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.1495\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eEplerenone (254 nm)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.0138\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.0135\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0139\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.0135\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.0133\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0137\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.0137\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.0136\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0140\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eMean\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.0137\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.0135\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.0139\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eSD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.000158\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e0.000158\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e0.000158\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e%RSD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e1.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e1.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003e1.14\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003eRobustness of the proposed method was evaluated by measuring derivative response at the selected wavelengths (232 nm for dapagliflozin and 254 nm for eplerenone) and at \u0026plusmn;\u0026thinsp;2 nm variations. Results are expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD (n\u0026thinsp;=\u0026thinsp;3) with %RSD values. Minimal variation in analytical response indicates that the method is robust and unaffected by small changes in wavelength.\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eDetection and Quantitation limits of the proposed method\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eName of Drug\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eLimit of Detection(\u0026micro;g/ml)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003eLimit of quantitation(\u0026micro;g/ml)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eDapagliflozin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.0153\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.0465\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eEplerenone\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e0.0380\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.0792\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\"\u003eLimits of detection (LOD) and quantitation (LOQ) for dapagliflozin and eplerenone were determined based on the standard deviation of the response and the slope of the calibration curve, in accordance with ICH guidelines. The low LOD and LOQ values indicate high sensitivity of the proposed method for the quantification of both analytes.\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\n \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003cbr\u003e\u003c/div\u003e\u0026nbsp;\u0026nbsp;\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eAssay results for Synthetic Mixture analysis\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e\n \u003cp\u003eTheoretical Amount (mg)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e\n \u003cp\u003eMeasured Amount (mg)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e\n \u003cp\u003eAssay value (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003eDapagliflozin\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003eEplerenone\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003eDapagliflozin\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003eEplerenone\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c5\"\u003e\n \u003cp\u003eDapagliflozin\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colname=\"c6\"\u003e\n \u003cp\u003eEplerenone\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e2.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e2.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c5\" morerows=\"2\" rowspan=\"3\"\u003e\n \u003cp\u003e98.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0057\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c6\" morerows=\"2\" rowspan=\"3\"\u003e\n \u003cp\u003e99.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.611\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e2.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e2.46\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colname=\"c1\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c2\"\u003e\n \u003cp\u003e2.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c3\"\u003e\n \u003cp\u003e0.99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colname=\"c4\"\u003e\n \u003cp\u003e2.48\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003eAssay of dapagliflozin and eplerenone in synthetic mixture was performed to evaluate the accuracy of the proposed method. Results are expressed as mean assay values\u0026thinsp;\u0026plusmn;\u0026thinsp;SD (n\u0026thinsp;=\u0026thinsp;3). The assay values close to 100% indicate the suitability of the method for accurate quantification of both analytes in combined formulations.\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eOverlay UV spectra showing the selection of analytical wavelength based on optimal absorbance and suitability for simultaneous estimation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePrecision\u003c/strong\u003e studies, including both intraday and interday precision Tabulated in Table no 3, produced low %RSD values, confirming the method\u0026apos;s reproducibility without requiring hazardous chemicals.\u003c/p\u003e\n\u003cp\u003eMinor, intentional variations in analytical conditions (\u0026plusmn;\u0026thinsp;2 nm wavelength changes) had minimal impact on results, demonstrating \u003cstrong\u003erobustness\u003c/strong\u003e and operational stability showed in Table no 4. The sensitivity parameters \u003cstrong\u003edetection limit (DL) and quantitation limit (QL)\u003c/strong\u003e showed in Table no 5 which indicate that the method can efficiently detect and quantify low analyte levels while maintaining environmental friendliness.