Comparative Analysis of the Effect of Dietary Supplementation with Fermented and Water-Extracted Leaf Extracts of Eucommia ulmoides on Egg Production and Egg Nutrition.

Although the water extract of Eucommia ulmoides leaf (WEE) promotes egg laying in hens, its palatability is poor. To improve the palatability of E. ulmoides leaf, probiotic fermentation was used, and fermented extract E. ulmoides leaf (FEE) was prepared using Lactiplantibacillus plantarum. The safety of FEE was investigated using a long-term toxicity test, and no oxidative damage, inflammatory reactions, or histological lesions were observed in the experimental rats receiving dietary supplementation of FEE at 200 mg/kg, suggesting that FEE is suitable for long-term feeding. Subsequently, dietary supplementation of FEE (group C) in comparison with dietary supplementation of WEE (group B), as well as a control (group A), was applied in the hen industry. Laying performance, egg quality, egg nutrition, egg flavor, and the gut microbiome were analyzed comparatively. Interestingly, the laying rate was observed to be four percentage points higher with dietary supplementation of FEE at 200 mg/kg compared with the control and two percentage points higher compared with the dietary addition of WEE at the same dosage. Simultaneously, a slight upregulation in daily feed consumption was determined in the FEE-supplemented group compared with the blank control and the WEE-supplemented group, indicating that the inclusion of FEE stimulated the hens' appetite. Moreover, variations in egg amino acids, fatty acids, and volatile components were obtained with either dietary addition, FEE or WEE, implying that dietary supplementation of the fermented and water-extracted E. ulmoides leaf extracts contributed to egg flavor change. Furthermore, variations in the gut microbiota were mediated by FEE, increasing the relative abundance of the genus Lactobacillus. These alterations in gut microbiota were tightly related to improved laying performance and egg flavor changes. Our results indicate that FEE is a better alternative feed additive in the hen industry than WEE.

Effect of dietary supplementation Ampelopsis grossedentata extract on growth performance and muscle nutrition of Megalobrama hoffmanni by gut bacterial mediation.

Nowadays, Megalobrama hoffmanni is a typical cultured fish in south China due to its resource decline in the Pearl River. Meanwhile, since antibiotics had been banned internationally, Chinese medical herbal plant serving as alternative to antibiotics has been adopted in aquaculture. In the present study, to ensure the health growth of M. hoffmanni, extract of traditional medical herbal plant Ampelopsis grossedentata was dietary supplemented and a series experiments were performed, including growth performance determination, physiological/biochemical detection, nutrition analysis, histology analysis, and 16S rRNA amplicon sequencing. Growth performance enhancement was determined since the weight gain rate (WGR), specific growth rate (SGR), and condition factor (CF) of M. hoffmanni increased as feeding inclusion A. grossedentata extract. Interestingly, the total content of muscle fatty acids ascended via supplementing A. grossedentata extract at middle level, in which group the activities of superoxide dismutase (SOD) and catalase (CAT) significantly increased and thus retarded the lipid peroxidation process (manifesting as malondialdehyde (MDA) content rising). Additionally, immune response and inflammatory reaction was stimulated in low and high level A. grossedentata extract added groups, indicating a suitable dosage of A. grossedentata extract benefited in safety production. Moreover, gut microbiota community varied hugely as daily supplementation A. grossedentata extract and the keystone species were tightly related to lipid transformation, which ultimately led to fatty acids composition variation. Our results confirmed that dietary supplementation A. grossedentata extract at the middle level (0.5‰, w/w) is suitable for serving as feed additive in healthful aquaculture of M. hoffmanni.

Effect of Fermented Artemisia argyi on Egg Quality, Nutrition, and Flavor by Gut Bacterial Mediation.

