Determination of bisphenol analogues in bottled water using deep eutectic solvent and magnetic multi-walled carbon nanotubes followed by ultra-high performance liquid chromatography coupled with tandem mass spectrometry.

Bisphenol analogues (BPs) are a class of typical environmental endocrine disruptors (EDCs) that have recently attracted increasing attention with regard to their potential effects on human health. The objective of this study was to develop a method using a magnetic soft material, which consisted of hydrophilic deep eutectic solvent (DES) and magnetic multi-walled carbon nanotubes (MMWCNTs), for the dispersive solid-phase extraction (d-SPE), coupled with ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), for the determination of the levels of nine BPs in bottled water. The hydrophilic DES enable the rapid dispersion of MMWCNTs when the material is injected rapidly into the sample solution using a pipette gun. This process can therefore be completed in a relatively short period of time, resulting in an efficient extraction. Under optimal conditions, the limit of detections (LODs) of the method were 0.0003-0.003 µg/L and the limit of quantifications (LOQs) were 0.001-0.01 µg/L. The relative standard deviations (RSDs) of the method were only 2.42-7.59 % for inter-day and 3.71-9.67 % for intra-day. The method demonstrated good reproducibilities and recoveries, rendering it suitable for the determination of BPs in large-volume water samples.

Lipidomic Profiling of Common Wheat Flours from 1891-2010.

Wheat lipids are a minor constituent of wheat, with an important influence on its processing properties. While breeding aimed to improve the protein composition of wheat flour, its influence on the lipid composition remains unknown. We therefore analyzed the lipidome of 60 different common wheat (Triticum aestivum) flours representing cultivars registered and grown in Germany from 1891 to 2010. Four different extraction techniques were tested before the application of a semiquantitative, untargeted UHPLC-MS/MS method. The measurements included 16 different lipid classes and 102 different lipid species. Based on the lipid profile, discrimination between old (registered between 1891 to 1950) and modern (1951 to 2010) cultivars was possible. While the lipid class composition remained constant, differences were due to variations within the class of triacylglycerols, with modern cultivars containing less unsaturated fatty acids than the older ones. Our results imply that improving the lipid class composition of common wheat is a promising target for further breeding.

Metabolomic analysis of HUVEC after Thermal denaturation UHPLC-MS/MS-based metabolomics.

Preserving denatured dermis has been shown to promote wound healing and improve skin appearance and function. Angiogenesis is crucial for the healing of burn wounds. However, the metabolic mechanisms underlying angiogenesis during burn recovery remain unclear. In this study, ultra-high performance liquid chromatography-mass spectrometry analysis revealed six distinct metabolites in a heat-denatured cell model. A bioinformatics approach was used to predict the differentially expressed metabolites, and four metabolic pathways closely related to trauma repair were identified. These pathways might play a significant role in the regression of thermally injured endothelial cells. We also found that increasing D-mannose level promoted the angiogenic activity of human umbilical vein endothelial cells in the heat-denatured cell model, enhancing cell proliferation, migration, and tube formation. In summary, these findings revealed changes in metabolites and metabolic pathways in thermally injured endothelial cells, and demonstrated that D-mannose could promote angiogenesis during the recovery of thermally injured endothelial cells.

A novel quantitative method for the determination of 10B-carrier boronophenylalanine in rat plasma by UHPLC-MS/MS and comparison with ICP-MS.

L-Boronophenylalanine (BPA), a widely used 10B carrier for clinical boron neutron capture therapy (BNCT), was quantified in rat plasma through a simple, effective and stable ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method. Chromatographic separation was performed on an ACQUITY UPLC HSS T3 (100 mm × 2.1 mm, 1.8 µm) column with the mobile phase of 0.5 % formic acid aqueous solution and acetonitrile. For the detector, the m/z ion pairs used for quantification were 209.1→120.1 for BPA and 210.1→120.1 for internal standard in a positive mode by electrospray ionization (ESI) using multiple reaction monitoring (MRM). The method is specific and robust with rare affection by endogenous substances in the matrix. A good linear relationship was observed over 80-80000 ng/mL (r2 = 0.9993). The values of inter- and intra-day accuracy and precision were within the acceptance criteria of ±15 %. BPA was found to be stable under different experimental conditions. This developed method was successfully applied on a pharmacokinetic experiment on Sprague-Dawley rats (intravenous injection, 125 mg/kg) and a comparation between UHPLC-MS/MS and ICP-MS for BPA was carried out.

