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Five putrescine and spermidine derivatives (1-5) together with five rotenoids (6-10) were isolated from a methanolic extract of the flowers of A. fruticosa that displayed promising inhibition of 76.0 ± 1.9% for AChE and 90.0 ± 4.0% for BuChE at a concentration of 1 mg/mL. Although the anticholinesterase activities of the isolated compounds did not reach that of galantamine, molecular docking revealed that all-trans-tri-p-coumaroylspermidine and trans-trans-cis-tri-p-coumaroylspermidine showed binding poses mimicking the known inhibitor galantamine and thus could serve as model molecules in future searches for new AChE and BuChE inhibitors.

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This study employed the MTT assay to assess the cytotoxicity of one flavan and two stilbene derivatives isolated from the false indigo-bush (Amorpha fruticosa L.) fruits: 5,7-dihydroxy-8-geranylflavanone (1), 2-carboxy-3,5-dihydroxy-4-geranylbibenzyl (2), and 2-carboxy-3-hydroxy-4-prenyl-5-methoxybibenzyl (3). The examined compounds reduced the survival of human cervical and colon tumour cells (HeLa, HT-29, HCT-116, and LS174) with IC50 values ranging from 10.55 to 147.09 µg/mL, except for 1, which did not affect LS174 cells within the tested concentrations. The highest activity was observed for 1 against HeLa cells, and 1 also exhibited the weakest effect against normal foetal lung fibroblasts (IC50 = 166.11 µg/mL), demonstrating good potency and selectivity. Stilbenes 2 and 3 proved efficacious, but lacked selectivity compared to 1. Our findings revealed the cytotoxicity of false indigo-bush constituents, justifying further mechanistic and in vivo investigations, particularly on 5,7-dihydroxy-8-geranylflavanone, which displayed considerable in vitro anticancer capacity and a potentially favourable safety profile.

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The extraction, isolation, structural characterisation and biological activities of polysaccharides from Amorpha fruticosa flowers were investigated. First, the crude polysaccharide AFP was extracted, and two major purified polysaccharide fractions AFP-2 and AFP-3 were isolated. The molecular weight and monosaccharide compositions of AFP-2 and AFP-3 were determined. Then the antioxidant activities of AFP, AFP-2 and AFP-3 were assessed by DPPH radical, ß-Carotene bleaching and hydroxyl radical assays. All three tested polysaccharides showed good antioxidant activity while AFP was the strongest one. The study also showed that AFP, AFP-2 and AFP-3 have good tyrosinase inhibition, moisture absorption and retention activities. The results will provide a helpful reference for the application of polysaccharide from Amorpha fruticosa flowers as a natural cosmetic ingredient.

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The WRKY gene family in plants regulates the plant's response to drought through regulatory networks and hormone signaling. AfWRKY20 (MT859405) was cloned from Amorpha fruticosa (A. fruticosa) seedlings using RT-PCR. The binding properties of the AfWRKY20 protein and the W-box (a DNA cis-acting element) were verified both in vivo and in vitro using EMSA and Dual-Luciferase activity assays. RT-qPCR detected that the total expression level of AfWRKY20 in leaves and roots was 22 times higher in the 30% PEG6000 simulated drought treatment compared to the untreated group. Under the simulated drought stress treatments of sorbitol and abscisic acid (ABA), the transgenic tobacco with the AfWRKY20 gene showed enhanced drought resistance at the germination stage, with significantly increased germination rate, green leaf rate, fresh weight, and root length compared to the wild-type (WT) tobacco. In addition, the superoxide dismutase (SOD) activity, chlorophyll content, and Fv/Fm ratio of AfWRKY20 transgenic tobacco were significantly higher than those of the WT tobacco under natural drought stress, while the malondialdehyde (MDA) content and 3,3'-diaminobenzidine (DAB) and nitroblue tetrazolium (NBT) staining levels were lower. The expression levels of oxidation kinase genes (NbSOD, NbPOD, and NbCAT) in transgenic tobacco under drought stress were significantly higher than those in WT tobacco. This enhancement in gene expression improved the ability of transgenic tobacco to detoxify reactive oxygen species (ROS). The survival rate of transgenic tobacco after natural drought rehydration was four times higher than that of WT tobacco. In summary, this study revealed the regulatory mechanism of AfWRKY20 in response to drought stress-induced ABA signaling, particularly in relation to ROS. This finding provides a theoretical basis for understanding the pathways of WRKY20 involved in drought stress, and offers genetic resources for molecular plant breeding aimed at enhancing drought resistance.

