Metabolomic insights into neurological effects of BDE-47 exposure in the sea cucumber Apostichopus japonicus.

The persistent organic pollutant 2,2',4,4'-Tetrabromodiphenyl ether (BDE-47), a prevalent congener among polybrominated diphenyl ethers (PBDEs), exhibits potent bioaccumulation and toxicity. Despite extensive research into the adverse effects of BDE-47, its neurotoxicity in sea cucumbers remains unexplored. Given the crucial role of the sea cucumber's nervous system in survival and adaptation, evaluating the impacts of BDE-47 is vital for sustainable aquaculture and consumption. In this study, we employed ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Triple-TOF-MS) to analyze metabolomic changes in neuro-related tissues of Apostichopus japonicus exposed to low (0.1 µg/L), medium (1.0 µg/L), and high (10.0 µg/L) BDE-47 concentrations. We identified significantly changed metabolites in each exposure group (87 in low, 79 in medium, and 102 in high), affecting a variety of physiological processes such as steroid hormone balance, nucleotide metabolism, energy metabolism, neurotransmitter levels, and neuroprotection. In addition, we identified concentration-dependent, common, and some other metabolic responses in the neuro-related tissues. Our findings reveal critical insights into the neurotoxic effects of BDE-47 in sea cucumbers and contribute to risk assessment related to BDE-47 exposure in the sea cucumber industry, paving the way for future neurotoxicological research in invertebrates.

Mechanochemical-Assisted Extraction and Biological Activity Research of Phenolic Compounds from Lotus Seedpod (Receptaculum Nelumbinis).

To explore the feasibility of the mechanochemical-assisted extraction (MCAE) of phenolic compounds from lotus seedpod (Receptaculum Nelumbinis), a single-factor experiment combined with response-surface methodology (RSM) was used to optimize the extraction process. The results showed the optimal extraction conditions as follows: Li2CO3 as a solid reagent (25%), an extraction time of 80 min, liquid/solid ratio of 42.8 mL/g, and extraction temperature of 80.7 °C; and the maximum value of total phenolic content (TPC) was 106.15 ± 1.44 gallic acid equivalents (GAE)/g dry weight (DW). Additionally, the 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) were 279.75 ± 18.71, 618.60 ± 2.70, and 634.14 ± 7.17 µmol TE/g, respectively. Ultra-high pressure liquid chromatography combined with triple-time-of-flight mass spectrophotometry (UPLC-Triple-TOF/MS) analysis identified eight phenolic compounds mainly consisting of polyphenols and flavonoids. Moreover, the phenolic compounds showed potent inhibitory effects on both α-amylase and α-glucosidase, with inhibition rates of over 80%. Furthermore, the results showed different degrees of inhibition activity against Bacillus subtilis, Staphylococcus aureus, and Escherichia coli, among which the inhibitory effect on the growth of B. subtilis was the best. This paper shows that the phenolic compounds have good biological activities, which provides a reference for the further exploitation of LSP.

Characterization of Ginsenosides from the Root of Panax ginseng by Integrating Untargeted Metabolites Using UPLC-Triple TOF-MS.

To compare the chemical distinctions of Panax ginseng Meyer in different growth environments and explore the effects of growth-environment factors on P. ginseng growth, an ultra-performance liquid chromatography-tandem triple quadrupole time-of-flight mass spectrometry (UPLC-Triple-TOF-MS/MS) was used to characterize the ginsenosides obtained by ultrasonic extraction from P. ginseng grown in different growing environments. Sixty-three ginsenosides were used as reference standards for accurate qualitative analysis. Cluster analysis was used to analyze the differences in main components and clarified the influence of growth environment factors on P. ginseng compounds. A total of 312 ginsenosides were identified in four types of P. ginseng, among which 75 were potential new ginsenosides. The number of ginsenosides in L15 was the highest, and the number of ginsenosides in the other three groups was similar, but it was a great difference in specie of ginsenosides. The study confirmed that different growing environments had a great influence on the constituents of P. ginseng, and provided a new breakthrough for the further study of the potential compounds in P. ginseng.

