Screening of anti-fatigue active ingredients of Eleutherococcus senticosus via spectrum-effect relationship based on factor analysis and LC-MS/MS.

ES contains compounds known to have significant anti-fatigue activity. In recent years, it has received extensive attention because it is efficient. However, its active ingredients on antifatigue effect are still unclear. This study attempts to establish the spectrum-effect relationship of ES antifatigue activity to screen the effective components. The results showed that the similarity of 15 ES fingerprints obtained by LC-MS/MS was 0.533-0.992, and the chemical structures of 22 common peaks were identified. The anti-fatigue activity of 15 batches of ES was characterized by forced swimming test of mice and quantified by CAFI, among which S4, S1 and S5 had better activity. 9 components (caffeic acid, 5-(4-O-ß-D-glucosylferoyl)-quinic acid, (±)13-HODE, isofraxidin, eleutheroside E, syringin, pinoresinol diglucoside or its isomer, 7,8-dihydrodehydrocarbinol alcohol-4-O-ß-D-glucoside, secoisolariciresinol-4-O-ß-D-glucoside) highly related to anti-fatigue activity may be the effective components of ES.

Ingenious application of ethylenediaminetetraacetic acid disodium to improve the extraction yield of psoralen in fig (Ficus carica L.) leaves.

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.

Seed oil of Rosa roxburghii Tratt against non-alcoholic fatty liver disease in vivo and in vitro through PPARα/PGC-1α-mediated mitochondrial oxidative metabolism.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD), characterized by hepatic steatosis and hepatocyte injury, is an obesity-induced metabolic dysregulation with few available therapeutic options. Enhancement of the mitochondrial function was considered as an effective treatment for NALFD. Unsaturated fatty acids (UFAs) have been shown to have beneficial effects on metabolic syndrome disease such as hyperlipidemia, coronary artery disease and cardiovascular diseases. The seed oil of Rosa roxburghii Tratt (ORRT) was of high quality in terms of its high amount of unsaturated fatty acids. However, the effects of ORRT on NALFD have not been reported so far. PURPOSE: The study aimed to evaluate the protective effects and molecular mechanism of ORRT for the treatment of NAFLD in vivo and in vitro. METHODS: The beneficial effects, especially improving the mitochondrial function, and the potential mechanism of ORRT on NAFLD were studied both in vivo and in vitro. Lipid levels were determined by triglyceride (TG), total cholesterol (TC), and Oil Red O staining. Oxidative stress and inflammation were assessed by detecting antioxidant enzyme activity, MDA content, and ELISA assay. Blood TG, TC, HDL-c and LDL-c levels were measured in HFD mice. Western blot analyses were used to determine the levels of the protein involved in fatty acid oxidation, oxidative metabolism, and mitochondria biogenesis and function. The mitochondrial membrane potential level was measured by JC-1 staining to teste the effect of ORRT on mitochondrial function in vitro. GW6471 (inhibitor of PPARα) was used to confirm the relationship between PPARα and PGC-1α. RESULTS: ORRT significantly restrained NAFLD progression by attenuating lipid accumulation, oxidative stress and inflammatory response. Furthermore, ORRT upregulated thermogenesis-related gene expressions, such as uncoupling protein 1 (UCP1) and p38 mitogen-activated protein kinase (p38 MAPK). The results showed that the expression of key genes involved in fatty acid oxidation (e.g., CPT-1α, ACADL, PPARα) and in mitochondrial biogenesis and function (e.g., TFAM, NRF1, PGC-1α, and COX IV) was significantly increased. Together with the observed MMP improvement, these findings suggested that ORRT activated the mitochondrial oxidative pathway. Additionally, GW6471 inhibited the ORRT on promoting the expression of PGC-1α, CPT-1α, and ACADL. In conclusion, ORRT possessed the potential to prevent lipid accumulation via the PPARα/PGC-1α signaling pathway, which could be developed as a natural health-promoting oil against NAFLD.

Quality evaluation of Acanthopanax senticosus via quantitative analysis of multiple components by single marker and multivariate data analysis.

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.

Ultrasonic/microwave-assisted extraction of polysaccharides from Camptotheca acuminata fruits and its antitumor activity.

