Effects of age and tissue of Juniperus sabina L. on its phytochemical characteristics, anti-cholinesterase, antidiabetes, and anti-drug resistant bacteria activities.

Juniperus sabina L. is used in the traditional Chinese medicine (TCM) system to prevent or treat various diseases. However, only the leaves and branches are used as medicinal parts. The aim of this study was to compare the chemical characteristics of different tissues (leaves, branches, stems, and roots) of J. sabina at different ages by HPLC-MS and to evaluate the biological activity (enzyme inhibition, anti-drug-resistant bacteria). Total phenol (TPC) and total lignan (TLC) contents in J. sabina were determined by Folin-Ciocalteu method and UV spectrophotometry, respectively. High levels of total phenols (87.16 mg GAE/g dry weight) and total lignans (491.24 mg PPT/g dry weight) were detected in fifteen annual J. sabina roots and current year leaves, respectively. Eleven compounds, of which six were phenolic compounds and five were lignans, were identified and quantified by HPLC/HPLC-MS. Statistical analysis showed that the distribution and content of the detected compounds showed considerable variation among ages and tissues, and that the current year leaves of fifteen annual J. sabina could be used as a potential application site for the source of podophyllotoxin. Acetylcholinesterase (AChE) inhibitory activity was found to be the highest on the extracts of fifteen annual J. sabina current year leaves (47.37 µg/mL), while the highest inhibition towards butyrylcholinesterase (BChE) was observed for the extracts of seven annual J. sabina previous year leaves (136.3 µg/mL). And the second annual J. sabina current year stem's extracts showed the best antidiabetic activity (anti-α-glucosidase, 62.59 µg/mL). In addition, the extracts of fifteen annual J. sabina roots (47.37 µg/mL) showed the highest anti-MRSA activity (31.25 µg/mL). Redundancy analysis (RDA) was conducted to clarify the factors affecting the biological activity of J. sabina, and its results showed that epicatechin and matairesinol showed positive promotion. This study provides a new perspective for understanding the chemical differences and comprehensive utilization of different tissues of J. sabina.

Juniperus sabina L. as a Source of Podophyllotoxins: Extraction Optimization and Anticholinesterase Activities.

Juniperus sabina L. (J. sabina) has been an important plant in traditional medicine since ancient times. Its needles are rich in podophyllotoxin, a precursor compound to anti-tumor drugs. However, no systematic research has been done on J. sabina as a source of podophyllotoxins or their biological action. Hence, extracts of podophyllotoxin and deoxypodophyllotoxin were the main optimization targets using the Box-Behnken design (BBD) and response surface methodology (RSM). The total phenol content and antioxidant activity of J. sabina needle extract were also optimized. Under the optimal process conditions (ratio of material to liquid (RLM) 1:40, 90% methanol, and ultrasonic time 7 min), the podophyllotoxin extraction rate was 7.51 mg/g DW, the highest level reported for Juniperus spp. distributed in China. To evaluate its biological potential, the neuroprotective acetyl- and butyrylcholinease (AChE and BChE) inhibitory abilities were tested. The needle extract exhibited significant anti-butyrylcholinesterase activity (520.15 mg GALE/g extract), which correlated well with the high levels of podophyllotoxin and deoxypodophyllotoxin. This study shows the potential medicinal value of J. sabina needles.

Sex-Related Differences on Chemical Composition, Anatomy, Histochemistry, and Biological Activities of Juniperus rigida.

