Journal of separation science最新文献

{"title":"Efficient and Symmetric Temperature Control in Capillary Electrophoresis II: Thermal Performance When Cooling Capillaries Are Tied Around Analytical Capillaries","authors":"Leonel Bortolotto Macedo,&nbsp;Cristian Bonatto,&nbsp;Tarso B. Ledur Kist","doi":"10.1002/jssc.70240","DOIUrl":"10.1002/jssc.70240","url":null,"abstract":"<p>Current commercially available capillary electrophoresis instruments lack centrosymmetric and efficient temperature control along the whole extend of the capillary. Here we studied the characteristics and thermal performance of a cooling system that consists of cooling capillaries (fused silica microtubes) which are tied around the complete extent of the analytical capillaries. Six cooling capillaries with only 75 µm inner diameter (320 µm outer diameter—without polyimide) were tied around the outer surface of 320 µm outer diameter analytical capillaries (also without polyimide). The tying process is detailed in a previous publication (<i>J Sep Sci</i>. 2025; 48: e70081. https://doi.org/10.1002/jssc.70081). The application of a pressure gradient of 0.25 bar/cm in the cooling liquid was enough to efficiently remove heat and control temperature. Very strong electric fields could be applied, producing very high and stable electric currents. Fields beyond 3500 Volts/cm were applied in a 50 µm inner diameter capillary filled with 20 mM sodium phosphate buffer solution at pH 7.20 and the coolant set at 25°C. This cooling system outperformed the two most used systems: forced air and recirculating liquid coolant in a tube with a capillary inside. The spatial steady-state temperature profiles of the systems were simulated by numerically solving the heat equation using a finite element method. The centrosymmetric temperature profile and efficiency of this cooling system was corroborated by these numerical results. An objective parameter indicating cooling asymmetry was introduced and used to quantify the superior performance of this new cooling system.</p>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 8","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/epdf/10.1002/jssc.70240","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144832899","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Efficient and Symmetric Temperature Control in Capillary Electrophoresis II: Thermal Performance When Cooling Capillaries Are Tied Around Analytical Capillaries","authors":"Leonel Bortolotto Macedo,&nbsp;Cristian Bonatto,&nbsp;Tarso B. Ledur Kist","doi":"10.1002/jssc.70240","DOIUrl":"10.1002/jssc.70240","url":null,"abstract":"<p>Current commercially available capillary electrophoresis instruments lack centrosymmetric and efficient temperature control along the whole extend of the capillary. Here we studied the characteristics and thermal performance of a cooling system that consists of cooling capillaries (fused silica microtubes) which are tied around the complete extent of the analytical capillaries. Six cooling capillaries with only 75 µm inner diameter (320 µm outer diameter—without polyimide) were tied around the outer surface of 320 µm outer diameter analytical capillaries (also without polyimide). The tying process is detailed in a previous publication (<i>J Sep Sci</i>. 2025; 48: e70081. https://doi.org/10.1002/jssc.70081). The application of a pressure gradient of 0.25 bar/cm in the cooling liquid was enough to efficiently remove heat and control temperature. Very strong electric fields could be applied, producing very high and stable electric currents. Fields beyond 3500 Volts/cm were applied in a 50 µm inner diameter capillary filled with 20 mM sodium phosphate buffer solution at pH 7.20 and the coolant set at 25°C. This cooling system outperformed the two most used systems: forced air and recirculating liquid coolant in a tube with a capillary inside. The spatial steady-state temperature profiles of the systems were simulated by numerically solving the heat equation using a finite element method. The centrosymmetric temperature profile and efficiency of this cooling system was corroborated by these numerical results. An objective parameter indicating cooling asymmetry was introduced and used to quantify the superior performance of this new cooling system.