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Cadmium (Cd) pollution has become aggravated during the past decades of industrialization, severely endangering human health through its entry into the food chain. While it is well understood that arbuscular mycorrhizal fungi (AMF) have a strong ability to regulate plant growth and Cd uptake, studies investigating how they affect soil Cd speciation and influence Cd uptake are limited. We designed a pot experiment comprising two AMF-inoculant groups (inoculation with Diversispora eburnea or no inoculation), three Cd concentration levels (0, 5, and 15 mg/kg), and two plant species (Lolium perenne and Amorpha fruticosa) to study the effect of AMF Diversispora eburnea on plant growth, Cd uptake, and Cd speciation in the soil. The results revealed that L. perenne exhibited higher productivity and greater Cd uptake than A. fruticosa, regardless of AMF D. eburnea inoculation. However, AMF D. eburnea significantly altered soil Cd speciation by increasing the proportion of exchangeable Cd and decreasing residual Cd, resulting in Cd enrichment in the plant root organs and the elimination of Cd from the polluted soils. Our experiments demonstrate that inoculating plants with AMF D. eburnea is an effective alternative strategy for remediating Cd-contaminated soil.

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Plants are often adversely affected by abiotic stresses such as drought, low temperature, and salinity during growth, and plant NAC-like transcription factors are involved in regulating growth and developmental processes in response to stresses such as drought and salinity. In this study, to investigate the function of AfNAC1, a co-expression network of AfNAC1 genes was constructed using gene expression data from the Chinese legume deciduous shrub, Amorpha fruticosa Linn. A 576 bp NAC transcription factor (AfNAC1 gene, MN180266) encoding 191 amino acids was isolated from Amorpha fruticosa seedlings by RT-PCR. qRT-PCR showed that the AfNAC1 gene was expressed in the roots, stems, leaves, and flowers of Amorpha fruticosa. However, drought stress significantly increased root expression, and the AfNAC1 protein was localized in the nucleus by green fluorescence detection. This study analyzed the drought resistance of overexpressing tobacco in depth. Under natural drought stress, the chlorophyll and antioxidant enzyme activities of overexpressing plants were significantly higher than those of wild-type (WT) plants, but the MDA content was lower than that of WT; after rehydration the Fv/Fm values of AfNAC1-overexpressing tobacco recovered faster than those of wild-type tobacco and rapidly reached the control levels; AfNAC1 may be involved in the regulation of the photosystem and indirectly in the regulation of the plant in response to drought stress.

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Increased mobility of people around the globe has facilitated transferring species to new environments, where some have found suitable conditions and even become invasive. False indigo-bush (Amorpha fruticosa L.) is a plant native to North America but has intentionally or unintentionally spread over the Northern Hemisphere, where it often becomes invasive. The plant is especially easily dispersed within the watersheds of large rivers, where seasonal flooding is regular. Seeds and other propagules are buoyant, and when the water recedes, new plants emerge, forming dense thickets where only a few other species can co-exist. In order to sustain native biodiversity, spread control is needed. However, mechanical control and eradication measures currently in use are labor demanding and costly, while application of herbicides is limited. On the other hand, the plant possesses a number of beneficial properties, such as phytochemical applications (medical and insecticidal effects), biocoenotic uses (honey plant, ornamental features), and ecosystem services (soil stabilization, provision of food for animals, and fiber and biomass for industry, e.g., nanocellulose). For the reasons above mentioned, the plant is considered quite controversial, and the paper discusses both aspects: potential detrimental effects when introduced to new habitats and its beneficial uses for human society. In addition, the paper presents alternative measures of spreading control (e.g., grazing) and argues that exploiting it for beneficial purposes might help spread control, thus covering the expenses of controlling its distribution.