Quantitative tracing of typical herbicides and their metabolites in sorghum agrosystems for fate tendency and cumulative risk.

Elucidating the combined exposure of agrochemicals is essential for safeguarding human health and agroecosystem safety. A rapid and high-sensitivity UHPLC-MS/MS method was developed for simultaneous quantification of nine compounds in sorghum by an assembly-line optimization process with a limit of quantitation of 0.001 mg/kg. The concentration variation of atrazine, quinclorac, fluroxypyr-meptyl and metabolites was reflected by terminal magnitudes of ≤0.0665 mg/kg. Additionally, atrazine was dealkylated to deethyl atrazine and desethyl desisopropyl atrazine at concentrations of 0.0014-0.0058 mg/kg during the sorghum harvest. Acceptable health hazardous of atrazine and quinclorac for all life cycle populations were comparatively assessed via deterministic and probabilistic models, in which atrazine gained an 83.55 % share of cumulative dietary risks. Rural residents had significantly higher risks than urban residents, and children were the most sensitive group. Despite the low health risks, combined exposure to herbicides and their metabolites should be continuously stressed, given their cumulative amplification effects.

Targeted metabolite and molecular profiling of carotenoids in rose petals: A step forward towards functional food applications.

Given the increasing consumer demand for natural and functional foods, rose petals offer a promising novel ingredient for food innovation, especially yellow and orange rose petals rich in carotenoids, making them ideal for food processing and color retention. Despite their potential, the metabolic profile of carotenoids in roses has not yet been fully explored. Therefore, the present study aimed to provide a comprehensive analysis of carotenoid metabolism in rose petals during three developmental stages. The results revealed that orange rose petals had the highest carotenoid content of 488.85 µg/g, with xanthophylls being identified as the primary carotenoid constituents (70.40 %) in roses for the first time. Furthermore, two genes, RhBCH1 and RhCCD4, were identified to be involved in the regulation of carotenoid biosynthesis in roses. Overall, this study demonstrates the enormous potential of rose petals as functional food ingredients, providing a theoretical basis for breeding high-carotenoid rose varieties.

Study on pharmacokinetic and tissue distribution of hyperin, astragalin, kaempferol-3-O-ß-D-glucuronide from rats with multiple administrations of Semen Cuscutae processed with salt solution with effect of treating recurrent spontaneous abortion.

Introduction: Semen Cuscutae is a traditional Chinese medicine (TCM) that tonifies the kidneys and prevents miscarriage. According to Chinese medicine theory, kidney deficiency is one of the main causes of recurrent spontaneous abortion (RSA). The previous studies showed that raw product of Semen Cuscutae (SP) and Semen Cuscutae processed with salt solution (YP) have ameliorative effects on RSA, and that YP is superior to SP. However, the active components of YP to ameliorate RSA remain unclear and require further studies. The objective of this study is to investigate the active components of YP in ameliorating RSA. Methods: First, a rat model of RSA was established using hydroxyurea in combination with mifepristone. Aqueous decoction of YP was given by gavage to rats. Second, pregnant rats were sampled on days 5, 7, 9, 10 and 12 during the modelling period. The content of Hyperin (HY), astragalin (AS) and kaempferol-3-O-ß-D-glucuronide (KA) in blood and liver, heart, spleen, lung and kidney tissues were detected by liquid chromatography-mass spectrometry (LC-MS). The pharmacodynamic indicators including progesterone (P), chorionic gonadotropin ß (ß-HCG), estradiol (E2), tumor necrosis factor-α (TFN-α), interleukin 4 (IL-4), and tryptophan (TRP) were measured by enzyme-linked immunosorbent assay (ELISA) Pearson's correlation analysis and grey relational analysis were used to establish the relationship between the pharmacodynamic indexes and chemical constituents. Results: The pharmacokinetic results showed that the area under curve (AUC) value of KA was the largest. The tissue distribution results showed that astragalin was widely distributed in liver, heart, spleen, lung and kidney in the RSA model rats, while HY was detected only in the uterus, and KA was detected only in the kidney. The pearson correlationl analysis showed that KA was significantly and positively correlated with the contents of E2, P, ß-HCG and TRP. Both AS and HY were significantly negatively correlated with the content of TNF-α, respectively. Discussion: This study reveals the pharmacokinetics and tissue distribution of KA, AS and HY in rats with RSA. It was elucidated that all three were involved in the regulation of progesterone levels and immune function. It initially revealed the mechanism of action of YP in enhancing the improvement of RSA, and it provided a theoretical basis for the quality assessment of YP.