Profile of Polyphenol Compounds of Five Muscadine Grapes Cultivated in the United States and in Newly Adapted Locations in China.

Polyphenol compositions and concentrations in skins and seeds of five muscadine grapes (cv. "Noble", "Alachua", "Carlos", "Fry", and "Granny Val") cultivated in the United States (Tallahassee-Florida, TA-FL) and South China (Nanning-Guangxi, NN-GX and Pu'er-Yunnan, PE-YN) were investigated, using ultra performance liquid chromatography tandem triple quadrupole time-of-flight mass spectrometry (UPLC Triple TOF MS/MS). Fourteen ellagitannins were newly identified in these muscadine grapes. The grapes grown in NN-GX accumulated higher levels of ellagic acid, methyl brevifolin carboxylate, and ellagic acid glucoside in skins, and penta-O-galloyl-glucose in seeds. In PE-YN, more flavonols were detected in skins, and higher contents of flavan-3-ols, ellagic acid, and methyl gallate were identified in seeds. Abundant seed gallic acid and flavonols were found among the grapes grown in TA-FL. Based on principal component analysis (PCA) of 54 evaluation parameters, various cultivars grown in different locations could be grouped together and vice versa for the same cultivar cultivated in different regions. This is the result of the interaction between genotype and environmental conditions, which apparently influences the polyphenol synthesis and accumulation.

Identification and comparative oridonin metabolism in different species liver microsomes by using UPLC-Triple-TOF-MS/MS and PCA.

Oridonin (ORI) is an active natural ent-kaurene diterpenoid ingredient with notable anti-cancer and anti-inflammation activities. Currently, a strategy was developed to identify metabolites and to assess the metabolic profiles of ORI in vitro using ultra-high-performance liquid chromatography-Triple/time-of-flight mass spectrometry (UPLC-Triple-TOF-MS/MS). Meanwhile, the metabolism differences of ORI in the liver microsomes of four different species were investigated using a principal component analysis (PCA) based on the metabolite absolute peak area values as the variables. Based on the proposed methods, 27 metabolites were structurally characterized. The results indicate that ORI is universally metabolized in vitro, and the metabolic pathway mainly includes dehydration, hydroxylation, di-hydroxylation, hydrogenation, decarboxylation, and ketone formation. Overall, there are obvious inter-species differences in types and amounts of ORI metabolites in the four species. These results will provide basic data for future pharmacological and toxicological studies of ORI and for other ent-kauranes diterpenoids. Meanwhile, studying the ORI metabolic differences helps to select the proper animal model for further pharmacology and toxicological assessment.

A pre-classification strategy based on UPLC-Triple-TOF/MS for metabolic screening and identification of Radix glehniae in rats.

Traditional Chinese Medicines (TCMs) have gained increasing popularity in modern society. However, the profiles of TCMs in vivo are still unclear owing to their complexity and low level in vivo. In this study, UPLC-Triple-TOF techniques were employed for data acquiring, and a novel pre-classification strategy was developed to rapidly and systematically screen and identify the absorbed constituents and metabolites of TCMs in vivo using Radix glehniae as the research object. In this strategy, pre-classification for absorbed constituents was first performed according to the similarity of their structures. Then representative constituents were elected from every class and analyzed separately to screen non-target absorbed constituents and metabolites in biosamples. This pre-classification strategy is basing on target (known) constituents to screen non-target (unknown) constituents from the massive data acquired by mass spectrometry. Finally, the screened candidate compounds were interpreted and identified based on a predicted metabolic pathway, well - studied fragmentation rules, a predicted metabolic pathway, polarity and retention time of the compounds, and some related literature. With this method, a total of 111 absorbed constituents and metabolites of Radix glehniae in rats' urine, plasma, and bile samples were screened and identified or tentatively characterized successfully. This strategy provides an idea for the screening and identification of the metabolites of other TCMs.

UPLC-Triple-TOF-MS-based serum metabonomic revealed the alleviating effect of QingYan Formula on perimenopausal syndrome rats.

