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Anthocyanins are a group of phytochemicals responsible for the purple or red color of plants. Additionally, they are recognized to have health promoting functions including anti-cardiovascular, anti-thrombotic, anti-diabetic, antimicrobial, neuroprotective, and visual protective effect as well as anti-cancer activities. Thus, consumption of anthocyanin supplement or anthocyanin-rich foods has been recommended to prevent the risk of development of chronic diseases. However, the low stability and bioavailability of anthocyanins limit the efficacy and distribution of anthocyanins in human body. Thus, strategies to achieve target site-local delivery with good bioavailability and controlled release rate are necessary. This review introduced and discussed the latest advanced techniques of designing lipid-based, polysaccharide-based and protein-based complexes, nano-encapsulation and exosome to overcome the limitation of anthocyanins. The improved bioavailability and controlled release of anthocyanins have great significance for gastrointestinal tract absorption, transepithelial transportation and cellular uptake. The techniques of applying different biocompatible materials and modifying the solubility of anthocyanins complex could achieve target site-local delivery with negligible degradation and good bioavailability in human body.

The kiwi berry (Actinidia arguta) is a new product on the market that expanding worldwide acceptance and consumption. This widespread interest has created an increasing demand to identify the nutritional and health benefits of kiwi berry. Many studies are being actively conducted to investigate the composition and health‐promoting effects of kiwi berry. In this study, the phytochemical content of free and bound fractions of eight kiwi berry varieties were systematically investigated in order to better understand the potential of this superfood crop. Nine phenolic monomers were identified and quantified by ultrahigh‐performance liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry and ultrahigh‐performance liquid chromatography‐PAD. Antioxidant activity was further determined via peroxyl radical scavenging capacity and cellular antioxidant activity assays. The free extracts had higher phytochemical contents and antioxidant activities than the corresponding bound extracts among the eight kiwi berry varieties. Bivariate Pearson's and multivariate correlation analyses showed that antioxidant activities were most related to the total phenolic, flavonoid, vitamin C, and phenolic acids contents. The results provide a theoretical basis for the selection of kiwi berry varieties and the utilization of functional foods., This study aimed to assess and summarize the phytochemical profile and antioxidant activities of eight common kiwi berry varieties in China, exploring their potential functional properties in foodstuffs. The results demonstrate the differences in phytochemical profiles and antioxidant activities among different kiwi berry varieties, which will help to form useful guidance for plant breeders, food scientists, and consumers to provide a theoretical basis for the selection of kiwi berry varieties and the development and utilization of functional foods.

Mulberry leaf is a common vegetable with a variety of beneficial effects, such as hypoglycemic activity. However, the underlying mechanism of its hypoglycemic effect have not been fully revealed. In this study, two flavonoid derivatives were isolated from mulberry leaves, a new geranylated flavonoid compound (1) and its structural analogue (2). The structures of compounds 1 and 2 were elucidated using spectroscopic analysis. To study the potential hypoglycemic properties of these compounds, their regulatory effects on protein tyrosine phosphatase 1B (PTP1B) were investigated. In comparison to oleanolic acid, compounds 1 and 2 showed significant inhibitory activities (IC50 = 4.53 ± 0.31 and 10.53 ± 1.76 μM) against PTP1B, the positive control (IC50 = 7.94 ± 0.76 μM). Molecular docking predicted the binding sites of compound 1 to PTP1B. In insulin-resistance HepG2 cell, compound 1 promoted glucose consumption in a dose-dependent manner. Furthermore, western blot and polymerase chain reaction analyses indicated that compound 1 might regulate glucose consumption through the PTP1B/IRS/PI3K/AKT pathway. In conclusion, geranylated flavonoids in mulberry leaves inhibite PTP1B and increase the glucose consumption in insulin-resistant cells. These findings provide an important basis for the use of mulberry leaf flavonoids as a dietary supplement to regulate glucose metabolism.

Malvidin-3-galactoside (Mv-3-gal) is the major anthocyanin monomer in blueberry anthocyanins. The compound is well-characterized by its anti-oncogenesis function in multiple organs including liver. In the current study, the mechanism driving the anti-hepatocellular carcinoma (HCC) function of Mv-3-gal was further explored by focusing on apoptosis and metastases pathways. HCC cell line Huh-7 was firstly administrated with Mv-3-gal of different concentrations and the effect of Mv-3-gal on cell proliferation, apoptosis, colony formation ability, metastasis potential as well as the activity of Akt/PTEN and MAPK pathways was assessed. The results showed that Mv-3-gal inhibited the cell proliferation and colony formation ability, induced cell cycle arrest and apoptosis in a dose-dependent manner. Regarding the metastasis potential, Mv-3-gal suppressed the migration and invasion potential of Huh-7 cells by regulating MMPs expression. Taken together, the findings highlighted the anti-HCC potent of Mv-3-gal, which was associated with the inhibition of Akt/PTEN, MAPK and MMP pathways.

Solanum rostratum is a worldwide malignant invasive weed, causing serious harm to the ecological environment and biodiversity. Strong chemical defense against herbivorous insects is supposed to be one of the successful invasive mechanisms of this exotic plant. However, the real defense components and their action mechanisms and distributions are still unknown. To address these problems, we bioassay-guided isolated compounds from the aerial part of S. rostratum and determined their structures using high-resolution electrospray ionization mass spectrometry, nuclear magnetic resonance, and electronic circular dichroism calculation. One new and seven known compounds were identified, and all of the isolates exhibited different levels of antifeedant activities, especially compounds 1 and 4. Consistently, compounds 1 and 4 displayed potent inhibitory effects on antifeedant-related enzymes (AchE and CarE). The action mechanisms of active compounds 1 and 4 were revealed by molecular docking and molecular dynamic simulation studies. Furthermore, the distributions of the active compounds in leaves, stems, and flowers were also analyzed by liquid chromatography-mass spectrometry.

Protein tyrosine phosphatase 1B (PTP1B) is an important target for type 2 diabetes. PTP1B inhibitors can reduce blood glucose levels by increasing insulin sensitivity. Anthocyanins often play a hypoglycemic effect, but the research about them have mainly focused on glucosidase. At present, the research about protein tyrosine phosphatase 1B (PTP1B) target is less, and the corresponding molecular mechanism is still unclear. Therefore, in this present study, anthocyanins isolated from blueberry were used to study the inhibitory activity on PTP1B. The isolated cyanidin-3-arabinoside (Cya-3-Ara) exhibited a better inhibitory activity with IC50 = 8.91 ± 0.63 μM, which was higher than the positive control (oleanolic acid, IC50 = 13.9 ± 1.01 μM), and the mechanism of PTP1B inhibition was reversible mixed pattern. The structure-activity relationship (SAR) between anthocyanins and PTP1B inhibition was investigated. The enzyme activity inhibition and molecular docking showed that anthocyanins had high selectivity for PTP1B inhibition. Further study showed that Cya-3-Ara could promote glycogen synthesis through ameliorating PTP1B-involved IRS-1/PI3K/Akt/GSK3β pathways. Cya-3-Ara could also be regarded as a synergistic inhibitor (CI ≤ 0.54) of oleanolic acid to obtain a better inhibitory effect on PTP1B. Taken together, our study clearly illustrates the SAR between anthocyanins and PTP1B inhibition and the mechanism of Cya-3-Ara in the insulin signaling pathway.

