Pub Date : 2025-11-12DOI: 10.1016/j.chroma.2025.466541
Dong Wook Seok , Seohyun Choi , Ji-Seon Jeong
We describe the simultaneous quantification of 14 underivatized amino acids (AAs) in various matrices using ion-pairing liquid chromatography–tandem mass spectrometry (LC-MS/MS) combined with isotope dilution mass spectrometry (IDMS) as a newly developed reference measurement procedure. The underivatized AAs were successfully separated on a reversed-phase column using mixed ion-pairing reagents of trifluoroacetic acid and heptafluorobutyric acid within 6 min through optimized gradient elution. The method was applied to different matrices, including buffer and plasma with appropriate sample treatments, followed by dilution with water to adjust the concentration range from 1 to 20 mg/kg, ensuring high measurement accuracy and precision. To facilitate exact-matched double IDMS, isotope-labeled AAs were added to the sample prior to treatment as internal standards. The optimized procedure was rigorously validated for linearity, sensitivity, accuracy, and precision, while its reliability was confirmed using commercially available certified reference materials for the AAs. Finally, the method was applied to certify AAs in candidate certified reference material batches. The optimized protocol proved to be suitable for the accurate quantification of 14 AAs in various matrices, and thus may serve as a reliable primary method for AA measurement across different applications.
{"title":"Simultaneous determination of 14 underivatized amino acids using ion-pairing chromatography and isotope dilution tandem mass spectrometry for a reference measurement procedure","authors":"Dong Wook Seok , Seohyun Choi , Ji-Seon Jeong","doi":"10.1016/j.chroma.2025.466541","DOIUrl":"10.1016/j.chroma.2025.466541","url":null,"abstract":"<div><div>We describe the simultaneous quantification of 14 underivatized amino acids (AAs) in various matrices using ion-pairing liquid chromatography–tandem mass spectrometry (LC-MS/MS) combined with isotope dilution mass spectrometry (IDMS) as a newly developed reference measurement procedure. The underivatized AAs were successfully separated on a reversed-phase column using mixed ion-pairing reagents of trifluoroacetic acid and heptafluorobutyric acid within 6 min through optimized gradient elution. The method was applied to different matrices, including buffer and plasma with appropriate sample treatments, followed by dilution with water to adjust the concentration range from 1 to 20 mg/kg, ensuring high measurement accuracy and precision. To facilitate exact-matched double IDMS, isotope-labeled AAs were added to the sample prior to treatment as internal standards. The optimized procedure was rigorously validated for linearity, sensitivity, accuracy, and precision, while its reliability was confirmed using commercially available certified reference materials for the AAs. Finally, the method was applied to certify AAs in candidate certified reference material batches. The optimized protocol proved to be suitable for the accurate quantification of 14 AAs in various matrices, and thus may serve as a reliable primary method for AA measurement across different applications.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1766 ","pages":"Article 466541"},"PeriodicalIF":4.0,"publicationDate":"2025-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145571026","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The majority of enzymological tasks are targeted at determining polar compounds representing substrates, products, related substances of enzymatic reactions, etc. Currently, they are still solved either by reversed-phase HPLC requiring preliminary derivatization step for increasing hydrophobicity, or even spectroscopy remaining non-selective toward the related sample compounds. Hydrophilic interaction liquid chromatography (HILIC) method for direct determination of polar analytes provides promising solutions for the modern biochemical tasks with much easier and faster analysis for hundreds and thousands of samples. The main focus in such investigations is choosing conditions for separating complex mixtures of diverse polar compounds which could be challenging in many cases. In this work, a simple and rapid HILIC method for determining enzymatic activity of Cysteine synthase A (CysK, EC 2.5.1.47) is developed. CysK is a pyridoxal-5'-phosphate phosphate dependent enzyme that catalyzes the formation of L-cysteine and acetate from O-acetyl-L-serine and sulfide. This reaction represents a critical step in the sulfur assimilation pathway, which plays a vital role in maintaining sulfur homeostasis and supporting cellular functions such as protein synthesis, antioxidant defense, and detoxification. CysK is one of the enzymes secreted by Lactobacilli while co-cultivating with Klebsiella, which can be used for cysteine synthesis and is also important for microbial vital activity. We obtained a new recombinant CysK from Limosilactobacillus reuteri LR1 by gene cloning and expression in E.coli cells. HILIC method development for determining the enzyme activity represented a challenge of separating two substrates, two products, and a buffer component in one sample. This task was solved by choosing both the column and elution conditions. Specific amide functionalized silica-based stationary phase obtained via the Ugi reaction was chosen among the three laboratory-designed HILIC columns. The impact of ion-exchange into the retention mechanism for the main components of the enzymatic reaction was assessed for determining separation conditions on the chosen column. The developed method allowed separating cysteine from all other peaks within 11 min under isocratic elution conditions. Limits of detection and quantification were 2 and 7 μM, respectively. Using the developed method, the reaction rate of CysK obtained from Limosilactobacillus reuteri LR1 was determined for the first time. The primary benefits of this method are rapidity of the analysis, direct determination avoiding derivatization step, and utilizing simple HPLC instrumentation with neither gradient elution nor advanced detection systems.