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In this study, we investigated the depigmentation effect of Amorpha fruticosa L. root extract (RE), an herbal medicine. A. fruticosa RE significantly induced depigmentation in α-MSH-treated B16F10 cells at noncytotoxic concentrations. Further, the RE decreased the protein levels of the melanosomal proteins Tyr and Pmel without decreasing their transcript levels. We found that MG132, a proteasome complex inhibitor, was unable to rescue the protein levels, but PepA/E-64D (a lysosomal enzyme inhibitor), 3-MA (a representative autophagy inhibitor), and ATG5 knockdown effectively rescued the protein levels and inhibited the depigmentation effect following RE treatment. Among rotenoids, amorphigenin composed in the RE was identified as a functional chemical that could induce depigmentation; whereas rapamycin, an mTOR inhibitor and a nonselective autophagy inducer, could not induce depigmentation, and amorphigenin effectively induced depigmentation through the degradation of melanosomal proteins. Amorphigenin activated AMPK without affecting mTOR, and knockdown of AMPK offset the whitening effect through degradation of melanosome proteins by amorphigenin. Results from this study suggested that amorphigenin can induce degradation of the melanosome through an AMPK-dependent autophagy process, and has the potential to be used as a depigmentation agent for the treatment of hyperpigmentation.

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The Obedska Bara Special Nature Reserve is one of the oldest protected areas in the world, also enlisted as an Important Bird Area, Ramsar and UNESCO world heritage site. False indigo bush (Amorpha fruticosa L.) represents an invasive alien species which is significantly deteriorating the biodiversity of the Obedska Bara Special Nature Reserve in Serbia. Mapping of A. fruticosa, using an unmanned aerial vehicle and GIS tools, showed that in meadows, pastures, ponds and wetland areas, A. fruticosa covered 85 ha or 11% of the area. However, coverage was uneven, and the most overgrown areas were some meadows (up to 35%), while flooded areas and areas where human impact is significant, as on pastures, were not so affected (1-3%). The most susceptible parts were forest edges. Active management practices, such as cattle grazing and topsoil tarping, and occasional moving, gave positive effects in A. fruticosa, but also other invasive terrestrial plant species spreading control in the reserve. This has also been confirmed by the UAV survey and mapping, which has proven to be an effective method for A. fruticosa monitoring over large areas.

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Despite increasing evidence indicating that invasive species are harming biodiversity, ecological systems and processes, impacts of multiple species invasion and their links with changes in plant and soil communities are inadequately documented and remain poorly understood. Addressing multiple invaders would help to ward against community-wide, synergistic effects, aiding in designing more effective control strategies. In this work, correlative relationships are examined for potential impacts of three co-occurring invasive plant species, Amorpha fruticosa, Fraxinus pennsylvanica, and Acer negundo, on soil conditions and native plant diversity. The research was conducted in riparian ecosystems and included the following treatments: (1) co-occurrence of the three invasive plant species, (2) occurrence of a single invasive species, and (3) control, i.e., absence of invasive species. Co-occurrence of three invasive plant species caused higher direct impact on soil properties, soil functioning, and native plant diversity. Soil in mixed plots (those populated with all three invaders) contained higher levels of nitrifying bacteria, organic matter, nitrogen, and carbon as well as lower carbon to nitrogen ratio as compared to single species invaded plots and control plots. Furthermore, native plant diversity decreased with invasive plants co-occurrence. Differences in soil conditions and lower native plant diversity revealed the interactive potential of multiple invasive species in depleting biodiversity and eroding soil functionality, ultimately affecting ecological and biogeochemical processes both below and above ground. Our results highlight the need to prevent the impact of multispecies invasion, suggesting that riparian ecosystems affected by co-occurring invaders should be prioritized for invasion monitoring and ecological restoration.