HPLC-MS/MS and ICP-MS for Evaluation of Mycotoxins and Heavy Metals in Edible Insects and Their Defatted Cakes Resulting from Supercritical Fluid Extraction.

Edible Insects (EIs) are an alternative source of bioactive compounds such as proteins or fatty acids and micronutrients as vitamins or minerals, thus showing potential to replace traditional foodstuffs in an economical and environmentally friendly way. Nonetheless, EIs can accumulate hazardous chemicals such as mycotoxins and heavy metals. The aim of the present study is to determine mycotoxins and heavy metal content in raw insect samples and those resulting products obtained after supercritical fluid extraction (SFE). Insect samples included Acheta domesticus (cricket) meal, Tenebrio molitor (mealworm) meal, Alphitobius diaperinus (buffalo worm), and Locusta migratoria (locust). For this purpose, a QuEChERS method followed by LC-MS/MS analysis was optimized with good results for the analysis of mycotoxins, principally Aflatoxins (AFs), Ochratoxin A (OTA), and Enniatins (ENNs). In contrast, heavy metals (As, Cd, Hg, Pb) were determined by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). The results obtained revealed that Locust was positive for AFG2 at a level of 115.5 µg/kg, and mealworm was only contaminated with OTA at 58.1 µg/kg. Emerging mycotoxins (ENNA, ENNA1, ENNB, and ENNB1) were detected at lower levels < 2.2 µg/Kg. Concerning heavy metals, limits exceeding regulation were detected for Cd in the insect species studied, with levels up to 219 µg/kg, and for Pb in crickets (100.3 µg/kg). Finally, the analysis of the post-extraction solids after SFE processing revealed that heavy metals remained in the resulting SFE cakes, while mycotoxins were detected at negligible levels (up to 1.3 µg/Kg).

Simultaneous and High-Throughput Analytical Strategy of 30 Fluorinated Emerging Pollutants Using UHPLC-MS/MS in the Shrimp Aquaculture System.

This study established novel and high-throughput strategies for the simultaneous analysis of 30 fluorinated emerging pollutants in different matrices from the shrimp aquaculture system in eastern China using UHPLC-MS/MS. The parameters of SPE for analysis of water samples and of QuEChERS methods for sediment and shrimp samples were optimized to allow the simultaneous detection and quantitation of 17 per- and polyfluoroalkyl substances (PFASs) and 13 fluoroquinolones (FQs). Under the optimal conditions, the limits of detection of 30 pollutants for water, sediment, and shrimp samples were 0.01-0.30 ng/L, 0.01-0.22 µg/kg, and 0.01-0.23 µg/kg, respectively, while the limits of quantification were 0.04-1.00 ng/L, 0.03-0.73 µg/kg, and 0.03-0.76 µg/kg, with satisfactory recoveries and intra-day precision. The developed methods were successfully applied to the analysis of multiple samples collected from aquaculture ponds in eastern China. PFASs were detected in all samples with concentration ranges of 0.18-0.77 µg/L in water, 0.13-1.41 µg/kg (dry weight) in sediment, and 0.09-0.96 µg/kg (wet weight) in shrimp, respectively. Only two FQs, ciprofloxacin and enrofloxacin, were found in the sediment and shrimp. In general, this study provides valuable insights into the prevalence of fluorinated emerging contaminants, assisting in the monitoring and control of emerging contaminants in aquatic foods.