\u003c/p\u003e\n\u003cp\u003eTable no 6 showed \u003cstrong\u003eAssay\u003c/strong\u003e results of the synthetic mixture further validated the approach, providing purity values of 98.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.0057% for Dapagliflozin and 99.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.611% for Eplerenone. Overall, the findings confirm that the developed first-order derivative UV method is precise, accurate, selective, and inherently green, supporting its suitability for routine eco-friendly analytical applications. Figure \u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e10\u003c/span\u003e shows the overall green analytical assessment of the UV spectrophotometric method evaluated using the AGREE software.\u0026rdquo;\u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eThe Successful development and validation of a green first-order derivative UV spectroscopic method are crucial for ensuring accurate, sustainable, and reliable quantification of drug combinations with overlapping spectral features. This technique inherently supports green analytical principles by eliminating the need for hazardous solvents and reducing overall resource consumption. The use of zero-crossing points significantly enhanced selectivity, allowing clear distinction between Dapagliflozin and Eplerenone without chemical separation steps. Carefully selected wavelengths based on derivative maxima and zero-crossing behavior provided optimal sensitivity for both analytes, contributing to strong analytical performance. The calibration data exhibited excellent linearity across the validated concentration ranges, fulfilling the expectations outlined in ICH guidelines. The high correlation coefficients confirm that the derivative approach maintains a proportional response, a key requirement for dependable quantitative analysis. Accuracy was further validated through recovery studies, which yielded values close to 100%, demonstrating that the method can accurately determine each analyte even in the presence of formulation excipients. These findings align with literature emphasizing accuracy as a cornerstone of pharmaceutical quality control. Precision, assessed through repeatability and intermediate studies, produced consistently low %RSD values, reinforcing the method's reproducibility. Robustness testing indicated that small, deliberate variations in analytical conditions\u0026mdash;such as minor wavelength shifts\u0026mdash;did not significantly impact results, demonstrating that the method is stable, practical, and suitable for routine use. The sensitivity parameters (DL and QL) reflected the method\u0026rsquo;s capability to detect and quantify low analyte levels, making it advantageous for impurity monitoring and low-dose formulations. Assay values consistent with pharmacopeial specifications further validate the applicability of this green derivative method for active pharmaceutical ingredient determination in synthetic mixtures. Overall, the method meets all essential ICH validation criteria, including specificity, linearity, accuracy, precision, robustness, and sensitivity. Its simplicity, minimal environmental impact, and reliable analytical performance make it a strong candidate for eco-friendly routine quality assessment. Future work may incorporate chemometric techniques to enhance spectral resolution and further broaden the scope of green UV derivative applications.\u003c/p\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eAn Eco-friendly reliable and straightforward simplified spectrum derivative for the first order of the UV for the method of spectrophotometry was developed and validated for the first time for the simultaneous measurement of Dapagliflozin and Eplerenone in a synthetic mixture. The method attained the ICH Q2 (R2) validation levels for the analysis of pharmaceuticals in the fields of accuracy, precision, specificity, and robustness. The method was reproducible, not interfering at the predetermined wavelengths. The method is suitable for routine pharmaceutical quality control due to the low sensitivity and the straightforward nature of the method to use.\u003c/p\u003e\n"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eACKNOWLEDGEMENTS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors sincerely thank to Alembic Research Center and Triveni chemicals for providing the pure drug samples and necessary resources that made this study possible.\u003cbr\u003e\u0026nbsp;We also express our gratitude to Parul University, Parul Institute of Pharmacy and Research, for offering laboratory facilities, encouragement, and continuous support throughout the research process. Finally, we appreciate all individuals who contributed to this work for their cooperation and valuable assistance.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor\u0026rsquo;s contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePrexa Tandel performed the experimental work and drafted the manuscript. Hiralben Mehta \u0026amp; Ratna Musale supervised the study and critically revised the manuscript. All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data generated or analyzed during this study are included in this published article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics declaration and consent to participate\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eOpen access funding provided by Parul University. None received.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAn, J., Niu, F. \u0026amp; Sim, J. J. Cardiovascular and kidney outcomes of spironolactone or eplerenone in combination with ACEI/ARBs in patients with diabetic kidney disease. \u003cem\u003ePharmacotherapy\u003c/em\u003e\u003cstrong\u003e41\u003c/strong\u003e, 998\u0026ndash;1008 (2021).\u003c/li\u003e\n \u003cli\u003eAbdel-Mooty, M. Albuminuria-lowering effect of dapagliflozin, eplerenone, and their combination in patients with chronic kidney disease: a randomized crossover clinical trial. \u003cem\u003eJ. Am. Soc. Nephrol.\u003c/em\u003e\u003cstrong\u003e33\u003c/strong\u003e, 1569\u0026ndash;1580 (2022).\u003c/li\u003e\n \u003cli\u003eNational Center for Biotechnology Information. PubChem compound summary for dapagliflozin (CID: 9887712). Available at: https://pubchem.ncbi.nlm.nih.gov/ (2025).\u003c/li\u003e\n \u003cli\u003eNational Center for Biotechnology Information. PubChem compound summary for eplerenone (CID: 443872). Available at: https://pubchem.ncbi.nlm.nih.gov/ (2025).