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A supramolecular solvent (SUPRAS)-based sample preparation method was proposed for the determination of lignans in Schisandra chinensis (S. chinensis) fruit. The optimal conditions were: proportion of hexafluoroisopropanol in SUPRAS: 35% (v/v), solid-to-liquid ratio: 20:1 mg/mL, and vortex time: 30 s. Additionally, simple, rapid, and sensitive high-performance liquid chromatography coupled with triple quadrupole mass spectrometry and multiple reaction monitoring was employed to simultaneously quantify seven lignans in S. chinensis. The validation showed that the linearity from 0.01 to 20.83 µg/mL was excellent (all R2 ≥ 0.99). The lowest limits of quantification were from 0.14 to 17.79 ng/mL. The relative standard deviation (RSD) of the intra-day retention time and peak area were from 0.12% to 0.21% and 2.35% to 6.41%. The RSD of retention time and peak area for the inter-day precision ranged from 0.17% to 0.32% and 3.83% to 7.39%. The average recoveries (80.21%–114.53%) demonstrated good accuracy for all analytes. The developed procedure was employed to determine lignans in processed and raw S. chinensis fruit. The lignans were concentrated in the seeds and the wine-processed material had the highest content. This study provides a reference for the analysis of S. chinensis fruit for lignans. [ABSTRACT FROM AUTHOR]

In this study, ethylenediaminetetraacetic acid (EDTA) disodium was first chosen as catalyst to convert psoralenoside (PO) to psoralen (PSO) for increasing the extraction yield of PSO. An efficient continuous system for synchronous transformation and extraction of PSO from fig leaves applying microwave-assisted EDTA disodium (MAE-EDTA) was developed. The optimal MAE-EDTA condition was obtained: EDTA disodium concentration of 0.07 mol·L−1, ethanol volume fraction of 56%, extraction time of 16 min, and extraction temperature of 70 °C by single factor experiments and response surface method (RSM). Under the optimal condition, the yield of PSO reached 27.24 mg·g−1. Compared with microwave-assisted ethanol extraction (MAE) and reflux extraction (RE), the yield of PSO by MAE-EDTA is 2.03-fold higher than RE and 1.70-fold higher than MAE. Therefore, MAE-EDTA is an efficient method for extracting PSO from fig leaves, and it might provide references for the extraction of PSO from other medicinal plants. [ABSTRACT FROM AUTHOR]

Water decoctions from the leaves of Taxus cuspidata are used in traditional Chinese medicine to treat cancer, suggesting that water soluble constituents from these leaves may possess anticancer properties. Interestingly, hydrophilic paclitaxel derivatives, as opposed to paclitaxel itself, can be detected by high pressure liquid chromatography in water decoctions from these leaves. The remainder extracts, which are free of paclitaxel and hydrophilic paclitaxel derivatives, from the T. cuspidata leaves were investigated for antitumor activity in vivo and in vitro for the first time in this study. EE80B, 7-xylosyl-10-deacetylpaclitaxel and 7-xylosyl-10-deacetylpaclitaxel C displayed the most antitumor activity in vivo. However, in vitro studies with tumor cell lines showed that EE80B had a significantly smaller antitumor effect than paclitaxel. We hypothesize that water decoctions from T. cuspidata leaves exhibit antitumor effects in vivo, which may be aided by the activation of specific host mechanisms (e.g. stimulation of antitumor immunity) which are not present in vitro. [ABSTRACT FROM AUTHOR]

Abstract: A method of high-performance liquid chromatography–tandem mass spectrometry (LC–MS-MS) has been developed for the trace analysis of paclitaxel and other six taxoids in three Taxus species including Taxus cuspidata, Taxus media and Taxus chinensis var. mairiei. Seven taxoids were separated using a gradient mode on an Eclipse XDB-C18 column (4.6×150mm; i.d., 5μm) at 20°C. The compound separations were detected by an API 3000 mass spectrometer equipped with a TurboIonSpray™ interface. The compounds were detected using electrospray ionization (ESI) in positive-ion mode and quantified by multiple-reaction monitoring (MRM) mode using the transition mass of m/z 567.4→445.4, 609.3→549.5, 944.9→286.4, 812.6→286.1, 832.8→264.1, 854.4→286.1, and 812.6→286.1 for 10-DAB III, baccatin III, 7-xyl-10-DAT, 10-DAT, cephalomannine, paclitaxel, and 7-epi-10-DAT, respectively. The ranges of limit of quantitation (LOQ) for taxoids were 32, 14, 26, 14, 20, 32, and 26ng/mL for 10-DAB III, baccatin III, 7-xyl-10-DAT, 10-DAT, cephalomannine, paclitaxel, and 7-epi-10-DAT, respectively. Linearity was confirmed over the whole calibration range (0.07–45, 0.058–37.5, 0.058–37.5, 0.056–36.3, 0.053–33.8, 0.057–37.5, and 0.06–38.8μg/mL for 10-DAB III, baccatin III, 7-xyl-10-DAT, 10-DAT, cephalomannine, paclitaxel, and 7-epi-10-DAT, respectively) with coefficients higher than 0.9903. The inter- and intra-day precision of taxoids ranged from 2.86% to 6.31% for retention time and ranged from 3.91% to 7.33% for peak area. The recovery rates of this method were higher than 94.32% for 10-DAB III, 94.68% for baccatin III, 93.65% for 7-xyl-10-DAT, 93.29% for 10-DAT, 92.91% for cephalomannine, 93.41% for paclitaxel, and 93.06% for 7-epi-10-DAT, respectively. [Copyright &y& Elsevier]