Quantification of tricyclic glycopeptide in human plasma by UHPLC-MS3 coupled with counter-extraction follow by protein precipitation to enhance sensitivity.

An ultra-high performance liquid chromatography tandem mass spectrometry cubed (UHPLC/MS3) assay coupled with protein precipitation and counter-extraction for detection of tricyclic glycopeptide vancomycin in human plasma was established and validated in this study. After protein precipitation and counter-extraction with dichloromethane, chromatographic separation of vancomycin and norvancomycin were performed on a reversed phase column (XBridge Peptide BEH C18 column, 2.1 × 100 mm I.D, 3.5 µm). The transition (parent ions â†’ fragment ions â†’ further fragment ions) at m/z 725.3 â†’ 144.1 â†’ 100.1 was used for quantification of vancomycin. The transition (parent ions â†’ fragment ions) at m/z 718.3 â†’ 144.2 was used for detection of norvancomycin. The linear range of the developed analytical method for quantification of vancomycin was 0.5-100 µg/mL (r = 0.9989). The range of intra- and inter-day precisions of the assay among low, medium and high concentrations is between 1.88 % and 6.33 %. The sensitivity of the analytical method was significantly improved by using MS3 technique as monitoring mode and counter-extraction with dichloromethane followed by protein precipitation as sample processing assay. The developed UHPLC/MS3 assay was successfully applied for clinical therapeutic drug monitoring (TDM) of vancomycin in 45 human plasma samples.

Investigation of the mechanism of Bark of Ailanthus altissima in the treatment of ulcerative colitis based on network pharmacology and experimental verification.

ETHNOPHARMACOLOGICAL RELEVANCE: The bark of Ailanthus altissima (Mill.) Swingle (BAA), a widely used Chinese medicinal herb in traditional remedies for bowel disorders, has yet to be explored in the context of ulcerative colitis (UC), and its therapeutic mechanisms remain unclear. AIM OF THE STUDY: This study integrated network pharmacology and experimental validation to investigate the effects and underlying mechanisms of BAA in treating UC. MATERIALS AND METHODS: First, UPLC-MS/MS analysis was employed to identify the chemical constituents of BAA. Network pharmacology was then applied to analyze the potential mechanisms of BAA based on these identified compounds. Lastly, a dextran sulfate sodium (DSS)-induced UC mouse model was utilized to assess BAA's therapeutic efficacy, with Western blotting performed to examine changes in protein expression within the key pathway influenced by BAA. RESULTS: UPLC-MS/MS and SwissADME analysis identified 223 active compounds in BAA. Network pharmacology suggested that the PI3K/AKT pathway may serve as a primary mechanism by which BAA exerts its anti-UC effects. In the DSS-induced UC mouse model, BAA significantly mitigated colonic injury, reduced DAI scores, and promoted weight recovery in mice. Additionally, BAA downregulated pro-inflammatory cytokines, including TNF-α, IL-1ß, and IL-6, thereby suppressing inflammatory responses in the colon. Western blot analysis further demonstrated that BAA primarily inhibited the PI3K/AKT pathway in UC mouse colon tissue. CONCLUSION: This study highlights that BAA effectively reduces colonic inflammation and preserves intestinal mucosal integrity, likely through the inhibition of PI3K/AKT pathway activity, positioning it as a potential treatment for UC.

[Determination of seven phenoxy carboxylic acid herbicides in water using dispersive solid phase extraction coupled with ultra-high performance liquid chromatography-tandem mass spectrometry based on metal-organic framework composite aerogel].