In the present study, an efficient ultrasonic/microwave-assisted extraction (UMAE) procedure for the polysaccharides from the fruit of Camptotheca acuminata (CAFP) was investigated and optimized. Under the optimum conditions (ratio of liquid to raw material 30 mL/g, microwave irradiation time of 20 min, microwave irradiation power of 570 W and a fixed ultrasonic power of 50 W obtained by the response surface analysis with Box-Behnken design, satisfactory yields of CAFP (6.81 ± 0.04%) were achieved. The development UMAE technique produced higher yields in a shorter time than conventional hot water extraction (HWE): 20 vs. 120 min. In addition, in vivo CAFP at suitable dose is effective on H22 murine hepatoma strains, and CAFP significantly inhibited the proliferation of human oral carcinoma KB, pancreatic carcinoma BXCP-3 and gastric carcinoma SGC-7901 cells in vitro, indicating CAFP might be suitable for nature antitumor therapeutic agent development.

Application of cavitation system to accelerate aqueous enzymatic extraction of seed oil from Cucurbita pepo L. and evaluation of hypoglycemic effect.

Cavitation-accelerated aqueous enzymatic extraction (CAEE) of seed oil from Cucurbita pepo was performed. An enzyme cocktail comprised of cellulose, pectinase and proteinase can work synergistically in releasing the oil. The CAEE extraction conditions were optimized by a Plackett-Burman design followed by a central composite methodology. A maximal extraction yield of 58.06% was achieved under optimal conditions of vacuum degree -0.07, enzyme amount 1.05% and extraction time 69min. As compared to soxhlet extraction (SE)-derived oil, CAEE-derived oil exhibited similar physical properties and better oxidation stability. In addition, chemical composition analyzing showed that the content of linoleic acid obtained by CAEE (47.67%) was higher than that of SE (44.51%). Moreover, the IC50 of oil obtained by CAEE and SE, as measured by α-amylase inhibition assay, were 40.68µg/mL and 45.46µg/mL. All results suggest that CAEE represents an excellent alternative protocol for production of oil from oil-bearing materials.

An effective homogenate-assisted negative pressure cavitation extraction for the determination of phenolic compounds in pyrola by LC-MS/MS and the evaluation of its antioxidant activity.

A novel extraction method, homogenate-assisted negative pressure cavitation extraction (HNPCE), was designed for the extraction and determination of the main phenolic compounds of Pyrola incarnata Fisch. by LC-MS/MS. The particle sizes and extraction yields in the process of homogenization were compared with conventional pulverization. The results showed that homogenization for less than 120 s could produce more suitable particle size powders for analyte extraction. The following NPCE parameters were optimized by a BBD test and under the optimal conditions, the maximum extraction yields of arbutin, epicatechin, hyperin, 2'-O-galloylhyperin and chimaphilin increased by 68.7%, 72.0%, 43.3%, 62.5% and 34.5% with respect to normal NPCE. The LC-MS/MS method was successfully applied for the quantification of five target compounds in pyrola, and the results of the precision test indicated a high accuracy of the present method for the quantification of the target compounds in pyrola. Furthermore, the antioxidant activities of the pyrola extracts were also determined. The results showed that pyrola had good antioxidant activities and it was a valuable antioxidant natural source.

Optimization of ionic liquid based simultaneous ultrasonic- and microwave-assisted extraction of rutin and quercetin from leaves of velvetleaf (Abutilon theophrasti) by response surface methodology.

An ionic liquids based simultaneous ultrasonic and microwave assisted extraction (ILs-UMAE) method has been proposed for the extraction of rutin (RU), quercetin (QU), from velvetleaf leaves. The influential parameters of the ILs-UMAE were optimized by the single factor and the central composite design (CCD) experiments. A 2.00 M 1-butyl-3-methylimidazolium bromide ([C4mim]Br) was used as the experimental ionic liquid, extraction temperature 60°C, extraction time 12 min, liquid-solid ratio 32 mL/g, microwave power of 534 W, and a fixed ultrasonic power of 50 W. Compared to conventional heating reflux extraction (HRE), the RU and QU extraction yields obtained by ILs-UMAE were, respectively, 5.49 mg/g and 0.27 mg/g, which increased, respectively, 2.01-fold and 2.34-fold with the recoveries that were in the range of 97.62-102.36% for RU and 97.33-102.21% for QU with RSDs lower than 3.2% under the optimized UMAE conditions. In addition, the shorter extraction time was used in ILs-UMAE, compared with HRE. Therefore, ILs-UMAE was a rapid and an efficient method for the extraction of RU and QU from the leaves of velvetleaf.