To improve the palatability of Artemisia argyi, fermented A. argyi (AAF) were prepared by Lactobacillus plantarum and Saccharomyces cerevisiae, which were used in the hen industry subsequently. Six hundred hens were randomly divided into three groups: control (A), dietary supplementation AAF at a low level (B), and dietary supplementation AAF at a high level (C). After feeding for four months, egg production, egg quality, egg nutrition, egg flavor, plasma biochemical parameters, intestinal histology, and microbiome of the gut contents were analyzed among the three tested groups. Interestingly, 5-6 percentage points elevation in the laying rates were observed in the AAF-supplemented groups in comparison to the control, accompanied with a 5 g increase in daily feed consumption. Since no alteration in egg/body weights was detected, laying performance enhancement was the main effect of dietary supplementation AAF. Meanwhile, the compositions of the egg amino acids and fatty acids changed as the feed inclusion AAF changed, e.g., His and linoleic acid decreased almost 0.1 and 0.5 g/100 g, respectively, while oleic acid increased almost 0.4 g/100 g. In addition, although no significant difference was detected (p > 0.05), the ß-diversity of the gut microbiota decreased as the diet addition of AAF decreased, and probiotics (Faecalibacterium, Prevotellaceae, Intestinimonas, and Lachnospiraceae) were the dominant keystone species under AAF treatments. These probiotics were well associated with the egg nutrition component variations based on the correlation analysis, as the Sankey plot showed. Furthermore, the results of headspace-gas chromatography-ion mobility spectrometry manifested that the egg volatile components varied (e.g., the contents of acetone, 4-methyl-3-penten-2-one, 1-hydroxy-2-propanone, ethyl acetate, ethyl octanoate, ethanol, and 2-butanol in the B and C groups were higher than in the A group) and separated clearly as daily supplementation AAF, indicating AAF hugely contributed to the egg flavor variation. Due to no significant differences noticed between the B and C groups, dietary supplementation AAF at a relative low level was enough to serve as a feed attractant in the hen industry for real feeding.

The effect of quercetin on diabetic nephropathy (DN): a systematic review and meta-analysis of animal studies.

Quercetin, a flavonoid possessing numerous biological activities, is reported to improve renal injury in diabetic animals. Here, the aim of this systematic review and meta-analysis is to assess the effect of quercetin on diabetic nephropathy and summarize its possible mechanisms. We searched in four databases PubMed, Web of Sciences (WOS), Cochrane and Embase from inception to May 2021 and ultimately included 20 animal studies in this review. A total of 12 outcome measurements including renal function indexes, oxidative stress biomarkers and inflammatory cytokines were extracted for meta-analysis using RevMan 5.4 software. Apart from creatinine clearance and uric acid with no significant difference, quercetin significantly decreased the levels of renal index, serum/plasma creatinine (SCr), blood urea nitrogen (BUN), urine protein, urine albumin, malondialdehyde (MDA), tumor necrosis factor (TNF)-α and interleukin (IL)-1ß, and increased superoxide dismutase (SOD) and catalase (CAT) activity. In short, quercetin improves renal function and attenuates the renal oxidative stress level and inflammatory response in DN animal models. Its possible action mechanisms include anti-oxidation, anti-inflammation, anti-fibrosis, and regulation of renal lipid accumulation.

Dietary supplementation Eucommia ulmoides extract at high content served as a feed additive in the hens industry.

Since numerous natural components in Eucommia ulmoides belong to phytoestrogen, its effect on hens production deserve more attention. To investigate the potential of E. ulmoides extract used as a feed additive, laying performance, egg quality, yolk cholesterol, yolk fatty acids, yolk fatty, yolk volatile components, albumen amino acids, plasma biochemical parameters, intestinal histology, and gut microbiota of hens (n = 120) were determined between basal diet (A) and dietary supplementation low (B), middle (C), and high (D) level E. ulmoides extract for 11 wk. When compared to A group, 2 percentage points elevation in laying rate was observed of D group. Significant up-regulation of immunoglobulin indexes and down-regulation of lipid related indexes in D group were also found if comparison with A group, suggesting that supplementation E. ulmoides extract at a relative high content benefited in immunity enhancing and blood-fat depressing. Meanwhile, obvious variation in albumen amino acids and yolk volatile compounds were inspected as dietary supplementation E. ulmoides extract, especially in D group, implied that the flavor of egg would change under high-level E. ulmoides extract treatment. Besides, villus height and villus height to crypt depth ratio of duodenum, jejunum, and ileum in D group were also significantly higher than that of in A group, indicating high-level E. ulmoides extract contributed to nutrient adsorption via intestinal histology changing. Moreover, the richness, diversity, and composition of gut microbiota in D group also significantly altered with a comparison of A group. These variation caused gut microbiota in D group major enriched in the KEGG pathway of insulin signing pathway, systemic lupus erythematosus, and bacterial invasion of epithelial cells, which were conducive to egg production elevation via facilitating nutrient adsorption, inflammation relieving, blood lipid amelioration, and insulin resistance alleviation. These results indicated that dietary supplementation E. ulmoides extract at high content could serve as a feed additive in the hens industry.