{"title":"Development of hydrophilic interaction liquid chromatography method for determining enzymatic activity of cysteine synthase A","authors":"A.V. Chernobrovkina , A.V. Gorbovskaia , N.Yu. Chikurova , E.K. Les , A.D. Efremova , E.S. Chichkanova , O.A. Shpigun , V.I. Tishkov , A.A. Pometun","doi":"10.1016/j.chroma.2025.466540","DOIUrl":"10.1016/j.chroma.2025.466540","url":null,"abstract":"<div><div>The majority of enzymological tasks are targeted at determining polar compounds representing substrates, products, related substances of enzymatic reactions, etc. Currently, they are still solved either by reversed-phase HPLC requiring preliminary derivatization step for increasing hydrophobicity, or even spectroscopy remaining non-selective toward the related sample compounds. Hydrophilic interaction liquid chromatography (HILIC) method for direct determination of polar analytes provides promising solutions for the modern biochemical tasks with much easier and faster analysis for hundreds and thousands of samples. The main focus in such investigations is choosing conditions for separating complex mixtures of diverse polar compounds which could be challenging in many cases. In this work, a simple and rapid HILIC method for determining enzymatic activity of Cysteine synthase A (CysK, EC 2.5.1.47) is developed. CysK is a pyridoxal-5'-phosphate phosphate dependent enzyme that catalyzes the formation of L-cysteine and acetate from O-acetyl-L-serine and sulfide. This reaction represents a critical step in the sulfur assimilation pathway, which plays a vital role in maintaining sulfur homeostasis and supporting cellular functions such as protein synthesis, antioxidant defense, and detoxification. CysK is one of the enzymes secreted by <em>Lactobacilli</em> while co-cultivating with <em>Klebsiella,</em> which can be used for cysteine synthesis and is also important for microbial vital activity. We obtained a new recombinant CysK from <em>Limosilactobacillus reuteri</em> LR1 by gene cloning and expression in <em>E.coli</em> cells. HILIC method development for determining the enzyme activity represented a challenge of separating two substrates, two products, and a buffer component in one sample. This task was solved by choosing both the column and elution conditions. Specific amide functionalized silica-based stationary phase obtained via the Ugi reaction was chosen among the three laboratory-designed HILIC columns. The impact of ion-exchange into the retention mechanism for the main components of the enzymatic reaction was assessed for determining separation conditions on the chosen column. The developed method allowed separating cysteine from all other peaks within 11 min under isocratic elution conditions. Limits of detection and quantification were 2 and 7 μM, respectively. Using the developed method, the reaction rate of CysK obtained from <em>Limosilactobacillus reuteri</em> LR1 was determined for the first time. The primary benefits of this method are rapidity of the analysis, direct determination avoiding derivatization step, and utilizing simple HPLC instrumentation with neither gradient elution nor advanced detection systems.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1766 ","pages":"Article 466540"},"PeriodicalIF":4.0,"publicationDate":"2025-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145601461","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-10DOI: 10.1016/j.chroma.2025.466489
Louis Schmidt, Jana Lemke, Phillip Schmiady, Tobias Keydel, Lukas Schulig, Andreas Link, Ulrike Garscha
Chiral analysis is an essential part of the development and quality control of enantiopure pharmaceuticals. Supercritical fluid chromatography (SFC) emerged as a powerful tool for the chiral separation of drugs, however, its implementation in the pharmacopoeia remains limited. In this study, we developed a rapid SFC-UV method for the impurity control of S-dapoxetine, enabling simultaneous chiral and achiral analysis in less than 20 min. The method was validated according to ICH Q2(R2) guidelines and demonstrated sufficient sensitivity, precision and accuracy down to 2.5 µg mL-1. A binary acid–base additive mixture was used to modify chemo- and stereoselectivity, achieving baseline separation of all relevant analytes. During method development, an unusual retention behaviour of basic analytes was observed, leading to a thermodynamic investigations using the Van ’t Hoff analysis. This revealed a rare case of entropy-driven retention, with strong acidic additives like trifluoroacetic acid leveraging the effect for basic analytes. Further investigations involving 15 diverse chiral amines, as well as molecular dynamic simulations, showed that isopropanol stabilizes the helical amylose backbone by reducing excessive flexibility, thereby increasing the enantioselective separation efficiency. Additionally, strong acids were found to form transient ion-pairs with basic analytes, which are favoured in the supercritical environment. The neutral complexes impact retention depending on the amine substitution pattern and may suppress polar and enhance hydrophobic interactions. These results underscore the potential application of combined additive systems to enhance SFC applications in the field of ionizable analytes.