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Metabolic syndrome is characterized by a variety of diagnostic criteria: obesity, dyslipidemia, type 2 diabetes, and arterial hypertension. They contribute to the elevated risk of cardiovascular morbidity and mortality. The potential for Amorpha fruticosa L. (Fabaceae) to improve diabetes and metabolic disease is promising, based on in vitro tests. This is why a further investigation of the species is needed. Additionally, a toxicity review in relation to safety revealed that to date, there are no published data regarding the toxicity of A. fruticosa towards humans. This species could provide abundant and cheap resources because it is an aggressive invasive plant that grows almost unrestrictedly. The objective of this study was to evaluate the acute toxicity of a purified extract of A. fruticosa (EAF), and to assess its antioxidant, antihypertensive, and antihyperglycemic activity in streptozotocin-induced diabetic spontaneously hypertensive rats (SHRs). The EAF was slightly toxic (LD50 = 2121 mg/kg, b.w.) when administered orally, and moderately toxic (LD50 = 316 mg/kg, b.w.) at intraperitoneal administration, both in mice. The oral administration of EAF (100 mg/kg) for 35 days to SHRs caused significant decreases in the systolic pressure, blood glucose levels, and MDA quantity. It also increased the hepatic level of the endogenous antioxidant GSH, not only in diabetic SHRs, but also in the control group. An additional potential benefit to human health might be conferred through the environmental management of A. fruticosa based on its large-scale use for medicinal purposes, as this aggressive invasive species brings problems to natural habitats in many European countries.

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BACKGROUND: Amorpha fruticosa L. is a deciduous shrub that is native to North America and has been introduced to China as an ornamental plant. In order to clarify the drought resistance characteristics of Amorpha fruticosa L. and excavate the related genes involved in drought resistance regulation pathway, the mechanism of drought resistance stress of Amorpha fruticosa L. was revealed by the changes of transcriptome of Amorpha fruticosa L. under drought stress.Through the changes of the transcriptome of Amorpha fruticosa L. under drought stress, the mechanism of anti-stress of Amorpha fruticosa L. could be revealed. METHODS: Different concentrations of polyethylene glycol-6000 (PEG-6000) was used to simulate drought stress, and transcriptomic analysis was used to reveal the changes of gene expression patterns in Amorpha fruticosa L. seedlings. RESULTS: Results showed that Amorpha fruticosa L. seedlings were seriously affected by PEG-6000. As for the differently expressed genes (DEGs), most of them were up-regulated. The additional Go and KEGG analysis results showed that DEGs were functionally enriched in cell wall, signal transduction and hormonal regulation related pathways. DEGs like AfSOD, AfHSP, AfTGA, AfbZIP and AfGRX play roles in response to drought stress. CONCLUSION: In conclusion, Amorpha fruticosa L. seedlings were sensitive to drought, which was different from Amorpha fruticosa L. tree, and the genes functions in drought stress responses via ABA-independent pathways. The up-regulation of Salicylic acid signal related DEGs (AfTGA and AfPR-1) indicated that Salicylic acid play a key role in response to drought stress in Amorpha fruticosa L.