Effects of dietary supplementation with Ampelopsis grossedentata extract on production performance and body health of hens.

To investigate the potential of Ampelopsis grossedentata extract used as a feed additive, laying performance, egg quality, yolk cholesterol, plasma biochemical parameters, intestinal histology, and gut microbiota of hens (n = 60) were determined between basal diet (CK) and dietary supplementation with A. grossedentata extract (RT) for 11 weeks. The laying rate in RT group was 6.3 percentage points higher than in CK group together with feed conversion rate decreasing. Significant upregulation of immunoglobulin indexes and downregulation of lipid-related indexes in RT group were also found in comparison with CK group, suggesting that dietary supplementation with A. grossedentata extract benefited in immunity enhancing and blood-fat depressing. Meanwhile, the villus height in duodenum and villus height to crypt depth ratio in duodenum and jejunum of RT group were significantly higher than that of CK group, indicating that dietary supplementation with A. grossedentata extract facilitated nutrient adsorption via intestinal histology changing. Moreover, the richness, diversity, and composition of gut microbiota in RT group significantly altered with a comparison of CK group, including beneficial bacterium and pathogenic bacterium, revealing that dietary supplementation with A. grossedentata extract could modify gut microbiota communities to affect intestinal adsorption and pathogen invasion. In addition, the lipid metabolism-related insulin signing pathway was significantly enriched by gut microbiota in RT group, which were conducive to egg production elevation via facilitating blood lipid amelioration and insulin resistance alleviation. These results provided a basis for A. grossedentata extract served as a feed additive in the hen industry.

Adsorption and desorption characteristics of polyphenols from Eucommia ulmoides Oliv. leaves with macroporous resin and its inhibitory effect on α-amylase and α-glucosidase.

BACKGROUND: Eucommia ulmoides Oliv. (EUO) was a traditional Chinese herb, its leaves were abundant in China, and polyphenol compounds were considered to be an important active ingredient in Eucommia ulmoides Oliv. leaves (EUOL). However, previous research mainly focused on compound identification and extraction process, there were few reported on the efficient enrichment process and biological activity evaluation of polyphenols in EUOL. METHODS: The adsorption and desorption characteristics of twelve different resins (HPD-100, HPD-300, HPD-600, D-3250, X-5, D-140, NKA-9, NKA-II, D-101, AB-8, S-8 and Polyamide) were investigated to develop an efficient method for the enrichment of polyphenol from EUOL, and the static adsorption, kinetics, isotherm and thermodynamics of the polyphenol from EUOL were analyzed. The eluted component was obtained through dynamic elution, and its main polyphenol compounds were detected by high-phase liquid chromatography (HPLC) and the inhibitory effects on the enzyme activity of α-amylase and α-glucosidase was also evaluated for different elution components. Meanwhile, the binding of main polyphenol compounds to enzyme was also evaluated. RESULTS: The selected resins (HPD-300, HPD-600, D-3250, X-5, D-140, NKA-9, D-101 and AB-8) showed adsorption patterns that fitted well to the pseudo second-order kinetics. The intra-particle diffusion model demonstrated that the diffusion of polyphenol compounds on these resins were divided into three processes. For HPD-300, HPD-600 and NKA-9, the Freundlich model better described the adsorption isotherm behavior than the Langmuir model, and the adsorption of polyphenol was a physical, exothermic, and spontaneous process. Subsequently, dynamic elution was performed yielding a higher polyphenol content in a 60% ethanol-water elution component, and it also exhibited a higher inhibitory effect on α-amylase and α-glucosidase activity. Furthermore, as the main polyphenol compounds, chlorogenic acid, rutin, quercetin and kaempferol were used to simulate the binding to the enzyme protein through molecular docking technology. The results showed that quercetin had a higher docking score for α-amylase, while rutin displayed superior binding to α-glucosidase. CONCLUSIONS: Therefore, polyphenols of EUOL could be enriched through macroporous resins and have the potential to be effective enzyme inhibitor.

Isolation of polyphenol compounds from olive waste and inhibition of their derivatives for α-glucosidase and α-amylase.