{"title":"Combined chiral–achiral supercritical fluid chromatography method for the impurity analysis of dapoxetine reveals insights in entropy-driven retention and acid-modulated selectivity","authors":"Louis Schmidt, Jana Lemke, Phillip Schmiady, Tobias Keydel, Lukas Schulig, Andreas Link, Ulrike Garscha","doi":"10.1016/j.chroma.2025.466489","DOIUrl":"10.1016/j.chroma.2025.466489","url":null,"abstract":"<div><div>Chiral analysis is an essential part of the development and quality control of enantiopure pharmaceuticals. Supercritical fluid chromatography (SFC) emerged as a powerful tool for the chiral separation of drugs, however, its implementation in the pharmacopoeia remains limited. In this study, we developed a rapid SFC-UV method for the impurity control of <em>S</em>-dapoxetine, enabling simultaneous chiral and achiral analysis in less than 20<!--> <!-->min. The method was validated according to ICH Q2(R2) guidelines and demonstrated sufficient sensitivity, precision and accuracy down to 2.5<!--> <!-->µg<!--> <!-->mL<sup>-1</sup>. A binary acid–base additive mixture was used to modify chemo- and stereoselectivity, achieving baseline separation of all relevant analytes. During method development, an unusual retention behaviour of basic analytes was observed, leading to a thermodynamic investigations using the Van ’t Hoff analysis. This revealed a rare case of entropy-driven retention, with strong acidic additives like trifluoroacetic acid leveraging the effect for basic analytes. Further investigations involving 15 diverse chiral amines, as well as molecular dynamic simulations, showed that isopropanol stabilizes the helical amylose backbone by reducing excessive flexibility, thereby increasing the enantioselective separation efficiency. Additionally, strong acids were found to form transient ion-pairs with basic analytes, which are favoured in the supercritical environment. The neutral complexes impact retention depending on the amine substitution pattern and may suppress polar and enhance hydrophobic interactions. These results underscore the potential application of combined additive systems to enhance SFC applications in the field of ionizable analytes.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1765 ","pages":"Article 466489"},"PeriodicalIF":4.0,"publicationDate":"2025-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145511378","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-10DOI: 10.1016/j.chroma.2025.466539
Maria Chiara Frondaroli , Susanna Della Posta , Eleonora D’Alessandro , Sara Maria Giannitelli , Marcella Trombetta , Alessandra Gentili , Monica Gherardi , Laura De Gara , Chiara Fanali
In this work, a matrix solid-phase dispersion extraction (MSPD) followed by a back-extraction method was used to determine 11 pesticides from hazelnut samples. An innovative dispersing material was prepared and a silica gel bulk was impregnated with a choline chloride:oxalic acid (molar ratio 1:1) deep eutectic solvent (DES).
The material was characterized via Fourier transform infrared spectroscopy, laser diffraction analysis, and scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy. Through a preliminary one-factor-at-a-time study and a final Box-Behnken Design optimization, the sample-to-dispersing material ratio, grinding time, and extraction solvent volume parameters were optimized, yielding the following values: 1:3.8 (w/w), 1.9 min, and 2.4 mL, respectively. Ultra high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis was used.
The method was validated according to SANTE 11,312/2021 guidelines. Recoveries were evaluated at three different spike levels (LOQ, 10LOQ, and MRL) and were all above 50 %. Precision and trueness intra- and interday were all below 20 %. Linearity was studied in the range of interest (LOQ–100 µg kg-1) with determination coefficients greater than 0.991. The limits of detection and quantification were in the range of 0.03–0.5 µg kg-1 and 0.1–1.7 µg kg-1, respectively.
The sustainability of the method was evaluated using AGREEprep, BAGI and ComplexMoGAPI software.