</p>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 8","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/epdf/10.1002/jssc.70240","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144833147","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Chemical Profiling of Anti-Inflammatory Constituents for Hongteng Baijiang Decoction Using Comprehensive Two-Dimensional Countercurrent Chromatography × High Performance Liquid Chromatography Coupled With Quadrupole Time-Of-Flight Mass Spectrometry","authors":"Hangjuan Lin,&nbsp;Wanshu Li,&nbsp;Yuli Gao,&nbsp;Bo Zhao,&nbsp;Bingqian Qin,&nbsp;Qiwen Shi,&nbsp;Chu Chu,&nbsp;Shengqiang Tong","doi":"10.1002/jssc.70245","DOIUrl":"10.1002/jssc.70245","url":null,"abstract":"<div>\u0000 \u0000 <p>Comprehensive two-dimensional reverse phase countercurrent chromatography (CCC)×high performance liquid chromatography (HPLC) was investigated for chemical profiling and identification of bioactive components with anti-inflammatory activities for a traditional Chinese medicine formula, Hongteng Baijiang decoction (HBD), for the first time. Anti-inflammatory activity of different extractions from HBD were evaluated. Two-dimensional contour plots for CCC×HPLC were obtained after comprehensive separation was completed. The total peak capacity of the two-dimensional system was evaluated, yielding approximately 782 peaks through comprehensive two-dimensional separation, which demonstrated a high degree of orthogonality and large separation space occupancy of 90.16%. Totally 207 chemical components were identified in ethyl acetate extract from HBD using liquid chromatography-quadrupole time-of-flight mass spectrometry based on the two-dimensional separation, with additional 70 compounds identified compared to that in one-dimensional HPLC separation. It was found that terpenoids were major bioactive components contributing to anti-inflammatory bioactivity in HBD. These findings highlight the potential of comprehensive two-dimensional chromatography (CCC×HPLC) as a powerful and promising method for separating and chemical profiling for complex traditional Chinese herbal formula.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 8","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144833208","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Five-Step Plan for Proposing a Balanced HPLC Method via Fishbone Diagram: Untangling the Synergy of Efficiency and Sustainability, Application on the Analysis of Isoniazid, Pyrazinamide, and Rifampicin in a Fixed Dose Combination","authors":"Rana W. Gaber,&nbsp;Amr M. Mahmoud,&nbsp;Sarah S. Saleh","doi":"10.1002/jssc.70241","DOIUrl":"10.1002/jssc.70241","url":null,"abstract":"<div>\u0000 \u0000 <p>A five-step plan is presented to propose an HPLC method with the aid of the fishbone diagram to balance efficiency and sustainability. The five steps include method development, method optimization, method validation, greenness assessment, and sustainability profiling. A sensitive, accurate, robust, and rapid HPLC method has been developed for simultaneous determination and separation of anti-tubercular medications: rifampicin (RIF), isoniazid (INH), and pyrazinamide (PYR) in a fixed dose combination (FDC). The separation was attained by using a stationary phase Kromasil C₁₈ column (250 mm, 4.6,5 µm), accompanied by a gradient mode of the mobile phase in which the first solution was made up by using acetonitrile and phosphate buffer in a ratio of 4:96 v/v, and the second solution consisted of phosphate buffer and acetonitrile in ratio 45:55 v/v. The method was optimized by applying a fractional factorial design (2²), where the flow rate was set at 1.5 mL/min, the injection volume was 10 µL, and the detection was performed at 238 nm at a temperature of 35°C. The retention time for INH, PYR, and RIF were 3.82, 5.36, and 8.72 min (±0.02), respectively. The method was validated per the ICH guidelines, and the proposed method was found to be accurate, robust, precise, and selective. A design of experiment was established to assess the robustness of the study, where the obtained results proved that the developed method is robust. Two greenness tools, namely moGAPI and AGREE, were used to evaluate the method's greenness. In addition, the method was subjected to sustainability profiling using the EVG framework, where the suggested method was compared to other reported methods. The proposed method was found to be more sustainable and green, with a low negative impact on the environment.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 8","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144833207","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Five-Step Plan for Proposing a Balanced HPLC Method via Fishbone Diagram: Untangling the Synergy of Efficiency and Sustainability, Application on the Analysis of Isoniazid, Pyrazinamide, and Rifampicin in a Fixed Dose Combination","authors":"Rana W. Gaber,&nbsp;Amr M. Mahmoud,&nbsp;Sarah S. Saleh","doi":"10.1002/jssc.70241","DOIUrl":"10.1002/jssc.70241","url":null,"abstract":"<div>\u0000 \u0000 <p>A five-step plan is presented to propose an HPLC method with the aid of the fishbone diagram to balance efficiency and sustainability. The five steps include method development, method optimization, method validation, greenness assessment, and sustainability