Protein tyrosine phosphatase 1B (PTP1B) has recently been identified as a potential target of Norathyriol. Unfortunately, Norathyriol is not a potent PTP1B inhibitor, which somewhat hinders its further application. Based on the fact that no study on the relationship of chemical structure and PTP1B inhibitory activity of Norathyriol has been reported so far, we attempted to perform structural optimization so as to improve the potency for PTP1B. Via structure-based drug design (SBDD), a rational strategy based on the binding mode of Norathyriol to PTP1B, we designed 26 derivatives with substitutions at the four phenolic hydroxyl groups of Norathyriol. By chemical synthesis and in vitro bioassay, we identified seven PTP1B inhibitors that were more potent than Norathyriol, of which XWJ24 showed the highest potency (IC50: 0.6 μM). We also found out that XWJ24 was a competitive inhibitor and showed the 4.5-fold selectivity over its close homolog, TC-PTP. Through molecular docking of XWJ24 against PTP1B, we highlighted the essential role of its hydrogen bond with Asp181 for PTP1B inhibition and identified a potential halogen bond with Asp48 that was not observed for Norathyriol. The current data indicate that our SBDD strategy is effective to discover potent PTP1B-targeted Norathyriol derivatives, and XWJ24 is a promising lead compound for further development.

The retinoic acid receptor-related orphan receptor-gamma-t (RORγt) is a promising therapeutic target for treatment of Th17 cell-mediated autoimmune diseases. Based on a scaffold hopping/conformational restriction strategy, a series of N-indanyl benzamides as novel RORγt inverse agonists was discovered. Exploration of structure-activity relationship on the piperazine ring, benzoyl moiety and cyclopentyl moiety of N-indanyl benzamides 2a and 2d led to identification of potent RORγt inverse agonists. Compound 5c with (S)-enantiomer was found having an IC50 of 153.7 nM in Fluorescence Resonance Energy Transfer (FRET) assay, and an IC50 of 47.1 nM in mouse Th17 cell differentiation assay, which represents a promising starting point for developing potent small molecule RORγt inverse agonists. Binding modes of the two enantiomers 5c and 5d in RORγt ligand binding domain were also discussed.

BACKGROUND Free fractions of different blackberry varieties' extracts are high in phenolic compounds with antioxidant activities. However, the phenolic profiles and antioxidant activities against peroxyl radicals of bound fractions of different blackberry varieties' extracts have not been previously reported. In addition, what the key antioxidant phenolic compounds are in free and bound fractions of blackberry extracts remain unknown. This study aimed to investigate the phenolic profiles and antioxidant activities of free and bound fractions of eight blackberry varieties' extracts and reveal the key antioxidant phenolic compounds by boosted regression trees. RESULTS Fifteen phenolics (three anthocyanins, four flavonols, three phenolic acids, two proanthocyanidins, and three ellagitannins) were identified in blackberry by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Ferulic acid, ellagic acid, procyanidin C1, kaempferol-O-hexoside, ellagitannins hex, and gallic acid were major bound phenolics. Bound fractions of eight blackberry varieties' extracts were high in phenolics and showed great antioxidant activity. Boosted regression trees analysis showed that cyanidin-3-O-glucoside and chlorogenic acid were the most significant compounds, contributing 48.4% and 15.9% respectively to the antioxidant activity of free fraction. Ferulic acid was the most significant antioxidant compound in bound fraction, with a contribution of 61.5%. Principal component analysis showed that Kiowa was the best among the eight varieties due to its phenolic profile and antioxidant activity. CONCLUSION It was concluded that blackberry varieties contained high amounts of bound phenolics, which confer health benefits through reducing oxidative stress. Ferulic acid was the key compound to explain the antioxidant activities of bound fractions. © 2021 Society of Chemical Industry.

The kiwi berry (Actinidia arguta) has been widely studied because of its rich phenolic, flavonoid, and vitamin C contents. Numerous reports have demonstrated that fruit peels contain higher phenolic content and antioxidant activity than that of flesh. In this study, the phytochemical content and antioxidant activities of peel and flesh extracts of six kiwi berries were analyzed from four regions (namely, Dandong, Benxi, Taian, and Tonghua) in China. The antioxidant activity was determined using the peroxyl radical scavenging capacity (PSC) and cellular antioxidant activity (CAA) assays. The phenolic, flavonoid, and vitamin C contents of kiwi berry peel were 10.77, 13.09, and 10.38 times richer than that of kiwi berry flesh, respectively. In addition, the PSC and CAA values of kiwi berry peel were higher than those of kiwi berry flesh. The analysis of the separation and contents of phenolics were performed by the high-performance liquid chromatography (HPLC)-diode-array detectormass spectrometry/mass (DAD-MS/MS) system, and the results illustrated that protocatechuic acid, caffeic acid, chlorogenic acid, and quinic acid were the major phenolic compounds. In conclusion, this study indicated that kiwi berry peel contains a rich source of antioxidants. These data are of great significance for the full development and utilization of kiwi berries in these four regions of China to produce nutraceutical and functional foods.

Blueberry anthocyanins are well known for their beneficial biological activities. However, the poor bioavailability of anthocyanins limits their functional capacity in vivo. Our current study aimed to detect the effects of α-casein on the absorption of blueberry anthocyanins and their metabolites in rats. Blueberry anthocyanins with and without α-casein were intragastrically administered to two groups of rats and their blood samples were collected within 24 h. Results illustrated that rapid absorption of anthocyanins was observed in the rat plasma, but their concentration was relatively low. With the complexation of α-casein, the maximum concentration (Cmax) of bioavailable anthocyanins and metabolites could increase by 1.5-10.1 times (P < 0.05 or P < 0.01). The promotional effect on the plasma absorption of malvidin-3-O-galactoside and vanillic acid was outstanding with the Cmax increasing from 0.032 to 0.323 and from 0.360 to 1.902 μg/mL, respectively (P < 0.01). Besides, the molecular docking models presented that anthocyanins could enter the structural cavity and interact with amino acid residues of α-casein, which was in accordance with the improved bioavailability of anthocyanins. Therefore, α-casein could assist more blueberry anthocyanins and their metabolites to enter blood circulation.