Sex-related differences on phenolic profiles, chemical composition of essential oils, anatomy, histochemistry and biological activities (antioxidant and antibacterial activities) of Juniperus rigida needles collected from Yijun and Fugu region were first studied. In two regions, female and male had similar contents of total phenolic and total flavonoid. 10 phenolic compounds were analyzed by RP-HPLC, amentoflavone content was significantly higher in female than male in Yijun, and chlorogenic acid content was significantly higher in female than male in Fugu. 30 compounds (over 0.5 %) were detected in the essential oils, and the total contents of female were lower than male in Yijun. This difference mainly comes from Germacrene D, which was about twice as high in male as in female. Male needles had significantly larger mechanical tissue and phloem in Yijun. Histochemical analysis indicated that the phenols were stored in epidermal cells, sponge tissue, endodermis cells, edge of resin duct, stomatal bands, and the flavonoids were stored in epidermal cells, endodermis cells, edge of resin duct, stomatal bands. No sex-related differences were found in histochemical analysis, antioxidant activities (ABTS, FRAP) and antibacterial activities (9 strains). This preliminary study provided a reference for production practice and theoretical research of J. rigida.

Quality evaluation of different varieties of Zanthoxylum bungeanum Maxim. peels based on phenolic profiles, bioactivity, and HPLC fingerprint.

This work aims to evaluate the quality of seven varieties of Zanthoxylum bungeanum peels based on phenolic compounds, bioactivity, and HPLC fingerprint combined with chemometrics analysis, and pick out the key chemical compounds. As a result, S2 (Fengxian Dahongpao) showed the highest content of total phenolic and rutin, as well as the strongest antioxidant activity, followed by S6 (Hancheng Shizitou). From HPLC fingerprint, eight common characteristic peaks were selected and proved to be effective in controlling the quality of Z. bungeanum peels with the total contribution of 92.775%. The similarity values of each sample ranged between 0.696 and 0.970. In addition, hyperoside was identified as another key chemical indicator for quality evaluation of Z. bungeanum peels by PCA. Accordingly, seven varieties of Z. bungeanum were classified into four groups, among which group 4 (Fengxian Dahongpao, S2) was considered as an outstanding variety, followed by group 1 (S5, S6, and S7) and group 3 (S3 and S4), and group 2 (Fugu late-maturing, S1) was the last one. These results will be helpful to establish an effective and comprehensive evaluation system of Z. bungeanum. PRACTICAL APPLICATION: This study not only provides another index for quality evaluation of Z. bungeanum peels, but it also expects to be a theoretical basis for high-quality germplasm cultivation and the development of Z. bungeanum peels as food additives, pharmaceutical, or health care products.

Chemical composition, antioxidant, antibacterial and cholinesterase inhibitory activities of three Juniperus species.

The chemical composition, antioxidant, antibacterial and cholinesterase inhibitory activities of three Juniperus species were studied. The contents of total phenolic and 10 phenolic compounds were highest in Juniperus rigida Sieb.et Zucc., of which catechin and cumaric acid were the predominant phenolic compounds, but were lowest in Juniperus sibirica Burgsd. GC-MS analysis showed the highest contents of essential oils were in J. rigida (92.61%), followed by Juniperus formosana Hayata (87.30%) and J. sibirica (84.89%). The a-pinene was the most dominant compound in J. rigida (23.99%) and J. formosana (9.71%), however, it has not been detected in J. sibirica. Ethanol extracts showed the higher radical scavenging capacity in ABTS, FRAP and DPPH assays than essential oils. The essential oils and ethanol extracts of J.sibirica showed the strong antibacterial activity against Salmonella typhimurium and Escherichia coli. Three Juniperus species showed certain acetylcholinesterase and butyrylcholinesterase inhibitions and J. formosana showed better cholinesterase inhibitory.

Chemical composition, antibacterial activity and related mechanism of valonia and shell from Quercus variabilis Blume (Fagaceae) against Salmonella paratyphi a and Staphylococcus aureus.