{"title":"Innovative dispersing material based on silica gel and deep eutectic solvents for the extraction of pesticides from hazelnuts followed by UHPLC-MS/MS analysis","authors":"Maria Chiara Frondaroli , Susanna Della Posta , Eleonora D’Alessandro , Sara Maria Giannitelli , Marcella Trombetta , Alessandra Gentili , Monica Gherardi , Laura De Gara , Chiara Fanali","doi":"10.1016/j.chroma.2025.466539","DOIUrl":"10.1016/j.chroma.2025.466539","url":null,"abstract":"<div><div>In this work, a matrix solid-phase dispersion extraction (MSPD) followed by a back-extraction method was used to determine 11 pesticides from hazelnut samples. An innovative dispersing material was prepared and a silica gel bulk was impregnated with a choline chloride:oxalic acid (molar ratio 1:1) deep eutectic solvent (DES).</div><div>The material was characterized via Fourier transform infrared spectroscopy, laser diffraction analysis, and scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy. Through a preliminary one-factor-at-a-time study and a final Box-Behnken Design optimization, the sample-to-dispersing material ratio, grinding time, and extraction solvent volume parameters were optimized, yielding the following values: 1:3.8 (w/w), 1.9 min, and 2.4 mL, respectively. Ultra high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis was used.</div><div>The method was validated according to SANTE 11,312/2021 guidelines. Recoveries were evaluated at three different spike levels (LOQ, 10LOQ, and MRL) and were all above 50 %. Precision and trueness intra- and interday were all below 20 %. Linearity was studied in the range of interest (LOQ–100 µg kg<sup>-1</sup>) with determination coefficients greater than 0.991. The limits of detection and quantification were in the range of 0.03–0.5 µg kg<sup>-1</sup> and 0.1–1.7 µg kg<sup>-1</sup>, respectively.</div><div>The sustainability of the method was evaluated using AGREEprep, BAGI and ComplexMoGAPI software.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1765 ","pages":"Article 466539"},"PeriodicalIF":4.0,"publicationDate":"2025-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145533934","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-08DOI: 10.1016/j.chroma.2025.466525
Jiong Li , Jinhua Yan , Qingwen Cao , Dan Wu , Weijie Lin , Mengna Jin
The widespread use of quaternary ammonium compounds (QACs) could pose certain risks to human like potential food contamination. In this study, we developed a method to quantitate 15 QACs commonly found in milk, yogurt, and powdered milk via combining dispersive solid-phase extraction (DSPE) followed by ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Sample preparation methods and liquid chromatography conditions were established and optimized based on the inherent attributes of sample types and characteristics of the analyzed compounds to ensure measuring accuracy with minimal interference. Specifically, samples were extracted with a solvent of ACN/MeOH (8:2, v/v) supplemented with 0.2 % formic acid, followed by clean-up with a purification agent comprising NH2-PSA (200 mg) and SAX (100 mg). The mobile phase, consisting of a mixture of ACN/MeOH (4:6, v/v) and an aqueous solution, both with 0.2 % formic acid, facilitated the analysis of 15 targeted QACs on an Atlantis BEH C18 AX column using gradient elution. Subsequently, the QACs were ionized in ESI+ mode and quantitatively analyzed using MRM. With optimized parameters, the limits of detection (LODs) ranged from 0.05 to 0.50 μg/kg, with the limits of quantification (LOQs) ranging from 0.10 to 1.00 μg/kg. Additionally, the average recoveries ranged from 85.6 % to 100.7 %, while the precision for intra-day and inter-day ranged from 0.5 % to 7.4 % and 0.6 % to 8.7 %, respectively. Therefore, the method developed in this study was featured by its simplicity, accuracy, reliability, and suitability for determining QACs in milk, yogurt, and powdered milk.