\u003c/li\u003e\n \u003cli\u003eJani, B. R., Shah, K. V. \u0026amp; Kapupara, P. P. Development and validation of UV spectroscopic first-derivative method for simultaneous estimation of dapagliflozin and metformin hydrochloride in synthetic mixture. \u003cem\u003eJ. Basic Appl. Sci.\u003c/em\u003e\u003cstrong\u003e4\u003c/strong\u003e, 1\u0026ndash;7 (2015).\u003c/li\u003e\n \u003cli\u003eAli, F., Khan, S., Jamil, S. \u0026amp; Siddiqui, F. A. UV\u0026ndash;visible first-order derivative spectrophotometric method development and validation for simultaneous estimation of amitriptyline hydrochloride and chlordiazepoxide in tablet dosage form. \u003cem\u003eAsian J. Chem.\u003c/em\u003e\u003cstrong\u003e28\u003c/strong\u003e, 2700\u0026ndash;2704 (2016).\u003c/li\u003e\n \u003cli\u003eErk, N. First-order derivative UV spectrophotometric method for simultaneous measurement of delapril and manidipine in tablets. \u003cem\u003eJ. Pharm. Biomed. Anal.\u003c/em\u003e\u003cstrong\u003e49\u003c/strong\u003e, 231\u0026ndash;236 (2009).\u003c/li\u003e\n \u003cli\u003eDarweesh, M. F., Sultan, M. A. \u0026amp; Al-Maiah, A. A. Advancement and validation of new derivative spectrophotometric method for individual and simultaneous estimation of diclofenac sodium and nicotinamide. \u003cem\u003eOrient. J. Chem.\u003c/em\u003e\u003cstrong\u003e34\u003c/strong\u003e, 1484\u0026ndash;1493 (2018).\u003c/li\u003e\n \u003cli\u003eRedasani, V. K., Patel, P. R., Marathe, D. Y. \u0026amp; Surana, S. J. Derivative UV spectrophotometry analysis of drugs in pharmaceutical formulations and biological samples: a review. \u003cem\u003eJ. Chil. Chem. Soc.\u003c/em\u003e\u003cstrong\u003e63\u003c/strong\u003e, 3864\u0026ndash;3871 (2018).\u003c/li\u003e\n \u003cli\u003eShah, J., Shah, S. A., Patel, T. \u0026amp; Patel, N. Simultaneous determination of moxifloxacin and cefixime by first- and ratio-first derivative ultraviolet spectrophotometry. \u003cem\u003eBMC Chem.\u003c/em\u003e\u003cstrong\u003e6\u003c/strong\u003e, 105 (2012).\u003c/li\u003e\n \u003cli\u003eInternational Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use. Validation of analytical procedures: Q2(R2). Available at: https://www.ich.org/page/quality-guidelines (2023).\u003c/li\u003e\n \u003cli\u003eSilva, C. F., Santos J\u0026uacute;nior, J. H., Fidelis, C. H. V., Santana, R. S. \u0026amp; Franqui, L. S. Integrating green chemistry, green analytical chemistry, and white analytical chemistry through the ESG perspective. \u003cem\u003eObserv. Econ. Latinoam.\u003c/em\u003e\u003cstrong\u003e23\u003c/strong\u003e, 148 (2023).\u003c/li\u003e\n \u003cli\u003eKumari, D., Alsayadi, Y. M. M. A. \u0026amp; Sharma, N. Exploratory analysis of recent advancement in green analytical chemistry application. \u003cem\u003eAnal. Methods Environ. Chem. J.\u003c/em\u003e\u003cstrong\u003e7\u003c/strong\u003e, 86\u0026ndash;114 (2024).\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Dapagliflozin, Eplerenone, Eco-friendly, Simultaneous Estimation, Synthetic Mixture","lastPublishedDoi":"10.21203/rs.3.rs-9264545/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9264545/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground:\u003c/strong\u003e Dapagliflozin and Eplerenone are commonly used together for the management of diabetes-related cardiovascular and renal disorders, necessitating a reliable analytical approach for their combined analysis.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eObjective:\u003c/strong\u003e The present work describes the development and validation of an environmentally benign first-order derivative UV-spectrophotometric method for the simultaneous determination of Dapagliflozin and Eplerenone in a synthetic mixture, in accordance with ICH Q2(R2) guidelines.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e First-order derivative spectra were employed to resolve spectral overlap between the two drugs. Dapagliflozin exhibited a zero-crossing point at 232 nm, while Eplerenone showed a zero-crossing at 254 nm, enabling selective quantification without mutual interference.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e The method demonstrated linearity over concentration ranges of 1–6 µg/mL for Dapagliflozin and 2.5–15 µg/mL for Eplerenone, with correlation coefficients approaching unity. Validation studies confirmed acceptable precision, accuracy, specificity, robustness, and sensitivity. No interference from excipients was observed, and the limits of detection and quantification were suitable for routine analysis.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion: \u003c/strong\u003eOverall, the proposed method is simple, accurate, cost-effective, and eco-friendly, making it suitable for routine quality control of pharmaceutical preparations containing these drugs.\u003c/p\u003e","manuscriptTitle":"Eco-Friendly First-Derivative UV-Spectrophotometric Method for Simultaneous Estimation of Dapagliflozin and Eplerenone in Bulk and Synthetic Mixture","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-04-10 06:12:23","doi":"10.21203/rs.3.rs-9264545/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"239024822655473506456745056492457568754","date":"2026-04-16T02:05:05+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"288388372178735561386686044055162430068","date":"2026-04-13T10:33:18+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"14866527928787489140524727055518723074","date":"2026-04-06T09:43:34+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-04-06T00:59:15+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-04-06T00:56:27+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2026-04-03T17:00:33+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-04-02T11:45:51+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2026-04-02T10:35:02+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"5cdb7c95-2c96-4780-9be9-5b58fd92b887","owner":[],"postedDate":"April 10th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":65913957,"name":"Physical sciences/Chemistry"},{"id":65913958,"name":"Biological sciences/Drug discovery"}],"tags":[],"updatedAt":"2026-04-10T06:12:23+00:00","versionOfRecord":[],"versionCreatedAt":"2026-04-10 06:12:23","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9264545","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9264545","identity":"rs-9264545","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}