BACKGROUND: Plant secondary metabolites and phytochemicals that exhibit strong bioactivities have potential to be developed as safe and efficient natural antimicrobials against food contamination and addressing antimicrobial resistance caused by the overuse of chemical synthetic preservative. In this study, the chemical composition, antibacterial activities and related mechanism of the extracts of the valonia and the shell of Quercus variabilis Blume were studied to determine its potential as a safe and efficient natural antimicrobial. METHODS: The phenolic compositions of valonia and shell extracts were determined by folin-ciocalteau colourimetric method, sodium borohydride/chloranil-based assay and the aluminium chloride method and then further identified by the reverse-phase HPLC analysis. The antibacterial activities of valonia and shell extracts were evaluated by the agar disk diffusion method and agar dilution method. The related antibacterial mechanism was explored successively by the membrane of pathogens effect, phosphorous metabolism, whole-cell proteins and the microbial morphology under scanning electron microscopy. RESULTS: The n-butanol fraction and water fraction of valonia along with n-butanol fraction of the shell contains enrich phenolics including ellagic acid, theophylline, caffeic acid and tannin acid. The n-butanol fraction and ethanol crude extracts of valonia exhibited strong antibacterial activities against Salmonella paratyphi A (S. paratyphi A) and Staphylococcus aureus (S. aureus) with the DIZ values ranged from 10.89 ± 0.12 to 15.92 ± 0.44, which were greater than that of the Punica granatum (DIZ: 10.22 ± 0.18 and 10.30 ± 0.21). The MIC values of the n-butanol fraction and ethanol crude extracts of valonia against S. paratyphi A and S. aureus were 1.25 mg/ml and 0.625 mg/ml. The related antibacterial mechanism of n-butanol fraction and ethanol crude extracts of valonia may be attributed to their strong impact on membrane permeability and cellular metabolism. Those extracts exhibited strong antibacterial activity according to inhibit the synthesis of bacterial proteins and seriously change morphological structure of bacterial cells. CONCLUSIONS: The n-butanol fraction and ethanol crude extracts of valonia had reasonably good antibacterial activities against S. paratyphi A and S. aureus. This study suggests possible application of valonia and shell as natural antimicrobials or preservatives for food and medical application.

Anatomical, Phytochemical, and Histochemical Study of Juniperus rigida Needles at Different Altitudes.

Needles of Juniperus rigida are used in Chinese traditional medicine for the treatment of brucellosis, dropsy, skin disease, and rheumatoid arthritis. This is the first study that reports anatomical structures of the J. rigida needles collected at different altitudes. The most common anatomical, phytochemical, and histochemical techniques and methods are used. The results show that anatomical structures and chemical composition change significantly at different altitudes. The main anatomical characters are significant xeromorphic structures (thick epidermis, hypodermis, and cuticle), a stomatal band, a developed vascular bundle, and a marginal resin duct. The xeromorphic structures become more pronounced with increasing altitude. The phytochemical and histochemical results demonstrate that the content of the main chemical compounds (phenols and terpenoids) basically increases at a higher elevation. Histochemical analysis localizes the phenols in epidermal cells, sponge tissue, endothelial layer cells, and stomatal bands, and the terpenoids in palisade tissue, sponge tissue, and the edge of the resin duct. This work reveals the relation between anatomy and chemistry in J. rigida needles, contributes to the quality control of its ethno-medicine, and provides the evidence to develop the commercial cultivation.

Polyphenols of Leaf, Litter and Soil of Pinus bungeana across China and Their Responses to Ecological Factors.

The importance of phenolic compounds for responding to various environmental conditions has been widely emphasized. However, the role of interactions between polyphenols and ecological factors, especially C, N, and P stoichiometry was little studied. Here, 15 sites across five provinces of Pinus bungeana in temperate regions of China were studied. The results showed that the higher values of total phenolic contents (TPC) of leaf and litter were distributed among the north distribution area of P. bungeana, lower values were in the south, whereas soil TPC were contrary to leaf and litter TPC. The stepwise regression, path analysis and decision index of path analysis for leaf TPC and ecological factors showed that altitude had the most direct impact on leaf TPC. Moreover, the principal determinants of leaf, litter and soil TPC were soil C/P ratios, longitude, and soil N/P ratios, respectively. In addition, the leaf, litter and soil TPC of P. bungeana were limited by soil C/N ratios, mean annual temperature, and soil P, respectively. Overall, our study provided evidence that ecological factors affected strongly the leaf, litter and soil TPC of P. bungeana.