{"title":"Determination of residual 15 quaternary ammonium compounds in dairy products by dispersive solid-phase extraction purification with ultra-high performance liquid chromatography-tandem mass spectrometry","authors":"Jiong Li , Jinhua Yan , Qingwen Cao , Dan Wu , Weijie Lin , Mengna Jin","doi":"10.1016/j.chroma.2025.466525","DOIUrl":"10.1016/j.chroma.2025.466525","url":null,"abstract":"<div><div>The widespread use of quaternary ammonium compounds (QACs) could pose certain risks to human like potential food contamination. In this study, we developed a method to quantitate 15 QACs commonly found in milk, yogurt, and powdered milk via combining dispersive solid-phase extraction (DSPE) followed by ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Sample preparation methods and liquid chromatography conditions were established and optimized based on the inherent attributes of sample types and characteristics of the analyzed compounds to ensure measuring accuracy with minimal interference. Specifically, samples were extracted with a solvent of ACN/MeOH (8:2, v/v) supplemented with 0.2 % formic acid, followed by clean-up with a purification agent comprising NH<sub>2</sub>-PSA (200 mg) and SAX (100 mg). The mobile phase, consisting of a mixture of ACN/MeOH (4:6, v/v) and an aqueous solution, both with 0.2 % formic acid, facilitated the analysis of 15 targeted QACs on an Atlantis BEH C<sub>18</sub> AX column using gradient elution. Subsequently, the QACs were ionized in ESI+ mode and quantitatively analyzed using MRM. With optimized parameters, the limits of detection (LODs) ranged from 0.05 to 0.50 μg/kg, with the limits of quantification (LOQs) ranging from 0.10 to 1.00 μg/kg. Additionally, the average recoveries ranged from 85.6 % to 100.7 %, while the precision for intra-day and inter-day ranged from 0.5 % to 7.4 % and 0.6 % to 8.7 %, respectively. Therefore, the method developed in this study was featured by its simplicity, accuracy, reliability, and suitability for determining QACs in milk, yogurt, and powdered milk.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1765 ","pages":"Article 466525"},"PeriodicalIF":4.0,"publicationDate":"2025-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145511421","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
We have developed a method, Vapour Pressure Purification (VPP), for extraction and purification of samples for gas chromatography. The automated procedure starts with a liquid/liquid extraction using a conventional organic solvent. After centrifugation, an aliquot (1 mL) is transferred to a new vial and moved to an automated dynamic headspace system, incorporating both a heater for the vial and a position for thermal desorption (TD) tubes. Here the solvent is first evaporated at low temperature, having an empty tube in the TD tube position. Immediately after dryness the TD tube is exchanged with a tube with Tenax TA and the temperature is increased to 150 °C. This enables extraction and quantitative transfer of the high boilers to the TD tube which is subsequently transferred to the thermal desorber and analysed using GC/MS.
The complete procedure, including vortex and centrifugation, may be automated, using up to 10 mL aqueous based samples and 2 – 4 mL solvent. This takes around 30 min. For more extreme liquid/liquid extraction ratios the extraction/centrifugation may be done manually and the collected solvent transferred to the autosampler vial. The method may be used for aqueous based- or food samples.
The method has so far been developed for three different extraction solvents: dichloromethane, methyl‑tert‑butyl‑ether and hexane.
{"title":"Principles for a purification tool for gas chromatography: Vapour Pressure Purification","authors":"Jens Glastrup, Rikke Barbro Bendixen, Rasmus Bøg Hansen","doi":"10.1016/j.chroma.2025.466538","DOIUrl":"10.1016/j.chroma.2025.466538","url":null,"abstract":"<div><div>We have developed a method, Vapour Pressure Purification (VPP), for extraction and purification of samples for gas chromatography. The automated procedure starts with a liquid/liquid extraction using a conventional organic solvent. After centrifugation, an aliquot (1 mL) is transferred to a new vial and moved to an automated dynamic headspace system, incorporating both a heater for the vial and a position for thermal desorption (TD) tubes. Here the solvent is first evaporated at low temperature, having an empty tube in the TD tube position. Immediately after dryness the TD tube is exchanged with a tube with Tenax TA and the temperature is increased to 150 °C. This enables extraction and quantitative transfer of the high boilers to the TD tube which is subsequently transferred to the thermal desorber and analysed using GC/MS.</div><div>The complete procedure, including vortex and centrifugation, may be automated, using up to 10 mL aqueous based samples and 2 – 4 mL solvent. This takes around 30 min. For more extreme liquid/liquid extraction ratios the extraction/centrifugation may be done manually and the collected solvent transferred to the autosampler vial. The method may be used for aqueous based- or food samples.</div><div>The method has so far been developed for three different extraction solvents: dichloromethane, methyl‑tert‑butyl‑ether and hexane.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1765 ","pages":"Article 466538"},"PeriodicalIF":4.0,"publicationDate":"2025-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145511404","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-07DOI: 10.1016/j.chroma.2025.466526
Xiaoqi Liu , Chenjia Jiang , Haodong Zhou , Yunfeng Yuan , Jinhua Zhu , Xiuhua Liu , Md.Zaved H. Khan , Minghua Lu
Synthetic phenolic antioxidants PG (propyl gallate), TBHQ (tert-butyl hydroquinone), BHA (butyl hydroxyanisole) are commonly used additives in edible oils, but excessive residues may pose health risks. Conventional detection methods suffer from high organic solvent consumption, complex pretreatment procedures, and environmental concerns. To address these limitations, this study developed a rapid, green and efficient analytical method combining deep eutectic solvent (DES)-based liquid-liquid microextraction (LLME) with high-performance liquid chromatography (HPLC) for the rapid determination of phenolic antioxidants in edible oils. Using an optimized hydrophilic DES system composed of choline chloride and 1,3-butanediol, (molar ratio 1:2), along with simplified extraction conditions (780 μL DES, 11 s vortex at room temperature), efficient extraction and enrichment of the target analytes were achieved. The method demonstrated excellent performance specifically for PG and TBHQ, exhibiting a wide linear range (1–400 μg/mL, R² > 0.9970), low limits of detection (0.12–0.28 μg/mL), satisfactory spiked recoveries (90.2 %-113 %), and good intra-day and inter-day precision (RSD < 5.0 %). Analysis of real samples demonstrated that the proposed method is highly sensitive, operationally simple, and environmentally friendly for the analysis of PG and TBHQ, offering a promising alternative for rapid monitoring and food safety regulation of these two antioxidants in edible oils. It should be noted that the current hydrophilic DES system did not achieve optimal extraction efficiency for BHA, highlighting the need for further optimization toward a comprehensive multi-analyte approach.
