HPLC法测定他达拉非片含量和有关物质

摘 要 目的： 建立他达拉非片含量测定及有关物质检查的方法。 方法: 采用高效液相色谱法。色谱柱：安捷伦ZORBAX Eclipse XDB C₈（250 mm×4.6 mm,5 μm） ;流动相：0.1% 三氟乙酸溶液-乙腈（65∶35）;流速：1.0 ml·min^-1;柱温：35℃;检测波长：285 nm,进样量20 μl。结果:在选定的色谱条件下,主成分与各杂质峰分离度良好。他达拉非质量浓度在20.13～201.30 μg·ml^-1范围内与峰面积有良好的线性关系(r=0.999 3);平均回收率为99.5%(RSD=1.1%,n=9) ;检出限为0.6 ng,定量限为2 ng。结论: 该方法专属性强、灵敏度高,可用于他达拉非片含量和有关物质测定。     

HPLC法测定盐酸头孢他美酯片的含量及有关物质

摘 要 目的： 建立盐酸头孢他美酯片含量测定及其有关物质检查的HPLC方法。方法: 色谱柱为Hypersil ODS2 C₁₈ (250 mm×4.6 mm,5 μm),流动相为0.005 mol·L^-1四丁基氢氧化铵溶液 乙腈（64∶36,用磷酸调节pH至4.5）,流速为1.0 ml·min^-1,检测波长为232 nm,柱温为30℃,进样量10 μl。结果: 头孢他美与其他有关检查物质能完全分离,其线性回归方程为Y=2.29×10⁴X－4.30×10⁴(r=0.999 9),表明头孢他美在51.01～510.07 μg·mL^-1的浓度范围内与峰面积线性关系良好。头孢他美平均加样回收率为99.60%,RSD为 0.84%（n=9）。结论:该方法简便、准确,专属性强,可用于盐酸头孢他美酯片的含量和有关物质测定。     

HPLC 法测定洛莫司汀-碘海醇复方脂质体的药物含量及包封率

目的：建立 RP-HPLC 法测定洛莫司汀-碘海醇复方脂质体中药物的含量及包封率。方法：使用 Diamonsil^TM（钻石）C₁₈ 色谱柱（200 mm × 4.6 mm,5 μm）,流动相为乙腈 - 水（65︰35）,柱温 25 ℃,体积流量 1.0 mL/min,检测波长为 230 nm,鱼精蛋白凝聚法分离游离药物,测定复方脂质体中洛莫司汀的含量及包封率;使用 Diamonsil ^TMC₁₈ 色谱柱（200 mm × 4.6 mm,5 μm）,流动相为甲醇 - 水（10︰90）,柱温 25 ℃,体积流量 1.0 mL/min,检测波长为 244 nm,鱼精蛋白凝聚法分离游离药物,测定复方脂质体中碘海醇的含量及包封率。结果：洛莫司汀与辅料及溶剂峰分离良好,在 1.0～20.0 μg/mL线性关系良好（r = 1.0, n = 5）,回收率为 99.0 %～101.0 %;碘海醇与辅料及溶剂峰分离良好,在 6.0～60.0 μg/mL线性关系良好（r = 0.999 9, n = 5）,回收率为99.0 %～101.0 %。结论：该方法准确、简单,可用于洛莫司汀-碘海醇复方脂质体含量及包封率的测定。     

HPLC法测定依达拉奉注射液中的有关物质

目的 建立高效液相色谱法测定依达拉奉注射液的有关物质。方法 采用Agilent SB-C₁₈（4.6 mm×250 mm,5 μm）色谱柱,流动相A为0.05 mol·L^-1磷酸二氢钾溶液（用三乙胺调节pH值至7.0）,流动相B为甲醇,梯度洗脱,流速为1.0 mL﹒min^-1,检测波长为243 nm,柱温30 ℃。结果 依达拉奉及各杂质均能有效分离,在相应的浓度范围峰面积与浓度呈良好的线性关系,相关系数均大于0.999,平均回收率在95%～105%范围内。结论 该方法专属性强,简便可靠、灵敏、准确,适用于依达拉奉注射液中的有关物质的测定。     

HPLC法测定盐酸赛庚啶片的含量及其有关物质

摘 要 目的： 建立高效液相色谱法测定盐酸赛庚啶片的含量及其有关物质。方法: 色谱柱：资生堂CAPCELL PAK C₁₈(250 mm×4.6 mm, 5 μm);流动相：甲醇-0.002 5mol·L^-1庚烷磺酸钠溶液（用磷酸调节pH至3.0）（60∶40）;检测波长：225 nm;流速：1.0 ml·min^-1;柱温：30℃;进样量：10 μl。结果: 盐酸赛庚啶在4.12～82.40 μg·mL^-1范围线性关系良好,r=1.000 0;平均加样回收率为99.2%（RSD=0.8%,n=9）。有关物质各杂质与盐酸赛庚啶主峰的分离度良好。结论:该方法操作简单、快捷、准确,可用于盐酸赛庚啶片的质量控制。     