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Puerol A (1) from Amorpha fruticosa showed highly potent inhibition against both monophenolase (IC50 = 2.2 µM) and diphenolase (IC50 = 3.8 µM) of tyrosinase. We tried to obtain a full story of enzyme inhibitory behavior for inhibitor 1 because the butenolide skeleton has never been reported as a tyrosinase inhibitor. Puerol A was proved as a reversible, competitive, simple slow-binding inhibitor, according to the respective parameters; k3 = 0.0279 µM-1 min-1 and k4 = 0.003 min-1. A longer lag-phase and a reduced static-state activity of the enzyme explained that puerol A had a tight formation of the complex with Emet. Dose-dependent inhibition was also confirmed by high-performance liquid chromatography (HPLC) analysis using N-acetyl-l-tyrosine as a substrate, which was completely inhibited at 20 µM. A high binding affinity of 1 to tyrosinase was confirmed by fluorescence quenching analysis. Moreover, puerol A decreased melanin content in the B16 melanoma cell dose-dependently with an IC50 of 11.4 µM.

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A greenhouse pot trial was conducted to investigate the effect of organic amendments combined with triple superphosphate on the bioavailability of heavy metals (HMs), Amorpha fruticosa growth and metal uptake from Pb-Zn mine tailings. Cattle manure compost (CMC), spent mushroom compost (SMC) and agricultural field soil (AFS) were applied to tailings at 5%, 10%, 20% and 30% w/w ratio, whereas sewage sludge (SS) and wood biochar (WB) were mixed at 2.5%, 5%, 10% and 20% w/w ratio. Triple superphosphate (TSP) was added to all the treatments at 4:1 (molar ratio). Amendments efficiently decreased DTPA-extracted Pb, Zn, Cd and Cu in treatments. Chlorophyll contents and shoot and root dry biomass significantly (p< 0.05) increased in the treatments of CMC (except T4 for chlorophyll b) and SMC, whereas treatments of SS (except T1 for chlorophyll a and b), WB and AFS (except T4 for chlorophyll a and b) did not show positive effects as compared to CK1. Bioconcentration factor (BCF) and translocation factor (TF) values in plant tissues were below 1 for most treatments. In amended treatments, soluble protein content increased, phenylalanine ammonialyase (PAL) and polyphenol oxidase (PPO) decreased, and catalase (CAT) activity showed varied results as compared to CK1 and CK2. Results suggested that A. fruticosa can be a potential metal phytostabilizer and use of CMC or SMC in combination with TSP are more effective than other combinations for the in situ stabilization of Pb-Zn mine tailings.

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BACKGROUND: Water availability and nutrient-status of soils play crucial roles in seedling establishment and plant survival in coal-spoiled areas worldwide. Restoration of spoils pertains to the application of proper doses of nutrients and water, and selection of particular plant species for efficient revegetation. This study aimed at examining the potential effects of different combinations of soil-water and fertilizers (nitrogen, N and phosphorus, P) on morpho-physiological and biochemical attributes of Amorpha fruticosa grown in coal-mined spoils. Three factors five-level central-composite-design with optimization technique response surface methodology (rsm) was used to optimize water irrigation and fertilizer application strategies. RESULTS: Our results revealed a strong correlation between experimental data and predicted values developed from the rsm model. The best responses of A. fruticosa in terms of plant height, stem diameter, root length, and dry biomass were observed under a high-water regime. Low-water regime caused a notable reduction in growth-associated parameters, and fertilization with either N or P did not show positive effects on those parameters, indicating that soil-water was the most influential factor for growth performance. Leaf water potential, gas-exchange parameters, and chlorophyll content significantly increased under high levels of soil-water, N and P, suggesting a synergistic effect of these factors for the improvement of photosynthesis-related parameters. At low soil-water contents and N-P fertilizer application levels, enhanced accumulation of malondialdehyde and proline indicated that A. fruticosa suffered from oxidative and osmotic stresses. Amorpha fruticosa also responded to oxidative stress by accelerating the activities of superoxide dismutase, catalase, and peroxidase. The effects of both fertilizers relied on soil-water, and fertilization was most effective under well-watered conditions. The maximum growth of A. fruticosa was observed under the combination of soil-water, N-dose and P-dose at 76% field capacity, 52.0 mg kg- 1 and 49.0 mg kg- 1, respectively. CONCLUSION: Our results demonstrate that rsm effectively designed appropriate doses of water and N-P fertilizer to restore coal-spoiled soils. Furthermore, A. fruticosa responded to low-water and fertilizer-shortage by upregulating defensive mechanism to avoid damage induced by such deficiencies. Finally, our findings provide effective strategies for revegetation of coal-contaminated spoils with A. fruticosa using appropriate doses of water and N-P fertilizers.

