反相高效液相色谱法同时测定藤三七不同部位中尿嘧啶和假鹰爪黄酮的含量

目的建立测定藤三七不同部位(包括珠芽、茎、叶和花)中尿嘧啶和假鹰爪黄酮含量的反相高效液相色谱法。方法采用Kromasil C18色谱柱(250 mm×4.6 mm,5μm),流动相:乙腈(A)-0.05%磷酸溶液(B),进行梯度洗脱,程序为:3%A(0～6 min),3%～60%A(6～8 min),60%～80%A(8～13 min),80%A(13～20 min);流速为1 mL.min-1;检测波长为254 nm;柱温:35℃。结果尿嘧啶在0.705～7.05μg.mL-1与峰面积呈良好线性关系,r=0.999 9(n=6),平均回收率(n=9)为99.2%(RSD=1.3%);假鹰爪黄酮在0.792～7.92μg.mL-1与峰面积呈良好线性关系r=0.999 9(n=6),平均回收率(n=9)为99.0%(RSD=1.2%)。结论该方法简单、准确,为藤三七药材质量评价提供了可靠依据。     

甘肃省不同产地萱草花蕾中芦丁、槲皮素、山柰酚的测定

目的:建立HPLC法同时测定萱草花蕾中芦丁、槲皮素、山柰酚3个黄酮类化合物的含量,并比较不同产地萱草中这3个化合物的含量。方法:采用Agilent SB-C18(4.6 mm×250 mm,5μm)色谱柱,以甲醇(A)-pH 3磷酸水溶液(B)为流动相,梯度洗脱(0～10 min,20%A;10～10.5 min,20%A→45%A;10.5～27.5 min,45%A;27.5～28 min,45%A→60%A;28～45min,60%A→20%A),流速0.7 mL.min-1,检测波长360 nm,柱温33℃,进样量20μL。结果:芦丁、槲皮素及山柰酚浓度分别在32.6～326μg.mL-1(r=0.9999)、1.06～10.6μg.mL-1(r=0.9997)和0.32～3.2μg.mL-1(r=0.9997)范围内线性关系良好;平均加样回收率(n=6)分别为99.53%(RSD=1.8%),96.96%(RSD=1.7%),98.75%(RSD=1.9%)。结论:该方法简便,操作简单,结果准确,重复性好,为评价萱草药材的质量提供了可靠的分析方法,可作为萱草中黄酮类化合物的含量测定方法。     

HPLC双波长梯度洗脱测定六味络通胶囊中绿原酸、阿魏酸、甘草苷的含量

目的:研究 HPLC 法同时测定六味络通胶囊中绿原酸、阿魏酸及甘草苷含量。方法:采用 Kromasil C_(18)柱(150 mm×4.6mm,5 μm)以乙腈(A)-2%醋酸水溶液(B)为流动相,梯度洗脱:0 min(6%A)→15 min(6%A)→18 min(12%A)→35 min(12%A)→40 min(50%A),二极管距阵检测,测定波长:绿原酸、阿魏酸用324 nm,甘草苷用276 nm。结果:绿原酸、阿魏酸、甘草苷分别在0.21～3.3μg、0.05～0.80μg、0.23～3.7μg范围内有良好的线性关系,r=0.9999。平均加样回收率(n=6)分别为98.6%,100.2%,98.3%。结论:操作简便,结果可靠,重复性好,可作为该产品质量控制的方法。     

RP-HPLC法测定狼毒中狼毒乙素和岩大戟内酯B的含量

目的:建立同时测定狼毒药材中狼毒乙素和岩大戟内酯B的HPLC定量分析方法。方法:采用Lichrospher C18(4.6 mm×250 mm,5μm)色谱柱,以乙腈(A)-水(B)为流动相梯度洗脱(0~5 min,40%A;5~10 min,40%A→50%A;10~30 min,50%A→55%A;30~40 min,55%A→60%A;40~45 min,60%A→70%A),流速1.0 mL.min-1;检测波长226 nm,柱温30℃。结果:狼毒乙素线性范围为0.005~0.863μg(r=0.9999),平均回收率为96.4%(RSD=1.4%,n=6);岩大戟内酯B线性范围为0.003~0.496μg(r=0.9999),平均回收率为96.0%(RSD=2.1%,n=6)。结论:本方法专属性强,结果准确,简便可行,可用于狼毒药材的质量控制。     