Simultaneous Enrichment and Separation of Four Flavonoids from Zanthoxylum bungeanum Leaves by Ultrasound-Assisted Extraction and Macroporous Resins with Evaluation of Antioxidant Activities.

Quercitrin, hyperoside, rutin, and afzelin are the dominant flavonoids compounds from Zanthoxylum bungeanum leaves, and they play major roles in the antioxidant activity. Macroporous adsorption resin (MAR) treatment, a simple, low-cost and efficient method, was combined with ultrasound-assisted extraction (UAE) to enrich and purify these four flavonoids from Z. bungeanum leaves efficiently. The optimal conditions for UAE based on Response Surface Methodology (RSM) were determined to be an ethanol concentration of 60%, leaves size of 40 mesh, temperature of 50 °C and ultrasonic power of 400 W with four flavonoids contents of 120.84 mg/g. After the extraction process, five kinds of MARs (D4020, D-101, NKA-9, AB-8, and X-5) were tested through static adsorption/desorption to enrich and purify the ultrasonic-assisted extracts, and D-101 was selected as the most suitable resin. The optimal adsorption conditions were 5 bed volumes (BV) of sample solution with an initial concentration of 7.5 mg/mL and pH 5.0. Meanwhile, the optimal desorption parameters were 5 BV each of deionized water and 30% ethanol, then 10 BV 70% ethanol, and a flow rate of 2 BV/hr. Under the optimized conditions, the contents of quercitrin, hyperoside, rutin, and afzelin increased by 276.39%, 187.46%, 221.81%, and 288.45%, respectively, and the recovery yields were 85.47%, 73.53%, 81.35%, and 65.06%. In addition, laboratory preparative-scale separation indicated that the preparative separation of four flavonoids was feasible and easy. Moreover, the antioxidant activities of the purified products were significantly increased after enrichment. In conclusion, all of the results indicated that these methods are highly efficient, low cost, environmentally friendly and easy to scale up. PRACTICAL APPLICATION: This study provided an environmentally friendly, rapid, and highly productive method for the extraction and purification of four active compounds from Zanthoxylum bungeanum leaves. The results can be used for the utilization of Z. bungeanum leaves as a kind of food supplement in an industrial setting.

Metabolite Profiles, Bioactivity, and HPLC Fingerprint of Different Varieties of Eucommia ulmoides Oliv.: Towards the Utilization of Medicinal and Commercial Chinese Endemic Tree.

Eucommia ulmoides Oliv. is widely regarded in China as a precious medicinal and commercial endemic tree. Due to cross-breeding or natural variation of E. ulmoides, the metabolite composition may vary significantly, making control of the medical quality difficult. In order to improve the rational development and utilization, the quality of seven varieties of E. ulmoides were evaluated based on metabolite profiles (total phenolic, total flavonoid, gutta-percha, aucubin, geniposidic acid, chlorogenic acid, geniposide, pinoresinol diglucoside, rutin, hyperoside, and astragalin), bioactivities (in vitro, in vivo antioxidant activities, and antibacterial activities) and HPLC fingerprint combined with chemometrics analysis. On this basis, the differences of medicinal parts (leaf and bark) were further carried out. For the traditional use of bark, Purple-leaf E. ulmoides was the most suitable. For the use of leaf, Qinzhong 1 and Purple-leaf E. ulmoides were appropriate. HPLC fingerprint analysis showed that significant differences in metabolite profiles exist among seven varieties of E. ulmoides. Combined with chemometrics analysis, seven varieties of E. ulmoides were divided into three groups from the use of leaf and bark. The analysis not only evaluated quality of seven varieties of E. ulmoides, but also could distinguish different varieties and different regions of origin. The results can provide theoretical basis for E. ulmoides resources utilization and cultivation of fine varieties.

