正交设计优选白花蛇舌草总黄酮的提取工艺及体外抗氧化性考察

摘 要 目的：优化白花蛇舌草总黄酮的提取工艺,检测其体外抗氧化活性。方法: 采用HPLC法检测山奈酚的含量;采用UV法检测总黄酮含量;采用正交设计法,以山奈酚提取率、总黄酮提取率为评价指标,对白花蛇舌草总黄酮的提取工艺进行优化;利用水杨酸法和DPPH法分别检测甲醇提取物清除·OH和DPPH·的活性。结果:最佳提取工艺为10倍量80%甲醇,超声时间15 min;1 ml甲醇提取物对DPPH·和·OH的清除率分别为73.89%和91.27%。结论:该提取工艺稳定可行,可作为白花蛇舌草总黄酮的提取工艺;甲醇提取物对·OH和DPPH·具有一定的清除作用,为开发相关的抗氧化性产品提供参考。     

银木总黄酮体外抗氧化活性研究

摘 要 目的： 对银木总黄酮的体外抗氧化活性进行研究。方法: 采用铁氰化钾还原法测定其还原能力,并测定其清除DPPH自由基（DPPH·）、羟基自由基（·OH）、超氧阴离子自由基（O₂-·）的能力。结果:银木总黄酮具有良好的还原力,较好地清除DPPH自由基（DPPH·）、羟基自由基（·OH）、超氧阴离子自由基（O₂-·）的能力,且随着浓度逐渐增加抗氧化作用呈递增趋势, 在银木总黄酮浓度为0.15mg·mL^-1时,对DPPH自由基（DPPH·）清除率高达86%、对羟基自由基（·OH）的清除率达到68%、对于超氧阴离子自由基（O₂-·）的清除率可以达到59%。结论: 银木总黄酮有较好的抗氧化活性,是一种天然的抗氧化剂,值得进一步研究开发。     

响应面法优化纤维素酶提取丹参多糖工艺

摘 要 目的： 采用响应面分析法优选丹参多糖的纤维素酶提取工艺。方法: 在单因素试验基础上,利用Design Expert 8.0.5软件釆用三因素三水平响应面分析法,以加酶量、酶解温度、酶解时间为考察因素,多糖提取率为响应值,进行回归分析,优选丹参多糖提取工艺条件。 结果: 最佳提取工艺为：加酶量0.5%,酶解温度65℃,提取时间120 min,模型相关系数R^{2＝0.979 5,拟合度较好。在该条件下,丹参多糖的提取率2.59 mg·g-1,与理论提取率(2.51 mg·g-1) 接近。 结论:该提取工艺客观可行、稳定合理,可为工业化生产提供理论依据。}     

Box-Behnken响应面法优化翻白草总黄酮的提取工艺

摘 要 目的：运用Box-Behnken响应面法优化翻白草中总黄酮的提取工艺。方法: 在单因素试验基础上,以提取时间、乙醇浓度以及料液比为考察因素,以翻白草总黄酮提取率为评价指标,利用 Box-Behnken 响应面法优化翻白草中总黄酮的提取工艺。结果: 提取的最佳工艺为：提取浓度为70%的乙醇,每次120 min,液料比为8∶1 ml·g^-1时,黄酮的提取率最高,总黄酮提取率预测值为3.86%,测定值为3.74%,其间相差0.12%。结论:使用Box-Behnken响应面法优化翻白草总黄酮的提取工艺科学可靠,可用于翻白草总黄酮的提取生产。     

离子色谱-电感耦合等离子体质谱联用测定琥珀酸亚铁片中三价铁和二价铁含量

摘 要 目的：建立测定琥珀酸亚铁片中三价铁和二价铁含量的方法。方法: 采用Dionex RFICTM保护柱(4mm×50mm),Dionex RFICM Ion PacRCS5A分析柱(4 mm×250 mm), 淋洗液：7 mmol·K^-1 吡啶二羧酸、66 mmol·K^-1氢氧化钾、5.6 mmol·K^-1硫酸二氢钾和74 mmol·K^-1甲酸的混合溶液;流速：1.5 ml·min^-1;柱温：30.0℃;进样体积：1.3 μl。结果: Fe³+在0.5～15 μg·ml^-1范围内线性良好（r=1.000 0）;Fe²+在25～200 μg·ml^-1范围内线性良好（r=1.000 0）; Fe³+的平均回收率为103.6%（RSD=2.7%, n=9）;Fe²+的平均回收率为98.3%（RSD=1.9%, n=9）。结论:该方法简便可靠,结果准确,可用于琥珀酸亚铁片的含量测定。     

响应面设计优化抗癌散总黄酮提取工艺

摘 要 目的： 采用响应面分析法对复方抗癌散总黄酮热回流提取工艺进行优化。方法: 以单因素试验选择了提取时间、乙醇浓度、料液比3个因素为自变量,总黄酮提取率为响应值,通过3因素3水平的响应面分析,确定最佳提取工艺条件。结果: 最佳工艺条件为乙醇浓度64.56%,料液比为1∶26.97,提取时间为2.29 h,提取温度为85℃,总黄酮提取率可达45.94 mg·g^-1。结论: 采用响应面分析法对复方抗癌散总黄酮提取条件优化合理可行,可为其工业生产提供工艺参数依据。     