The application of blueberry anthocyanins (ANs) was limited due to their low in-process stability and bioavailability. In our study, the stability and antioxidant capacity of ANs before and after adding bovine serum albumin (BSA) were examined by simulating various processing, storage (light, sucrose, and vitamin C (Vc)), and in vitro simulated digestion parameters. For this purpose, pH-differential method, high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), peroxyl scavenging capacity assay, and cellular antioxidant assay were conducted. BSA at different concentrations, specifically at 0.15 mg/mL, inhibited the degradation of ANs and the loss of antioxidant capacity. The results suggest that BSA has a positive effect on ANs.

The molecular interactions of xylitol (XY) with α-casein (α-CN) and κ-casein (κ-CN) at pH 7.4 as a function of temperature (298, 308, and 318 K) were characterized by multispectral techniques and molecular docking. The fluorescence results showed that XY strongly quenched the intrinsic fluorescence of α- and κ-CN by static quenching, as well as the presence of a single binding site for XY on both proteins with a binding constant value of ∼105 L/mol. The binding affinity of both proteins for XY decreased with increasing temperature, and Van der Waals forces, hydrogen bonding and protonation were the key forces in the interactions. The addition of XY altered the polarity of the microenvironment of proteins and changed their secondary structure from ordered to disordered. The molecular docking results showed that XY had different binding sites to α- and κ-CN, with several amino acids involved in the binding processes.

The current study is aimed to evaluate the combined effect of thermosonication (TS) and high hydrostatic pressure (HHP) on enzyme activities (polyphenolase and peroxidase), microbial load and phenolic compounds (phenols, flavonoids, and anthocyanins) of blueberry juice. Blueberry juice has been treated with TS (40 kHz and 240 W) at different temperatures (25 °C and 45 °C) for 15 mins with subsequent different HHP (200, 400 and 600 MPa) for 5 mins at room temperature. The results revealed that a combined use of HHP of 400 MPa and 600 MPa with TS at 45 °C not only reduced microorganisms below 1 logCFU/mL, but also significantly inactivated enzymes. The treatments also increased the phenolic compounds, peroxyl radical scavenging capacity (PSC), and DPPH free radical scavenging activity to a higher level without causing any changes in soluble solids and pH. Therefore, the combination of HHP and TS can be used as a novel alternative nonthermal technology to improve the nutritional qualities of blueberry juice, which produces a desirable, healthy juice for consumers.

The current study investigated the positive effects of blueberry anthocyanin-rich extracts (BAE) on either peripheral or hippocampal antioxidant defensiveness and established the connection of the improved antioxidant status with the altered fatty acid species and gut microbiota profile. High-fat diet-induced oxidative stress in C57BL/6 mice was attenuated by BAE administration, which was reflected by strengthened antioxidant enzymes, alleviated hepatic steatosis, and improved hippocampal neuronal status. Serum lipidomics analysis indicated that the fatty acid species were altered toward the elevated unsaturated/saturated ratio, along with phospholipid species toward enriched n-3 polyunsaturated fatty acid compositions. The modulated antioxidant pattern could be attributed to the increased bacteria diversity, stimulated probiotics (Bifidobacterium and Lactobacillus) and short-chain fatty acid (SCFA) producers (Roseburia, Faecalibaculum, and Parabacteroides) improved by anthocyanins and their metabolites, which improved the colon environment, characterized by promoted SCFAs, restored colonic mucosa, and reorganized microbial structure. Thus, anthocyanin-rich dietary intervention is a promising approach for the defensiveness in human oxidative damage and neurodegeneration.

Blueberry anthocyanin-rich extract (BAE) was supplemented to high-fat diet (HFD)-fed mice to investigate sphingolipid metabolism modulating factors involved in the attenuated hyperinsulinemia and hyperlipidemia. A BAE-containing diet effectively controlled food intake and liver weight and significantly attenuated insulin resistance triggered by a HFD. Higher BAE (200 mg/kg of body weight) administration performed more efficiently in the improvement of hepatic steatosis and adipocyte hypertrophy, together with distinct suppressions in serum triacylglycerol and cholesterol in total and species. Serum lipid compositions revealed 200 mg/kg of BAE supplementation remarkably suppressed ceramide accumulation. Consistently, genes encoding enzymes associated with sphingomyelin conversion and ceramide de novo synthesis were modulated toward a healthy direction for restrained sphingolipid accumulation. Further, the inhibited mRNA expressions of protein phosphatase 2A and protein kinase Cζ involved in blocking Akt phosphorylation connected the controlled ceramides with the restored insulin sensitivity.

Cyanidin-3-glucoside (C3G) is an abundant anthocyanin that exhibits antimicrobial, cardioprotective, and hepatoprotective effects. We applied C3G to silica-injured mice to investigate the efficacy of dietary C3G on lung inflammation and the underlying action mechanism as this information is currently unavailable. We found that C3G treatment significantly alleviated lung inflammation and reduced inflammatory cell infiltration into the lungs. It also reduced the pro-inflammatory cytokine levels in bronchoalveolar lavage fluid. Flow cytometry analysis revealed that C3G treatment ameliorated pulmonary helper and cytotoxic T cell immune responses by decreasing cytokine levels. Regulatory T cells and IL-17A producing-regulatory T cells were also decreased by C3G treatment. Furthermore, C3G effectively suppressed SP-induced activation of the STAT1 and STAT3 signaling pathways of lung tissues. These results demonstrated that dietary C3G could alleviate SP induced lung injury in mice and support the use of C3G as functional food to modulate pulmonary immune responses to silica particles.

Anthocyanin, a natural antioxidant, is reported to have cytotoxicity against cancer cells; however, the mechanism remains unclear. The aim of the present study was to investigate the mechanism by which malvidin-3-galactoside (M3G), the prominent anthocyanin in blueberry, suppresses the development of hepatocellular carcinoma. In vitro, M3G suppressed the proliferation, polarization, migration, and invasion activities of HepG2 cells by regulating the protein expression of cyclin D1, cyclin B, cyclin E, caspase-3, cleaved caspase-3, Bax, p-JNK, and p-p38, activating phosphatase and tensin homologue deleted on chromosome 10 (PTEN), accompanied by a decrease in the p-AKT level, and lowering the protein expression levels of MMP-2 and MMP-9. In vivo, M3G promoted the apoptosis of liver tumor cells, as determined by immunohistochemistry (cleaved caspase-3, Ki-67, PTEN, and p-AKT), a terminal deoxynucleotidyl transferase dUTP nick end labeling assay, and hematoxylin-eosin staining. Overall, these results suggest that M3G, as an adjuvant ingredient or nutritional supplement, may be beneficial for liver cancer prevention and the modulatory mechanism seems to be associated with inhibition of proliferation, apoptosis, migration, and invasion-related pathways.