{"title":"Determination of synthetic phenolic antioxidants in edible oils: A vortex-assisted hydrophilic DES-LLME/HPLC approach with focus on PG and TBHQ","authors":"Xiaoqi Liu , Chenjia Jiang , Haodong Zhou , Yunfeng Yuan , Jinhua Zhu , Xiuhua Liu , Md.Zaved H. Khan , Minghua Lu","doi":"10.1016/j.chroma.2025.466526","DOIUrl":"10.1016/j.chroma.2025.466526","url":null,"abstract":"<div><div>Synthetic phenolic antioxidants PG (propyl gallate), TBHQ (tert-butyl hydroquinone), BHA (butyl hydroxyanisole) are commonly used additives in edible oils, but excessive residues may pose health risks. Conventional detection methods suffer from high organic solvent consumption, complex pretreatment procedures, and environmental concerns. To address these limitations, this study developed a rapid, green and efficient analytical method combining deep eutectic solvent (DES)-based liquid-liquid microextraction (LLME) with high-performance liquid chromatography (HPLC) for the rapid determination of phenolic antioxidants in edible oils. Using an optimized hydrophilic DES system composed of choline chloride and 1,3-butanediol, (molar ratio 1:2), along with simplified extraction conditions (780 μL DES, 11 s vortex at room temperature), efficient extraction and enrichment of the target analytes were achieved. The method demonstrated excellent performance specifically for PG and TBHQ, exhibiting a wide linear range (1–400 μg/mL, R² > 0.9970), low limits of detection (0.12–0.28 μg/mL), satisfactory spiked recoveries (90.2 %-113 %), and good intra-day and inter-day precision (RSD < 5.0 %). Analysis of real samples demonstrated that the proposed method is highly sensitive, operationally simple, and environmentally friendly for the analysis of PG and TBHQ, offering a promising alternative for rapid monitoring and food safety regulation of these two antioxidants in edible oils. It should be noted that the current hydrophilic DES system did not achieve optimal extraction efficiency for BHA, highlighting the need for further optimization toward a comprehensive multi-analyte approach.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1765 ","pages":"Article 466526"},"PeriodicalIF":4.0,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145501446","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-07DOI: 10.1016/j.chroma.2025.466530
Luisa Raspini e Silva , Bruna Espíndola da Silva , Carolina Henkes Inamassu , Camila Marchioni
Synthetic cannabinoids (SCs) are the largest and most diverse class of new psychoactive substances (NPS) and continue to account for the highest number of newly emerging compounds on the market. The vast chemical diversity, complex metabolism, low concentrations in biological samples, and rapid emergence of new NPS make it challenging for forensic methods to keep pace. In this scoping review, the recent advances and challenges of the SCs analysis in biological samples are described. Published studies from January 2018 to August 2025 were searched in 8 databases, and after screening and exclusions, the selected 33 articles had their data tabulated, summarized, and analyzed. The main characteristics of the groups of SCs analyzed and the potential use of each biological sample were described, indicating gaps in the literature that still need to be explored. Challenges regarding the use of reference standards for SCs and their metabolites are discussed. Well-established and emerging sample preparation protocols, along with chromatographic separation techniques, are reviewed with a focus on their advantages, limitations, and challenges. The application of Green Analytical Chemistry (GAC) principles and White Analytical Chemistry (WAC) within these approaches is also explored. Finally, the review addresses future directions, emerging challenges, and current trends in the analysis of synthetic cannabinoids.
