酸水解HPLC测定桑叶中槲皮素及山柰酚含量

目的 建立桑叶中水解黄酮苷元(槲皮素、山柰酚)的含量测定方法,并考察湖州地区不同品种、不同采摘时间、不同种植基地桑叶中上述成分的含量差异。方法 粉碎的桑叶样品经甲醇-盐酸混合溶液提取水解后,用反相高效液相色谱法,以Zorbax SB C₁₈为分离柱,甲醇-0.2%磷酸(55∶45)为流动相,流速1.0 mL·min^-1,检测波长370 nm。结果 经方法学验证,本方法各项指标均符合中药成分含量测定要求;不同品种、不同种植基地、不同采集时间桑叶中上述2种成分含量均有较大变化;同一基地、同一品种,冬桑叶中上述2成分的含量明显高于其他季节采集的桑叶。结论 水解后经HPLC测定桑叶中槲皮素、山柰酚含量方法可用于桑叶药材质量控制。桑叶中水解槲皮素及山柰酚含量受多种因素影响。     

酸水解提取HPLC法测定不同来源莲须中槲皮素和山柰酚的含量

目的:研究莲须的酸水解提取条件及其水解苷元槲皮素和山柰酚的HPLC含量测定方法,并分析不同来源莲须中上述成分的含量差异。方法:莲须样品经体积分数为25%的盐酸-甲醇溶液(1∶4)水解提取后,采用高效液相色谱法测定其水解液中槲皮素和山柰酚的含量。色谱柱为Thermo Scientific Syncronis C18柱(250 mm×4.6 mm,5μm),流动相为甲醇-0.012 5%磷酸溶液(60∶40),等度洗脱,流速1.0 m L·min-1,检测波长370 nm,柱温30℃。结果:方法学考察结果表明,槲皮素和山柰酚进样量分别在0.013 99~0.279 8μg(r=0.999 9)和0.028 70~0.574 2μg(r=0.999 9)范围内与其对应的峰面积线性关系良好;它们的低、中、高3个加入水平的平均回收率(n=3)分别为102.4%(RSD=1.9%)和100.1%(RSD=1.7%)、101.2%(RSD=0.87%)和96.0%(RSD=0.18%)、101.8%(RSD=1.7%)和95.5%(RSD=1.6%)。15批不同来源的莲须样品中槲皮素和山柰酚含量差异较大,分别为0.07~0.76 mg·g-1和0.24~2.40 mg·g-1。其中1号样品(福建福州1)中槲皮素和山柰酚的含量最高,而14号样品(福建永春)中槲皮素和山柰酚的含量最低。结论:酸水解提取HPLC法可用于莲须药材中槲皮素和山柰酚含量的测定;15批不同来源莲须的含量测定结果为进一步修订莲须的质量标准提供了实验依据。     

RP-HPLC测定桑叶、桑枝和桑花中槲皮素和山奈酚的含量

目的:建立 HPLC 法测定桑叶及桑枝、桑花中的槲皮素和山奈酚含量。方法:先用甲醇-25%盐酸(4:1)混合液水解桑叶、桑枝及桑花中的黄酮类成分成槲皮素和山奈酚,并采用 HPLE 法测定槲皮素和山奈酚含量。色谱柱 Shim-ODS(150 mm×4.6 mm,5 μm),流动相为甲醇-0.4%磷酸溶液(70:30);流速为1 mL·min~(-1);检测波长为368 nm。结果:槲皮素和山奈酚的线性范围分别为0.025～0.15μg(r=0.9999),0.05～0.3μg(r=0.9998),平均回收率分别为96.2%及95.3%。5月份桑叶中槲皮素和山奈酚含量较高,桑花中山奈酚含量较高,8月桑叶中槲皮素和山奈酚含量较低。结论:此法简单、重现性好,可用于桑叶、桑枝及桑花中槲皮素和山奈酚含量的测定。     

UPLC测定桑叶中的槲皮素和山柰酚

目的 采用超高效液相色谱法测定桑叶提取物中槲皮素和山柰酚的含量.方法 采用Agilent Ecilipse C18色谱柱(50 mm× 2.1 mm,1.8-Micron),以乙腈-水(含0.1％甲酸)(35∶65)为流动相,流速为0.4 mL· min-1,柱温为25℃,检测波长370 nm,以外标法定量,进样量2.0 μL.结果 槲皮素和山柰酚0.01 ～1.50 mg·mL-1与峰面积的线性关系良好,平均回收率分别为100.11％、101.92％,RSD分别为0.10％、0.16％.结论 该测定方法快速,准确,可用于桑叶提取物中槲皮素和山柰酚的含量测定.     