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Floodplains are prone to plant invasions, which increase their roughness and decrease their flood conveyance capacity. In recent decades, extremely high floods have occurred in the Tisza River (Hungary) without an increase in discharge. This could be partly explained by land cover changes, as plough fields and pastures have been replaced by forest plantations and invasive plants have become widespread in the Tisza River floodplain. The aims of the present research were (1) to evaluate long-term land cover changes from the point of view of floodplain roughness, (2) to calculate vegetation density with and without the invasive shrub Amorpha fruticosa, and (3) to model (HEC-RAS) the flood conveyance in the case of unmanaged and managed vegetation (eliminating invasive plants). The study was carried out at three floodplain sections of the Tisza and Maros rivers, Hungary. In the eighteenth century, wetlands (61-93%) covered the studied floodplain areas, but as a result of mid-nineteenth-century channel regulation works, pastures and plough fields (42-72%) became widespread, and riparian forests (8-19%) appeared. In the late twentieth century, poplar plantations (43-86%) replaced pastures and plough fields and provided a perfect habitat for invasive plants. As a result of these land cover changes, the mean vegetation roughness of the floodplains increased from 0.021-0.032 (1783) to 0.066-0.092 (2017). However, at-site measurements indicate considerably higher vegetation roughness values (0.093-0.134) when the invasive Amorpha is also considered. Invasive species clearance could decrease the vegetation roughness by 86%. Based on our modelled data, peak flood stages could be decreased by 13-34 cm after the clearance of invasive plants. However, these values are influenced by the floodplain slope and characteristics of the modelled flood wave. The management of longer floodplain sections would have a considerable effect on flood stages, while the clearance of smaller patches would not have this effect.

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A concise, unified total synthesis of the two prenylated aromatic polyketides amorfrutins A and C, which exhibit various medicinally important biological profiles such as antimicrobial, PPARγ modulating and quorum sensing inhibitory activities, has been achieved from commercially available 3,5-dimethoxybenzaldehyde in 38% and 10% overall yields through nine and ten steps, respectively. The key transformation for the synthesis of amorfrutin A was the Claisen rearrangement of a mono-O-(1,1-dimethylallyl)resorcinol derivative to install the C3-prenyl substituent, while that for the synthesis of amorfrutin C was the double Claisen rearrangement of a di-O-(1,1-dimethylallyl)resorcinol derivative to introduce the two prenyl groups at the C3 and C5 positions all at once.

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From May to October in 2017, the sap flow, water consumption and the effects of environmental factors on the sap flow of Amorpha fruticosas with different stem diameters under different water and salt conditions were examined with the packaged sap flow measuring system in a secondary saline-alkali land of Ningxia Yellow River irrigation area. The sap flow rate showed a broad peak curve with no obvious phenomenon of 'midday break' in sunny days and a multi-peak curve in cloudy, overcast and rainy days. Weak sap flow was found at night. In the growing season (May - October), total sap flow of A. fruticosas with basal stem diameters of 13, 16 and 22 mm were 138.14, 206.06, 370.11 kg, respectively. The water consumption was largest in June and July, accounting for about 50% of the whole growing season, followed by May and August, and lowest in September and October. At both 0.5 h and day scales, photosynthetically active radiation (PAR) was the dominant meteorological factor affecting the sap flow. In the first growth stage (May 13th - August 19th), soil water in shallow layer (0-40 cm) had significant effect on the sap flow of A. fruticosas with three different diameters, and soil salt in shallow layer had significant inhibitory effect only on A. fruticosa with the diameter of 13 mm. In the second growth stage (August 20th - October 10th), soil water and soil salt had no significant effect on A. fruticosas with three different diameters. In summary, the A. fruticosas with diameters <13 mm was not suitable for planting in saline soil (electrical conductivity (EC)>2 dS·m-1), and individuals with diameters >16 mm could grow well in saline soil (EC=5 dS·m-1). Moreover, reasonable irrigation should be carried out in the vigorous growth stage of the A. fruticosas.