Phenoxy carboxylic acid herbicides (PCAs) are difficult to degrade and, thus, pose significant threats to the environment and human health. The limit for 2,4-dichlorophenoxyacetic acid is 30 µg/L in China's standards for drinking water quality, 70 µg/L in the United States' drinking water standards, and 30 µg/L in the World Health Organization's guidelines for drinking water quality. Therefore, the development of an effective detection method for trace PCAs in water is a crucial endeavor. Metal-organic frameworks (MOFs) are novel porous materials that possess advantages such as a large specific surface area, adjustable pore size, and abundant active sites. They exhibit excellent adsorption capability for various compounds. However, the applications of MOFs as adsorbents are limited. For example, the process of isolating powdered MOFs from aqueous solutions is laborious, and microporous MOFs exhibit limited surface affinity, which decreases their mass transfer efficiency in the liquid phase. MOF crystals can be embedded in a substrate to overcome these limitations. Aerogels are obtained by drying hydrogels, which are hydrophilic polymers with a three-dimensional crosslinked network structure. Spongy aerogel materials exhibit unique structural properties such as high porosity, large pore volume, ultralow density, and easy tailorability. When MOFs are combined with an aerogel, their efficient and selective adsorption properties are preserved. In addition, MOF aerogels exhibit a hierarchical porous structure, which enhances the affinity and mass transfer efficiency of the MOF for target molecules. At present, MOF aerogels are primarily prepared by freeze-drying or using supercritical carbon dioxide. These drying processes require significant amounts of energy and time. Hence, the development of greener and more efficient methods to prepare skeleton aerogels is urgently needed. In this study, we prepared an environment-friendly aerogel at ambient temperature and pressure without the use of specialized drying equipment. This ambient-dried MOF composite aerogel was then used for the dispersive solid phase extraction (DSPE) of seven PCAs from environmental water, followed by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The key parameters affecting the efficiency of DSPE, including the extraction conditions, ratio of MIL-101(Fe)-NH2 to sodium alginate, pH of the aqueous samples, extraction time, ionic strength (salinity), and elution conditions, such as the elution solvent ratio, elution time, and elution volume, were investigated to obtain optimal extraction efficiency. The adsorbent could adsorb the target contaminants within 12 min, and the analytes could be completely desorbed within 30 s by elution with 4 mL of 1.5% (v/v) formic acid in methanol solution. The water samples could be analyzed without pH adjustment. The main adsorption mechanisms were electrostatic interactions and π-π conjugation. Thus, a new method based on MOF aerogels coupled with UHPLC-MS/MS was developed for the determination of the seven PCA residues in water. The calibration curves for the seven PCAs showed good linearity (r2≥0.9986), with limits of detection (LODs) and quantification (LOQs) ranging from 0.30 to 1.52 ng/L and from 1.00 to 5.00 ng/L, respectively. Good intra- and inter-day precision values of 6.5%-17.1% and 7.4%-19.4%, respectively, were achieved under low (8 ng/L), medium (80 ng/L), and high (800 ng/L) spiking levels. The developed method was applied to the detection of PCAs in surface water, seawater, and waste leachate, and the detected mass concentrations ranged from 0.6 to 19.3 ng/L. Spiked recovery experiments were conducted at mass concentrations of 8, 80, and 800 ng/L, and the recoveries ranged from 61.7% to 120.3%. The proposed method demonstrates good sensitivity, precision, and accuracy, and has potential applications in the detection of trace PCAs in environmental water.

An Ultra-Fast Green UHPLC-MS/MS Method for Assessing the In Vitro Metabolic Stability of Dovitinib: In Silico Study for Absorption, Distribution, Metabolism, Excretion, Metabolic Lability, and DEREK Alerts.