HPLC法测定三七药材中3种皂苷的含量

目的采用高效液相色谱法同时测定三七中三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1的含量.方法采用luna C18(2)分析柱;以(A)乙腈-(B)0.05%磷酸二元线性梯度洗脱:0～8 min(20%A∶80%B),8～20 min(20%A～40%A)∶(80%B～60%B),20～30 min(40%A～20%A)∶(60%B～80%B),检测波长203 nm.结果测得三七皂苷R1、人参皂苷Rg1、人参皂苷Rb1的回收率分别是(97.7±1.6)%、(98.6±1.1)%、(99.2±0.8)%.线性范围分别是:三七皂苷R112～150 μg·mL-1(r=0.9990)、人参皂苷Rg140～500 μg·mL-1(r=0.999 4)、人参皂苷Rb144～550 μg·mL-1(r=0.999 7).结论采用高效液相色谱梯度洗脱法能将多种皂苷很好地分离检测,方法准确可靠,重复性好,结果稳定.     

HPLC法测定香附丸(水丸)中芍药苷的含量

目的:建立HPLC法对香附丸(水丸)中芍药苷的含量测定方法.方法:采用HPLC法.色谱柱:Alltech Apollo C18色谱柱(4.6 mm×250 mm,5μm),流动相:A为乙腈,B为0.1%磷酸溶液,梯度洗脱程序为0 min(14%A),21 min(86%A),41 min(14%A);检测波长:230 nm;流速为1.0 mL·min-1;柱温为30℃.结果:芍药苷在0.123 6～1.854μg内呈良好的线性关系,回归方程为Y=1.06×106X-2.33×103,r=1.000 0;平均回收率为99.07%(RSD=0.35%).结论:本法准确度高,重复性好,可作为本制剂的质量控制方法.     

RP-HPLC同时测定大卫颗粒中绿原酸、黄芩苷、连翘苷、黄芩素和汉黄芩素的含量

目的:建立RP-HPLC法同时测定大卫颗粒中绿原酸、黄芩苷、连翘苷、黄芩素和汉黄芩素的含量。方法:采用Diamon-sil C18(200 mm×4.6 mm,5μm)色谱柱,以乙腈(A)-0.05%磷酸溶液(B)为流动相进行梯度洗脱(0～10 min,10%A→15%A;10～15 min,15%A→25%A;15～25 min,25%A→28%A;25～35 min,28%A→35%A),柱温35℃,流速1.0 mL.min-1,检测波长230 nm,进样量20μL。结果:绿原酸、黄芩苷、连翘苷、黄芩素和汉黄芩素5个成分质量浓度分别在2.80～28.0μg.mL-1(r=0.9992)、25.71～257.1μg.mL-1(r=0.9994)、0.71～7.1μg.mL-1(r=0.9995)、0.81～8.1μg.mL-1(r=0.9990)、0.50～5.0μg.mL-1(r=0.9992)的范围内与峰面积呈良好的线性关系;方法的平均回收率(n=9)分别为100.0%,99.0%,99.0%,99.9%,99.4%。结论:所建立的高效液相色谱测定法简便、准确,重复性好,专属性强,可用于大卫颗粒的含量测定质量控制。     