The galloyl catechins contributing to main antioxidant capacity of tea made from Camellia sinensis in China.

Total polyphenol content, catechins content, and antioxidant capacities of green, dark, oolong, and black teas made from Camellia sinensis in China were evaluated. The total polyphenol content of 20 samples of tea was in the range of 7.82-32.36%. Total catechins content was in the range of 4.34-24.27%. The antioxidant capacity of tea extract was determined by the oxygen radical absorbance capacity (ORAC) test and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging test. Total polyphenol content, catechins content, and antioxidant capacity decreased in the following order: green > oolong > black > dark tea. A positive correlation existed between the antioxidant capacity and total polyphenol content or catechins content (R (2) = 0.67-0.87). The antioxidant capacities of five major catechins (epigallocatechin gallate (EGCG), epicatechin gallate (ECG), epicatechin, epigallocatechin, and catechin) were determined by online HPLC DPPH radical-scavenging; the antioxidant activity of tea was mainly attributed to the esterified catechins (EGCG or ECG).

Variation of active constituents and antioxidant activity in pyrola [P. incarnata Fisch.] from different sites in Northeast China.

The variation of antioxidant activity and active components in pyrola [Passiflora incarnata Fisch.] from eight sites in Northeast China were investigated. Total phenolic and flavonoid contents were determined and varied within the range of 39.66-181.48 mg/g and 2.47-22.11 mg/g, respectively. Antioxidant activities were determined by scavenging activity against DPPH and ABTS, by a reducing power test and by a ß-carotene-linoleic acid bleaching test. The IC50 of Tahe samples determined by the DPPH test was 0.106±0.006 mg/mL which was very close to that of Vc (0.076±0.004 mg/mL). The Tahe samples had good antioxidant activity. Principal component activity analysis indicated that the Tahe samples of P. incarnata had the highest potential antioxidant properties, and may be a valuable antioxidant natural resource in the northeast of China.

Development of sample preparation method for eleutheroside B and E analysis in Acanthopanax senticosus by ionic liquids-ultrasound based extraction and high-performance liquid chromatography detection.

An ionic liquids-based ultrasonic-assisted extraction (ILUAE) method was successfully developed for extracting eleutheroside B and E from Radix Acanthopanax senticosus. Thirteen 1-alkyl-3-methylimidazolium ionic liquids with different cations and anions were investigated and 1-butyl-3-methylimidazolium bromide ([C4mim]Br) solution was selected as the solvent. The conditions for ILUAE, including the ionic liquid concentration, soaking time, ultrasonic power, ultrasonic time, solid-liquid ratio and number of extraction cycles, were optimized. With the proposed method, the energy consumption time was reduced to 30 min, whereas conventional method requires about 4h. The proposed method had good recovery (97.96-103.39%) and reproducibility (RSD, n=5; 3.3% for eleutheroside B, 4.6% for eleutheroside E). ILUAE was an efficient, rapid and simple sample preparation technique that showed high reproducibility and was environmental friendly.

Aqueous enzymatic process assisted by microwave extraction of oil from yellow horn (Xanthoceras sorbifolia Bunge.) seed kernels and its quality evaluation.

In this study, aqueous enzymatic process (AEP) assisted by microwave extraction (ME) of oil from yellow horn (Xanthoceras sorbifolia Bunge.) seed kernel was investigated. Central composite design (CCD) and response surface methodology (RSM) were used to optimise an enzyme cocktail (cellulase, hemicellulase, pectinase) for AEP. The main factors of ME were also studied. A maximal oil extraction yield of 55.8% was achieved under optimal conditions. Moreover, scanning electron microscope (SEM) was applied to characterise the extraction process. Analysing chemical composition of the extracted oil by GC-MS showed that the content of unsaturated fatty acids by this emerging method (91.18%) was similar to that by conventional organic solvent extraction (88.76%). In addition, the main physicochemical properties and antioxidant activities of yellow horn oil were measured to evaluate its quality. The present research supported necessary data for the green extraction method of edible oil in food industry.

The phytoestrogenic compound cajanol from Pigeonpea roots is associated with the activation of estrogen receptor α-dependent signaling pathway in human prostate cancer cells.