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Amorfrutins with a 2-hydroxybenzoic acid core structure are promising natural PPARγ agonists with potent antidiabetic activity. Owing to the complex matrix and low concentration in botanical material, the identification of unknown amorfrutins remains a challenge. In the present study, a combined application of UPLC-Q-TOF-MS and UPLC-QqQ-MS was developed to discover unknown amorfrutins from fruits of Amorpha fruticosa. First, reference compounds of amorfrutin A (AA), amorfrutin B (AB), and 2-carboxy-3,5-dihydroxy-4-geranylbibenzyl (AC) were analyzed using UPLC-Q-TOF-MS to reveal the characteristic fragment ions and the possible neutral loss. Second, the extract of A. fruticosa was separated and screened by UPLC-QqQ-MS using neutral loss scan to find out suspect compounds associated with the specified neutral fragment Δm/z 44. Third, the extract was re-analyzed by UPLC-Q-TOF-MS to obtain the exact mass of quasi-molecular ion and fragment ions of each suspect compound, and to subsequently calculate their corresponding molecular formulas. Finally, according to the molecular formula of suspect compound and its fragment ions and comparing with literature data, structure elucidation of four unidentified amorfrutins was achieved. The results indicated that the combination of QqQ-MS neutral loss scan and Q-TOF-MS molecular formula calculation was proven to be a powerful tool for unknown natural product identification, and this strategy provides an effective solution to discover natural products or metabolites of trace content. Graphical Abstract ᅟ.

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Amorpha fruticosa L. is a Chinese folk medicine and rich in polyphenols. Fifteen known compounds were isolated and identified from the leaves of A. fruticosa L. They are tephrosin (1), 6a,12a-dehydrodeguelin (2), vitexin (3), afrormosin-7-O-ß-d-glucopyranoside (4), 2″-O-α-l-rhamnopyranosyl isovitexin (5), rutin (6), chrysoeriol (7), 7-O-methylluteolin (8), trans-p-coumaric acid (9), 2-benzyl-4,6-dihydroxybenzoic acid-4-O-ß-d-glucopyranoside (10), formononetin (11), quercetin (12), apigenin (13), ß-sitosterol (14), and ß-daucosterol (15). Compounds 3, 4, 5, and 7-9 were isolated from A. fruticosa L. for the first time. Cytotoxicity of individual compounds 3-10 and 90% ethanol extract against human cancer cell lines HCT116 and HepG2 were reported. The results suggested that compounds 7 and 8, and the crude extract exhibited inhibitory effects on human cancer cell line HCT116, at concentrations of 100 µg/mL, 5 µg/mL, and 25 µg/mL at <60% of cell viability rate, respectively. In addition, a valid high-performance liquid chromatography diode array detector method was established to quantitatively analyze compounds 1-12 in the leaves of A. fruticosa L., which was harvested at three different stages of maturity from May 20 to August 10, 2014. The results demonstrated that contents were greatly influenced by the maturity. Total amounts of the analytical constituents gradually increased from May 20 to August 10, with the values ranging from 10.86 mg/g to 18.84 mg/g, whereas bioactive compounds 7 and 8 presented the opposite variation trend. The results of this study may provide data for further study and comprehensive utilization of A. fruticosa L.

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