Synergistic effects and related bioactive mechanisms of Potentilla fruticosa Linn. leaves combined with green tea polyphenols studied with microbial test system (MTS).

Previous research found Potentilla fruticosa leaf extracts (PFE) combined with green tea polyphenols (GTP) showed obvious synergistic effects based on chemical mechanisms. This study further confirmed the synergy of PFE + GTP viewed from bioactivities using the microbial test system (MTS). The MTS antioxidant activity results showed the combination of PFE + GTP exhibited synergistic effect and the ratio 3:1 showed the strongest synergy, which were in accordance with the results in H2O2 production rate. The combination of PFE + GTP promoted CAT and SOD enzyme activity and their gene expression especially at the ratio 3:1. Therefore, the synergism of PFE + GTP may be due to the promotion of CAT and SOD genes expression which enhanced the CAT and SOD enzyme activities. These results confirmed the synergy of PFE + GTP and could provide theoretical basis to produce a compounded tea made of a mixture of leaves from Potentilla species.

Plants as highly diverse sources of construction wood, handicrafts and fibre in the Heihe valley (Qinling Mountains, Shaanxi, China): the importance of minor forest products.

BACKGROUND: Chinese rural communities living among species-rich forests have little documentation on species used to make handicrafts and construction materials originating from the surrounding vegetation. Our research aimed at recording minor wood uses in the Heihe valley in the Qinling mountains. METHODS: We carried out 37 semi-structured interviews in seven villages. RESULTS: We documented the use of 84 species of plants. All local large canopy trees are used for some purpose. Smaller trees and shrubs which are particularly hard are selectively cut. The bark of a few species was used to make shoes, hats, steamers and ropes, but this tradition is nearly gone. A few species, mainly bamboo, are used for basket making, and year-old willow branches are used for brushing off the chaff during wheat winnowing. CONCLUSIONS: The traditional use of wood materials documented suggests that some rare and endangered tree species may have been selectively cut due to their valuable wood, e.g. Fraxinus mandshurica and Taxus wallichiana var. chinensis. Some other rare species, e.g. Dipteronia sinensis, are little used and little valued.

Allelopathic Effects, Physiological Responses and Phenolic Compounds in Litter Extracts of Juniperus rigidaSieb. et Zucc.

The allelopathic effects of Juniperus rigida litter aqueous extract (LE) on wheat and Pinus tabuliformis were studied, as well as the physiological responses to the extract. High concentration LE (0.10 g Dw/ml) significantly inhibited the seed germination and seedling growth in receptor plants. The chlorophyll content and root activity in the wheat seedlings were reduced significantly across all treatments; however, those were more prominently reduced at high concentration (0.10 g Dw/ml) but received little stimulation at low concentration (0.025 g Dw/ml) in P. tabuliformis. The content of malonaldehyde (MDA) increased with increasing concentrations of LE, except at 0.025 g Dw/ml. Activities of antioxidant enzymes (POD, CAT and SOD) in receptor plants were all significantly inhibited at high concentrations but stimulated at low concentrations. These results demonstrate that the aqueous extract from J. rigida litter has allelopathic potential. Various phenolic compounds were identified in litter aqueous extract and litter ethanol extract by HPLC. The phenolic compound content in the aqueous extract was significantly lower than that in the ethanol extract. Chlorogenic acid and podophyllotoxin were the predominant phenolic compounds in both types of litter extracts. These findings suggest that the seed germination and seedling growth of P. tabuliformis and wheat would be inhibited when planted near large amounts J. rigida litter.

Indigenous Uses and Pharmacological Activity of Traditional Medicinal Plants in Mount Taibai, China.

This study was carried out to investigate the indigenous use and pharmacological activity of traditional medicinal plants of Mount Taibai, China. Pharmacological data were collected by conducting informal interviews with local experienced doctors practicing traditional Chinese medicine and via open-ended questionnaires on villagers. We conclude that the residents of Mt. Taibai possess rich pharmacological knowledge. This study may help identify high-value traditional medicinal plant species, promote economic development associated with local medicinal plants, and increase awareness from government departments.