Olive waste was used as a sustainable resource because it contained a variety of valuable compounds. The polyphenols active fraction from enrichment by microporous resin and extraction with ethyl acetate were analysed by different chromatographic methods. A total of 14 polyphenolic compounds were isolated and identified by structure elucidation. Based on the above obtained compounds, tyrosol was selected as a characteristic polyphenol and participated in transesterification reaction to synthesise ß-ketoester using Yb(OTf)3. Then the Biginelli reaction with benzaldehyde, urea and ketoester (1:1.2:1.2) was performed at 90 °C for 3.0 h under the acidic condition. In addition, the ß-ketoester prepared using tyrosol with benzyl had a greater inhibitory effect on α-glucosidase and α-amylase, and the inhibition of enzyme activity for 3, 4-dihydropyrimidinone derivatives prepared using abovementioned ß-ketoester was improved significantly. Meanwhile, fluorine-containing dihydropyrimidinone derivatives were considerable inhibitors for both enzymes.

Dietary supplementation with the extract from Eucommia ulmoides leaves changed epithelial restitution and gut microbial community and composition of weanling piglets.

This study was conducted to compare the effects of Eucommia ulmoides leaves (EL) in different forms (EL extract, fermented EL, and EL powder) with antibiotics on growth performance, intestinal morphology, and the microbiota composition and diversity of weanling piglets. Compared to the control group, the antibiotics and EL extract significantly increased the average daily gain and decreased the feed: gain ratio as well as the diarrhea rate (P < 0.05). The EL extract significantly decreased the crypt depth and increased the ratio of villus height to crypt depth (P < 0.05), while the fermented EL group did the opposite (P < 0.05). The crypt depth in the antibiotics group was of similar value to the EL extract group, and was lower than the fermented EL and EL powder groups (P < 0.05). Compared to the control and antibiotics groups, the jejunul claudin-3 mRNA expression and the concentrations of total VFA, Chao 1, and ACE were significantly augmented in the EL extract group of piglets (P < 0.05). The EL extract groups also showed elevated Shannon (P < 0.05) and Simpson (P = 0.07) values relative to the control and antibiotics groups. At the phylum level, the EL extract group exhibited a reduced abundance of Bacteroidetes and an enhanced abundance of Firmicutes. At the genus level, the abundance of Prevotella was augmented in the EL extract group. Moreover, compared with the antibiotic group, the acetate concentration was enhanced in the EL extract and fermented EL groups. Overall, dietary supplementation with the EL extract, but not the fermented EL or EL powder, improved growth performance, jejunul morphology and function, as well as changed colonic microbial composition and diversity, which might be an alternative to confer protection against weanling stress in weanling piglets.

Molecularly Imprinted Polymers-Coated CdTe Quantum Dots for Highly Sensitive and Selective Fluorescent Determination of Ferulic Acid.

Ferulic acid (FA), an important phenolic acid, is widely distributed in higher plants and presents many pharmacological effects. Therefore, sensitive determination of FA in complex matrix is necessary. Molecularly imprinted polymers-coated CdTe quantum dots (CdTe-QDs@MIPs) exhibited incomparable advantages because of their combination of excellent selectivity of MIPs and high sensitivity of QDs. Here, a fluorescent probe based on CdTe-QDs@MIPs was successfully fabricated for selective and sensitive determination of FA. MIPs shell was obtained by the reverse microemulsion method using FA, 3-(aminopropyl) triethoxysilane (APTES), and tetraethyl orthosilicate (TEOS), as template, functional monomer, and crosslinker. In optimal conditions, the fluorescence CdTe-QDs@MIPs sensor exhibited fast response (within only 3 min), high sensitivity (limit of detection, LOD at 0.85 µg/l), excellent linear ranges (2-100 µg/l) with a correlation coefficient of 0.9996, and distinguished selectivity for FA. Satisfactory recoveries from 91.8% to 110.3% were achieved with precisions below 6.6% for FA analysis in real pineapple juice and apple juice by developed CdTe-QDs@MIPs. The fluorescence results coincided well with those obtained by high-performance liquid chromatography (HPLC). It could be concluded that the resultant CdTe-QDs@MIPs offered a new way for rapid and sensitive analysis of FA in the complex matrix.

Online extraction-HPLC-FRAP system for direct identification of antioxidants from solid Du-zhong brick tea.