• A microdialysis-HPLC was applied in detecting allelochemicals for the first time. • An in-situ detected method without damaging rhizosphere structure was established. • Detection limits were 1.23 µg/g for five phenolic acids. Microdialysis is an important technology in the field of medicine, and with the popularization and development of microdialysis technology, it has been gradually applied to the field of ecology. In this study, microdialysis-HPLC as a new and environmentally friendly in situ technique was developed and applied for the first time to detect phenolic acid allelochemicals in Rehmannia glutinosa rhizosphere. A response surface analysis (RSM) was used to optimize the microdialysis in situ based on the Box–Behnken design (BBD) experiment. The optimum dialysis conditions were obtained as follows: the perfusion solution was a NaCl solution of 200 mmol/L with pH 4, and the dialysis temperature was 37 °C. Under these conditions, the content of phenolic acids in the rhizosphere 10, 20 and 30 cm away from the root was analyzed using high-performance liquid chromatography (HPLC) detection combined with the standard curve of phenolic acid addition. Compared to traditional methods, microdialysis-HPLC is a simple and environmentally friendly method for the in situ detection of phenolic acids in rhizosphere. [ABSTRACT FROM AUTHOR]

Flavonoids are important metabolites of pigeon pea in relation to its stress resistance. However, the molecular basis and regulatory mechanisms of flavonoids in pigeon pea remain unclear. Methyl jasmonate (MeJA) is a signaling molecule associated with biosynthesis of flavonoids. In this study, after exogenous treatment of 10 mg/L MeJA, infection of pathogenic fungi to pigeon pea was alleviated and the content of flavonoids was increased. Results of gene expression and metabolic changes that were respectively analyzed by transcriptome sequencing and high performance liquid chromatography (HPLC) showed that (1) concentrations of various flavonoids, such as genistein, apigenin, vitexin and biochanin A were significantly up-regulated; (2) 13675 differentially expressed genes were produced, mainly enriched in signal transduction and isoflavone biosynthesis pathways: (3) the expression levels of key synthase genes (CcI2′H, CcHIDH, Cc7-IOMT) in the flavonoid biosynthesis pathway were significantly up-regulated; (4) Overexpression of CcbHLH35 significantly induced upregulation of flavonoid synthase genes and accumulation of genistein, vitexin and apigenin. Our findings reveals the pivotal roles of MeJA in synthesis and functioning of flavonoids in pigeon pea, which provide a basis for further studies on flavonoid-mediated defense responses. • MeJA alleviated the pathogenic fungi-induced damages in pigeon pea. • MeJA stimulated the biosynthesis of genistein, apigenin, vitexin and biochanin A. • Cc I2′H , Cc HIDH and Cc7-IOMT were induced to be significantly up-regulated by MeJA. • CcbHLH35 played a regulatory role in flavonoid biosynthesis. [ABSTRACT FROM AUTHOR]

In this study, novel and green natural deep eutectic solvents (NADESs) as extraction media, are presented for the extraction and determination of phenolics in Cajanus cajan leaves by ultra performance liquid chromatography. Fourteen different NADES systems were investigated for the extraction of phenolics with diverse polarity in C. cajan leaves. Experiments with different extraction solvents and multivariate data analysis demonstrated that the NADES composed of choline chloride and maltose possessed excellent extractability for both polar and weak polar compounds compared to conventional solvents. The influential parameters of microwave-assisted extraction procedure with NADES were examined and optimized. The optimized conditions were as follows: extraction solvent 20% of water in choline chloride/maltose (1/2), extraction temperature 60 °C, liquid/solid ratio 30:1 mL/g and irradiation time 12 min. Moreover, a fast and sensitive UPLC method was firstly developed for the comprehensive analysis of fourteen phenolics in C. cajan leaves. This method was performed within 40 min, and exhibited satisfactory linearity, precision and recovery. These results indicate that the proposed method represents an excellent alternative for the extraction and quantification of phenolics in plant materials. [ABSTRACT FROM AUTHOR]