Background and Objectives: Dovitinib (DVB) is a pan-tyrosine kinase inhibitor (TKI) that can be administered orally. In September 2023, the FDA granted Oncoheroes approval to proceed with an Investigational New Drug (IND) application for dovitinib. This application is intended for the treatment of relapsed or advanced juvenile solid tumors, namely, osteosarcoma. Materials and Methods: The target of the present study was to develop a rapid, green, accurate, and sensitive UHPLC-MS/MS method for measuring DVB levels in human liver microsomes (HLMs). The validations of the HLMs were performed via the established UHPLC-MS/MS approach, as stated in the US FDA reported guidelines for the standards of bioanalytical method validation protocol. The StarDrop in silico software package (version 6.6), which involves the DEREK and WhichP450 in silico modules, was used to check the DVB structure for hazardous alerts and metabolic instability. The DVB and encorafenib (EFB), internal standard, and chromatographic peaks were successfully separated using a reversed phase column (an Eclipse Plus Agilent C8 column) and an isocratic mobile phase. The production of DVB parent ions was accomplished by utilizing the positive ionization mode of an ESI source. The identification and measurement of DVB daughter ions were conducted using the MRM mode. Results: The inter-day accuracy and precision exhibited a spectrum of values in the range of -0.56% to 9.33%, while the intra-day accuracy and precision showcased a range of scores between 0.28% and 7.28%. The DVB calibration curve showed a linear relationship that ranged from 1 to 3000 ng/mL. The usefulness of the currently validated UHPLC-MS/MS method was approved by the lower limit of quantification (LLOQ) of 1 ng/mL. The AGREE findings demonstrate that the UHPLC-MS/MS method had a noteworthy degree of ecological greenness. The in vitro half-life (t1/2) and intrinsic clearance (Clint) of DVB were calculated to be 15.48 min and 52.39 mL/min/kg, respectively, which aligned with the findings from the WhichP450 software (version 6.6). Conclusions: Via the usage of in silico software, it has been observed that making small changes to the structure of the aryl piperazine ring and quinolinone moieties, or replacing these groups in the drug design process, shows potential for enhancing the metabolic safety and stability of newly developed derivatives compared to DVB.

Characterization of etomidate and emerging analogs in human hair using UHPLC-MS/MS and confirmation in real forensic cases.

The regulation of etomidate, a widely used drug for anesthesia induction and short surgical procedures, has led to the emergence of etomidate analogs such as metomidate, propoxate, and isopropoxate. This study introduces and validates a simple, rapid, and cost-effective ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the determination and quantification of etomidate, etomidate acid, metomidate, propoxate, and isopropoxate in human hair. Using a five-minute gradient elution on a Phenomenex Kinetex Biphenyl column, the method achieves low limits of quantification (LOQs), ranging from 5 to 20 pg/mg. Method validation confirms robust linear calibration curves for the target substances in hair samples within the range of LOQs to 1000 pg/mg, with average correlation coefficients (r) all exceeding 0.999. The method also achieves acceptable intra-day and inter-day precision (<15 %) and trueness(bias, -10.9 % to 14.4 %). The matrix effect ranges from 46.9 % to 94.2 %, and the extraction recovery rate ranges from 84.5 % to 105 %. When applied to authentic cases, this method successfully identified 56 positive samples. The concentrations of etomidate, etomidate acid, metomidate, propoxate, and isopropoxate in positive samples ranged from 27 to 1.5 × 105 pg/mg, 21 to 1.5 × 103 pg/mg, 18 to 8.7 × 104 pg/mg, 25 to 1.6 × 104 pg/mg, and 22 to 5.0 × 105 pg/mg, respectively.

Effect of the addition of cocoa sweatings and time of fermentation on flavor compounds and sensory perception of 100% roasted cocoa liquor.

Fermentation is critical for producing high-quality cocoa, yet its kinetics and resulting chemical and sensory outcomes are poorly understood and thus difficult to manage. Cocoa sweatings (CS), the liquid runoff produced early during fermentation and typically drained off, may beneficially affect fermentation outcome when added back into the fermenting mass. Here, we report how back-addition of CS affects composition and sensory perception of roasted cocoa liquor after 5, 6, and 7 days of fermentation. Cocoa liquor (= 100% chocolate) made from beans fermented for 5 days with the addition of CS were similar in sensory perception to those fermented for 7 days without added CS. Twenty-one flavor compounds showed similar patterns to the sensory results: In the beans fermented with CS, these compounds remained at similar levels after 5, 6, and 7 days of fermentation, while the same compounds significantly changed in the samples fermented conventionally, without CS addition. These results suggest a link between changes in flavor composition and sensory differences in roasted cocoa. Future work is needed to reveal the mechanism of flavor stabilization throughout fermentation resulting from the back-addition of CS. PRACTICAL APPLICATION: Roasted cocoa liquor fermented with cocoa sweating (CS) is sensorily similar when fermented for 5 or 7 days and produces cocoa that is sensorily similar to traditionally fermented cocoa in shorter time (5 days vs. 7 days). The addition of CS seems to stabilize 21 flavor compounds throughout fermentation mimicking changes in sensory perception. The back-addition of CS could help standardize cocoa fermentation as indicated by more consistent temperature evolution.