Du-zhong brick tea (DBT), a new post-fermented functional tea, is marketed in China for its prevention of diseases from oxidative stress. However, antioxidant compounds in DBT are not clear. Here, a novel online extraction-HPLC-ferric reducing antioxidant power (OLE-HPLC-FRAP) has been developed to screen antioxidants in DBT. For OLE, guard column is packed with DBT (1.0 mg), then compounds are extracted directly by HPLC mobile phase for HPLC-FRAP analysis without sample pretreatment procedures. Eleven antioxidants have been screened and identified, three of which are detected in Du-zhong for the first time. Contributions of eleven antioxidant compounds to total antioxidant activity are evaluated using chlorogenic acid as antioxidant marker. Results indicate that the developed OLE-HPLC-FRAP system is a simple and powerful tool to analyze antioxidant compounds from solid complex mixtures, which also contribute to the enhancement of the value of DBT as a good functional tea.

[Rapid identification of flavonoid compounds in Pericarpium Citri Reticulatae by online extraction-high performance liquid chromatography-diode array detection-quadrupole time-of-flight mass spectrometry].

A simple method was developed for the rapid extraction, separation and identification by online extraction (OLE) coupled with high performance liquid chromatography-diode array detection-quadrupole time-of-flight mass spectrometry (HPLC-DAD-QTOF-MS). For OLE strategy, a guard column filled with Pericarpium Citri Reticulatae powder (2.0 mg) was connected to manual injection valve by replacing the sample loop. Components were directly extracted and transferred to HPLC-DAD-QTOF-MS system by mobile phases without any extra operation. In comparison with offline heat-reflux extraction, OLE strategy presented higher extraction efficiency. A total of twenty-four flavonoid compounds were detected from Pericarpium Citri Reticulatae according to their retention times, UV spectra, accurate molecular masses, and fragment ions in MS/MS spectra. Neoeriocitrin, limocitrin-3-O-(3-hydroxy-3-methylglutarate)-glucoside and its isomer were discovered for the first time. It is concluded that the developed method offers new perspectives for the rapid separation and identification of natural products.

ß-Hydroxy-ß-methylbutyrate modulates lipid metabolism in adipose tissues of growing pigs.

BACKGROUND: The effects and roles of the leucine (Leu) metabolite ß-hydroxy-ß-methylbutyrate (HMB) in lipid metabolism in adipose tissues of pigs are still unknown. OBJECTIVES: This study was conducted to investigate the effects of excess Leu versus HMB on growth, carcass traits, and lipid metabolism in adipose tissues of growing pigs. METHODS AND RESULTS: Compared to control, the Leu/HMB group significantly increased/reduced weight of total fat mass, respectively, with a concurrent increase of serum adiponectin concentration (P < 0.05). Moreover, dietary HMB supplementation regulated the expression of genes involved in adipose tissue function, accompanied by increases/decreases in the phosphorylation of AMPKα/mTOR in perirenal adipose tissue, respectively (P < 0.05). Serum IL-15 concentration and the mRNA abundance of IL-15, PGC-1α, and NRF-1 were also increased in the HMB group (P < 0.05). CONCLUSIONS: HMB supplementation can regulate adipose tissue function including fatty acid oxidation, lipolysis, and adipokine secretion. These effects may be partly mediated by AMPKα-mTOR pathway and associated with mitochondrial biogenesis, the AMPK-PGC-1α axis, and myokines secreted by muscle tissues.

Direct from solid natural products to pure compounds in a single step: Coupling online extraction with high-speed counter-current chromatography.

Extraction is the most important step in the purification of bioactive compounds from natural products. This study introduces a simple online extraction strategy coupled with high-speed counter-current chromatography for efficient extraction and purification of bioactive components from solid natural products. For online extraction strategy, 1.0 g of ground Mangnolia officinalis or Piper nigrum was loaded into a guard column, which was then positioned on the manual injection valve instead of the sample loop. Bioactive components were directly extracted by the mobile phase of high-speed counter-current chromatography, and then transferred into high-speed counter-current chromatography for purification. In addition, the compatibility of the developed methodology for direct purification of bioactive components from fresh M. officinalis was successfully demonstrated. Obviously, in comparison with traditional offline heat-reflux extraction, online extraction avoided the instrument, time, solvent, and energy consumption, and purified two phenolic compounds (honokiol and magnolol) from M. officinalis and three alkaloids (piperyline, piperine, and piperanine) from P. nigrum with high extraction efficiency. The superiority of the developed methodology is to establish an easy, rapid, and efficient technique for the purification of a wide variety of bioactive components from solid natural products.

