HPLC法测定复方硫酸氢氯Ⅱ比格雷片中硫酸氢氯吡格雷和阿司匹林的含量

目的 建立复方硫酸氢氯吡格雷片中硫酸氢氯吡格雷和阿司匹林含量测定的HPLC法.方法 色谱柱为氰基柱,流动相为甲醇-水-三乙胺(体积比为500∶500∶2,磷酸调节pH值至3.8),流速为1 mL·min-1,检测波长为235 nm,柱温为35℃,进样量为20 μL.结果 在该色谱条件下,硫酸氢氯吡格雷与阿司匹林可达到较好分离,硫酸氢氯吡格雷在12.0～100.0 mg·L-1内质量浓度与峰面积呈良好的线性关系(r=0.999 5).阿司匹林在38.4～320.0 mg·L-1内质量浓度与峰面积呈良好的线性关系(r=0.999 7).硫酸氢氯吡格雷与阿司匹林的平均回收率分别为(99.7±1.80)%(n=9)和(100.3±0.53)%(n=9).结论 HPLC法适用于复方硫酸氢氯吡格雷片中硫酸氢氯吡格雷和阿司匹林的含量测定.     

HPLC法测定复方硫酸氢氯吡格雷片中硫酸氢氯吡格雷和阿司匹林的含量

目的建立复方硫酸氢氯吡格雷片中硫酸氢氯吡格雷和阿司匹林含量测定的HPLC法。方法色谱柱为氰基柱,流动相为甲醇-水-三乙胺(体积比为500∶500∶2,磷酸调节pH值至3.8),流速为1 mL.min-1,检测波长为235 nm,柱温为35℃,进样量为20μL。结果在该色谱条件下,硫酸氢氯吡格雷与阿司匹林可达到较好分离,硫酸氢氯吡格雷在12.0~100.0 mg.L-1内质量浓度与峰面积呈良好的线性关系(r=0.999 5)。阿司匹林在38.4~320.0 mg.L-1内质量浓度与峰面积呈良好的线性关系(r=0.999 7)。硫酸氢氯吡格雷与阿司匹林的平均回收率分别为(99.7±1.80)%(n=9)和(100.3±0.53)%(n=9)。结论HPLC法适用于复方硫酸氢氯吡格雷片中硫酸氢氯吡格雷和阿司匹林的含量测定。     

硫酸氢氯吡格雷的有关物质UPLC-MS测定方法的研究

目的研究建立硫酸氢氯吡格雷有关物质的UPLC-MS测定方法,考察其国产原料药有关物质。方法选用ULTRON ES-OVM手性色谱柱(15cm×4mm,5μm),流动相:0.03mol/L的醋酸氨溶液-乙腈(78:22),流速:0.8 mL/min,柱温:30℃,进样量:10μl。结果硫酸氢氯吡格雷的高温降解产物(杂质A,分子式:C15H14ClNO2S,分子量:307.8;杂质C,分子式:C16H16ClNO2S.H2SO4,分子量:419.90)及各已知杂质可有效分离。结论该方法稳定可靠,经与国外产品图谱对比,国产硫酸氢氯吡格雷的杂质少,质量较高。     

硫酸氢氯吡格雷的有关物质UPLC—MS测定方法的研究

目的研究建立硫酸氢氯吡格雷有关物质的UPLC-MS测定方法,考察其国产原料药有关物质。方法选用ULTRON ES-OVM手性色谱柱（15cm×4mm,5μm）,流动相：0.03mol/L的醋酸氨溶液-乙腈（78：22）,流速：0.8 mL/min,柱温：30℃,进样量：10μl。结果硫酸氢氯吡格雷的高温降解产物（杂质A,分子式：C15H14ClNO2S,分子量：307.8;杂质C,分子式：C16H16ClNO2S.H2SO4,分子量：419.90）及各已知杂质可有效分离。结论该方法稳定可靠,经与国外产品图谱对比,国产硫酸氢氯吡格雷的杂质少,质量较高。     

两种晶型硫酸氢氯吡格雷片稳定性的比较

目的建立高效液相色谱法测定硫酸氢氯吡格雷片中的有关物质,并对2种晶型片剂的稳定性进行考察。方法采用ULtron ES-OVM手性色谱柱(150 mm×4.6 mm,5μm),流动相:0.01 mol.L-1磷酸二氢钾溶液-乙腈(体积比为75∶25),流速:1.0 mL.min-1,进样量:10μL,检测波长:220 nm,柱温:25℃。结果在该色谱条件下,杂质A、杂质B的第一个异构体(杂质B1)、杂质C质量浓度分别在0.3～3.0、0.45～4.50、1.4～14.0 mg.L-1内与峰面积呈良好的线性关系,硫酸氢氯吡格雷与杂质B1之间的分离度大于2.5。稳定性试验表明,在高温条件下,两种晶型硫酸氢氯吡格雷片中杂质B、C含量明显增加,晶型I杂质A略有增加;在高湿条件下2种晶型均易产生杂质A;在光照条件2种晶型的杂质均未明显增加。结论高温是影响2种晶型硫酸氢氯吡格雷片稳定性的主要因素,2种晶型中Ⅱ型硫酸氢氯吡格雷片稳定性更佳。     