The study aimed to investigate the relieving effect of QingYan Formula (QYF) in treating perimenopausal syndrome. A combination of metabonomic analysis and in vitro pharmacodynamic experiments was employed to achieve this objective.Over a period of 12 weeks, ovariectomized (OVX) rats were orally administered QYF's 70 % ethanol extract or estradiol valerate (EV). The results demonstrate that QYF restored the estrous cycle of ovariectomized rats and exhibited significant estrogenic activity, as indicated by reversal of uterine and vagina atrophy, improvement of serum estradiol level and decrease of serum luteinizing hormone(LH) level. Additionally, QYF administration effectively reduced high bone turnover and repaired trabecular microstructure damage. Metabonomic analysis of the OVX rats treated with QYF revealed the identification of 55 different metabolites in the serum, out of which 35 may be potential biomarkers. QYF could regulate the disturbed metabolic pathways including the Biosynthesis of unsaturated fatty acids, arachidonic acid metabolism, bile secretion, and steroid hormone biosynthesis. PI3KCA, SRC, and MAPK3 are potential therapeutic targets for QYF therapeutic effects. These findings support the efficacy of QYF in alleviating perimenopausal syndrome and regulating lipid metabolic disorders in OVX rats.

Phenolic composition, antioxidant, cytotoxic activities and cardioprotective effect of hydroalcoholic extract from aerial-parts of Hypericum attenuatum Fisch. ex Choisy.

This study investigated phenolic metabolites, antioxidant, cytotoxic and cardioprotective effects of the hydroalcoholic extract from the aerial parts of Hypericum attenuatum Fisch. ex Choisy. The total phenolic and flavonoid contents of the extract were 132.40 ± 2.06 mg GAE/g and 101.46 ± 1.47 mg QE/g respectively. The extract exhibited antioxidant activities with an EC50 value against DPPH radical of 0.099 ± 0.03 mg/mL and a FRAP value of 1.22 ± 0.086 mmol/L Fe2+. The extract could protect H9c2 cardiomyoblasts from the injury of H2O2, while it restored the H9c2 cell viability to 82.69 ± 2.33% at 100 µg/mL. The extract possessed cytotoxicity on MGC803, C666-1 and SW620 cells with IC50 values of 69.77 ± 2.43 µg/mL, 74.97 ± 1.08 µg/mL and 58.91 ± 1.81 µg/mL, respectively. Moreover, it could promote apoptosis of the tested cancer cells. This research provided useful information for the utilization of H. attenuatum as herbal medicine.

Optimization of subcritical water extraction, UPLC-triple-TOF-MS/MS analysis, antioxidant and α-glucosidase inhibitory activities of anthocyanins from Nitraria sibirica Pall. fruits.

This study aimed to optimize subcritical water extraction process, characterize chemical composition and investigate the biological activities of Nitraria sibirica Pall. (NSP) anthocyanin. Overall, the optimization was achieved under following conditions: extraction temperature 140 °C, extraction time 45 min and flow rate 7 mL/min with the extraction yield of 1.075 mg/g. 3 cyanidin, 3 petunidin, 1 delphinidin and 1 pelargonidin compounds were identified in the anthocyanic extract from NSP via UPLC-Triple-TOF-MS/MS. NSP anthocyanin exhibited better DPPH free-radical scavenging activity than ascorbic acid. It displayed superior α-glucosidase inhibition activity, which was ∼14 times higher than that of acarbose. Moreover, enzyme kinetics results indicated that NSP anthocyanin behaved as a reversible, mixed-type inhibitor. Molecular docking and molecular dynamics simulation results revealed that NSP anthocyanin interacted with α-glucosidase mainly via van der Waals forces, hydrogen bond and possessed fairly stable configuration. Therefore, NSP anthocyanin is a promising α-glucosidase inhibitor for diabetes mellitus.

Characterization of alkaloids and phenolics in Nitraria roborowskii Kom. fruit by UHPLC-triple-TOF-MS/MS and its sucrase and maltase inhibitory effects.