In the present study, the main natural estrogen-agonist/antagonist from Pigeonpea roots was studied by the estrogen receptor α-dependent signaling pathway in human prostate cancer cell. First, the natural products with estrogenic activity in Pigeonpea roots were screened by pER8-GFP transgenic Arabidopsis, and cajanol (5-hydroxy-3-(4-hydroxy-2-methoxyphenyl)-7-methoxychroman-4-one) was confirmed as the active compound. Further study showed that cajanol significantly arrested the cell cycle in the G1 and G2/M phase and induced nuclei condensation, fragmentation and the formation of apoptotic bodies. Western blotting showed that cajanol modulated the ERα-dependent PI3K pathway and induced the activation of GSK3 and CyclinD1 closely following the profile of PI3K activity. Based on above results, we proposed a mechanism through which cajanol could inhibit survival and proliferation of estrogen-responsive cells (PC-3 cells) by interfering with an ERα-associated PI3K pathway, following a process that could be dependent of the nuclear functions of the ERα. Above all, we conclude that cajanol represents a valuable natural phytoestrogen source and may potentially be applicable in health food industry.

Dihydroquercetin (DHQ) induced HO-1 and NQO1 expression against oxidative stress through the Nrf2-dependent antioxidant pathway.

Dihydroquercetin (DHQ) is a well-known antioxidant agent. In the present investigation, we reported for the first time that DHQ stimulates the expression of phase II detoxifying enzymes through the Nrf2-dependent signaling pathway. The IC50 values of DHQ for reduction of 2,2-diphenyl-1-picrylhydrazol (DPPH), reducing power assay, lipid peroxidation assay, and xanthine oxidase inhibition were 5.96, 4.31, 2.03, and 13.24 µM, respectively. DHQ possessed considerable protective activity from oxidative DNA damage. A luciferase reporter assay also demonstrated that DHQ-activated signaling resulted in the increased transcriptional activity of Nrf2 through binding to the ARE (antioxidant response element) enhancer sequence. Furthermore, Western blotting and luciferase assay revealed DHQ activated ERK1/2, Akt, and JNK signaling pathways, subsequently leading to Nrf2 nuclear translocation. DHQ upregulated the Nrf2-related antioxidant genes heme oxygenase-1 (HO-1), NAD(P)H quinone oxidoreductase-1 (NQO1), and glutamate-cysteine ligase modifier subunits. Inhibition of Nrf2 by siRNA reduced DHQ-induced upregulation of these antioxidant genes.

UV-induced changes of active components and antioxidant activity in postharvest pigeon pea [Cajanus cajan (L.) Millsp.] leaves.

In this study, the effect of UV irradiation (UV-A, UV-B, and UV-C) on phytochemicals, total phenolics, and antioxidant activity of postharvest pigeon pea leaves was evaluated. The response of pigeon pea leaves to UV irradiation was phytochemical specific. UV-B and UV-C induced higher levels of phytochemicals, total phenolics, and antioxidant activity in pigeon pea leaves compared with UV-A. Furthermore, UV-B irradiation proved to possess a long-lasting effect on the levels of phenolics and antioxidant activity. After adapting for 48 h at 4 °C following 4 h UV-B irradiation, total phenolics and antioxidant activity were approximately 1.5-fold and 2.2-fold increased from 39.4 mg GAE/g DM and 15.0 µmol GAE/g DM to 59.1 mg GAE/g DM and 32.5 µmol GAE/g DM, respectively. These results indicate that UV irradiation of pigeon pea leaves can be beneficial in terms of increasing active components and antioxidant activity.

Optimization of shikonin homogenate extraction from Arnebia euchroma using response surface methodology.

An efficient homogenate extraction technique was employed for extracting shikonin from Arnebia euchroma. The homogenate extraction procedure was optimized and compared with other conventional extraction techniques. The proposed method gave the best result with the highest extraction efficiency in the shortest extraction time. Based on single-factor experiments, a three-factor-three-level experimental design has been developed by Box-Behnken design. The optimal conditions were 78% ethanol as solvent, homogenate extraction time of 4.2 min, 10.3 liquid to solid ratio and two extraction cycles. Moreover, the proposed method was validated by stability, repeatability and recovery experiments. The developed homogenate extraction method provided a good alternative for the extraction of shikonin from A. euchroma. The results indicated that the proposed homogenate extraction was a convenient, rapid and efficient sample preparation technique and was environmental friendly. Furthermore, homogenate extraction has superiority in the extraction of thermally sensitive compounds from plant matrices.

Development of sample preparation method for isoliquiritigenin, liquiritin, and glycyrrhizic acid analysis in licorice by ionic liquids-ultrasound based extraction and high-performance liquid chromatography detection.