profiling. A sensitive, accurate, robust, and rapid HPLC method has been developed for simultaneous determination and separation of anti-tubercular medications: rifampicin (RIF), isoniazid (INH), and pyrazinamide (PYR) in a fixed dose combination (FDC). The separation was attained by using a stationary phase Kromasil C₁₈ column (250 mm, 4.6,5 µm), accompanied by a gradient mode of the mobile phase in which the first solution was made up by using acetonitrile and phosphate buffer in a ratio of 4:96 v/v, and the second solution consisted of phosphate buffer and acetonitrile in ratio 45:55 v/v. The method was optimized by applying a fractional factorial design (2²), where the flow rate was set at 1.5 mL/min, the injection volume was 10 µL, and the detection was performed at 238 nm at a temperature of 35°C. The retention time for INH, PYR, and RIF were 3.82, 5.36, and 8.72 min (±0.02), respectively. The method was validated per the ICH guidelines, and the proposed method was found to be accurate, robust, precise, and selective. A design of experiment was established to assess the robustness of the study, where the obtained results proved that the developed method is robust. Two greenness tools, namely moGAPI and AGREE, were used to evaluate the method's greenness. In addition, the method was subjected to sustainability profiling using the EVG framework, where the suggested method was compared to other reported methods. The proposed method was found to be more sustainable and green, with a low negative impact on the environment.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 8","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144833151","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Chemical Profiling of Anti-Inflammatory Constituents for Hongteng Baijiang Decoction Using Comprehensive Two-Dimensional Countercurrent Chromatography × High Performance Liquid Chromatography Coupled With Quadrupole Time-Of-Flight Mass Spectrometry","authors":"Hangjuan Lin,&nbsp;Wanshu Li,&nbsp;Yuli Gao,&nbsp;Bo Zhao,&nbsp;Bingqian Qin,&nbsp;Qiwen Shi,&nbsp;Chu Chu,&nbsp;Shengqiang Tong","doi":"10.1002/jssc.70245","DOIUrl":"10.1002/jssc.70245","url":null,"abstract":"<div>\u0000 \u0000 <p>Comprehensive two-dimensional reverse phase countercurrent chromatography (CCC)×high performance liquid chromatography (HPLC) was investigated for chemical profiling and identification of bioactive components with anti-inflammatory activities for a traditional Chinese medicine formula, Hongteng Baijiang decoction (HBD), for the first time. Anti-inflammatory activity of different extractions from HBD were evaluated. Two-dimensional contour plots for CCC×HPLC were obtained after comprehensive separation was completed. The total peak capacity of the two-dimensional system was evaluated, yielding approximately 782 peaks through comprehensive two-dimensional separation, which demonstrated a high degree of orthogonality and large separation space occupancy of 90.16%. Totally 207 chemical components were identified in ethyl acetate extract from HBD using liquid chromatography-quadrupole time-of-flight mass spectrometry based on the two-dimensional separation, with additional 70 compounds identified compared to that in one-dimensional HPLC separation. It was found that terpenoids were major bioactive components contributing to anti-inflammatory bioactivity in HBD. These findings highlight the potential of comprehensive two-dimensional chromatography (CCC×HPLC) as a powerful and promising method for separating and chemical profiling for complex traditional Chinese herbal formula.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 8","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144832809","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Measurement of Total and Free Cefiderocol Concentrations in Human Plasma by UHPLC-MS/MS: Application to Patients With Difficult-To-Treat Gram-Negative Bacterial Infections","authors":"Raúl Rigo-Bonnin,&nbsp;Cristina El Haj,&nbsp;Eva Benavent,&nbsp;Judith Poblet-Florentin,&nbsp;Paula Pons-Oltra,&nbsp;Jaime Esteban,&nbsp;Oscar Murillo","doi":"10.1002/jssc.70237","DOIUrl":"https://doi.org/10.1002/jssc.70237","url":null,"abstract":"<div>\u0000 \u0000 <p>Cefiderocol is an innovative siderophore cephalosporin against some of the most difficult-to-treat Gram-negative bacterial infections. As with other β-lactam antibiotics, therapeutic drug monitoring of cefiderocol in specific clinical scenarios could improve patient outcomes, minimize toxicity, and maximize treatment effectiveness. Thus, we aimed to develop and validate an ultra-high-performance liquid chromatography-tandem mass spectrometry procedure for measuring total and free cefiderocol concentrations in human plasma. Protein precipitation was used to extract patient