HPLC测定愈创木酚磺酸钾的有关物质

目的 建立HPLC测定愈创木酚磺酸钾有关物质的方法。方法 采用InertSustain C₁₈色谱柱（4.6 mm×250 mm，5 μm），以乙腈-0.02 mol·L^-1磷酸盐缓冲液（20∶80）为流动相，流速1 mL·min^-1，柱温35℃，检测波长279 nm。结果 愈创木酚磺酸钾的2个异构体能得到有效分离，主成分与杂质的分离度良好。主成分及其各杂质在各自浓度范围内呈现良好的线性关系（r>0.999）；异构体Ⅲ、愈创木酚和杂质Ⅰ的平均加样回收率分别为101.35%，100.78%，99.45%，RSD分别为0.7%，0.7%，1.4%。结论 该方法快速、准确、专属性强，可用于愈创木酚磺酸钾原料药的有关物质检查。     

HPLC法测定长春西汀注射液的有关物质

摘 要 目的：建立测定长春西汀注射液有关物质的方法。方法: 采用高效液相色谱法。色谱柱InertSustain C₁₈(250 mm×4.6 mm,5 μm),检测波长：280 nm;流动相：0.2 mol·L^-1醋酸铵溶液-乙腈（40∶60）,流速：1.0 ml·min^-1;柱温：30 ℃,进样量：10 μl。结果: 长春西汀主峰与各杂质峰均能良好的分离。杂质A、B、C和D分别在0.276～5.520,0.283～5.660,0.269～5.380,0.282～5.640 μg·ml^-1浓度范围内具有良好的线性关系,r分别为1.000 0,1.000 0,0.999 9,0.999 9。平均回收率分别为100.5%,100.7%,100.04%,99.9%（RSD分别为1.35%,0.99%,1.13%,1.10%,n=9）。结论:该方法具有较高的专属性、灵敏度、和精密度,能够有效控制长春西汀注射液中的有关物质。     

HPLC法测定依非韦伦片中三个特定杂质的含量

目的:建立测定依非韦伦片中三种难分离的特定杂质含量的方法。方法:采用Waters Symmetry C₁₈柱(4.6 mm×250 mm,5μm),以水-乙腈-三氟乙酸为流动相梯度洗脱,检测波长为250 nm,流量为1.5 mL·min^-1。结果:三种特定杂质峰与主峰间的分离度良好,最低检测限为0.5 ng,依非韦伦浓度在0.125~2.5μg·mL^-1范围内与峰面积呈良好的线性关系(r=0.9999,n=6)。结论:本方法简便,专属性强,可用于依非韦伦片中三种特定杂质的控制。     

HPLC法测定醋甲唑胺片的含量和有关物质

摘 要 目的： 建立高效液相色谱法测定醋甲唑胺片的含量和有关物质。方法: 色谱柱为Symmetry C₁₈（150 mm×4.6 mm,5 μm）;流动相为乙腈-0.1 mol·L^-1醋酸钠（pH4.5）（20∶80）;流速:1.0 ml·min^-1;检测波长:252 nm;柱温:30℃;进样量:20 μl。结果: 醋甲唑胺浓度在10.0～80.0 μg·mL^-1范围内峰面积值呈良好的线性关系（r=0.999 7）,平均加样回收率为99.56%,RSD=0.95%(n=9)。测得3批样品杂质含量分别为0.25%,0.21%,0.23%。结论: 本法简便、准确,专属性好,精密度高,专属性好,可用于醋甲唑胺片的含量和有关物质测定。     

HPLC-CAD测定羟基脲胶囊含量及有关物质

目的 建立测定羟基脲胶囊含量及有关物质检查的HPLC-CAD方法。方法 采用Phenomenx Luna^® NH₂柱（250 mm×4.6 mm，10 μm，100Ǻ），柱温为40℃；以乙腈-水（82：18）为流动相，流速为1.0 mL·min^-1，检测器为电雾式检测器，雾化温度50℃，进样量10 μL。结果 在选定的色谱条件下，主峰与各杂质峰均能良好分离。采用外标法计算羟基脲胶囊的含量，采用主成分自身对照法计算脲的含量；羟基脲和脲分别在0.304 8~1.270 0 mg·mL^-1（r=0.999 8）、0.241 1~1.004 6 mg·mL^-1（r=1.000 0）内与峰面积呈良好线性关系。脲的检测限和定量限分别为2.82 ng和8.46 ng。结论 本方法操作简便，专属性强，结果可靠，可用于羟基脲胶囊的含量及有关物质的测定。     

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.