硫酸氢氯吡格雷杂质D的合成

目的 合成硫酸氢氯吡格雷杂质D。方法 以右旋邻氯苯甘氨酸为原料,经过氨基保护、与(R)-2-氯扁桃酸甲酯成酯、脱保护后得到(2R)-(2-氯苯基)-[(2S)-(2-氯苯基)-(2-氨基)乙酸酯基]乙酸甲酯,再与2-(2噻吩)乙醇对甲苯磺酸酯发生亲核取代反应后,经成盐、环合反应得到硫酸氢氯吡格雷杂质D。结果 设计合成杂质D路线总收率为54%,杂质D纯度在99.0%以上。结论 合成杂质D产品纯度较高,可用作硫酸氢氯吡格雷杂质研究对照品。     

复方硫酸氢氯吡格雷阿司匹林双层片的制备和溶出度测定

目的建立复方硫酸氢氯吡格雷阿司匹林双层片的制备和溶出度测定方法,通过比较自制品和市售产品的体外溶出行为,考察其有效性。方法制备复方硫酸氢氯吡格雷双层片,采用反相高效液相色谱法测定含量。色谱条件:色谱柱为Ultimate XB-C18柱(250mm×4.6mm,5μm),流动相为乙腈-水-磷酸(体积比40∶60∶2),检测波长为235nm,流速为1.0mL·min-1;采用桨法,以pH值2.0盐酸缓冲液为溶出介质,转速为75 r·min-1测定溶出度;采用f2因子比较法,考察体外溶出行为。结果硫酸氢氯吡格雷和阿司匹林的质量浓度在20~200mg·L-1内与峰面积具有良好的线性关系(r=0.999 9)。硫酸氢氯吡格雷与阿司匹林的平均回收率分别为100.2%和99.8%。自制品和市售品的溶出曲线一致。结论制备方法简单易行。所建立的溶出度测定方法专属、高效、简便,可用于测定复方硫酸氢氯吡格雷阿司匹林双层片的溶出度。     

用液相色谱-质谱联用法同时测定人血浆中阿司匹林和氯吡格雷的浓度

目的：建立液相色谱-质谱联用（LC-MS）法同时测定人血浆中阿司匹林和氯吡格雷的浓度。方法 ：以苯甲酸为内标,色谱柱为Kromasil C18柱（150mm×4.6mm,5μm）;流动相为甲醇-0.2%乙酸溶液（60∶40）,流速为0.8ml/min。采用正离子选择性离子监测模式进行检测,参比离子m/z分别为322.1（氯吡格雷）、178.9（阿司匹林）和121.1（苯甲酸）。结果：阿司匹林和氯吡格雷分别在10~5000ng/ml和1~200ng/ml的浓度范围内呈良好线性关系（r≥0.996 8）,低、中、高浓度阿司匹林（40、800、4000ng/ml）和氯吡格雷（4、40、160ng/ml）的日内、日间RSD均〈15%,提取回收率为82.4%~92.4%（n=5）,稳定性良好。结论：本方法准确、灵敏,可同时测定人血浆中阿司匹林和氯吡格雷的浓度。     

阿司匹林联合硫酸氢氯吡格雷口服治疗脑梗死的有效性和安全性研究

目的分析阿司匹林联合硫酸氢氯吡格雷治疗脑梗死的应用效果。方法选择50例在我院进行救治的脑梗死患者进行研究,根据患者入院时间的顺序将50例患者划分为观察组和对照组,每组25例;两组均用阿司匹林治疗,观察组加入硫酸氢氯吡格雷。结果疗效及神经功能缺损评分对比,观察组均明显占优(P<0.05)。结论阿司匹林联合硫酸氢氯吡格雷治疗脑梗死整体效果良好,值得借鉴。     

大鼠口服硫酸氢氯吡格雷的多晶型吸收特点研究

本文采用多种分析检测技术对硫酸氢氯吡格雷的4种晶型物质进行了表征,并采用高效液相色谱(high performance liquid chromatogram,HPLC)技术测定大鼠血浆中硫酸氢氯吡格雷及其代谢产物的含量,研究了大鼠口服硫酸氢氯吡格雷4种不同晶型的吸收特点。结果显示大鼠灌胃给予硫酸氢氯吡格雷后血浆中可以检测到反应吸收过程的代谢产物,硫酸氢氯吡格雷4种晶型的代谢产物曲线下面积有显著性差异。故认为硫酸氢氯吡格雷的不同晶型物质表现出不同的药代动力学特征,提示晶型物质状态不同可能会影响药物的体内作用。     

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.     

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.     

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.     

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.