HPLC梯度法测定复方三七维康胶囊中三七皂苷R1、人参皂苷Rb1、Rg1的含量

摘 要 目的:建立HPLC梯度法同时测定复方三七维康胶囊中三七皂苷R₁、人参皂苷R_b1、R_g13种皂苷的含量。方法: 色谱柱：大连Spherisorb C¹⁸分析柱(200 mm×4.6 mm,5 μm);流动相：乙腈-水,梯度洗脱;流速:1.0 ml·min^-1;检测波长 203 nm。结果:三七皂苷R₁、人参皂苷R_g1和R_b1分别在1.06～18.55 μg（r=0.997 3）、2.02～35.35 μg（r=0.998 2）和2.02～35.35 μg（r=0.998 2）之间线性关系良好。平均回收率三七皂苷R₁为100.8%（RSD=1.53%,n=5）,人参皂苷 R_g1为98.9%（RSD=1.87%,n=5）,人参皂苷R_b1为99.7%（RSD=1.90%,n=5）。结论:HPLC梯度洗脱法能够将多种皂苷很好地分离检测,该方法准确可靠,重复性好,可用于控制其质量。     

HPLC-PAD法测定硫酸依替米星注射液有关物质及含量

目的：建立硫酸依替米星注射液有关物质及含量的反相高效液相色谱-脉冲安培电化学(HPLC-PAD)法。方法：采用十八烷基硅烷键合硅胶为填充剂(4.6mm×250mm,5μm或效能相当的色谱柱),柱温35℃。以0.2mol.L_^-1三氟乙酸溶液(含0.05%五氟丙酸,1.5g.L_^-1无水硫酸钠,0.8%(V/V)的50%氢氧化钠溶液,用50%氢氧化钠溶液调节pH值至3.5)-乙腈(95：5)为流动相,流速为每分钟1.0ml。柱后加碱(50%氢氧化钠溶液1→25,流速每分钟0.5ml)。用积分脉冲安培电化学检测器检测。结果：依替米星在0.075~50μg.mL_^-1(r =0.9996)内线性关系良好;单个最大杂质的重复性试验的RSD(n =6)为1.4%,总杂质的重复性试验的RSD(n =6)为1.7%,含量测定的重复性试验的RSD(n =6)为0. 8%;方法的检测限(S/N =3)为75ng.mL_^-1,定量限(S/N =10)为250ng.mL_^-1;供试品溶液在12 h内稳定性良好。结论：经方法学验证,该方法可以用于硫酸依替米星注射液有关物质及含量的测定。     

反相高效液相色谱法测定重组人干扰素α1b多剂量注射液中间甲酚含量

目的: 建立测定重组人干扰素α1b多剂量注射液中间甲酚含量的高效液相色谱(HPLC)检测方法,并对方法进行验证。方法: 采用反相高效液相色谱(RP-HPLC)法,选用Venusil, ASB-C18(5μm,4.6mm×150mm)色谱柱,流动相为0.2mol.L_^-1硫酸盐缓冲液(pH=2.3)-乙腈(74﹕26),流速1mL.min_^-1,进样量20μL,检测波长270nm,柱温40℃,等度洗脱。对方法进行专属性、线性、准确度、精密度、重复性、检测限、定量限等一系列验证。结果: 该方法在间甲酚含量为100.0～300.0μg.mL_^-1范围内线性良好,r_²值为1.000。相关辅料及干扰素α1b对间甲酚检测无干扰。该方法检测高、中、低3种浓度9组样品回收率(n=9)在96.2%～101.5%;精密度 RSD(n=9)为0.78%;定量下限为4.3mg.mL_^-1,检测下限为1.4mg.mL_^-1。对 6批次重组人干扰素α1b多剂量注射液中间甲酚含量均在其标示量的90～110%之间,质量合格。结论: 建立的RP-HPLC方法专属性良好,精密度、准确性高,可用于检测重组人干扰素α1b多剂量注射液中的间甲酚含量。     

HPLC同时测定鱼腥草芩蓝合剂中3种活性成分的含量

目的 建立HPLC测定鱼腥草芩蓝合剂中绿原酸、黄芩苷和黄芩素的含量。方法 采用Kromasil-100 C₁₈色谱柱(4.6?mm×250 mm,5 μm),流动相为甲醇-0.2%磷酸梯度洗脱,流速为1.0 mL·min^-1,检测波长为322 nm,柱温为35 ℃。结果 绿原酸、黄芩苷和黄芩素线性范围分别为0.022 4~1.12 μg(R²=0.999 7),0.045 8~2.29 μg(R²=0.999 5)和0.031 8~ 1.59?μg(R²=0.999 9);3种成分的平均回收率分别为99.45%(RSD=1.4%),99.37%(RSD=2.0%)和97.90%(RSD=1.7%)。结论 本方法快速简便,精密度高,稳定性和重复性好,可用于鱼腥草芩蓝合剂中绿原酸、黄芩苷和黄芩素3种成分的定量分析和药品质量控制。     