Synergistic effects and related bioactive mechanism of Potentilla fruticosa L. leaves combined with Ginkgo biloba extracts studied with microbial test system (MTS).

BACKGROUND: Potentilla fruticosa, also called "Jinlaomei" and "Gesanghua", is widely used as folk herbs in traditional Tibetan medicine in China to treat inflammations, wounds, certain forms of cancer, diarrhoea, diabetes and other ailments. Previous research found P. fruticosa leaf extract (C-3) combined with Ginkgo biloba extracts (EGb) showed obvious synergistic effects in a variety of oxidation systems. The aim of the present study was to further confirm the synergy of P. fruticosa combined with EGb viewed from physiological bioavailability and explore the related bioactive mechanism behind the synergism. METHODS: The microbial test system (MTS) was adopted to evaluate the related bioactive mechanism. The synergistic effects were evaluated by isobolographic analysis. The H2O2 production rate and antioxidant enzyme (Catalase (CAT), Peroxidase (POD), Superoxide dismutase (SOD), Glutathione peroxidase (GSH-PX)) activities were determined by the colorimetric method. Enzyme gene (CAT, SOD) expression was measured by real time-PCR. RESULTS: The MTS antioxidant activity results showed the combination of C-3 + EGb exhibited synergistic effects especially at the ratio 5:1. Components of isorhamnetin and caffeic acid in C-3 and EGb displayed strong antioxidant activities on MTS and their combination also showed significant synergy in promoting H2O2 production. The combinations of C-3 + EGb and isorhamnetin + caffeic acid promoted CAT and SOD enzyme activities and the ratio 1:1 exhibited the strongest synergy while no obvious promotion on POD and GSH-PX enzyme activities was found. Both combinations above promoted gene expression of CAT and SOD enzymes and the ratio 1:1 exhibited the strongest synergy. CONCLUSIONS: Antioxidant activity results in MTS further confirmed the significant synergy of C-3 combined with EGb and isorhamnetin combined with caffeic acid. The synergy of C-3 combined with EGb may be attributed to the combination of isorhamnetin + caffeic acid, which promoted CAT and SOD enzyme gene expression and further promoted the enzyme activities in E. coli. This study could further provide rational basis for optimizing the physiological bioavailability of P. fruticosa by using natural and safe antioxidants in low doses to produce new medicines and functional products.

Chemical composition, antibacterial activity and related mechanism of the essential oil from the leaves of Juniperus rigida Sieb. et Zucc against Klebsiella pneumoniae.

ETHNOPHARMACOLOGICAL RELEVANCE: Juniperus rigida is used as Tibetan and Mongolian medicine in China for the treatment of rheumatoid arthritis, nephritis, brucellosis and other various inflammatory diseases. AIM OF THE STUDY: To evaluate antibacterial potential of essential oils from J. rigida leaves against Klebsiella pneumoniae and to examine its possible related mechanisms. The study was undertaken in order to scientifically validate the traditional use of J. rigida. MATERIALS AND METHODS: The essential oil was extracted from the leaves of J. rigida by supercritical CO2 fluid extraction technology. Chemical composition of essential oils was analyzed by gas chromatography-mass spectrometry (GC-MS). The antibacterial activity was evaluated against 10 bacteria by the paper disc diffusion method. The minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values of the essential oil were estimated by agar dilution method. The antibacterial mechanism was evaluated by growth curve, the integrity of cell membrane, the SDS-PAGE of protein patterns and scanning electron microscope (SEM). RESULTS: 61 components were identified from the essential oil. Caryophyllene (13.11%) and α-Caryophyllene (11.72%) were found to be the major components. The antibacterial activities of the essential oil were screened and compared against 10 bacteria. The essential oil showed good antibacterial activity against K. pneumoniae, with the biggest diameters of inhibition zones (DIZ) (16.00±0.25mm) and the lowest MIC and MBC values of 3.125mg/mL. The increase in proteins, 260nm absorbing materials of bacterial cells suspension indicated that the cytoplasmic membranes were broken by the essential oil. The SDS-PAGE of bacterial proteins demonstrated that the essential oil could damage bacterial cells through the destruction of cellular proteins. Scanning electron microscopy (SEM) showed that the essential oil damaged the morphology of cell wall and membrane. CONCLUSIONS: The essential oil of J. rigida has potential antibacterial activities against K. pneumoniae. The antibacterial mechanism is the essential oil causing the irreversible damage to the cell wall and membrane, leading to the leakage of proteins and 260nm absorbing materials (DNA and RNA). Further phytochemical and pharmacological studies are required for proper scientific validation of the folk use of this plant species.

