二苯乙烯苷特性及DPPH自由基清除活性

讨论何首乌中二苯乙烯苷晶体的理化性质及清除DPPH自由基活性.从何首乌中分离结晶得到二苯乙烯苷晶体,并运用液-质(LC-MS)、核磁(NMR)、X-射线衍射、差示扫描量热分析(DSC)、热重分析(TGA)等技术分析晶体性质及稳定性,对照Vc、BHT、白藜芦醇测定了其清除DPPH自由基的活性.试验表明二苯乙烯苷晶体不存在同质多晶现象,仅有一种针状晶体存在形式,且不含结晶水,熔点为151.33℃.且其晶体具有良好的稳定性.其清除自由基活性依次为:Vc IC505.38μg/mL,BHT IC506.20μg/m l,二苯乙烯苷IC508.06μg/mL,白藜芦醇IC509.21μg/mL.从何首乌提取物中可以通过结晶得到单一针状、稳定的二苯乙烯苷晶体,且该晶体具有良好的清除DPPH自由基功能.     

超微粉碎-微波双水相萃取红花龙胆中环烯醚萜苷类

以红花龙胆全草为原料,采用超微粉碎和微波控温辅助双水相法提取了其中的环烯醚萜苷类。通过单因素实验和正交实验确定了最佳提取工艺;考察了所得环烯醚萜苷类的总抗氧化作用及其清除1,1-二苯基-2-三硝基苯肼自由基(DPPH·)的能力。结果表明,最佳提取工艺为:在30.0 g〔无水乙醇7.8 g(质量分数26%),磷酸氢二钾(K2HPO4)3.0 g (质量分数10%)〕乙醇/K2HPO4双水相体系中,红花龙胆干粉粒度为400目,微波温度(70±1)℃,微波时间110 s,红花龙胆干粉加入量3.5 g,得率为5.48%,相比传统提取工艺热回流得率(3.61%)显著提高。此法能抑制试材内源性水解酶活性且中和植酸(红花龙胆中的酸性物质)来降低溶出环烯醚萜苷类发生聚合反应的几率,有效解决了红花龙胆中环烯醚萜苷类需富集才能检测到的问题;总环烯醚萜苷类的抗氧化能力在0.2～0.6 g/L内强于2,6-二叔丁基-4-甲基苯酚(BHT),对DPPH·的清除能力在0.2～1.0 g/L内强于BHT。     

天然植物中的甜糖苷和抑甜糖苷

着重介绍了来源于植物组织中的两种活性不同的糖苷,即甜糖苷和抑甜糖苷的特性以及研究现状。     

Determination of steroidal saponins in different organs of yam (Dioscorea pseudojaponica Yamamoto)

Yams (Dioscorea spp.) are perennial trailing rhizome plants. Steroidal saponins, furostanol and spirostanol glycosides are the marked functional compounds in yams. In this investigation, a C18 solid phase extraction method was developed for yam saponins purification. The contents of saponins in various organs of yam (Dioscorea pseudojaponica Yamamoto) were also determined. Results showed that the recoveries of yam saponins extracted by the developed method were about 99.48–100.08% when the saponins (each saponin weighed 0.20, 0.50 and 1.00 mg) passing through the C18 cartridge. The extractive method could efficiently reduce the interferences from impurities in yam saponin extracts prior to HPLC analysis. The recoveries of added saponins in different yam organs were 98.34–99.92% for tuber flesh, 95.98–98.89% for tuber cortex, 97.89–99.44% for rhizophor, 93.82–98.01% for leaf and 93.87–97.65% for vine, respectively. The yam tuber cortex had the highest amount of saponins (582.53 μg/g dw), which was higher than that existed in the tuber flesh (227.86 μg/g dw) about 2.55 times. The contents of saponins in the rhizophor, leaf and vine of yam were 29.39, 24.41 and 23.96 μg/g dw, respectively.     

Separation and determination of amygdalin and unnatural neoamygdalin in natural food supplements by HPLC-DAD

ABSTRACT

This work is focused on separation and determination of amygdalin and its unnatural form neoamygdalin in natural food supplements. Reversed-phase high-performance liquid chromatography with a high-stability silica-based column with C₁₈ functional group has been used for solving this problem. The effect of the mobile phase composition as well as the column temperature on the separation of the amygdalin epimers has been investigated. Isocratic elution using a mobile phase composed of 0.05% aqueous formic acid and acetonitrile achieved the required separation within 17 min. Under optimum chromatographic conditions, the developed method was validated and was applied for the determination of amygdalin epimers in natural food supplements containing apricot or peach kernels. A simple extraction method using methanol as an extractant supported by an ultrasonic bath was used with recovery in the range of 94.8% to 104.3%. The limit of detection and limit of quantification values for R-amygdalin were 0.13 mg/L and 0.40 mg/L, respectively. The developed method proved to be precise with the intra-day and inter-day relative standard deviation values less than 2.23%.     