Fingerprinting alkaloids for traceability: Semi-untargeted UHPLC-MS/MS approach in raw lupins as a case study.

Lupin seeds are recognized for their nutritional value and potential health benefits, but they contain also a significant amount of alkaloids, an anti-nutritive class of compounds, which vary significantly in composition within and between species due to environmental factors. In this study, we developed a predictive multi-experiment approach using ultra-high performance liquid chromatography coupled with triple quadrupole with linear ionic trap tandem mass spectrometry (UHPLC-QqQ-LIT-MS/MS) for comprehensive alkaloid profiling and geographical classification of Lupinus albus L. samples originating from four different Italian regions. Six targeted quinolizidine alkaloids were detected and 21 other alkaloids were putatively identified. Hierarchical Cluster Analysis (HCA) and partial least squares discriminant analysis (PLS-DA) were applied to explore the data structure and successfully classify samples according to their geographical origin. The data demonstrate the efficacy of the developed approach in providing valuable insights in alkaloid profiles of lupin seeds and their potential as markers for geographical traceability.

Development and validation of a green analytical method for determining fourteen bisphenols in bee pollen by ultra-high-performance liquid chromatography-tandem mass spectrometry.

A new analytical method was developed and validated to determine fourteen bisphenols (A, B, C, E, F, M, P, S, Z, AF, AP, BP, FL, PH) in bee pollen using ultra-high-performance liquid chromatography-tandem mass spectrometry. Two different sample treatments were proposed and evaluated: one based on the QuEChERS (quick, easy, cheap, effective, rugged & safe) approach and the other utilizing microextraction with a supramolecular solvent (SUPRAS). In both cases, average analyte recovery ranged between 71 % and 114 %, and the matrix effect was between -45 % and +5 %, although it was not significant when using the QuEChERS-based method (<±20 %). The environmental impact of both sample treatments was assessed using different analytical metrics, with both procedures classified as environmentally friendly, though slightly better results were obtained for SUPRAS. The method was fully validated, showing that the QuEChERS approach had better overall performance, particularly regarding sensitivity and matrix effect. Consequently, the QuEChERS methodology was applied to determine bisphenols in thirty bee pollen samples from different Spanish regions. Residues of three bisphenols (M, P, and S) were detected, although only bisphenol S was quantified in several samples at low concentration levels (<7 µg kg-1), which is below the established specific migration limit (SML; 50 µg kg-1). However, regarding human health, the estimated daily intake, target hazard quotient, and hazard index assessed were higher than acceptable limits, suggesting a potential risk for human consumers.

Bisphenols in daily clothes from conventional and recycled material: evaluation of dermal exposure to potentially toxic substances.

Given the increasing concern about chemical exposure from textiles, our study examines the risks of dermal exposure to bisphenol A (BPA), bisphenol S (BPS), bisphenol B (BPB) and bisphenol F (BPF) from conventional and recycled textiles for adults, aiming to obtain new data, assess exposure, and evaluate the impact of washing on bisphenol levels. A total of 57 textile samples (33 from recycled and 24 from conventional material) were subjected to ultrasound-assisted extraction (UAE) followed by ultra-high performance liquid chromatography with tandem mass spectrometry analysis (UHPLC-MS/MS). The BPA and BPS concentrations varied widely (BPA: < 0.050 to 625 ng/g, BPS: 0.277-2,474 ng/g). The median BPA content in recycled textiles (13.5 ng/g) was almost twice as high as that of 7.66 ng/g in conventional textiles. BPS showed a median of 1.85 ng/g in recycled textiles and 3.42 ng/g in conventional textiles, indicating a shift from BPA to BPS in manufacturing practices. Simulated laundry experiments showed an overall reduction in bisphenols concentrations after washing. The study also assessed potential health implications via dermal exposure to dry and sweat-wet textiles compared to a tolerable daily intake (TDI) of 0.2 ng/kg bw/day for BPA set by the European Food Safety Authority (EFSA). Exposure from dry textiles remained below this threshold, while exposure from wet textiles often exceeded it, indicating an increased risk under conditions that simulate sweating or humidity. By finding the widespread presence of bisphenols in textiles, our study emphasises the importance of being aware of the potential risks associated with recycling materials as well as the benefits.

