GC-MS/MS法测定甲磺酸卡莫司他原料药中N-亚硝基二甲胺

目的建立GC-MS/MS法测定甲磺酸卡莫司他原料药中的亚硝胺类基因毒性杂质N-亚硝基二甲胺(NDMA)。方法采用Agilent DB-SELECT 624UI色谱柱(30 m×0.32 mm×1.80μm)色谱柱,离子源为EI源,采用多反应监测模式,以m/z 74→44.2为定量离子,m/z 74→42.2为定性离子,进行数据采集。结果 NDMA在12.571 2～100.569 6 ng/mL线性良好(r=0.997 6),检测限质量浓度为5.028 5 ng/mL,平均回收率为107.4%,RSD值为2.5%。结论该方法操作简单,灵敏度高,专属性和重复性好,可用于甲磺酸卡莫司他原料药中NDMA的检测。     

LC-MS/MS法测定Wistar大鼠组织中银杏内酯B的浓度

目的建立LC-MS/MS法测定大鼠组织中银杏内酯B的浓度,并将此方法应用于银杏内酯B注射液在Wistar大鼠体内的组织分布研究。方法色谱柱:AQVASIL C18柱(100 mm×2.0 mm I.D.,5μm),流动相:乙腈-水-甲酸(体积比为60.0∶40.0∶0.4),流速:0.4 mL.min-1,离子源为电喷雾电离(electrospray ionization,ESI)源,负离子化方式,以选择反应监测(selective reaction monito-ring,SRM)方式进行扫描定量,用于定量分析的离子反应分别为m/z 423→m/z 367(银杏内酯B)和m/z 444→m/z 138(格列吡嗪)。测定了18只Wistar大鼠静脉注射给予银杏内酯B(给药剂量为3.75 mg.kg-1)后大鼠体内各组织和脏器中银杏内酯B的质量浓度。结果测定大鼠组织匀浆液中银杏内酯B的线性为1.0～500.0μg.L-1,定量下限为1.00μg.L-1。低、中、高3个质量浓度质控样品的日内和日间精密度分别小于6.4%和14.5%,准确度为97.3%～103.6%。银杏内酯B在心、肝、脾、肺、肾、胰、胃、小肠、脑、肌肉、脂肪、子宫、卵巢和睾丸14种组织中(除脑组织6 h时间点外)均被检出。银杏内酯B在大鼠各组织中分布广泛,自各组织中消除也较快。结论该方法适用于银杏内酯B注射液在Wistar大鼠体内的组织分布研究。     

高效液相色谱法测定银杏内酯B滴丸的含量

目的建立银杏内酯B滴丸中银杏内酯B的含量测定方法。方法采用HPLC,选择Ec lipseXDB-C18(150mm×4.6mm,5μm)色谱柱,以甲醇-0.1%磷酸溶液(33.3∶66.7)为流动相;流速:1.0mL.m in-1;检测波长:225nm。结果银杏内酯B在0.2650~4.240mg.mL-1范围内线性关系良好(r=1.000);平均加样回收率为99.6%(RSD=1.3%)。结论此方法简便、快速、准确,可作为银杏内酯B滴丸的含量测定方法。     

盐酸美金刚缓释胶囊在比格犬体内的药动学研究

目的建立LC-MS/MS法研究盐酸美金刚缓释胶囊在比格犬体内的药动学特征。方法采用Agilent Extend-C18色谱柱(30 mm×4.6 mm,1.8μm);流动相A为0.1%甲酸,流动相B为甲醇,流动相B比例为65%,等度洗脱2.5 min;体积流量为1 mL/min,柱温40℃;进样量5μL。采用电喷雾离子源(ESI),多级反应监测(MRM)模式,以正离子化方式进行检测,用于定量分析的离子反应分别为m/z 180.0→107.0(盐酸美金刚)、m/z 268.0→116.0(内标琥珀酸美托洛尔)。将6只比格犬随机分为两组,采用两种制剂双周期交叉给药方法,分别ig盐酸美金刚缓释胶囊受试制剂或参比制剂1粒。采用LC-MS/MS法测定比格犬体内盐酸美金刚血药浓度。采用DAS ver 2.1.1软件进行数据分析并计算出相关药动学参数。结果盐酸美金刚的线性范围为1.25～125 ng/mL,定量下限为1.25 ng/mL,日内、日间精密度RSD值均小于6.5%。单次ig受试制剂和参比制剂后,比格犬血浆中盐酸美金刚的Cmax分别为(59.99±14.78)、(64.79±16.26)ng/mL,tmax分别为(7.83±2.64)、(7.33±3.5)h,t1/2分别为(10.84±2.04)、(11.39±1.31)h,AUC0-t分别为(9.99±3.3)、(11.02±2)μg·h/mL,AUC0-∞分别为(10.63±3.42)、(11.72±2.13)μg·h/mL。结论采用LC-MS/MS方法操作简单、准确度、专属性高,可用于盐酸美金刚缓释胶囊的药动学研究,盐酸美金刚受试制剂在比格犬体内药动学行为与参比制剂相似。     