重组人促卵泡激素氧化亚基分析方法研究

目的:研究建立重组人促卵泡激素(rhFSH)氧化亚基分析方法,制备稳定的氧化亚基分析用系统适用性溶液。方法:通过加入过氧化氢氧化破坏重组人促卵泡激素,后加入甲硫氨酸终止该氧化作用,制备系统适用性溶液。采用RP-HPLC法分离分析重组人促卵泡激素氧化亚基。使用C₄色谱柱(250 mm×4.6 mm, 5μm);流动相A为0.2 mol·L^-1磷酸二氢钾缓冲液(pH 2.5),流动相B为80%乙腈;流速1 mL·min^-1;柱温30℃;检测波长210 nm;梯度洗脱(0～10 min, 82%A→74%A;10～25 min, 74%A→60%A;25～26 min, 60%A→20%A;26～56 min, 20%A;56～56.1 min, 20%A→82%A;56.1～75 min, 82%A)。结果:加入甲硫氨酸可有效终止氧化作用,制备得到稳定的系统适用性溶液。优化建立的氧化亚基分析方法专属性良好,重复性(RSD=0.22%)、色谱柱耐用性(RSD=3.21%)、36 h内溶液稳定性(RSD=0.65%)结果良好。9...     

蒲地灌肠液定性定量方法的研究

目的为保证蒲地灌肠液质量,建立其质量标准。方法采用薄层色谱法对紫花地丁、当归、香附、白芍进行鉴别;RP-HPLC法测定蒲地灌肠液中秦皮乙素与芍药苷的含量。采用Kromasil C18色谱柱(250 mm×4.6 mm,5μm),流动相为0.1%磷酸水溶液(A)-乙腈(B),梯度洗脱0~18 min,88%A;18~25 min,88%~80%A;25~35 min,80%A;35~40 min,80%~88%A;40~45min,88%A;流速为1.0 m L·min~(-1),检测波长为230 nm,柱温为30℃,进样量5μL。结果紫花地丁、当归、香附和白芍薄层色谱鉴别方法专属性强;秦皮乙素和芍药苷线性范围分别为4.24~67.84μg·m L~(-1)(r=0.9995)和8~320μg·m L~(-1)(r=0.9995);平均加样回收率分别为99.9%(RSD=1.9%)和100.9%(RSD=1.9%)。结论该方法简单、准确、重复性好,能有效地控制该制剂的质量。     

丹参药材脂溶性成分的HPLC指纹图谱

目的建立丹参药材的高效液相指纹图谱分析方法。方法采用Kromasil C18(4.6 mm×250 mm,5μm)色谱柱,以体积分数为0.02%磷酸乙腈(A)与体积分数为0.02%磷酸水(B)梯度洗脱,线性梯度洗脱程序为:0~15 min,20%~30%A;15~18 min,30%~50%A;18~35 min,50%~60%A;35~60 min,60%~80%A;60~75 min,80%~100%A;75~100 min,100%A。流速为1.0 mL.min-1,柱温为40℃,检测波长280 nm,以丹参酮ⅡA为参照物。结果通过对11批不同产地丹参药材的测定,标定了14个共有峰,并通过“中药指纹图谱计算机辅助相似度软件”,计算出其相似度均在0.9以上。结论该方法准确可靠,重现性好,为更好地控制丹参内在质量提供了科学依据。     

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.

     

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.     

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.     

Gender-related symptoms and correlates of alcohol dependence among men and women with a lifetime diagnosis of alcohol use disorders

This study explored gender-related symptoms and correlates of alcohol dependence in a crosssectional study of 150 men and 150 women with a lifetime diagnosis of alcohol use disorders (AUD). Participants were recruited in equal numbers from treatment settings, correctional centres and the general community. Standardized measures were used to determine participants' use of substances, history of psychiatric disorders and psychosocial stress, their sensation seeking and family history of substance use and mental health disorders. Multivariate analyses were used to detect patterns of variables associated with gender and the lifetime severity of AUD. Men had a longer history of severe AUD than women. Women had similar levels of alcohol dependence and medical and psychological sequelae as men, despite 6 fewer years of AUD. More women than men had a history of severe psychosocial stress, severe dependence on other substances and antecedent mental health problems, especially mood and anxiety disorders. There were differences in family history of alcohol-related problems approximating same-gender aggregation. The severity of a lifetime AUD was predicted by its earlier age at onset and the occurrence of other disorders, especially anxiety, among both men and women. The limitations in the generalizability of these findings due to sample idiosyncrasies are discussed.     

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.