Influence of Environmental Factors on the Active Substance Production and Antioxidant Activity in Potentilla fruticosa L. and Its Quality Assessment.

Environmental factors may influence types and contents of active substances. This study investigated the influence of environmental factors on the active substance contents and antioxidant activity of Potentilla fruticosa L. from different regions of China. Also, HPLC fingerprint similarity analysis (SA) coupled with hierarchical cluster analysis (HCA) and discriminant analysis (DA) were further introduced for the accurate classification and quality assessment of P. fruticosa. The results showed that altitude was significantly and negatively correlated to the content of tannin (P < 0.05). Annual sunshine duration and altitude were significantly and positively correlated to the flavonoids content, rutin content and antioxidant activity (P < 0.05). Annual mean temperature was significantly and negatively correlated to the content of total phenolics, while altitude was significantly and positively correlated to the content of total phenolics (P < 0.05). Eight samples were unambiguously separated into three groups. Two types of discriminant functions with a 100% discrimination ratio were constructed. All data consistently supported the conclusion that P. fruticosa produced from Kangding, Sichuan Province had high quality among all samples, therefore, Kangding in Sichuan Province with favorable environmental conditions is recommended as a preferable production location.

Wild food plants and fungi used in the mycophilous Tibetan community of Zhagana (Tewo County, Gansu, China).

BACKGROUND: The aim of the study was to investigate knowledge and use of wild food plants and fungi in a highland valley in the Gannan Tibetan Autonomous Region on the north-eastern edges of the Tibetan Plateau. METHODS: Field research was carried out in four neighbouring villages in a mountain valley of the Diebu (Tewo) county, surrounded by spruce forests. The study consisted of 30 interviews with single informants, or group interviews (altogether 63 informants). Apart from collecting voucher specimens, we also identified fungi using DNA barcoding. RESULTS: We recorded the use of 54 species of vascular plants. We also recorded the use of 22 mushroom taxa, which made up the largest category of wild foods. Fruits formed the largest category of food plants, with 21 species, larger than the wild greens category, which consisted of 20 species eaten after boiling or frying and 7 as raw snacks. We also recorded the alimentary use of 10 species of edible flowers and 3 species with underground edible organs. On average, 20.8 edible taxa were listed per interview (median - 21). The most listed category of wild foods was green vegetables (mean - 7.5 species, median - 8 species), but fruits and mushrooms were listed nearly as frequently (mean - 6.3, median - 6 and mean - 5.8, - median 6 respectively). Other category lists were very short, e.g., flowers (mean - 1.3, median - 1) and underground edible parts (mean - 0.7, median - 1). Wild vegetables are usually boiled and/or fried and served as side-dishes, or their green parts are eaten as snacks during mountain treks (e.g., peeled rhubarb shoots). Wild fruits are mainly collected by children and eaten raw, they are not stored for further use. The most widely used wild staple foods are Potetilla anserina roots, an important ceremonial food served on such occasions as New Year or at funerals. They are boiled and served with sugar and butter. The most important famine plants remembered by people are the aerial bulbils of Persicaria vivipara. Flowers are used as children's snacks - their nectar is sucked. CONCLUSIONS: The number of wild taxa eaten in the studied valley is similar to that of other Tibetan areas. The structure of wild food plant taxa is also very typical for Tibetan speaking areas (e.g., the use of rhubarb shoots, Potentilla anserina, Persicaria vivipara). The studied community show a high level of mycophilia.