Nitraria roborowskii Kom (NRK), with high economic and ecological value, is mainly distributed in the Qaidam Basin, China. However, research on its chemical components and bioactivities is still rare. In this study, its chemical constituents (52) including 10 ß-carboline alkaloids, nine cyclic peptides, three indole alkaloids, five pyrrole alkaloids, eight phenolic acids and 17 flavonoids were identified tentatively using UPLC-triple-TOF-MS/MS. Notablely, one new ß-carboline alkaloid and five new cyclic peptides were confirmed using MS/MS fragmentation pathways. In addition, experiments in vitro indicated that NRK-C had strong maltase and sucrase inhibitory activities (IC50 of 0.202 and 0.103 mg/mL, respectively). Polysaccharide tolerance experiments confirmed NRK-C (400 mg/kg) was associated with decreased postprandial blood glucose (PBG) in diabetic mice. These results suggested that NRK fruit might be used as a functional ingredient in food products.

Hypoglycaemic activity of the anthocyanin enriched fraction of Lycium ruthenicum Murr. Fruits and its ingredient identification via UPLC-triple-TOF-MS/MS.

Lycium ruthenicum Murr. is mainly distributed in the northwest region of China and its berries are rich in anthocyanin. This study evaluated the hypoglycaemic activity of the anthocyanin-enriched fraction (AEF) of L. ruthenicum Murr. on α-glucosidase in vivo and in vitro. Overall, 10 anthocyanins were identified via UPLC-Triple-TOF-MS/MS. The AEF exhibited strong inhibitory activity against α-glucosidase, with an IC50 value of 4.468 mg/mL. It behaved as a reversible, mixed-type inhibitor. Molecular docking and dynamic results indicated that the compounds in AEF interacted with enzymes primarily through van der Waals and hydrogen bond and the complex system was stable. The postprandial blood glucose and area under the curve of diabetic mice was significantly decreased by AEF in the carbohydrate tolerance experiments. The results indicate that the AEF from L. ruthenicum Murr. berries could be as a promising food supplement for managing blood sugar levels in patients with diabetes mellitus.

Optimized extraction, enrichment, identification and hypoglycemic effects of triterpenoid acids from Hippophae rhamnoides L pomace.

The Hippophae rhamnoides L. pomace was generated in the production process for juice, wine of food industry. To expand the application of pomace, the extraction process optimization, enrichment and identification of triterpene acids were performed in this study. The extraction yield was 14.87% under optimal ultrasound-assisted extraction techniques performed via response surface methodology. The extract was subsequently purified to obtain the triterpenoid acid enrichment fraction (TPF) with the content of 75.23% ± 1.45%. 13 triterpenoid acids were identified via UPLC-Triple-TOF MS/MS and further semi-quantified through comparison with triterpenoid acid standards. TPF exhibited a strong inhibitory effect on α-glucosidase with IC50 value of 5.027 ± 0.375 µg/mL, as determined via enzyme inhibition experiment and molecular docking. Additionally, the TPF significantly reduced postprandial glucose levels, as revealed via carbohydrate tolerance tests, as well as ameliorate serum lipid profiles. Therefore, pomace may be a promising resource of functional food components with therapeutic and commercial values.

Deep eutectic solvent on extraction of flavonoid glycosides from Dendrobium officinale and rapid identification with UPLC-triple-TOF/MS.

In this study, ultrasonic-assisted deep eutectic solvent (DES) method was used to extract flavonoids from Dendrobium officinale. The mechanism of DESs extraction differences was investigated by physicochemical characterization and molecular dynamics simulation experiments. Moreover, flavonoids from Dendrobium officinale were rapidly identified and quantitatively analyzed by UPLC-Triple-TOF/MS without removing DES. The results showed choline chloride-lactic acid (Chol-Lac) had the best extraction effect among forty groups of DESs, the extraction yield was up to 35.23 ± 2.12 mg/g. It was speculated that the tight hydrogen bond structure was the key to the better extraction ability of Chol-Lac. Meanwhile, the viscosity was the main physical parameter reflecting the hydrogen bond structure of DES. Fourteen flavonoid glycosides were identified, among which the content of rutin (1.94 ± 0.24 mg/g) was the highest. And the results of cluster analysis showed that different DESs had great differences in the extraction of flavonoid glycosides.

Effects of soda water on blood lipid and metabolic profiling of urine in hyperlipidemia rats using UPLC/Triple-TOF MS.