samples. Equilibrium dialysis preceded sample preparation for measuring free cefiderocol concentrations. An Acquity-UPLC-BEH (2.1 mm × 100 mm id, 1.7 µm) column was selected for analyte separation, using 0.1% (v/v) formic acid in water and acetonitrile as mobile phases and working in a gradient containing. The cefiderocol and its internal standard ([²H₈-cefiderocol) were detected by electrospray ionization mass spectrometry in positive and multiple reaction monitoring modes, using transitions of 752.2→285.3/468.2 and 760.2→293.3/468.2, respectively. Analysis time was 3.5 min per run. Precisions, absolute relative biases, normalized-matrix effect, and normalized recoveries were ≤13.9%, ≤14.2%, 95.7%–104.1%, and 95.9%–104.8%, respectively. Linearity was observed between 0.50 and 65.0 mg/L with linear regression coefficients greater than 0.9984. No significant interferences and carry-over were observed. The UHPLC-MS/MS procedures we present here could be suitable for clinical research purposes and hold promise for routine analysis. Their versatility would allow for application in the therapeutic drug monitoring of cefiderocol in subjects with difficult-to-treat Gram-negative bacterial infections to guide the treatment and adjust the daily dose of this new β-lactam antibiotic.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 8","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144811080","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a Supercritical Fluid Extraction Process for Preparation of Phytosteroids Rich Extract From Tinospora cordifolia Stem and Determination of Content of Three Phytosteroids Using a Validated Ultra-High Performance Liquid Chromatography Method","authors":"Rohan Sarkar,&nbsp;Satyanshu Kumar,&nbsp;Raghuraj Singh","doi":"10.1002/jssc.70246","DOIUrl":"https://doi.org/10.1002/jssc.70246","url":null,"abstract":"<div>\u0000 \u0000 <p><i>Tinospora cordifolia</i> is an integral part of traditional medicine, rich in secondary metabolites of various chemical classes including phytosteroids. These phytosteroids possess cholesterol-lowering property; however, systematic studies are lacking regarding preparation of extract enriched with such bioactive compounds. In this study, supercritical fluid extraction (SFE) method was developed to obtain three phytosteroids (ecdysterone, stigmasterol, and β-sitosterol) enriched extract from <i>T. cordifolia</i> stem. The phytosteroid content was quantified using validated ultra-high performance liquid chromatography method. Separation of analytes was achieved using C-18 column (150 mm × 4.6 mm, 5.0 µm) as stationary phase and water–acetonitrile mixture (80:20) as mobile phase with detection at 242 nm (range of 190–350 nm; photodiode array detector). The limit of detection was 0.25, 0.23, and 0.39 µg/mL for ecdysterone, stigmasterol, and β-sitosterol respectively, whereas, limit of quantification was 0.85, 0.78, and 1.25 µg/mL for ecdysterone, stigmasterol, and β-sitosterol respectively. Accuracy of recovery was 80%–109%, with precision (RSD) of &lt; 2 for all three targeted analytes. For SFE, under optimized condition i.e., automated back pressure = 100 bar, CO₂ flow = 30 g/min, modifier flow = 4 g/min, extraction and collection vessel temperature = 60°C and 45°C, respectively, and extraction time = 120 min; phytosteroid (ecdysterone + stigmasterol + β-sitosterol) content was obtained as 319.36 µg/g which was significantly higher than its content in extract (108.04 µg/g) obtained by refluxing with 50% aqueous ethanol as extraction solvent. Method performance and greenness of SFE was compared with ultrasound-assisted extraction. Result showed 2.06-fold increase in extract yield in case SFE than ultrasound-assisted extraction. Greenness score calculated for SFE and ultrasound-assisted extraction was 0.65 and 0.59, respectively. <i>T. cordifolia</i> SFE extract can be key component in developing herbal products to address obesity related problems. Thus, the developed extraction method can be recommended to herbal industries for preparation of effective medicines with diverse therapeutic applications.