Chemical investigation of the roots of Croton crassifolius led to the isolation of five pyran-2-one derivatives, including two brand new compounds (1–2), one new natural product (3) and two known compounds (4–5). Their structures and absolute configurations were established by spectroscopic analyses as well as comparison between the calculated optical rotation (OR) values with the experimental data. Interestingly, the new compound 1 showed an unusual negative chemical shift at H-11. It is well known that negative chemical shift values of 1H NMR spectrum are extremely rare in natural products. Such a negative chemical shift of 1H NMR spectrum was reproduced by density functional theory (DFT) calculations and explained by the shielding effect from the pyran-2-one ring over the hydrogen atom in the 3D conformations. Then, MTT assay was applied to evaluate the cytotoxicity of the isolated compounds (1–5) against two liver cancer cell lines (HepG2 and MHCC97H). The results suggested that compound 1 displayed the highest cytotoxicity with an IC50 value of 9.8 μM against HepG2 cells. Moreover, there was no obvious cytotoxicity of compounds 1–5 on normal liver cell line LO2. Furthermore, the mechanism of apoptosis induction in compound 1-treated HepG2 cells was investigated. The results showed that compound 1 could induce apoptosis via p53-mediated Ras/Raf/ERK suppression in HepG2 cells.

A mangiferin aglycon derivative J99745 has been identified as a potent xanthine oxidase (XOD) inhibitor by previous in vitro study. This study aimed to evaluate the hypouricemic effects of J99745 in experimental hyperuricemia mice, and explore the underlying mechanisms. Mice were orally administered 600 mg/kg xanthine once daily for 7 days and intraperitoneally injected 250 mg/kg oxonic acid on the 7th day to induce hyperuricemia. Meanwhile, J99745 (3, 10, and 30 mg/kg), allopurinol (20 mg/kg) or benzbromarone (20 mg/kg) were orally administered to mice for 7 days. On the 7th day, uric acid and creatinine in serum and urine, blood urea nitrogen (BUN), malondialdehyde (MDA) content and XOD activities in serum and liver were determined. Morphological changes in kidney were observed using hematoxylin and eosin (H&E) staining. Hepatic XOD, renal urate transporter 1 (URAT1), glucose transporter type 9 (GLUT9), organic anion transporter 1 (OAT1) and ATP-binding cassette transporter G2 (ABCG2) were detected by Western blot and real time polymerase chain reaction (PCR). The results showed that J99745 at doses of 10 and 30 mg/kg significantly reduced serum urate, and enhanced fractional excretion of uric acid (FEUA). H&E staining confirmed that J99745 provided greater nephroprotective effects than allopurinol and benzbromarone. Moreover, serum and hepatic XOD activities and renal URAT1 expression declined in J99745-treated hyperuricemia mice. In consistence with the ability to inhibit XOD, J99745 lowered serum MDA content in hyperuricemia mice. Our results suggest that J99745 exerts urate-lowering effect by inhibiting XOD activity and URAT1 expression, thus representing a promising candidate as an anti-hyperuricemia agent., Graphical abstract Mangiferin aglycon derivative J99745 possessed potent xanthine oxidase (XOD) inhibition in vitro. In hyperuricemic mice, J99745 reduced serum urate and MDA content, enhanced fractional excretion of uric acid, and attenuated kidney damage. J99745 might represent a promising antihyperuricemia agent by inhibiting XOD activity and urate transporter 1 expression.fx1

In this study, we tested the in vitro efficacy of a graphene oxide-chitooligosaccharide (GO-COS) complex developed to protect blueberry anthocyanins (An) from degradation by various physicochemical factors and the digestive process. We prepared a GO-COS complex to adsorb An and protect them from the destructive effects of their ambient environment. The complex protected the An under various temperature, pH, light, oxidant, and reductant conditions. We evaluated An content and composition in a simulated digestive system using the pH differential method and the high performance liquid chromatography-mass spectrometry (HPLC-MS). The GO-COS carrier stabilized An in the intestine and protected their peroxyl radical-scavenging capacity. Additionally, we observed a dose-response relationship between An content and cellular antioxidant activity, and simultaneous improvement of An bioavailability when the An were encapsulated in the complex. The complex inhibited HepG2 cell proliferation at the tested dose range. This study provides valuable information for stability of An-rich products.

A pH-sensitive food freshness indicator film (PS-CS-BA) was fabricated with potato starch (PS) as film-forming substrate, blueberry anthocyanins (BA) as an indicator, and chondroitin sulfate (CS) as an enhancer. The results exhibited that the presence of CS significantly altered water solubility and improved mechanical properties of films, in particular enhanced pH and ammonia responsiveness. FT-IR and XRD results associated with the enhanced pH sensitivity and colorimetry with the ionic complex between the sulfate group of CS and anthocyanin flavylium ion. When applied in monitoring shrimp freshness at 4 °C, the PS–CS–BA film indicated sensitively with vision recognizable color changes from original pink to light grey and finally to grayish-green, which highly correlated with the deterioration indexes of the total volatile basic nitrogen (TVB-N), total viable count (TVC), and pH values of shrimp. The results provided a novel and highly-sensitive intelligent packaging strategy by the co-pigmentation of CS for freshness monitoring.

Anthocyanins (ACNs) have attracted considerable research attention because of their excellent biological activities, but their low stability and bioavailability limit their applications. The objective of this study was to prepare nanocomplexes by the self-assembly of casein (CA) and carboxymethyl cellulose (CMC) to encapsulate and stabilize ACNs. Three types of nanocomplexes (Complex Ⅰ, Ⅱ, and Ⅲ) were formed, with some differences in particle size, polydispersity index (PDI), and encapsulation efficiency (EE) due to the different adding sequences of the raw materials. Complex III (CA-CMC-ACNs) had the smallest size (209.9 nm) and PDI (0.327), while Complex II (CMC-ACNs-CA) had the highest EE (44.23%). Scanning electron microscopy (SEM) revealed that the ACNs-loaded nanocomplexes had a coarser spherical structure than the empty nanocomplexes. Fourier-transform infrared (FTIR) spectroscopy and differential scanning calorimetry (DSC) confirmed that ACNs-loaded nanocomplexes were formed by electrostatic interactions and hydrogen bonding. The nanocomplexes, particularly Complex II, effectively attenuated the degradation of ACNs under light, thermal, or ascorbic acid treatment. Complex II also showed the highest ACNs retention rate, bioaccessibility, and antioxidant capacity after simulated digestion. These results emphasized that the addition of raw materials in different sequences is an important factor influencing the particle size and EE of nanocomplexes, which in turn affected their protection of ACNs. This study provided great potential for the applicable value in the development of stabilizing anthocyanins.