Comprehensive Analysis of Cetuximab Critical Quality Attributes: Impact of Handling on Antigen-Antibody Binding.

BACKGROUND/OBJECTIVES: Cetuximab, formulated in Erbitux® (5 mg/mL), is a therapeutic monoclonal antibody (mAb) widely used in several cancer treatments. Currently, there is insufficient knowledge about the behavior of cetuximab with regard to the risk associated with its routine handling or unintentional mishandling in hospitals. Forced degradation studies can simulate these conditions and provide insights into the biophysical and biochemical properties of mAbs. METHODS: In this study, we conducted a deep physicochemical and functional characterization of the critical quality attributes of cetuximab in control samples and under controlled degraded conditions, including freeze-thaw cycles, heat, agitation, and light exposure. To achieve this purpose, we used a set of proper analytical techniques, including CD, IT-FS, DLS, SE/UHPLC-UV, UHPLC-MS/MS, and ELISA, to check functionality based on antigen-antibody binding. RESULTS: The results revealed that light exposure was the stress stimuli with the greatest impact on the drug product, leading to the formation of non-natural oligomers, fragmentation, and oxidation of methionine residues. Additionally, cetuximab (Erbitux®, 5 mg/mL) showed a tendency to aggregate when submitted to 60 °C for 1 h. In terms of functionality, cetuximab (Erbitux®, 5 mg/mL) samples were found to be affected when subjected to freeze-thaw cycles, 60 °C (1 h), and when exposed to light (daylight with room temperature excursion and accelerated light exposure). CONCLUSIONS: Thus, we suggest that Erbitux® (5 mg/mL) should be shielded from these environmental conditions, as they compromise both the safety and efficacy of the drug product.

Metabolic and Pharmacokinetic Profiling Studies of N, N-Dimethylaniline-Heliamine in Rats by UHPLC-Q-Orbitrap MS/MS.

Cardiovascular disease is the first cause of death worldwide and kills more people each year than any other cause of death. N, N-dimethylaniline-heliamine (DH), a synthetic tetrahydroisoquinoline alkaloid, has shown notable antiarrhythmic activity. However, the metabolic processes and pharmacokinetic characteristics of DH in rats have not been studied. This study aims to identify its metabolites, as well as develop and validate a rapid and efficient bioanalytical method for quantifying DH in rat plasma over a wide range of concentrations. Its metabolites were characterized in silico, in vitro, and in vivo. A series of 16 metabolites were identified, of which 12 were phase I metabolites and 4 were phase II metabolites. A low probability of DH binding to DNA, protein, and glutathione is predicted by the in silico model. The main metabolic processes of DH were demethylation, dehydrogenation, glucuronidation, and sulfation. Concentration-time profiles were generated by analyzing the plasma, and the outcomes were analyzed via non-compartmental analysis to identify the pharmacokinetic parameters. Among the detected parameters were the volume of distribution, estimated at 126,728.09 ± 56,867.09 mL/kg, clearance at 30,148.65 ± 15,354.27 mL/h/kg, and absolute oral bioavailability at 16.11%. The plasma distribution volume of DH was substantially higher than the overall plasma volume of rats, which suggests that DH has a specific tissue distribution in rats. This study suggests that DH is appropriately bioavailable and excreted via a variety of routes and has low toxicity.