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 8","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144811081","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Measurement of Total and Free Cefiderocol Concentrations in Human Plasma by UHPLC-MS/MS: Application to Patients With Difficult-To-Treat Gram-Negative Bacterial Infections","authors":"Raúl Rigo-Bonnin,&nbsp;Cristina El Haj,&nbsp;Eva Benavent,&nbsp;Judith Poblet-Florentin,&nbsp;Paula Pons-Oltra,&nbsp;Jaime Esteban,&nbsp;Oscar Murillo","doi":"10.1002/jssc.70237","DOIUrl":"10.1002/jssc.70237","url":null,"abstract":"<div>\u0000 \u0000 <p>Cefiderocol is an innovative siderophore cephalosporin against some of the most difficult-to-treat Gram-negative bacterial infections. As with other β-lactam antibiotics, therapeutic drug monitoring of cefiderocol in specific clinical scenarios could improve patient outcomes, minimize toxicity, and maximize treatment effectiveness. Thus, we aimed to develop and validate an ultra-high-performance liquid chromatography-tandem mass spectrometry procedure for measuring total and free cefiderocol concentrations in human plasma. Protein precipitation was used to extract patient samples. Equilibrium dialysis preceded sample preparation for measuring free cefiderocol concentrations. An Acquity-UPLC-BEH (2.1 mm × 100 mm id, 1.7 µm) column was selected for analyte separation, using 0.1% (v/v) formic acid in water and acetonitrile as mobile phases and working in a gradient containing. The cefiderocol and its internal standard ([²H₈-cefiderocol) were detected by electrospray ionization mass spectrometry in positive and multiple reaction monitoring modes, using transitions of 752.2→285.3/468.2 and 760.2→293.3/468.2, respectively. Analysis time was 3.5 min per run. Precisions, absolute relative biases, normalized-matrix effect, and normalized recoveries were ≤13.9%, ≤14.2%, 95.7%–104.1%, and 95.9%–104.8%, respectively. Linearity was observed between 0.50 and 65.0 mg/L with linear regression coefficients greater than 0.9984. No significant interferences and carry-over were observed. The UHPLC-MS/MS procedures we present here could be suitable for clinical research purposes and hold promise for routine analysis. Their versatility would allow for application in the therapeutic drug monitoring of cefiderocol in subjects with difficult-to-treat Gram-negative bacterial infections to guide the treatment and adjust the daily dose of this new β-lactam antibiotic.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 8","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144811078","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a Supercritical Fluid Extraction Process for Preparation of Phytosteroids Rich Extract From Tinospora cordifolia Stem and Determination of Content of Three Phytosteroids Using a Validated Ultra-High Performance Liquid Chromatography Method","authors":"Rohan Sarkar,&nbsp;Satyanshu Kumar,&nbsp;Raghuraj Singh","doi":"10.1002/jssc.70246","DOIUrl":"10.1002/jssc.70246","url":null,"abstract":"<div>\u0000 \u0000 <p><i>Tinospora cordifolia</i> is an integral part of traditional medicine, rich in secondary metabolites of various chemical classes including phytosteroids. These phytosteroids possess cholesterol-lowering property; however, systematic studies are lacking regarding preparation of extract enriched with such bioactive compounds. In this study, supercritical fluid extraction (SFE) method was developed to obtain three phytosteroids (ecdysterone, stigmasterol, and β-sitosterol) enriched extract from <i>T. cordifolia</i> stem. The phytosteroid content was quantified using validated ultra-high performance liquid chromatography method. Separation of analytes was achieved using C-18 column (150 mm × 4.6 mm, 5.0 µm) as stationary phase and water–acetonitrile mixture (80:20) as mobile phase with detection at 242 nm (range of 190–350 nm; photodiode array detector). The limit of detection was 0.25, 0.23, and 0.39 µg/mL for ecdysterone, stigmasterol, and β-sitosterol respectively, whereas, limit of quantification was 0.85, 0.78, and 1.25 µg/mL for ecdysterone, stigmasterol, and β-sitosterol respectively. Accuracy of recovery was 80%–109%, with precision (RSD) of &lt; 2 for all three targeted analytes. For SFE, under optimized condition i.e., automated back pressure = 100 bar, CO₂ flow = 30 g/min, modifier flow = 4 g/min, extraction and collection vessel temperature = 60°C and 45°C, respectively, and extraction time = 120 min; phytosteroid (ecdysterone + stigmasterol + β-sitosterol) content was obtained as 319.36 µg/g which was significantly higher than its content in extract (108.04 µg/g) obtained by refluxing with 50% aqueous ethanol as extraction solvent. Method performance and greenness of SFE was compared with ultrasound-assisted extraction. Result showed 2.06-fold increase in extract yield in case SFE than ultrasound-assisted extraction. Greenness score calculated for SFE and ultrasound-assisted extraction was 0.65 and 0.59, respectively. <i>T. cordifolia</i> SFE extract can be key component in developing herbal products to address obesity related problems. Thus, the developed extraction method can be recommended to herbal industries for preparation of effective medicines with diverse therapeutic applications.</p>\u0000 </div>","PeriodicalId":17098,"journal":{"name":"Journal of separation science","volume":"48 8","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144811079","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}