4-O-Methyl Modifications of Glucuronic Acids in Xylans Are Indispensable for Substrate Discrimination by GH67 α-Glucuronidase from Bacillus halodurans C-125

A GH67 α-glucuronidase gene derived from Bacillus halodurans C-125 was expressed in E. coli to obtain a recombinant enzyme (BhGlcA67). Using the purified enzyme, the enzymatic properties and substrate specificities of the enzyme were investigated. BhGlcA67 showed maximum activity at pH 5.4 and 45 °C. When BhGlcA67 was incubated with birchwood, oat spelts, and cotton seed xylan, the enzyme did not release any glucuronic acid or 4-O-methyl-glucuronic acid from these substrates. BhGlcA67 acted only on 4-O-methyl-α-D-glucuronopyranosyl-(1→2)-β-D-xylopyranosyl-(1→4)-β-D-xylopyranosyl-(1→4)-β-D-xylopyranose (MeGlcA³Xyl₃), which has a glucuronic acid side chain with a 4-O-methyl group located at its non-reducing end, but did not on β-D-xylopyranosyl-(1→4)-[4-O-methyl-α-D-glucuronopyranosyl-(l→2)]-β-D-xylopyranosyl-(1→4)-β-D-xylopyranosyl-(1→4)-β-D-xylop- yranose (MeGlcA³Xyl₄) and α-D-glucuronopyranosyl-(l→2)-β-D-xylopyranosyl-(1→4)-β-D-xylopyranosyl-(1→4)-β-D-xylopyranose (GlcA³Xyl₃). The environment for recognizing the 4-O-methyl group of glucuronic acid was observed in all the crystal structures of reported GH67 glucuronidases, and the amino acids for discriminating the 4-O-methyl group of glucuronic acid were widely conserved in the primary sequences of the GH67 family, suggesting that the 4-O-methyl group is critical for the activities of the GH67 family.     

Development and Validation of a HPLC Method for Determination of Flavone Glycoside-Camellianin B in Cephalotaxus Sinensis

A simple,rapid and sensitive method for the quantification of camellianin B in Cephalotaxus sinensis(C.sinensis)a natural plant with anti-hyperglycemic effect,was developed and validated by reversed phase liquid chromatography.Chromatographic separation was achieved on a C18 column and an isocratic elution was carried out at a flow-rate of 1.0 mL·min~(-1) with the acetonitrile-water containing0.1%formic acid(19∶81,v/v).The detection wavelength was set at 330 nm.The calibration curve was linear over the concentration range of 0.25-50μg·mL~(-1) with correlation coefficients larger than0.999 5.The limit of detection and limit of quantification were 0.09μg·mL~(-1)and 0.25μg·mL~(-1),respectively.The precisions and accuracy for all samples were acceptable.The validated method has been successfully applied for the quantification of camellianin B in C.sinensis harvested in different months and may also be used as the quality evaluation of this herb medicine.     

Phytochemicals in Chinese Chive (Allium tuberosum) Induce the Skeletal Muscle Cell Proliferation via PI3K/Akt/mTOR and Smad Pathways in C2C12 Cells

Chinese chive (Allium tuberosum) is a medicinal food that is cultivated and consumed mainly in Asian countries. Its various phytochemicals and physiological effects have been reported, but only a few phytochemicals are available for skeletal muscle cell proliferation. Herein, we isolated a new compound, kaempferol-3-O-(6″-feruloyl)-sophoroside (1), along with one known flavonoid glycoside (2) and six amino acid (3–8) compounds from the water-soluble fraction of the shoot of the Chinese chive. The isolated compounds were identified using extensive spectroscopic methods, including 1D and 2D NMR, and evaluated for their proliferation activity on skeletal muscle cells. Among the tested compounds, newly isolated flavonoid (1) and 5-aminouridine (7) up-regulated PI3K/Akt/mTOR pathways, which implies a positive effect on skeletal muscle growth and differentiation. In particular, compound 1 down-regulated the Smad pathways, which are negative regulators of skeletal muscle growth. Collectively, we suggest that major constituents of Chinese chive, flavonoids and amino acids, might be used in dietary supplements that aid skeletal muscle growth.     

Discovery of Phenolic Glycoside from Hyssopus cuspidatus Attenuates LPS-Induced Inflammatory Responses by Inhibition of iNOS and COX-2 Expression through Suppression of NF-κB Activation

It seems quite necessary to obtain effective substances from natural products against inflammatory response (IR) as there are presently clinical problems regarding accompanying side effects and lowered quality of life. This work aimed to investigate the abilities of hyssopuside (HY), a novel phenolic glycoside isolated from Hyssopus cuspidatus (H. cuspidatus), against IR in lipopolysaccharide (LPS)-induced RAW 264.7 cells and mouse peritoneal macrophages. The results indicated that HY could reduce nitric oxide (NO) production and inhibit the production and secretion of pro-inflammatory mediators including tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in LPS-stimulated macrophages. Moreover, data from the immunofluorescence study showed that HY suppressed nuclear translocation of nuclear factor-kappa B (NF-κB) upon LPS induction. The Western blot results suggested that HY reversed the LPS-induced degradation of IκB (inhibitor of NF-κB), which is normally required for the activation of NF-κB. Meanwhile, the overexpression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) diminished significantly with the presence of HY in response to LPS stimulation. On the other hand, HY had a negligible impact on the activation of mitogen-activated protein kinase (MAPK) pathways. Moreover, an in silico study of HY against four essential proteins/enzymes revealed that COX-2 was the most efficient enzyme for the interaction, and binding of residues Phe179, Asn351, and Ser424 with HY played crucial roles in the observed activity. The structure analysis indicated the typical characterizations with phenylethanoid glycoside contributed to the anti-inflammatory effects of HY. These results indicated that HY manipulated its anti-inflammatory effects mainly through blocking the NF-κB signal transduction pathways. Collectively, we believe that HY could be a potential alternative phenolic agent for alleviating excessive inflammation in many inflammation-associated diseases.