反相高效液相色谱法测定蒺藜类3种水解黄酮苷元的含量

目的：研究蒺藜、中亚滨藜黄酮类成分的含量情况。方法：采用反相高效液相色谱法,以槲皮素、山柰酚、异鼠李素为对照品,测定蒺藜、中亚滨藜果实等２１个样品中上述３种水解黄酮苷元的含量。色谱柱Ｈｙｐｅｒｓｉｌ－ＯＤＳ,流动相为甲醇－０４％磷酸（３７∶６３）,流速１ｍＬ·ｍｉｎ－１,检测波长３６０ｎｍ。２种浓度槲皮素加样回收率分别为９４０９％、９８３２％,ＲＳＤ分别为１０％、１５％,ｎ＝３。结果：蒺藜中以槲皮素为母核的黄酮类成分含量最高,中亚滨藜中以异鼠李素为母核的含量最高;各个不同部位以叶中黄酮类成分的含量最高,茎次之,果实更次之;不同采集时间蒺藜地上部分黄酮类成分的含量以６月份采集的样品最高。结论：蒺藜中黄酮类成分以６月份采集的地上部分含量最高,且通过比较,主要黄酮类成分的含量是主要皂苷类成分含量的１５倍左右,应对蒺藜中黄酮类成分加以研究利用。中亚滨藜含有较大量以异鼠李素为母核的黄酮类成分,亦值得进行研究开发。     

卧龙沙棘果实中多酚类成分的含量测定

目的 测定卧龙沙棘果实中多酚类成分的含量.方法 采用比色法测定总酚和总黄酮的含量;采用UPLC法测定槲皮素、山柰酚和异鼠李素的含量.结果 5批沙棘果实中总酚、总黄酮、槲皮素、山柰酚、异鼠李素的含量分别为29.80 ～38.80、17.60 ～27.40、0.38 ～2.01、0.17 ～2.00、0.49 ～4.53 mg·g-1.结论 卧龙沙棘果实中多酚类成分的含量较高,值得进一步研究与开发.     

桑叶中绿原酸和主要黄酮苷含量的RP—HPLC测定

目的:考察不同品种、不同采摘时间和不同种植基地桑叶中绿原酸、芦丁和异槲皮苷的含量差异。方法:采用反相高效液相色谱法,以 Zorbax SB C_(18)(250 mm×4.6 mm,5μm)为分离柱,90%乙腈(A)-0.2%磷酸溶液(B)为流动相进行梯度洗脱[0 min,A-B(12:88);10 min,A-B(16:84);20 min,A-B(20:80);30 min,A-B(25:75)],流速1.0 mL·min~(-1),检测波长358 nm。结果:不同种植基地的桑叶中绿原酸、芦丁和异槲皮苷的含最有一定的差异;不同品种和采摘时间的桑叶中上述三成分含量差异较大。结论:桑叶中绿原酸、芦丁、异槲皮苷的含量与种植基地、采摘季节、品种等有关,其中尤以季节和品种影响较大。     

一测多评法测定槐花中3种黄酮类成分的含量

目的建立一测多评法(QAMS)用于测定槐花中芦丁,山柰酚?3?O?芸香糖苷及槲皮素 3种黄酮类成分的含量。方法芦丁作为内标物,计算芦丁与槲皮素和山柰酚 ?3?O?芸香糖苷的相对校正因子,分别采用 QAMS和外标法测定 40批槐花药材中以芦丁,山柰酚 ?3?O?芸香糖苷和槲皮素的含量,比较两种测定方法之间的差异。结果 QAMS与外标法所测得的槐花中芦丁、山柰酚 ?3?O?芸香糖苷和槲皮素的含量分别为(9.98±1.39)%、(10.08±1.38)%,差异无统计学意义(t＝0.324,P＝0.747)。结论 QAMS准确可行,可用于槐花中 3种黄酮类成分测定。     

HPLC-一测多评法测定黄精及其饮片中6种成分的含量

目的:建立黄精及其饮片中薯蓣皂苷元、5-羟甲基糠醛、香草酸、芦丁、槲皮素及山柰酚6种成分的一测多评(QAMS)法。方法:采用高效液相色谱(HPLC)外标法同时测定黄精及其饮片中6种成分的含量[色谱柱为Diamonsil-C_(18);流动相为乙腈-水(梯度洗脱);5-羟甲基糠醛、香草酸、芦丁、槲皮素及山柰酚检测波长为254 nm(0～60 min),薯蓣皂苷元检测波长为202 nm(60～75min);流速为1.0 mL/min;柱温为30℃]。以香草酸为内标,分别计算出其余5种成分的相对校正因子(RCFs)并进行耐用性考察,并采用相对保留法对待测成分色谱峰进行准确定位,然后根据RCFs计算黄精药材中其他5种成分的含量,并与外标法测定结果进行比较。以黄精饮片进行方法验证,分别采用QAMS法和外标法测定其中6种成分的含量,比较两种方法含量测定差异。结果:HPLC方法学考察结果均符合相关要求。在线性范围内,薯蓣皂苷元、5-羟甲基糠醛、芦丁、槲皮素及山柰酚的RCFs分别为0.195、0.025、0.263、0.345、0.075,在不同试验条件下RCFs重现性良好。外标法和QAMS法两种方法测得的黄精药材及其饮片中6种成分的含量差异均无统计学意义(P>0.05)。结论:建立的QAMS法可用于同时测定黄精药材及其饮片中6种成分含量。     

银杏叶提取物注射液中槲皮素、山柰酚、异鼠李素及总黄酮醇苷的HPLC法测定

建立了高效液相色谱法测定银杏叶提取物注射液中的槲皮素、山柰酚、异鼠李素及总黄酮醇苷,评价了注射液的质量.色谱柱为Diamonsil C18柱,流动相为甲醇-0.4％磷酸(55∶45),检测波长360 nm.银杏叶提取物注射液水解前后,其主要组分槲皮素、山柰酚和异鼠李素分离良好,线性范围宽,平均加样回收率水解前分别为100.2％、100.3％和100.2％,RSD分别为0.64％、1.14％和0.40％水解后分别为100.0％、100.7％和100.3％,RSD分别为0.79％、0.98％和1.00％.     

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.