HPLC法同时检测茶叶中八种儿茶素

建立一种茶叶中多组分儿茶素的HPLC测定方法。将茶叶用80℃热水超声提取20min,提取液离心分离后,采用Kromasil C18(250mm×4.6mm,5μm)色谱柱分离,以水-乙腈-磷酸(49.95:50:0.05,V:V)为流动相A,水-乙腈-磷酸(95.45:4.5:0.05,V:V)为流动相B,进行梯度洗脱,流速为0.5mL/min,柱温25℃,紫外检测波长231nm。在给定的浓度范围内,各儿茶素组分的峰面积与其相应浓度呈现良好的线性相关,R2>0.9950。添加已知儿茶素含量的回收试验结果显示,各组分儿茶素的平均回收率为94.8%～110.8%,相对标准偏差≤2%。     

HPLC法测定不同种类茶叶咖啡因含量

目的采用高效液相色谱法测定8种茶叶中咖啡因的含量。方法采用Ultimate XB-C18（4.6×250mm,5μm）色谱柱,流动相：乙腈-0.2%磷酸水溶液,梯度洗脱,流速1.0mL·min-1,检测波长为280nm,进样量20μL。结果在所选条件下,样品中待测组分与其它组分分离良好（R〉1.5）,咖啡因对照品在10-100μg·mL-1质量浓度范围内线性关系良好,r=0.9998;回收率为98.8%-102.5%,RSD=1.6%（n=6）。结论该方法操作简便,结果可靠,适合用于茶叶中咖啡因定量分析。     

苦丁茶老叶中槲皮素和山柰素的含量测定

目的:建立苦丁茶老叶中槲皮素和山柰素含量的HPLC测定方法。方法:色谱柱为Nova-Pak C18柱(3.9mm×300mm,4μm)柱,以甲醇-0.4%磷酸溶液(50∶50)为流动相,流速为0.7mL.m in-1,检测波长为360nm。结果:测定组分槲皮素、山柰素与其他组分的色谱峰基线分离;槲皮素加样回收率平均值为99.24%,RSD为1.60%,山柰素加样回收率平均值为98.77%,RSD为2.11%。结论:该方法样品处理简单,准确度高,分离效能好,适合于苦丁茶老叶中槲皮素和山柰素含量的测定。     

反相高效液相色谱法测定银杏叶中聚戊烯醇类化合物含量

目的:HPLC/外标法测定银杏叶中聚戊烯醇类化合物(PPAs,PPs)含量。方法:银杏叶样品10 g,加入石油醚100mL,室温下超声萃取45 min,共4次。提取物硅胶柱层析(硅胶100～140目,柱15 mm×350 mm),用石油醚-乙酸乙酯(90:10)洗脱。样品溶液采用HPLC分析:Inertsil ODS-3柱(4.6 mm×250 mm),柱温40℃,异丙醇-甲醇-正己烷-水(250:125:75:10)为流动相,流速1 mL·min~(-1),紫外检测器,检测波长215 nm。结果:聚戊烯醇类化合物中各组分基线分离,可同时分析银杏叶中PPAs、PPs含量,PPAs(或 PPs)进样量在0.26～5.2μg范围内与峰面积呈线性关系,PPAs、PPs平均加样回收率分别为98.8％和97.4％,RSD分别为1.8％和2.4％。结论:样品处理简单,结果稳定、准确,可作为银杏叶中聚戊烯醇类化合物的测定方法。     

ICP—MS法测定茶叶中微量金属元素含量

目的:建立了茶叶和茶叶提取物中微量金属含量的测定方法。方法:样品经微波消解后,以铟为内标,用电感耦合等离子体质谱(ICP-MS)测定样品中10种微量金属元素。结果:方法快速、准确、检测限低。对茶树叶标准物质(GBW08513)测定结果满意。结论:ICP-MS 法是测定茶叶中微量金属元素的有效分析方法,内标法使得测定方法更加完善、可靠。     

RP-HPLC法测定患者血浆中异烟肼及乙酰异烟肼的浓度

目的建立高效液相色谱法测定患者血浆中异烟肼和乙酰异烟肼的浓度。方法色谱柱为Inertsil ODS-SP(4.6 mm×150 mm,5μm),以甲醇-0.05 mol/L磷酸二氢钾(三乙胺调pH=5.6)为流动相,柱温:40℃,流速:1.0 mL/min,进样体积:20μL,检测波长:261 nm。结果三组分在线性范围内与峰面积线性关系良好(r2>0.99),日内、日间精密度RSD<10%,平均回收率为80%¹20%。结论本方法专属性好,样品处理简单,符合生物样品分析要求。     

液相色谱-串联质谱法研究升麻提取物在大鼠体内的药动学

目的:建立同时测定大鼠血浆中升麻4组分,升麻素、升麻亭、7,8-二脱氢-27-脱氧升麻亭和25-O-甲基升麻醇-3-β-D-半乳糖苷的液相色谱-串联质谱方法。方法:采用20(S)-人参皂苷Rg3为内标,血浆样品经固相萃取处理后,在负离子条件下多反应监测(MRM)方式进行扫描。结果:血浆中4种组分的线性范围为0.1～500 ng.mL-1,最低定量下限为0.1 ng.mL-1。方法的日内和日间精密度(RSD)均小于11.6%,样品提取回收率大于89.5%。结论:该方法灵敏、快速、准确,可用于大鼠血浆中升麻提取物有效成分含量的测定。     

茶叶下脚料中表没食子儿茶素没食子酸酯含量测定研究

目的:建立茶叶下脚料中表没食子儿茶素没食子酸酯(EGCG)的HPLC含量测定方法.方法:采用HPLC法,以C18柱(250 mm×4.6 mm,5 μm)为色谱柱,以乙腈-水-冰醋酸(14:84:2)为流动相,流速:1.0 ml·min-1,检测波长:276 nm.结果:EGCG在1.66～8.30 μg范围内线性关系良好,平均回收率为100.26%(RSD=1.41%).结论:该方法简便,准确,重复性好,可作为茶叶下脚料中EGCG的含量测定方法.     

HPLC-ELSD在大豆活性组分提取过程控制中的应用

目的:建立高效液相色谱法-蒸发光散射检测器(HPLC-ELSD)对大豆活性组分含量的检测方法.方法:采用Hypersil C8柱(150mm×4.0mm,10μm),流动相为乙腈-0.001mol·L-1的乙酸水溶液.流速:0.7mL·min-1.结果:在一个色谱过程中同时测定大豆中3种活性组分.结论:本法简便、准确、专属性强,为工艺控制和质量控制提供有效的分析方法.     

高效液相色谱法测定普洱熟沱茶叶中咖啡因的含量

目的 采用高效液相色谱法测定了普洱熟沱中咖啡因的含量.方法 采用大连依利特Sinochrom ODS-BP(4.6mm×250mm,5μm)色谱柱,流动相甲醇:水(60∶40),流速1.0(mL·min-1),检测波长为275 nm,柱温为25℃.结果 在所选条件下,样品中待测组分与其他组分分离良好(R＞1.5),通过重复性试验和精密度试验,验证了方法的稳定性和可靠性.在本实验范围内,咖啡因峰面积与含量之间具有良好的线性关系r2 =0.9981,普洱熟沱中咖啡因的回收率为93.03％ ～ 100.0％,RSD=2.79％.结论 本文采用高效液相色谱法测定普洱熟沱中咖啡因的含量,供人们选购茶叶参考,同时可为工厂制备相关产品提供依据.     

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.     

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.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

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.