The interaction of proteins with anthocyanins (ACNs) gives rise to significant improvements in their functional properties. To explore the interactions between whey protein isolate (WPI)/bovine serum albumin (BSA) and blueberry ACNs, light (5000 Lux, 7 days), heat (85 °C, 2 h), sucrose (0.25 mg/mL), and vitamin C (0.15 mg/mL) were used to accelerate the degradation of ACNs. Through the observation of color changes and variations in the free radical scavenging rates, WPI and BSA were found to protect the ACNs to varying degrees. Moreover, WPI/BSA exhibited increased solubilities, foaming and emulsifying properties, and changed thermal stabilities and protein structures, in addition to decreased surface hydrophobicities and zeta potentials. Furthermore, the addition of WPI/BSA increased the number of ACN monomers in a simulated digestion system, as determined by high-performance liquid chromatography-mass spectrometry. Therefore, by studying the interactions between proteins and ACNs, we can provide a strategy and theoretical basis for determining the steady-state of ACNs in blueberry products.

The nuclear receptor retinoic acid receptor-related orphan receptor gamma-t (RORγt) is a transcription factor regulating Th17 cell differentiation and proliferation from naive CD4+ T cells. Since Th17 cells have demonstrated the antitumor efficacy by eliciting remarkable activation of CD8+ T cells, RORγt agonists could be applied as potential small molecule therapeutics for cancer immunotherapy. Based on the previously reported RORγt agonist 1 and its resolved co-crystal structure, a series of new tertiary amines were designed, synthesized and biologically evaluated, yielding optimal moieties with improved chemical properties and biological responses. The combination of these optimal moieties resulted in identification of novel RORγt agonists such as 8b with further elevated RORγt agonism responses at a target-based level as well as in cell-based assays, which provided some structural knowledge for further optimization of RORγt agonists as small molecule therapeutics for cancer immunotherapy.

In our efforts to find potential agents for the treatment of Alzheimer's disease in naturally occurring compounds, a systematic investigation for the active constituents of Flemingia philippinensis was carried out. Four new prenylated isoflavones, philippinone A-D (1-4), together with six known analogues (5-10), were obtained from the roots of Flemingia philippinensis. Their structures were established through extensive physical and spectroscopic data analysis. All the isolates were evaluated for their inhibitory effect of self-induced Aβ aggregation among which compound 5 showed significant Aβ aggregation inhibitory ability with the inhibitory rate of 70.56%. The results of molecular docking experiment for compounds 1 and 6 corresponded to that of the thioflavin-T assay. Moreover, the results further clarified the effects of different substituent group of tested compounds on the Aβ aggregation inhibition. A preliminary structure-activity relationship is further discussed. Our results suggested that the isoflavonoids may mitigate the progression of AD and compounds 2 and 5 may be a candidate in the treatment of AD.

Aflatoxin B1 (AFB1) causes oxidative stress and hepatocyte apoptosis through its epoxidized metabolite AFBO, which is catalyzed by CYP450 enzymes. Ferulic acid (FA) is a phenolic acid commonly found in plants and is known for its antioxidant capacity. However, the role of FA in AFB1-induced liver injury is still elusive. In this study, rats were exposed to AFB1 and simultaneously treated with FA for 30 days. The results showed that I) FA alleviated the histopathological changes induced by AFB1, inhibited the elevation of serological indexes induced by AFB1, and reduced the production of AFBO in liver. II) AFB1-induced increase in CYP450 expression was significantly reduced by FA. The molecular docking results of FA and CYP2A6 showed high fitness score and interaction. III) FA obviously inhibited the production of MDA, and significantly activated the Nrf2/GST pathway and antioxidant enzymes (SOD and GST). IV) AFB1-induced hepatocyte apoptosis, the high expression of p53, bax, cyt-c, caspase-9, caspase-3, and the low expression of bcl-2 were all restored by FA. It has been suggested from these results that FA proved effective against AFB1-induced liver damage in rats via inhibiting CYP450 enzyme, promoting antioxidant pathway Nrf2/GST, activating antioxidant enzymes (SOD and GST), and regulating the mitochondrial pathway.

Hepatocellular carcinoma (HCC) is the most common type of liver cancer, and treatment options for HCC are limited. In addition, the discovery of new natural compounds with anti-hepatocarcinoma activity is attracting increasing attention. For this reason, phytochemical investigation of Croton crassifolius led to the isolation of 17 diterpenoids, including three new clerodane diterpenoids, named crassifolius A-C (1-3), along with 14 known ones (4-17). Their structures were established by 1D, 2D NMR, HR-ESI-MS, detailed calculated electronic circular dichroism (ECD) spectra and the assistance of quantum chemical predictions (QCP) of 13C NMR chemical shifts. The cytotoxicities of all these compounds against human liver cancer lines (HepG2 and Hep3B) were determined. Among them, compound 1 exhibited good cytotoxicity with IC50 value of 17.91μM against human liver tumor cells Hep3B. Following further studies of the anti-tumor mechanism of compound 1-induced cell growth inhibition, we found that compound 1 caused apoptotic cell death in Hep3B cells by detecting morphologic changes and Western blotting analysis.

Cheese lacks essential fatty acids (EFAs). Delta 12 fatty acid desaturase (FADS12) is a critical enzyme required for EFA biosynthesis in fermentation of the predominant strains of cheese. Previously, we identified the FADS12 gene and characterized its function for the first time in Geotrichum candidum, a dominant strain used to manufacture soft cheese with white rind. In this study, we analyzed the molecular mechanism of FADS12 function by swapping domains from Mortierella alpina and G. candidum that had, respectively, high and low oleic acid conversion rates. The results revealed three regions that are essential to this process, including regions from the end of the second transmembrane domain to the beginning of the third transmembrane domain, from the end of the third transmembrane domain to the beginning of the fourth transmembrane domain, and from the 30-amino acid from the end of the sixth transmembrane domain to the C-terminal end region. Based on our domain swapping analyses, nine pairs of amino acids including H112, S118, H156, Q161, K301, R306, E307, A309 and S323 in MaFADS12 (K123, A129, N167, M172, T302, D307, I308, E310 and D324 in GcFADS12) were identified as having a significantly effect on FADS12 catalytic efficiency, and linoleic acid and its analogues (12,13-cyclopropenoid fatty acid) were found to inhibit the catalytic activity of FADS12 and related recombinant enzymes. Furthermore, the molecular mechanism of FADS12 inhibition was analyzed. The results revealed two allosteric domains, including one domain from the N-terminal region to the beginning of the first transmembrane domain and another from the 31st amino acid from the end of the sixth transmembrane domain to the C terminus. Y4 and F398 amino acid residues from MaFADS12 and eight pairs of amino acids including G56, L60, L344, G10, Q13, S24, K326 and L344 in MaFADS12 (while Y66, F70, F345, F20, Y23, Y34, F327 and F345 in GcFADS12) played a pivotal role in FADS12 inhibition. Finally, we found that both allosteric and active sites were responsible for the catalytic activity of FADS12 at various temperatures, pH, and times. This study offers a solid theoretical basis to develop preconditioning methods to increase the rate at which GcFADS12 converts oleic and linoleic acids to produce higher levels of EFAs in cheese.