HPLC法同时测定野豌豆中芦丁、金丝桃苷和槲皮素的含量

〗摘 要 目的： 建立野豌豆药材中芦丁、金丝桃苷和槲皮素三个活性成分的HPLC含量测定方法。 方法: 采用Agilent ZORBAX Eclipse XDB C₁₈色谱柱(250 mm×4.6 mm,5 μm),流动相为乙腈 1‰磷酸水溶液,梯度洗脱,流速:0.8 ml·min^-1,检测波长:370 nm,柱温：30℃。结果:芦丁、金丝桃苷和槲皮素的线性范围分别为4.090～130.940 μg ·ml^-1(r=0.999 9)、4.600～147.200 μg ·ml^-1(r=0.999 9)和0.810～25.780 μg·ml^-1(r=0.999 8);平均回收率分别为103.45%（RSD=1.25%）、98.96%（RSD=1.77%）和102.88%（RSD=0.84%）（n=6）。结论: 本方法稳定,重复性好,操作简单,可用于野豌豆药材的质量控制。     

HPLC同时测定白花蛇舌草中熊果酸、异熊果酸和对香豆酸的含量

目的 建立HPLC测定白花蛇舌草中熊果酸、异熊果酸和对香豆酸的含量。方法 采用 Diamonsil C₁₈ ODS (4.6 mm×250 mm,5 μm),流动相为乙腈-15 mmol·L^-1 NH₄Ac(12∶88,pH 3.5),流速为1.0 mL·min^-1,检测波长为308 nm,柱温为35 ℃。结果 熊果酸、异熊果酸和对香豆酸线性范围分别为0.075 7~2.271 μg(R²=0.999 6),0.010 3~0.309 μg (R²=0.999 2)和0.020 2~0.606 μg(R²=0.999 6);3种成分测定的平均回收率分别为99.9% (RSD=0.7%),99.0% (RSD=1.7%)和101.1% (RSD=0.7%)。结论 本法简便、准确度高、稳定性和重复性好,可用于白花蛇舌草中指标性成分定量分析和质量控制。     

甘肃产马尾连药材质量标准提升研究

目的 完善和提高马尾连药材质量标准,为进一步开发马尾连药材提供基础。方法 在市场调研的基础上,采集和收集样品,并查阅馆藏标本,进行对比研究,掌握当前市场商品情况;利用薄层色谱法对马尾连进行定性鉴别,同时建立HPLC法测定马尾连药材中盐酸小檗碱的含量测定方法。结果 甘肃产马尾连主要来源于贝加尔唐松草Thalictrum baicalense Turez.的干燥根;薄层色谱分离良好,斑点清晰;盐酸小檗碱在5.0～160.0μg?mL_^-1范围内线性关系良好,精密度、重复性、稳定性试验 RSD均小于1.8%,平均加样回收率为101.99%(RSD=2.53%,n=6)。结论 本研究所建立方法简单、准确,可用于马尾连药材的质量控制。     

HPLC同时测定蒙药材黄花铁线莲中4种黄酮含量

目的 通过对蒙药材黄花铁线莲中4种黄酮的HPLC含量测定,建立黄花铁线莲质量控制方法。方法 采用YMC C₁₈色谱柱（4.6 mm×250 mm,5 μm）,甲醇-0.04%磷酸水溶液为流动相,梯度洗脱,检测波长350 nm,柱温25℃,流速1.0 mL·min^-1。结果 木犀草苷、金丝桃苷、芦丁和槲皮素分别在0.021 37~0.427 4,0.010 85~0.217 0,0.021 41~0.428 2,0.017 46~0.349 2 μg内与峰面积线性关系良好;精密度、重复性、稳定性的RSD均< 2%,平均回收率为97.4%~102.3%。结论 该方法简便、准确、可靠,重复性好,可用于蒙药材黄花铁线莲的质量评价与控制。     

HPLC法测定炉甘石硫洗剂中苯酚的含量

摘 要 目的:建立HPLC法测定炉甘石硫洗剂中苯酚含量的方法。方法: 采用Eclipse XDB-C₁₈(150 mm×4.6 mm,5 μm)色谱柱,流动相：甲醇-水（40∶60）,流速1.0 ml ·min^-1,检测波长270 nm。结果:苯酚在0.543 5～5.435 0 μg范围内线性关系良好,（r=0.999 9）;平均回收率为99.3%,RSD=0.71%(n=6)。结论:本方法简便、准确、重复性好,可用于该制剂的质量控制。     

参茸天麻酒的质量标准研究和探讨

目的:完善参茸天麻酒的质量控制标准。方法:采用薄层色谱法对方中主要药味天麻、枸杞子、何首乌、人参和五味子展开鉴别考察,采用高效液相色谱法测定君药天麻中天麻素的含量。结果:薄层鉴别斑点清晰、分离度好、专属性强且重复性良好。天麻素含量测定方法操作简单、准确,天麻素的线性范围9.76μg.ml_^-1～97.6μg.ml_^-1(r=0.9999);精密度、稳定性、重复性试验的RSD<1.0%;加样回收率为99.7%(RSD=1.5%,n=6)。结论:定性、定量方法准确、简单,能有效控制参茸天麻药酒的质量。     

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.