An ionic liquid-based ultrasonic-assisted extraction (ILUAE) method had been used for the effective extraction of isoliquiritigenin (IQ), liquiritin (LQ) and glycyrrhizic acid (GA) from licorice. The ionic liquids with different cations and anions were investigated in this work and 0.5 M 1-butyl-3-methylimidazolium bromide solution was selected as solvent. In addition, the technical parameters including soaking time, solid-liquid ratio, ultrasonic power and time were optimized. Compared with the conventional solvent extraction, the proposed approach exhibited higher efficiency, which indicated the ILUAE was an efficient, rapid and simple sample preparation technique. There was no degradation of the target analytes had been observed at the optimum conditions which was evidenced by the stability studies performed with standard of IQ, LQ and GA. The proposed method also showed high reproducibility and was environmental friendly.

Effect of corilagin on membrane permeability of Escherichia coli, Staphylococcus aureus and Candida albicans.

Corilagin is a member of polyphenolic tannins. Its antimicrobial activity and action mechanism against Escherichia coli, Staphylococcus aureus and Candida albicans were investigated through membrane permeability. Crystal violet staining determination, outer membrane (OM) and inner membrane (IM) permeability, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and atomic force microscopy (AFM) were used as methods for our investigation. The minimum inhibitory concentrations were 62.5, 31.25 and 62.5 µg/mL for E. coli, S. aureus and C. albicans, respectively. Crystal violet results and SDS-PAGE of supernatant proteins showed that corilagin dose-dependently affected membrane permeability of E. coli and C. albicans but not of S. aureus. OM and IM permeability assays revealed comparable results for E. coli. By using AFM, we demonstrated extensive cell surface alterations of corilagin-treated E. coli and C. albicans. SDS-PAGE of precipitated proteins revealed possible targets of corilagin, i.e. Fib, Sae R, Sar S in S. aureus and Tye 7p in C. albicans. In conclusion, corilagin inhibited the growth of E. coli and C. albicans by disrupting their membrane permeability and that of S. aureus by acting on Fib, Sae R and Sar S but not on membrane integrity.

Content and color stability of anthocyanins isolated from Schisandra chinensis fruit.

In this work, a multivariate study based on Box-Behnken Design was used to evaluate the influence of three major variables affecting the performance of the extraction process of Schisandra chinensis anthocyanins. The optimum parameters were 5.5 h extraction time; 1:19 solid-liquid ratio and 260 r/min stirring rate, respectively. The extraction yield of anthocyanins was 29.06 mg/g under the optimum conditions. Moreover, many factors on the impact of heating, ultrasound, microwave treatment and ultraviolet irradiation on content and color stability of anthocyanins from Schisandra chinensis fruit were investigated. The results show that thermal degradation reaction of anthocyanins complies with the first order reaction kinetics, and the correlation coefficient is greater than 0.9950 at 40-80°C. Ultrasound and microwave treatment has little effect on the stability of anthocyanins, and the extraction time of ultrasound and microwave should be no more than 60 min and 5 min, respectively. The anthocyanins degradation effect of UVC ultraviolet radiation is greater than UVA and UVB; after 9 h ultraviolet radiation, the anthocyanins content degradation of UVC is 23.9 ± 0.7%, and the ∆E* was changed from 62.81 to 76.52 ± 2.3. Through LC-MS analysis, the major composition of Schisandra chinensis anthocyanins was cyanidin-3-O-xylosylrutinoside.

Ultrasound-assisted extraction of carnosic acid and rosmarinic acid using ionic liquid solution from Rosmarinus officinalis.

Ionic liquid based, ultrasound-assisted extraction was successfully applied to the extraction of phenolcarboxylic acids, carnosic acid and rosmarinic acid, from Rosmarinus officinalis. Eight ionic liquids, with different cations and anions, were investigated in this work and [C(8)mim]Br was selected as the optimal solvent. Ultrasound extraction parameters, including soaking time, solid-liquid ratio, ultrasound power and time, and the number of extraction cycles, were discussed by single factor experiments and the main influence factors were optimized by response surface methodology. The proposed approach was demonstrated as having higher efficiency, shorter extraction time and as a new alternative for the extraction of carnosic acid and rosmarinic acid from R. officinalis compared with traditional reference extraction methods. Ionic liquids are considered to be green solvents, in the ultrasound-assisted extraction of key chemicals from medicinal plants, and show great potential.