HPLC-ELSD法测定白果中银杏内酯A、银杏内酯B和银杏内酯C

目的建立HPLC-ELSD法同时测定白果药材中银杏内酯A、银杏内酯B和银杏内酯C 3个萜杏内酯成分。方法采用Agilent 5TC C18色谱柱(250 mm×4.6 mm,5μm),以甲醇–四氢呋喃–水(25∶10∶65)为流动相,体积流量1 m L/min,柱温30℃,蒸发光散射检测器检测,对照品进样量为10、20μL,供试品溶液进样量为5~20μL。结果银杏内酯A在0.37~5.92μg、银杏内酯B在2.8~44.8μg、银杏内酯C在0.65~10.4μg时线性关系良好。银杏内酯A、银杏内酯B、银杏内酯C平均回收率分别为92.40%、95.03%、91.29%,RSD值分别为2.75%、2.06%、2.88%。结论本法操作简单,重复性好,为白果药材的质量控制提供了依据。     

高效液相色谱法测定银杏总内酯注射液中银杏总内酯的含量

目的:采用HPLC法测定银杏总内酯注射液中总内酯的含量.方法:采用Dikma Diamosn型色谱柱(5μm,4.6×200 mm),流动相为甲醇一水(35:65),流速为1.0 mL/mim,示差折光检测器,柱温为30C.结果:白果内酯在0.206～2.060mg/mL(r=0.9998,n=5),银杏内酯C在0.056～0.560mg/mL(r=0.9996,n=5),银杏内酯A在0.150～1.500 mg/mL,(r=0.9994,n=5),银杏内酯B在0.102～1.020 mg/mL范围内与峰面积积分值线性关系良好.结论:本方法测定结果准确,操作简便,灵敏度高,重复性好,可用于本品的质量控制.     

HPLC-CAD法测定银杏叶片中萜类内酯的含量

目的建立银杏叶片中萜类内酯的含量测定方法。方法采用HPLC-CAD法测定白果内酯、银杏内酯A、银杏内酯B和银杏内酯C的含量。使用Agilent TC C_(18)色谱柱(250 mm×4.6 mm,5μm),以正丙醇-四氢呋喃-水(1∶15∶84)为流动相,流速1.0 mL·min~(-1),柱温40℃,采用电喷雾检测器进行检测。结果各萜类内酯在线性范围内与峰面积线性关系良好,相关系数为0.9997～1.000,平均回收率(n=6)在97.3%～99.1%,RSD在1.6%～2.5%。结论该方法灵敏度高,精密度、重复性、准确度均良好,可作为银杏叶片中萜类内酯的含量测定方法。     

HPLC-ELSD法测定注射用银杏二萜内酯中银杏内酯A、B、C、K

目的建立测定注射用银杏二萜内酯中银杏内酯A、B、C、K的HPLC-ELSD法。方法采用HPLC-ELSD法,Phenomenex Luna C18色谱柱(250 mm×4.6 mm,5μm),流动相为四氢呋喃–甲醇–水,梯度洗脱;体积流量为1.0 mL/min;柱温为30℃;漂移管温度为105℃,氮气体积流量为3.0 L/min。结果注射用银杏二萜内酯中银杏内酯A、B、C、K分别在1.70~8.50μg(r=0.999 7)、2.45~12.5μg(r=0.999 8)、0.30~1.50μg(r=0.999 7)、0.25~1.25μg(r=0.999 7)呈良好的线性关系;平均回收率分别为101.03%、100.86%、100.78%、101.02%,RSD值分别为0.87%、0.29%、1.30%、1.34%。结论该方法简便准确,重复性好,可用于注射用银杏二萜内酯中银杏内酯A、B、C、K的测定。     

银杏酮酯分散片萜类内酯的溶出度试验

目的建立银杏酮酯分散片萜类内酯的溶出度测定方法.方法以萜类内酯为指标,采用高效液相色谱法-蒸发光检测法进行检测,采用ZORBAX RX-C18色谱柱(4.6mm×150 mm,5nμm);以水-甲醇-四氢呋喃(752010)为流动相;流速1.0mL·min-1.结果白果内酯、银杏内酯A、银杏内酯B、银杏内酯C的线性范围分别为1.519～7.595、1.087～6.522、1.01～3.03、1.056～3.168μg,平均回收率分别为99.55%(RSD为1.87%);99.79%(RSD为1.94%);99.78%(RSD为2.38%);98.21%(RSD为2.54%).结论该方法重复性较好,可作为银杏酮酯分散片的溶出度测定方法.     

LC-MS/MS法测定Beagle犬血浆中右旋酮洛芬的浓度

目的:建立LC-MS/MS法测定Beagle犬血浆中右旋酮洛芬的浓度。方法:血浆样品采用乙腈沉淀法处理,取上清液进样。以乙腈(每500 mL中加入甲酸10μL)-水(80∶20,v/v)为流动相;用Zorbax Eclipse SB-C18柱(5μm,150 mm×4.6 mm)色谱柱分离;以布洛芬为内标。流速0.5 mL·min-1;进样量25μL;柱温为室温。质谱检测采用ESI正离子模式,扫描方式为多反应监测,扫描离子对m/z右旋酮洛芬253.0/209.0,布洛芬204.8/160.8。结果:右旋酮洛芬的线性范围为1.0～1000 ng·mL-1,线性关系良好(r=0.9979),最低定量下限为1.0 ng·mL-1,绝对回收率在100.0%～104.3%之间,变异系数均小于6%。结论:该方法预处理快速简单,灵敏度高,所建立的LC-MS/MS法准确、可靠,可用于犬血浆中右旋酮洛芬的浓度测定及药代动力学研究。     

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.     

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.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

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.