The effects of a natural soda water (Shi Han Quan, SHQ) on hyperlipidemia and the changes of urine metabolic profiling by metabolomics techniques were investigate. Thirty six Wistar rats weighing 160-200 g were divided into control group, hyperlipidemia (HL) group, and hyperlipidemia + SHQ water (SHQ) group. The metabolites in urine were determined using ultra high performance liquid chromatography-triple-time of flight-mass spectrometry (UPLC/Triple-TOF MS). At the end of 1 month and 3 months, the total glyceride (TG) level was significantly lower in SHQ group compared to HL group. There was no significantly difference in total cholesterol (TC) levels in HL group compared with SHQ group. The results showed that dinking SHQ water can improve the TG, but with no effects on TC. After drinking SHQ water for 3 months, the rats in different groups could be classified into different clusters according to the metabolites in urine. Total 15 important metabolites were found and correlated with disturbance of amino acid, phospholipid, fatty acid and vitamin metabolism, which suggested the changes of metabolism in the body and possible mechanism by which SHQ improved the TG. These findings provide a new insight for the prevention and control of hyperlipidemia.

Application of UPLC-Triple TOF-MS/MS metabolomics strategy to reveal the dynamic changes of triterpenoid saponins during the decocting process of Asian ginseng and American ginseng.

Triterpenoid saponins are the main bioactive components contributed to the nutritional value of ginseng, and different process conditions will affect their content and quality. To study the holistic characterization and dynamic changes of triterpenoid saponins in Asian ginseng (ASG) and American ginseng (AMG) during soaking and decoction, a UPLC-Triple TOF-MS/MS-based metabolomics strategy was used to characterize and discover differential saponin markers. In total, 739 triterpenoid saponins (including 225 potential new saponins) were identified from ASG and AMG in untargeted metabolomics. Based on PCA and OPLS-DA, 51 and 48 saponin markers were screened from soaked and decocted ASG and AMG, respectively. Additionally, targeted metabolomics analysis and HCA of 22 ginsenoside markers suggested that decoction of ASG and AMG for 2 h to 4 h could significantly increase the contents of rare ginsenosides (G), such as G-Rg3, G-Rg5, G-F4. This study provides a scientific insight that high boiling combined with simmering enriches ASG and AMG extracts with rich rare ginsenosides that are more beneficial to human health.

Acidified glycerol as a one-step efficient green extraction and preservation strategy for anthocyanin from blueberry pomace: New insights into extraction and stability protection mechanism with molecular dynamic simulation.

In present work, green and efficient glycerol solvent system was coupled with pulse-probe ultrasonication for one-step extraction and preservation of anthocyanin from blueberry pomace. Under optimal conditions (40 min, 174 W, 18.6 mL/g, 20% of glycerol fraction), extraction yield was 23.07 ± 0.09 mg C3GE/g DW. The extracted anthocyanins were characterized by UPLC-Triple-TOF/MS and 10 anthocyanins compounds were tentatively identified. Stability of anthocyanins influenced by solvents were evaluated in varying temperature, pH and light exposure conditions, demonstrating higher stability of anthocyanins in glycerol solvent system than methanol one. Furthermore, mechanism of high efficiency extraction and stability of anthocyanin using glycerol were investigated by quantum chemical calculation with molecular dynamic simulation. Larger solvent accessible surface area (127.16 nm2), hydrogen bonds number (228.16) and hydrogen bonds lifetime (4.35 ps), and lower intermolecular interaction energy (-1080.48 kJ/mol) between anthocyanin and glycerol were responsible for better extraction and preservation of anthocyanins using glycerol system.

UPLC-Triple-TOF/MS characterization of phenolic constituents and the influence of natural deep eutectic solvents on extraction of Carya cathayensis Sarg. peels: Composition, extraction mechanism and in vitro biological activities.