Magnetic Porous Molecularly Imprinted Polymers Based on Surface Precipitation Polymerization and Mesoporous SiO2 Layer as Sacrificial Support for Efficient and Selective Extraction and Determination of Chlorogenic Acid in Duzhong Brick Tea.

Magnetic porous molecularly imprinted polymers (MPMIPs) for rapid and efficient selective recognition of chlorogenic acid (CGA) were effectively prepared based on surface precipitation polymerization using CGA as template, 4-vinylpyridine (4-VP) as functional monomer, and mesoporous SiO2 (mSiO2) layer as sacrificial support. A computational simulation by evaluation of electronic binding energy is used to optimize the stoichiometric ratio between CGA and 4-VP (1:5), which reduced the duration of laboratory trials. The porous MIP shell and the rid of solid MIPs by magnet gave MPMIPs high binding capacity (42.22 mg/g) and fast kinetic binding (35 min). Adsorption behavior between CGA and MPMIPs followed Langmuir equation and pseudo-first-order reaction kinetics. Furthermore, the obtained MPMIPs as solid phase adsorbents coupled with high performance liquid chromatography (HPLC) were employed for selective extraction and determination of CGA (2.93 ± 0.11 mg/g) in Duzhong brick tea. The recoveries from 91.8% to 104.2%, and the limit of detection (LOD) at 0.8 µg/mL were obtained. The linear range (2.0⁻150.0 µg/mL) was wide with R² > 0.999. Overall, this study provided an efficient approach for fabrication of well-constructed MPMIPs for fast and selective recognition and determination of CGA from complex samples.

Online extraction-high performance liquid chromatography-diode array detector-quadrupole time-of-flight tandem mass spectrometry for rapid flavonoid profiling of Fructus aurantii immaturus.

Chemical profiling of natural products by high performance liquid chromatography (HPLC) was critical for understanding of their clinical bioactivities, and sample pretreatment steps have been considered as a bottleneck for analysis. Currently, concerted efforts have been made to develop sample pretreatment methods with high efficiency, low solvent and time consumptions. Here, a simple and efficient online extraction (OLE) strategy coupled with HPLC-diode array detector-quadrupole time-of-flight tandem mass spectrometry (HPLC-DAD-QTOF-MS/MS) was developed for rapid chemical profiling. For OLE strategy, guard column inserted with ground sample (2 mg) instead of sample loop was connected with manual injection valve, in which components were directly extracted and transferred to HPLC-DAD-QTOF-MS/MS system only by mobile phase without any extra time, solvent, instrument and operation. By comparison with offline heat-reflux extraction for Fructus aurantii immaturus (Zhishi), OLE strategy presented higher extraction efficiency perhaps because of the high pressure and gradient elution mode. A total of eighteen flavonoids were detected according to their retention times, UV spectra, exact mass, and fragmentation ions in MS/MS spectra, and compound 9, natsudaidain-3-O-glucoside, was discovered in Zhishi for the first time. It is concluded that the developed OLE-HPLC-DAD-QTOF-MS/MS system offers new perspectives for rapid chemical profiling of natural products.

Use of an online extraction liquid chromatography quadrupole time-of-flight tandem mass spectrometry method for the characterization of polyphenols in Citrus paradisi cv. Changshanhuyu peel.

Chemical profiling of natural products by high performance liquid chromatography (HPLC) was critical for understanding of their clinical bioactivities, and sample pretreatment steps have been considered as a bottleneck for analysis. Currently, concerted efforts have been made to develop sample pretreatment methods with high efficiency, low solvent and time consumptions. Here, a simple and efficient online extraction (OLE) strategy coupled with HPLC-diode array detector-quadrupole time-of-flight tandem mass spectrometry (HPLC-DAD-QTOF-MS/MS) was developed for rapid chemical profiling. For OLE strategy, guard column inserted with ground sample (2 mg) instead of sample loop was connected with manual injection valve, in which components were directly extracted and transferred to HPLC-DAD-QTOF-MS/MS system only by mobile phase without any extra time, solvent, instrument and operation. By comparison with offline heat-reflux extraction of Citrus paradisi cv. Changshanhuyu (Changshanhuyu) peel, OLE strategy presented higher extraction efficiency perhaps because of the high pressure and gradient elution mode. A total of twenty-two secondary metabolites were detected according to their retention times, UV spectra, exact mass, and fragmentation ions in MS/MS spectra, and nine of them were discovered in Changshanhuyu peel for the first time to our knowledge. It is concluded that the developed OLE-HPLC-DAD-QTOF-MS/MS system offers new perspectives for rapid chemical profiling of natural products.