Cheese lacks essential fatty acids (EFAs). Delta 12 fatty acid desaturase (FADS12) is a critical enzyme required for EFA biosynthesis in fermentation of the predominant strains of cheese. Previously, we identified theFADS12gene and characterized its function for the first time inGeotrichum candidum, a dominant strain used to manufacture soft cheese with white rind. In this study, we analyzed the molecular mechanism ofFADS12function by swapping domains fromMortierella alpinaandG. candidumthat had, respectively, high and low oleic acid conversion rates. The results revealed three regions that are essential to this process, including regions from the end of the second transmembrane domain to the beginning of the third transmembrane domain, from the end of the third transmembrane domain to the beginning of the fourth transmembrane domain, and from the 30-amino acid from the end of the sixth transmembrane domain to the C-terminal end region. Based on our domain swapping analyses, nine pairs of amino acids including H112, S118, H156, Q161, K301, R306, E307, A309 and S323 in MaFADS12 (K123, A129, N167, M172, T302, D307, I308, E310 and D324 in GcFADS12) were identified as having a significantly effect onFADS12catalytic efficiency, and linoleic acid and its analogues (12,13-cyclopropenoid fatty acid) were found to inhibit the catalytic activity ofFADS12and related recombinant enzymes. Furthermore, the molecular mechanism ofFADS12inhibition was analyzed. The results revealed two allosteric domains, including one domain from the N-terminal region to the beginning of the first transmembrane domain and another from the 31stamino acid from the end of the sixth transmembrane domain to the C terminus. Y4 and F398 amino acid residues fromMaFADS12and eight pairs of amino acids including G56, L60, L344, G10, Q13, S24, K326 and L344 in MaFADS12 (while Y66, F70, F345, F20, Y23, Y34, F327 and F345 in GcFADS12) played a pivotal role inFADS12inhibition. Finally, we found that both allosteric and active sites were responsible for the catalytic activity ofFADS12at various temperatures, pH, and times. This study offers a solid theoretical basis to develop preconditioning methods to increase the rate at whichGcFADS12converts oleic and linoleic acids to produce higher levels of EFAs in cheese.

Most of the previous in vitro digestion treatments were conducted directly to whole grains without extraction of free phenolics, thus the bioaccessible phenolics contained both free phenolics that survived the digestion and digested phenolics released by digestion. However, the profiles of digested phenolics released by digestion remain unknown. This study was designed to investigate the phytochemical contents, peroxyl radical scavenging capacities (PSCs), and cellular antioxidant activities (CAAs) of free, digested, and bound fractions of whole grains. Total phenolic contents of whole grains were highest in digested fraction, followed by free and bound fractions. The predominant phenolics were 12 phenolic acids and one flavonoid, which mostly existed in bound forms, then in digested and free forms. The digested phenolics bound to proteins were in conjugated form. The bound fractions had the highest PSCs, followed by free and digested fractions. CAAs were highest in bound fractions, followed by digested and free fractions.

This study aimed to quantitatively screen and compare the chemical compounds including protein, lipid, sucrose, total phenolics (TPC), and total titratable acidity (TTA) along with the fatty acid profile of green coffee beans from various geographical origins (China, Indonesia, Ethiopia, Kenya, Guatemala, Honduras, Brazil, and Colombia). Meanwhile, partial least squares discriminant analysis (PLS-DA) was used to investigate whether the analyzed parameters’ data can be successfully applied to differentiate the geographical origins of green coffee beans. The levels of protein, lipid, sucrose, TTA, and TPC in all samples were in the range of 13.06–15.98 g 100 g−1, 12.88–16.29 g 100 g−1, 7.30–11.45 g 100 g−1, 231.43–361.21 mL 0.1 mol L−1 NaOH 100 g−1, and 37.56–50.57 mg GAE g−1, respectively. The fatty acid composition was dominated by C18:2, C16:0, C18:1, and C18:0. The major fatty acid group was unsaturated fatty acids (USFA), accounting for 51.66–54.28% of total fatty acids. Statistical analysis showed significant differences in the chemical and fatty acid composition of green coffee beans from eight geographical origins (P＜0.05). Moreover, the PLS-DA results revealed excellent discrimination between the different geographical origins of green coffee beans, highlighting lipid, C24:0, C22:0, C18:3, C17:0, C18:0, C20:0, C16:0, and protein as discriminatory features.

Thermal processing (TP) and high hydrostatic pressure (HHP) are two important puree processing methods. In this study, the polyphenol oxidase (PPO) and β-glucosidase activities, chromatic values, peroxide radical scavenging capacities (PSCs), cellular antioxidant activities (CAAs), and anthocyanin profiles were evaluated in blueberry puree following TP and HHP treatments. Nine anthocyanins were identified and cyanidin glycosides were the most abundant compounds in the blueberry puree sample. Petunidin-3-O-arabinoside, malvidin-3-O-galactoside, and malvidin-3-O-glucoside concentrations increased at temperatures of 70–90 °C (TP) and a pressure of 300 MPa (HHP). The highest total anthocyanin concentration (503.5 μg/mL) and PSC (13.45 µg VCE/mL) were observed following the TP (90 °C) treatment. Furthermore, a positive correlation was observed between the anthocyanin content and PSC (R2 = 0.655, P

This research established an optimized method for the extraction and enrichment of flavonoids from R. corchorifolius fruit. Under the optimized extraction conditions, 12 flavonoids (1–12) were isolated, of which six (2–4, 9–10, 12) were obtained from R. corchorifolius for the first time. Compound 4 showed significant α-glucosidase (4.96 μM) and α-amylase (8.04 μM) inhibitory effects. Molecular modeling revealed that compound 4 exhibits strong binding with the active sites of α-glucosidase and α-amylase. Lineweaver-Burk plot analysis and surface plasmon resonance revealed the possible dynamic binding mechanism of the flavonoids with α-glucosidase and α-amylase. The enriched flavonoids and compound 4 showed significant hypoglycemic effects in mice administered a high dose of glucose. In this study, a variety of flavonoids with hypoglycemic activity were found for the first time, revealing the rich chemical composition of R. corchorifolius fruit and highlighting the potential value of R. corchorifolius fruit flavonoids as dietary supplements.