• Abscisic acid (ABA) enhanced drought resistance of pigeon pea. • The content of flavonoids increased after the application of ABA. • Abscisic acid (ABA) regulates the up-regulation of flavonoid synthesis genes. • MYB and bHLH TFs may play a central regulatory role in flavonoid biosynthesis. Abscisic acid (ABA) plays an important role in drought resistance of plants. Plants subjected to drought stress often produce a large number of secondary metabolites. More and more studies indicate that secondary metabolites are directly related to plant stress resistance. However, there are few reports about the influence of ABA regulating secondary metabolism on drought. In our study, we found that as a pioneer tree species in karst landforms, the drought resistance can also be improved by ABA. ABA can reduce the stomatal opening of pigeon pea under drought stress and reduce the influx of H+ from the root tip. At the same time, we found that in ABA-treated pigeon pea, flavonoid metabolites accumulate in large amounts, especially genistein, genistin and pterostilbene. Through transcriptome sequencing analysis, it was found that after ABA treatment, flavonoids biosynthesis pathways changed significantly; among them, the key enzyme genes CHS, HIDH and ROMT in the biosynthesis pathway of genistein, genistin and pterostilbene were significantly up-regulated. According to the co-expression analysis in the transcriptome data, we screened that multiple MYB and bHLH transcription factors may be involved in the regulation of flavonoid accumulation. And the biosynthetic genes contained binding sites of related transcription factors. Through RT-PCR, we further analyzed the expression of MYB, bHLH and related biosynthetic genes after ABA treatment at different times, and found that they were all regulated by ABA. By High Performance Liquid Chromatography (HPLC) analysis, we found the increase of genistein, genistin and pterostilbene content in pigeon pea treated with ABA under drought stress. Through hairy root transgenic analysis, We found that CcMYB114 regulate drought resistance by modulating the expression of CcHIDH1 and CcHIDH3, and then regulating genistein and genistin accumulation in pigeon pea. This study provides evidence to analyze the regulation of ABA in pigeon pea on the metabolism of flavonoids and thus affect its drought resistance. [ABSTRACT FROM AUTHOR]

Blue honeysuckle (Lonicera caerulea var. edulis) berry has been used in folk medicine for the treatment of bacterial infections, gastrointestinal disorders, and metabolic diseases. There is evidence to support its pharmacological effects in improving diabetes, fatty liver, and obesity. To investigate the effect of blue honeysuckle berry extract (BHBE) on lipid accumulation in adipocytes and the underlying mechanism. High-performance liquid chromatography (HPLC) coupled with mass spectrometry (MS) was applied to analyze the polyphenolic compounds in BHBE. 3T3-L1 cells were used to induce into adipocytes. Oil Red O staining combined with triglyceride (TG) content determination were carried out to evaluate intracellular lipid accumulation. Western blot was used to determine the expression of lipogenic enzymes and transcription factors. Real-time PCR was used to determine the expression of lipolytic enzymes and adipocyte markers. The primary polyphenols in BHBE are flavonoids (mainly flavonols and anthocyanins). BHBE dose-dependently inhibited lipid accumulation in adipocytes by reducing the expression of fatty acid synthase (FAS) and increasing the phosphorylation level of acetyl-CoA carboxylase (ACC). Moreover, BHBE was found to promote the phosphorylation of AMP-activated protein kinase (AMPK) and further reduce the expression of lipogenic transcription factors (PPARγ, C/EBPα, and SREBP-1c), while the selective AMPK inhibitor attenuated the suppressive effect of BHBE on lipogenesis. In addition, BHBE increased the expression of beige adipocyte markers (Cd137 and Tmem26) and uncoupling protein 1 (UCP1) without affecting the expression of brown adipocyte markers (Ebf3 and Eva1). BHBE inhibits lipid accumulation in adipocytes by suppressing lipogenesis via AMPK activation as well as by promoting beiging of adipocytes, which supports the anti-obesity potential of blue honeysuckle berry. [Display omitted] • Blue honeysuckle berry extract inhibited lipid accumulation in adipocytes. • Blue honeysuckle berry extract suppressed lipogenesis via AMPK activation. • Blue honeysuckle berry extract promoted beiging of adipocytes. [ABSTRACT FROM AUTHOR]

• Quantitative study on eight components of Acanthopanax senticosus (AS). • Factor analysis (FA) and hierarchical cluster analysis (HCA) process the data. • Qualitative analysis of AS based on fingerprint and similarity analysis (SA). A comprehensive method for the evaluation of Acanthopanax senticosus (AS) was established by the quantitative analysis of multiple components by single marker (QAMS), fingerprint, similarity analysis (SA), hierarchical cluster analysis (HCA), and factor analysis (FA) based on high performance liquid chromatography (HPLC). A total of 27 common peaks were identified in the standard fingerprint of 20 batches of AS from different regions in China, of which 8 peaks were identified as protocatechuic acid, syringin, chlorogenic acid, caffeic acid, eleutheroside E, hyperoside, isofrqxidin, and acacetin, and the concentrations of these eight components were determined simultaneously by QAMS. The results showed that the QAMS method was effective and feasible compared with the external standard method (ESM) (RD < 3.3 %, P ≤ 0.01). Sample 1 (S1) was used as the reference chromatogram, the similarity of other samples was between 0.765 and 0.968. Through HCA, AS could be mainly divided into two production areas, the north Liaoning (including Liaoning) and the south Liaoning areas. Furthermore, FA showed that the quality of AS in the north Liaoning area was better than that in the south Liaoning area. In summary, the method established in this study can comprehensively and systematically evaluate quality differences in AS samples, and may be used to help to improve the quality control of AS. [ABSTRACT FROM AUTHOR]