{"title":"Synthetic cannabinoids: a critical review of challenges and advances in biological sample preparation","authors":"Luisa Raspini e Silva , Bruna Espíndola da Silva , Carolina Henkes Inamassu , Camila Marchioni","doi":"10.1016/j.chroma.2025.466530","DOIUrl":"10.1016/j.chroma.2025.466530","url":null,"abstract":"<div><div>Synthetic cannabinoids (SCs) are the largest and most diverse class of new psychoactive substances (NPS) and continue to account for the highest number of newly emerging compounds on the market. The vast chemical diversity, complex metabolism, low concentrations in biological samples, and rapid emergence of new NPS make it challenging for forensic methods to keep pace. In this scoping review, the recent advances and challenges of the SCs analysis in biological samples are described. Published studies from January 2018 to August 2025 were searched in 8 databases, and after screening and exclusions, the selected 33 articles had their data tabulated, summarized, and analyzed. The main characteristics of the groups of SCs analyzed and the potential use of each biological sample were described, indicating gaps in the literature that still need to be explored. Challenges regarding the use of reference standards for SCs and their metabolites are discussed. Well-established and emerging sample preparation protocols, along with chromatographic separation techniques, are reviewed with a focus on their advantages, limitations, and challenges. The application of Green Analytical Chemistry (GAC) principles and White Analytical Chemistry (WAC) within these approaches is also explored. Finally, the review addresses future directions, emerging challenges, and current trends in the analysis of synthetic cannabinoids.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1765 ","pages":"Article 466530"},"PeriodicalIF":4.0,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145501401","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-07DOI: 10.1016/j.chroma.2025.466537
Elisabetta Santarelli , Vicky Caponigro , Paolo Benedetti , Massimiliano Frattoni , Piero Ciccioli , Pietro Campiglia , Andrea Manni , Carlo Crescenzi , Ettore Guerriero
Active carbon fibers (ACFs) were evaluated as alternative sorbent materials for thermal desorption (TD) tubes in the analysis of volatile organic compounds (VOCs). Five commercially available ACFs, in both felt and fabric formats, were compared with conventional multi-bed tubes (MBTs) based on granular carbon adsorbents, with a focus on sampling from moisture-rich matrices, such as exhaust gas and exhaled breath. Sampling was conducted under both dry and saturated (100 % relative humidity) conditions using standard gas mixtures containing 39 VOCs from EPA TO-14 and 33 ozone precursors from the Photochemical Assessment Monitoring Station (PAMS) list. TD–gas chromatography–mass spectrometry (TD-GC–MS) was employed for compound analysis, with method parameters adapted for each sorbent type. ACFs demonstrated superior adsorption performances than MBTs, particularly for very volatile compounds (VVOCs) such as Freon-12, chloromethane, vinyl chloride, bromomethane, chloroethane, which are typically poorly retained by conventional TD tubes. ACF-F-20 achieved recovery rates above 80 % for 49 compounds and over 90 % for 37 compounds, even under humid conditions. In storage experiments, while most analytes remained stable for at least 7 days at 4 °C, highly volatile compounds such as chloromethane and bromomethane showed lower recoveries (34.2 % and 45.1 %, respectively). Application of exhaled breath analysis in one exemplary sample from healthy volunteer identified 19 VOCs, with 10 exhibiting significantly higher concentrations when sampled using ACF-T-20b compared to conventional MBTs. Notably, ACF-T-20b enabled enhanced capture of very volatile species, including methanol, propane, methylglyoxal, and hydroxylamine. These results demonstrate the potential of ACFs to improve VVOCs and VOCs sampling in high-humidity environments, broaden the detectable compound range, and reduce analytical cost and complexity in environmental and biomedical applications.
{"title":"Innovative active carbon fibers-based multipurpose thermal desorption tubes for the analysis of volatile organic compounds","authors":"Elisabetta Santarelli , Vicky Caponigro , Paolo Benedetti , Massimiliano Frattoni , Piero Ciccioli , Pietro Campiglia , Andrea Manni , Carlo Crescenzi , Ettore Guerriero","doi":"10.1016/j.chroma.2025.466537","DOIUrl":"10.1016/j.chroma.2025.466537","url":null,"abstract":"<div><div>Active carbon fibers (ACFs) were evaluated as alternative sorbent materials for thermal desorption (TD) tubes in the analysis of volatile organic compounds (VOCs). Five commercially available ACFs, in both felt and fabric formats, were compared with conventional multi-bed tubes (MBTs) based on granular carbon adsorbents, with a focus on sampling from moisture-rich matrices, such as exhaust gas and exhaled breath. Sampling was conducted under both dry and saturated (100 % relative humidity) conditions using standard gas mixtures containing 39 VOCs from EPA TO-14 and 33 ozone precursors from the Photochemical Assessment Monitoring Station (PAMS) list. TD–gas chromatography–mass spectrometry (TD-GC–MS) was employed for compound analysis, with method parameters adapted for each sorbent type. ACFs demonstrated superior adsorption performances than MBTs, particularly for very volatile compounds (VVOCs) such as Freon-12, chloromethane, vinyl chloride, bromomethane, chloroethane, which are typically poorly retained by conventional TD tubes. ACF-F-20 achieved recovery rates above 80 % for 49 compounds and over 90 % for 37 compounds, even under humid conditions. In storage experiments, while most analytes remained stable for at least 7 days at 4 °C, highly volatile compounds such as chloromethane and bromomethane showed lower recoveries (34.2 % and 45.1 %, respectively). Application of exhaled breath analysis in one exemplary sample from healthy volunteer identified 19 VOCs, with 10 exhibiting significantly higher concentrations when sampled using ACF-T-20b compared to conventional MBTs. Notably, ACF-T-20b enabled enhanced capture of very volatile species, including methanol, propane, methylglyoxal, and hydroxylamine. These results demonstrate the potential of ACFs to improve VVOCs and VOCs sampling in high-humidity environments, broaden the detectable compound range, and reduce analytical cost and complexity in environmental and biomedical applications.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1765 ","pages":"Article 466537"},"PeriodicalIF":4.0,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145522615","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-07DOI: 10.1016/j.chroma.2025.466529
Xin Zhang, Juanjuan Feng, Baoen Su, Haidi Zhang, Yana Luo, Min Sun
Owing to high specific surface area and excellent chemical stability, carbon materials as stationary phases demonstrated significant advantages. However, their practical application was limited by high cost and complex synthesis process. In this work, the low-cost petroleum asphalt (PA) was successfully transformed into a stationary phase for high-performance liquid chromatography. Through a facile coating way, the PA was immobilized onto porous silica for producing a new stationary phase. As the concentration of asphalt in the coating solution was increased, the carbon content of the stationary phases rose from 6.67 % (SiO2@PA-1) to 18.80 % (SiO2@PA-3), leading to the enhanced selectivity toward polycyclic aromatic hydrocarbons (PAHs). The separation selectivity (α=1.16–1.62) of PAHs by SiO2@PA-3 column was superior to that by C18 column (α=1.00–1.49), attributed to the π-stacking and hydrophobic methanism from PA phase. The SiO2@PA-3 column separately achieved baseline separation of some alkylbenzenes and tetrabromobisphenol A derivatives. Notably, it also effectively separated several isomers of PAHs, phenols, and anilines, with a column efficiency reaching 38,260 plates m−1. Then, a HPLC-DAD method with a linear range (0.33–12.5 mg L−1) and detection limits (0.10–0.20 mg L−1), was developed for determining tetrabromobisphenol A derivatives in water samples. This research proposed a cost-effective and facile strategy for the development of stationary phase.
{"title":"Petroleum asphalt-coated silica as the stationary phase for liquid chromatography","authors":"Xin Zhang, Juanjuan Feng, Baoen Su, Haidi Zhang, Yana Luo, Min Sun","doi":"10.1016/j.chroma.2025.466529","DOIUrl":"10.1016/j.chroma.2025.466529","url":null,"abstract":"<div><div>Owing to high specific surface area and excellent chemical stability, carbon materials as stationary phases demonstrated significant advantages. However, their practical application was limited by high cost and complex synthesis process. In this work, the low-cost petroleum asphalt (PA) was successfully transformed into a stationary phase for high-performance liquid chromatography. Through a facile coating way, the PA was immobilized onto porous silica for producing a new stationary phase. As the concentration of asphalt in the coating solution was increased, the carbon content of the stationary phases rose from 6.67 % (SiO<sub>2</sub>@PA-1) to 18.80 % (SiO<sub>2</sub>@PA-3), leading to the enhanced selectivity toward polycyclic aromatic hydrocarbons (PAHs). The separation selectivity (α=1.16–1.62) of PAHs by SiO<sub>2</sub>@PA-3 column was superior to that by C<sub>18</sub> column (α=1.00–1.49), attributed to the π-stacking and hydrophobic methanism from PA phase. The SiO<sub>2</sub>@PA-3 column separately achieved baseline separation of some alkylbenzenes and tetrabromobisphenol A derivatives. Notably, it also effectively separated several isomers of PAHs, phenols, and anilines, with a column efficiency reaching 38,260 plates m<sup>−1</sup>. Then, a HPLC-DAD method with a linear range (0.33–12.5 mg L<sup>−1</sup>) and detection limits (0.10–0.20 mg L<sup>−1</sup>), was developed for determining tetrabromobisphenol A derivatives in water samples. This research proposed a cost-effective and facile strategy for the development of stationary phase.</div></div>","PeriodicalId":347,"journal":{"name":"Journal of Chromatography A","volume":"1765 ","pages":"Article 466529"},"PeriodicalIF":4.0,"publicationDate":"2025-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145501441","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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