The processing stability and antioxidant capacity of blueberry anthocyanins (ANs) in the presence of whey protein isolate (WPI) were examined. WPI was found to enhance both the stability and antioxidant activity of ANs during processing and simulated in vitro digestion, especially at a concentration of 0.15 mg·mL−1. Fluorescence and ultraviolet-visible absorption spectroscopy showed that ANs were primarily stabilized by hydrophobic forces between WPI and malvidin-3-O-galactoside (M3G), the major anthocyanin monomer. Circular dichroism and Fourier-transform infrared spectroscopy confirmed that the structure of WPI changed and the microenvironments of certain amino acid residues were modulated by non-covalent binding to M3G; furthermore, fewer α-helices and more β-sheets were formed. Molecular docking studies revealed that WPI, especially immunoglobulin (IgG), contributed the most to ANs stability via hydrogen bonds and hydrophobic forces according to molecular docking scores (−141.30 kcal/mol). These results provided an important fundamental basis for improving the stabilities of ANs in milk systems.

Chokeberry pomace contains abundant polymeric proanthocyanidins with low antioxidant activity and bioaccessibility. In this study, polymeric proanthocyanidins from chokeberry pomace were converted into anthocyanidins with higher antioxidant activity. Optimal conversion conditions were determined by full factorial design. The composition of anthocyanidin was analyzed by TOF-MS and UPLC. In addition, the antioxidant activity of the conversion products was evaluated by PSC and CAA assays. The results showed that the yield of anthocyanidins was 11.59 mg/g when the pomace was treated with 95% ethanol concentration, 50:1 liquid-solid ratio, 4% hydrochloric acid and 0.2 mg/mL ammonium ferric citrate concentration at 80 °C for 30 min. The newly generated anthocyanidins were identified as cyanidin, which accounting for 51.3% of the total anthocyanidins. The antioxidant activity values of PSC and CAA increased 2.81 and 4.77 folds respectively. The proposed method provides a new strategy for preparation of antioxidants from naturally abundant polymeric ones.

This study aimed to examine the interaction mechanism between xylitol (XY) and whey protein isolate (WPI) using multispectral techniques and molecular docking. Additionally, we investigated the effect of XY on WPI functionality using the method of fluorescent probe, high-speed dispersion and differential scanning calorimetry. The fluorescence quenching results such as quenching constants, binding constants and thermodynamic parameters showed strong susceptibility to interacting of WPI and its fractions to XY and the sequence was: α-lactalbumin (α-La) > bovine serum albumin (BSA) > β-lactoglobulin (β-Lg). Docking results revealed that XY was bound to the residues of aromatic cluster II in α-La, the hydrophobic cavity of β-Lg and the subdomain IIA of BSA through hydrogen bonding and van der Waals forces, resulting in conformational changes in secondary structures of proteins, which converted α-helix to β-turn and random coils. Further, XY increased thermal stability and emulsifying properties and reduced surface hydrophobicity and zeta-potential of WPI.

This study investigates the effects of bovine serum albumin (BSA) on blueberry anthocyanins and their interaction. Findings showed that BSA could protect blueberry anthocyanins against degradation and retain their antioxidant activity at an ideal concentration of 0.15 mg/mL under three deteriorating treatments: illumination, vitamin C + illumination, and sucrose + illumination. The fluorescence and UV absorption spectra showed that malvidin-3-o-galactoside (M3G), the major monomer in blueberry anthocyanins, led to a static quenching of BSA and the binding site of M3G to BSA was approximately one. Further, the interaction was a spontaneous process with electrostatic interactions being the main force. CD spectra and synchronous fluorescence spectra presented alterations in the secondary structure and microenvironment of Trp and Tyr residues of BSA, respectively, upon interaction with M3G. Finally, molecular docking analysis showed that M3G mainly bound the II and III domains of BSA by hydrogen bonds and electrostatic interaction. In conclusion, our study highlights the protective effects of BSA on the stability and anti-oxidant activity of blueberry anthocyanins and their interaction analysis.

Scope This study investigates the modulatory effects of Lonicera caerulea L. polyphenols (LCPs) on the intestinal environment and lipopolysaccharide (LPS)-induced liver injury via the nuclear factor erythroid-2-related factor 2/heme oxygenase-1 (HO-1)/NQO1 and mitogen-activated protein kinase (MAPK) pathways in a rat model of oxidative stress damage (OSD). Methods and results To examine the prebiotic properties of LCP, a model of high-fat-diet-induced OSD is established using Sprague Dawley rats. In the colon, treatment with LCP for 8 weeks ameliorates enhanced intestinal permeability (glucagon-like peptide-2 content and occludin protein increase, whereas claudin-2 protein decreases), intestinal inflammation (levels of pro-inflammatory cytokines, such as tumor necrosis factor-α, interleukin-6, cyclooxygenase-2, and nuclear factor kappa-B p65 (NF-κB p65), decrease), and intestinal OSD (through regulation of the Nrf2/HO-1/NQO1 pathway). Moreover, LCP alleviates LPS-induced liver injury by suppressing the nuclear translocation of NF-κB p65 and activation of the MAPK signaling pathway. Additionally, Bacilli, Lactobacillales, Lactobacillaceae, Lactobacillus, Akkermansia, Actinobacteria, Proteobacteria, Rothia, and Blautia are found to be the key intestinal microbial taxa related to intestinal OSD and LPS-induced liver injury in rats. Conclusion LCP treatment potentially modulates the intestinal environment and alleviates liver injury by suppressing oxidative-stress-related pathways and altering the composition of the intestinal microbiota.

Helicobacter pylori (HP) contributes to the development of gastrointestinal disorders. Cyanidin-3-O-glucoside (C3G) is a major anthocyanin in human diets. In the current study, the effects and associated mechanism of C3G on HP lipopolysaccharide (LPS)-induced symptoms on human gastric epithelial cell (HGEC) cells were explored. The cells were exposed to LPS and C3G, and cellular viability, apoptotic rate, inflammation level, and toll-like receptor (TLR)-mediated nuclear factor-kappa B (NF-κB) pathway activity were measured. C3G suppressed the abnormal DNA synthesis and induced apoptosis in LPS-treated HGEC cells. C3G also reduced inflammation in LPS-treated HGEC cells. Although C3G exposure reduced the expressions of TLR2 and TLR4, it had no influence on TLR5. C3G deactivated TLR-mediated NF-κB signaling. The effects of C3G were stronger than anthocyanin extract but weaker than curcumin. The protective effects of C3G against HP LPS-induced injuries in gastric epithelial cells were associated with the deactivation of TLR2- and TLR4-mediated NF-κB signaling.

A novel and practical copper-catalyzed strategy for intermolecular cyanosulfonylation and cyanotrifluoromethylation of unactivated alkenes in water-containing solvents is described. The methodology developed provides an efficient and convenient access to a variety of β-sulfonyl nitriles and β-trifluoromethyl nitriles, which would have wide applications in chemical and pharmaceutical industries.