Synthesis and characterization of hollow porous molecular imprinted polymers for the selective extraction and determination of caffeic acid in fruit samples.

Hollow porous molecular imprinted polymers (HPMIPs) were prepared using caffeic acid (CA) as template, 4-vinylpyridine (4-VP) as functional monomer, and MCM-48 as sacrificial support. Large surface area (562.7m2/g) of HPMIPs resulted in high binding capacity (27.26mg/g) and fast kinetic binding (40min) in comparison with solid MIPs. Equilibrium data fitted well to Freundlich equation, and adsorption process with multi-diffusion mechanisms could be described by pseudo-second order model. Selectivity of HPMIPs was favorable. Finally, HPMIPs were successfully used to rapidly and selectively extract CA from fruits with a relatively satisfactory recovery (87.1-101.3%). Coupling with HPLC, contents of CA in kiwifruit, apple, papaya and waxberry were less than 1.0µg/g fresh fruit. Results indicated the superiority of HPMIPs in the separation field.

High-capacity hollow porous dummy molecular imprinted polymers using ionic liquid as functional monomer for selective recognition of salicylic acid.

The existence of strong intramolecular hydrogen bond in salicylic acid (SA) weakens its intermolecular hydrogen bonding with functional monomer, then it is a challenge work to fabricate molecularly imprinted polymers (MIPs) for SA recognition with high capacity and good selectivity. Here, hollow porous dummy MIPs (HPDMIPs) were prepared using benzoic acid (BA) as dummy template, ionic liquid (i.e. 1-vinyl-3-methylimidazolium chloride) as functional monomer, and MCM-48 as sacrificial support. Factors that affected adsorption, such as type of template and porogen, mole ratio of template-functional monomer-cross-linker and type of binding solvent, were optimized in detail. Multiple strong interactions between SA and ionic liquid in HPDMIPs deduced higher binding capacity (29.75mg/g), imprinting factor (5.61) and selectivity than any previously reported MIPs by traditional or surface imprinting technology. The large surface area (543.9m2/g) with hollow porous structure resulted in faster kinetic binding (25min). The equilibrium data fitted well to Freundlich equation and the adsorption process could be described by pseudo-second order model. Finally, HPDMIPs were successfully applied to selectively extract and enrich SA from Actinidia chinensis with a relatively high recovery (84.6-94.5%).

Boronate affinity-based surface molecularly imprinted polymers using glucose as fragment template for excellent recognition of glucosides.

Rapid and efficient extraction of bioactive glycosides from complex natural origins poses a difficult challenge, and then is often inherent bottleneck for their highly utilization. Herein, we propose a strategy to fabricate boronate affinity based surface molecularly imprinted polymers (MIPs) for excellent recognition of glucosides. d-glucose was used as fragment template. Boronic acid, dynamic covalent binding with d-glucose under different pH conditions, was selected as functional monomer to improve specificity. Fe3O4 solid core for surface imprinting using tetraethyl orthosilicate (TEOS) as crosslinker could control imprinted shell thickness for favorable adsorption capacity and satisfactory mass transfer rate, improve hydrophilicity, separate easily by a magnet. Model adsorption studies showed that the resulting MIPs show specific recognition of glucosides. The equilibrium data fitted well to Langmuir equation and the adsorption process could be described by pseudo-second order model. Furthermore, the MIPs were successfully applied for selective extraction of three flavonoid glucosides (daidzin, glycitin, and genistin) from soybean. Results indicated that selective extraction of glucosides from complex aqueous media based on the prepared MIPs is simple, rapid, efficient and specific. Moreover, this method opens up a universal route for imprinting saccharide with cis-diol group for glycosides recognition.