Natural deep eutectic solvent (NADES) has received increasing interest as a green alternative to traditional organic solvents for efficient extraction of bioactive compounds from natural sources. In this study, phytochemicals in Carya cathayensis Sarg. peels extracted with Choline chloride-Malic acid (ChCl-MA) were identified using UPLC-Triple-TOF/MS. Effect of NADES on phenolic composition, antioxidant properties and inhibition of α-glucosidase and α-amylase were evaluated. Furthermore, extraction mechanism caused by different solvents were investigated by quantum chemical calculation combined with molecular dynamic simulation. A total of 29 phytochemicals were identified, and catechin, procyanidin B1, 2,3-dihydroxybenzoic acid, pinocembrin, procyanidin B3, myricetrin were the most abundant compounds. The extract using ChCl-MA exhibited the highest phenolic compounds content, antioxidant capacity, and α-glucosidase and α-amylase inhibition activities. Larger solvent accessible surface area, more hydrogen bonds between ChCl-MA and extract, longer lifetime of the hydrogen bonds, and lower intermolecular interaction energy account for higher extraction efficiency of ChCl-MA.

Characterization and quantification of ginsenosides from the root of Panax quinquefolius L. by integrating untargeted metabolites and targeted analysis using UPLC-Triple TOF-MS coupled with UFLC-ESI-MS/MS.

The root of Panax quinquefolius L. (RPQ) is considered as an important functional food and rich in bioactive components, ginsenosides. To comprehensively characterize ginsenosides and evaluate the quality of RPQ from different sources, UPLC-Triple TOF-MS coupled with UFLC-ESI-MS/MS was applied to untargeted metabolites and targeted analysis for the first time. In untargeted metabolites analysis, a total of 225 ginsenosides were identified from RPQ using UPLC-Triple TOF-MS combined with SWATH data-independent strategy. Furthermore, the contents of 39 targeted ginsenoside markers in 14 RPQ samples were analyzed by a rapid and sensitive UFLC-ESI-MS/MS method. In addition, the results of chemometric analysis showed the quality of American RPQ was distinguished from that of Chinese RPQ according to the amount of targeted ginsenosides. This newly developed approach provides a powerful tool for enriching the diversity of saponins database and assessing the quality of RPQ, which can be further extended to other ginseng products and functional foods.

Effects of the Oxygen Content and Light Intensity on Milk Photooxidation Using Untargeted Metabolomic Analysis.

Photooxidation is the main cause of milk quality deterioration during processing and transportation. Oxygen and light are responsible for dairy milk photooxidation. The objective of this study was to determine the characteristic metabolites after photooxidation and how they are affected by oxygen exposure and light intensity. Ultra-performance liquid chromatography coupled with triple time-of-flight mass spectrometry and multivariate data analysis were used for the high-throughput evaluation of milk photooxidation. Four products were identified as biomarkers: uric acid, riboflavin, lumichrome, and indole-3-carboxaldehyde. Afterward, the effects of oxygen content and light intensity on the biomarkers were investigated, and a sensory evaluation was performed. Both oxygen exposure and light intensity affected the contents of photooxidation biomarkers in milk samples. The sensory score correlated well with the oxygen content but not with the light intensity. The untargeted metabolomic method was an effective tool to identify biomarkers for milk photooxidation evaluation.

Comparative lipid profile of four edible shellfishes by UPLC-Triple TOF-MS/MS.

An ultra performance liquid chromatography-Triple time of flight mass spectrometry (UPLC-Triple TOF-MS/MS) method were established to characterize the lipid profiles in four shellfish species. More than 600 lipid molecular species belonging to 14 classes were detected. Phospholipids (PLs) were predominant in Chlamys farreri (54.9%) and glycerolipids (GLs) were dominant in Ostrea gigas (51.6%). PLs that contained polyunsaturated fatty acids (PUFAs) such as PC (16:0/20:5), PC (16:0/22:6) and PE (18:0/22:6) were the main molecular species. Especially, the percentage of sphingolipids (SLs) in four shellfishes is considerable (18.8-38.6%), the characterization of their special long-chain base (LCB) structure (mainly d19:3) and N-acyl group (mainly 16:0) was realized. Several SL subclasses with low abundance in four shellfish species, such as ceramide 2-aminoethylphosphonate (CAEP) and deoxy-ceramide (DeoxyCer), were also detected. These active lipids identified by this method have potential value in revealing the nutritional value of shellfishes and serving as biomarkers for distinguishing different shellfishes.