Microbial transformation is a remarkable tool for the structural modification of bioactive natural compounds converting them into more valuable biologically active derivatives. In order to obtain the bioactive metabolites of methyl cyperenoate, it was biotransformed using Cunninghamella elegans AS 3.2028 to afford eight new derivatives (1–8) including six new hydroxylated metabolites (2–6, 8), one new oxidation metabolite (7), along with one unusual new dehydration metabolite (1). Metabolite 1 and metabolite 7 with unsaturated ring C were found in patchoulane sesquiterpene for the first time. In addition, the biotransformation time-course of methyl cyperenoate by Cunninghamella elegans AS 3.2028 was described and the possible transformation pathway was also proposed on the basis of structural analyses and biotransformation time-courses. It was noteworthy that the major biotransformation reactions only occurred in ring C of methyl cyperenoate. All the biotransformation metabolites were tested in vitro for antithrombotic activity by measuring the platelet count, together with the partial thromboplastin time (APTT), the thrombin time (TT) and the prothrombin time (PT). The biotransformation derivatives 1–3 exhibited potent antithrombotic activity in vitro. Furthermore, the active compounds 1–3 were tested for their in vivo antithrombotic activity using a transgenic zebrafish model and compound 1 exhibited good antithrombotic activity in this model. Interestingly, transformation derivative 1 displayed potent antithrombotic activity both in vivo and in vitro compared with its precursor (the original compound), suggesting that it could be a promising agent for further preclinical evaluation for preventing abnormal blood clotting.

Phytochemical investigation of Croton crassifolius roots afforded five sesquiterpenes (1–5), including two new sesquiterpenes 6S-hydroxy-cyperenoic acid (1) and crassifterpenoid A (5), together with three known compounds (2–4). The structures of the new compounds were determined by comprehensive spectroscopic methods, and their absolute configurations were determined by quantum chemical ECD calculation. Crassifterpenoid A (5) is the first germacrane-type sesquiterpene isolated from C. crassifolius, which enriched the diversity of chemical constituents in Croton crassifolius. In addition, the cytotoxicities of all compounds against human liver cancer lines HepG2 and Hep3B were determined, but none showed significant activity.

Introduction Portulaca oleracea L. (P. oleracea, purslane) is an edible plant that is widely distributed around the world, and flavonoids are its main bioactive constituents. Therefore, the detection of flavonoids is very important for a better understanding of its pharmacological actions and to monitor the product quality control of P. oleracea. Objective To develop a rapid method to extract and determine 26 bioflavonoids in P. oleracea, based on microwave extraction (MWE) and triple quadrupole-linear ion trap mass spectrometry. Methods The optimal conditions of MWE for the extraction of flavonoids from P. oleracea involved the use of methanol as the extraction solvent, a microwave power of 300 W, an extraction time of 450 s, and a solvent-to-solid ratio of 30 mL/g. The samples were analysed using an ultra-performance liquid chromatograph coupled with a triple quadrupole-linear ion trap mass spectrometer (UPLC–MS/MS) system. Results The calibration curves of all 26 analytes showed good linearity (r ≥ 0.999) and the intra- and interday precisions and repeatability were all within required limits. The mean recoveries measured at three concentrations were higher than 94.2%, with RSDs lower than 2.94% for the targets. Conclusion The established MWE/UPLC–MS/MS method is a rapid and effective method for quality evaluation of P. oleracea from different production regions and different harvest periods. Copyright © 2014 John Wiley & Sons, Ltd.

A rapid and reliable microwave extraction and the triple quadrupole-linear ion trap mass spectrometry method was developed and validated for the determination of eight alkaloids in Portulaca oleracea L. The optimal microwave extraction (MWE) condition was performed at 60 °C for 12 min with ethanol-water (70:30, v/v) as the extracting solvent, and the solvent to solid ratio was 30:1. The alkaloids were first detected simultaneously by electrospray ionization tandem mass spectrometry under positive-negative conversion multiple reaction monitor ((+/-)MRM) technique. With investigating three different columns, samples were separated in only 8 min on a Waters ACQUITY UPLC HSS T3 (50 × 2.1 mm(2), 1.8 μm) column using acetonitrile and formic acid-water solution as a mobile phase with a flow rate at 0.2 mL/min. All calibration curves showed good linearity (r>0.999) within the test ranges. The method developed was validated with acceptable sensitivity, intra- and inter-day precision, reproducibility, and extraction recoveries. It was successfully applied to the determination of eight alkaloids in Portulaca oleracea L. from different sources and different harvest periods. The method also provide a reference for extraction and determination of alkaloids in other complex systems.

Materials developed from Broussonetia papyrifera have been widely used in food and paper industries, but this root bark of the plant has been overlooked despiteits potentially economic values. Thus, the aim of the present study was to develop the root bark of B. papyrifera as a source of bioactive compounds. Chemical investigation of the bark of B. papyrifera led to the isolation of nine flavonoids derivatives (1-9) and two coumarins (10-11), which includes five new compounds (1–4 and 11). The structures of isolates were elucidated based on spectroscopic analyses. All the isolates were evaluated for their biological effects including tyrosinase inhibition, cytotoxicity, and anti-inflammatory activity. Compound 3 showed inhibition activity against tyrosinase with an IC50 value of 9.29 μM (positive control, kojic acid: 30.56 μM). Molecular docking was used to support their inhibition activities which indicated that hydrogen bond formation played an important role in the ligand-enzyme complex. In addition, compound 4 exhibited cytotoxic activity against three cancer cell lines (NCI-H1975, HepG2, and MCF-7) and the structure-activity relationship of flavonoids (1-9) were established. Results from anti-inflammatory assay showed that compound 1 inhibited the production of IL-2 in Jurkat cells induced by phytohemagglutinin (PHA) and phorbol 12-myristate 13-acetate (PMA), with immunosuppressive activity. Overall, our study suggests that flavonoids from root bark of B. papyrifera can be used as a valuable source of bioactive compounds and support its use as a crop benefits to local economic.

Two new alkaloids named (3R)-3,5-bis(3-methoxy-4-hydroxyphenyl)-2,3-dihydro-2(1H)-pyridinone (1) and 1,5-dimethyl-6-phenyl-1,2-dihydro-1,2,4-triazin-3(2H)-one (2), together with two known compounds (7'R)-N-feruloyl normetanephrine (3) and N-trans-feruloyl tyramine (4) were isolated from the air-dried aerial parts of Portulaca oleracea L. Their structures and configurations were elucidated by spectroscopic methods including 1D NMR, 2D NMR, and HR-MS techniques. In addition, compounds 1-4 were tested for in vitro cytotoxic activities against human lung (K562 and A549) and breast (MCF-7 and MDA-MB-435) cancer cell lines.