玛卡多糖体外抗氧化及对H2O2诱导SH-SY5Y细胞氧化损伤的保护作用

摘 要 目的：对玛卡多糖的体外抗氧化活性进行研究。方法: 通过构建化学反应体系,测定0.18～5.00 mg·ml-1浓度范围内玛卡多糖对羟自由基（·OH）、氧自由基（O2-· ）、DPPH自由基的清除作用,测定其总抗氧化能力。另建立H2O2诱导人神经瘤母细胞（SH SY5Y）氧化损伤模型,SH SY5Y细胞分为对照组、H2O2模型组、玛卡多糖给药组（25, 50, 100 μg·ml-1）。给药组细胞处理：各浓度玛卡多糖分别预保护SH SY5Y细胞24 h后,用300 μmol·L-1 H2O2作用6 h。MTT法测定SH SY5Y各组细胞存活率。结果:玛卡多糖在0.18～5.00 mg·ml-1浓度范围内随着浓度的增加对·OH,O2-·,DPPH· 抗氧化作用有递增趋势,在浓度为2.50 mg·ml-1时,对DPPH·清除率高达60.36%,对O2-·清除率达到34.66%,对·OH清除率达到36.01%,总抗氧化能力相当于FeSO4标准液浓度0.179 μmol。MTT法测得玛卡多糖给药组浓度为25,50,100 μg·ml-1时细胞活力分别达到（61.71±2.45）%,（61.73±4.51）%,（60.98±3.12）%,细胞活力显著高于模型组（P<0.05）。结论:玛卡多糖有较好的抗氧化活性,是一种天然的抗氧化剂,值得进一步研究开发。     

空心柴胡不同部位中柴胡皂苷a、d和总黄酮的含量比较

摘 要 目的：比较空心柴胡不同部位中柴胡皂苷a、d和总黄酮的含量,为临床合理使用药用部位提供参考。方法: 柴胡皂苷a、d含量测定采用HPLC法：色谱柱Inertsil C18柱（250 mm×4.6 mm,5 μm）;流动相：乙腈-水（40∶60）,流速：1.0 ml·min^-1;检测波长：210 nm,柱温：40 ℃,进样量：10 μl。总黄酮的测定采用比色法：检测波长为500 nm。结果: 柴胡皂苷a、柴胡皂苷d和柴胡总黄酮分别在0.21～1.26 mg·ml^-1(r=0.999 6)、0.25～1.51 mg·ml^-1(r=0.999 7)和4.00～25.00 μg·ml^-1(r=0.999 6)范围内与峰面积呈良好的线性关系;平均回收率分别为99.27%(RSD=2.15%,n=6)、99.4%(RSD=2.14%,n=6)和99.03%(RSD=1.34%,n=6）。空心柴胡根中柴胡皂苷a、d含量分别为0.31%、0.50%,其他部位含量较低;总黄酮在花中含量高达8.48%,在叶、茎、根中依次降低。结论:空心柴胡地上部分富含黄酮类化合物,根中柴胡皂苷含量较高,临床使用带根全草比较合理。     

酶法-超声提取黄精总黄酮及其抗氧化活性研究

目的 研究酶法-超声提取黄精中总黄酮（total flavonoids from Polygonatum sibirici,TFPS）的最佳工艺及总黄酮的体外抗氧化活性。方法 以总黄酮含量为指标,采用单因素和正交试验对纤维素酶用量、乙醇浓度、液料比和超声时间进行考察,优化工艺条件并考察总黄酮对DPPH自由基、ABTS⁺自由基的清除能力和亚铁离子螯合活性。结果 最佳提取条件：纤维素酶用量为0.75%,乙醇浓度为40%,液料比为20 mL·g^-1,超声时间30 min,总黄酮含量为1.595%。体外抗氧化试验表明,黄精总黄酮对DPPH自由基、ABTS⁺自由基有很强的清除能力,IC₅₀分别为27.55,11.47 μg·mL^-1,对亚铁离子的螯合活性较强,IC₅₀为32.26 μg·mL^-1。结论 酶法-超声提取工艺具有提取效率高、稳定性及重复性好等优点,可用于黄精中总黄酮的提取,在该工艺下提取的总黄酮具有较强的抗氧化活性。     

不同炮制方法对麦麸中化学成分的影响

摘 要 目的：分别用高效液相色谱法和紫外可见分光光度法测定湖北麦麸及其炒制品中阿魏酸和总黄酮的含量,探讨不同炮制方法对麦麸中阿魏酸和总黄酮含量的影响。方法: 采用SHIMADZU C18色谱柱（250 mm×4.6 mm,5 μm）,流动相为乙腈 0.1%磷酸水溶液（16∶84）等度洗脱,流速1.0 ml·min^-1,检测波长320 nm,柱温35℃;采用紫外可见分光光度法,在（258±2）nm处对不同炮制品中总黄酮含量进行测定。结果: 麦麸的生品、炒黄品、炒焦品、过焦品、蜜炙品中阿魏酸的含量分别为2.07,2.33,1.68,1.11,0.83 μg·g-1,总黄酮含量分别为5.96,7.15,9.73,13.61,9.42 mg·g-1。结论: 根据麦麸炒制程度的加大,阿魏酸含量呈下降趋势,总黄酮含量呈增加趋势。     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.