Extraction and Purification of Quercitrin, Hyperoside, Rutin, and Afzelin from Zanthoxylum Bungeanum Maxim Leaves Using an Aqueous Two-Phase System.

In this study, an aqueous two-phase system (ATPS) based on ethanol/NaH2 PO4 was developed for the extraction and purification of quercitrin, hyperoside, rutin, and afzelin from Zanthoxylum bungeanum Maxim leaves. These 4 flavonoids were 1st extracted from dried Z. bungeanum leaves using a 60% ethanol solution and subsequently added to the ATPS for further purification. The partition behavior of the 4 flavonoids in ATPS was investigated. The optimal ATPS conditions were: 29% (w/w) NaH2 PO4 , 25% (w/w) ethanol concentration, 1% (w/w) added amount of leaf extracts, no pH adjustment, and repeated 1 h extractions at 25 °C. Under the optimal conditions for the 10 g ATPS, the absolute recovery of quercitrin, hyperoside, rutin, and afzelin reached 90.3%, 83.5%, 92.3%, and 89.1%, respectively. Compared to the 60% ethanol extracts, the content of quercitrin (44.8 mg/g), hyperoside (65.6 mg/g), rutin (56.4 mg/g), and afzelin (6.84 mg/g) in the extracts increased by 49.9%, 38.8%, 45.6%, and 36.8% respectively. The extracts after ATPS also exhibited stronger antioxidant activities, the 2,2-diphenyl-1-picrylhydrazyl IC50 value (10.5 µg/mL) decreased by 41.8%, and the 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt value (966 µmol Trolox/g) and ferric reducing power value (619 µmol Trolox/g) increased by 29.8% and 53.7%, respectively. Furthermore, scale-up experiments indicated that a larger scale experiment was feasible for the purification of the 4 flavonoids.

Synergistic Effects of Potentilla fruticosa L. Leaves Combined with Green Tea Polyphenols in a Variety of Oxidation Systems.

Potentilla fruticosa L. leaves are widely used as tea in China, with many commercial "Jinlaomei" teas available in markets. It has been confirmed to possess significant antioxidant activity than that of butylated hydroxytoluene. In this study, the synergistic effects of P. fruticosa leaves extracts (PFE) combined with green tea polyphenols (GTP) were studied to elucidate their use in combination and find specific combinations with least concentrations that enhance the antioxidant activity. Isobolographic analysis indicated that the combination of PFE and GTP demonstrated extensive synergism (22/28 of the tests showed significant synergy) with 3:1 (PFE:GTP) exhibiting the best synergistic effect. Chemical compositions and content of 7 phenolic compounds in PFE, GTP, and their mixtures were evaluated by reverse-phase-high-performance liquid chromatography. While chemical composition did not seem to change after the combination, as no new peaks appeared in the chromatogram, and no existing peaks disappeared. However, the content of (+)-catechin, (-)-epigallocatechin (EGC), and , (-)-epigallocatechin gallate (EGCG) changed. Besides, antioxidant interactions of extracts and compounds were evaluated, EGC with hyperoside exhibited the greatest synergistic effect and the combination of 3:1 exhibited the strongest synergism (DPPH γ = 0.86, ABTS γ = 1.12, FRAP γ = 1.16). Therefore, interaction of phytochemicals may be one reason for the synergistic effects in PFE + GTP, with EGC + hyperoside likely playing an important role. This report provides a theoretical basis for the concomitant use of P. fruticosa blended with GTP, which can be effectively used as a compounded tea, dietary supplements, and substituent of synthetic antioxidant.