UPLC-MS/MS法测定奥美沙坦酯中7个亚硝胺类基因毒性杂质

目的 建立超高效液相色谱-串联质谱法（UPLC-MS/MS）测定奥美沙坦酯中7个基因毒性杂质：N-亚硝基二甲胺、N-亚硝基-4-甲基-4-氨基丁酸、N-亚硝基二乙胺、N-亚硝基乙基异丙基胺、N-亚硝基二异丙胺、N-亚硝基二丙胺、N-亚硝基二丁胺。方法 采用Agilent poroshell PFP （100 mm×2.1 mm,2.7 μm）色谱柱;流动相为0.1%甲酸水溶液（A）-甲醇（B）梯度洗脱;体积流量0.4 mL/min,柱温40℃;采用APCI离子源正离子扫描,多反应监测（MRM）模式下,对7个基因毒性杂质同时进行定量检测。结果 各杂质质量浓度在1~100 ng/mL内具有良好线性关系,r>0.995;低、中、高3个浓度的加样回收率（n=3）为83%~117%,RSD值为0.8%~4.1%,平均加样回收率为87%~106%;检测限范围为0.02~0.19 ng/mL,定量限为0.06~0.65 ng/mL。4批奥美沙坦酯样品中均未检出杂质。结论 该方法灵敏度高,专属性强,可用于测定奥美沙坦酯原料药中7个亚硝胺类杂质,为奥美沙坦酯的质量控制提供参考。     

顶空气相色谱法测定药用复合膜中乙酸丙酯和环戊酮含量

目的建立测定药用复合膜中残留溶剂乙酸丙酯和环戊酮含量的顶空气相色谱法。方法色谱柱为安捷伦DB-WAXETR柱(30m×0.32mm,1.00μm),顶空平衡温度为100℃,平衡时间为60min,分流比为20∶1,进样口温度为200℃,选择程序升温(色谱柱初始温度为55℃保持5min,以5℃/min的速率升至150℃保持3min,再以60℃/min的速率升至220℃保持3min),检测器为火焰离子化检测器,温度为220℃。结果乙酸丙酯和环戊酮的线性范围为0.5~10.0μg/mL,相关系数均大于0.999;乙酸丙酯和环戊酮的检测限分别为0.0009,0.0030mg/m2(n=6),回收率分别为110.30%,99.52%(n=6)。结论该方法简单、快速、准确,可用于药用复合膜中乙酸丙酯和环戊酮的含量测定。     

GC顶空进样测定注射用葛根素中残留溶剂乙醇

目的建立Gc法检查葛根素中残留溶剂乙醇的方法。方法采用GC法测定,色谱条件为：采用DB-624毛细管柱为色谱柱（30m×0．32mm,0．25μm）,柱温40℃保持10min,20℃／min升至180℃;检测器为氢火焰离子化检测器（FID）,检测器温度为300℃;进样口温度为220℃。结果乙醇的检测限为9．3508μg·mL^-1。结论本法简便、准确、灵敏度高,适用于葛根素中乙醇的测定。     

顶空气相色谱法测定头孢泊肟酯中三乙胺的残留量

目的 建立头孢泊肟酯原料药中三乙胺残留量的测定方法。方法 采用顶空气相色谱法，FID 检测器，色谱柱 为Elite-5-AMINE 石英毛细管柱(30m×0.32mm, 1.5μm)，柱温采用程序升温，初始柱温为50℃，保持10min，再以20℃ /min 的速率升至200℃，保持5min，进样口温度为200℃，检测器温度为250℃，载气为氮气，流速为2.0mL/min，进样体积为 1.0mL，分流比为5:1，平衡温度为80℃，平衡30min。结果 在选定的色谱条件下，三乙胺的分离度在35 以上，能够得到良 好分离，0.531~10.617μg/mL 范围内线性关系良好，r=0.9992；检测限为0.0265μg/mL，定量限为0.0796μg/mL；平均回收率为 100.16%(RSD=3.38%, n=9)。结论 该方法简便灵敏、结果准确、重复性好，适用于头孢泊肟酯中三乙胺残留量的检测。     

顶空气相色谱法测定注射用甲磺酸吉米沙星中基因毒性杂质

目的：建立测定注射用甲磺酸吉米沙星中甲磺酸烷基酯类基因毒性杂质的顶空气相色谱法。方法：以Supelco 毛细管柱（30m×250μm,0．25μm）为色谱柱,程序升温：氮气为载气,流速为0．5mL·min-1：采用微池电子捕获检测器,检测器温度为250℃：顶空进样,平衡温度为60℃,平衡时间为30min,进样口温度为110℃,进样量为1mL;分流比为1：10。结果：基因毒性杂质甲磺酸甲酯、甲磺酸乙酯和甲磺酸异丙酯在限度为20％～120％的范围内线性关系良好,定量限分别为0．0100、0．2500和1．2500μg·L-1．检测限分别为0．0020、0．0100和0．2500μg·L-1,平均回收率分别为99．20％、99．45％和99．98％（n=9）。结论：该方法准确、简便,适用于甲磺酸吉米沙星中基因毒性杂质的检测及限度控制。     

枸橼酸坦度螺酮中间体中潜在基因毒性杂质1，4-二溴丁烷的控制研究

目的 建立枸橼酸坦度螺酮原料药中间体中潜在基因毒性杂质1,4-二溴丁烷的控制检测方法。方法 采用高效气相色谱（GC）法,使用色谱柱Agilent DB-624（30 m×320 μm,1.8 μm）,升温程序：初始柱温95℃,保持2 min,以20℃/min升温至220℃,保持5 min。进样口温度250℃,检测器温度250℃,体积流量3 mL/min;分流比1∶1;进样量1 μL。进行专属性、线性范围、灵敏度、稳定性、重复性、回收率、耐用性方法学考察。结果 1,4-二溴丁烷在0.109 5～1.094 8 μg/mL内峰面积与质量浓度线性关系良好（r=0.998 6）;检测限为0.109 5 μg/mL,定量限为0.273 7 μg/mL,平均回收率为116.2%,方法学考察均符合要求。各批次样品中1,4-二溴丁烷的质量分数均小于0.002 5%。结论 建立的方法专属性强、灵敏度高、测定结果准确、操作简单,可用于枸橼酸坦度螺酮原料药中间体中1,4-二溴丁烷的检测。     

GC法检测布洛芬注射液中杂质F

目的建立气相色谱法测定布洛芬注射液中的杂质F。方法采用气相色谱法,色谱柱：DB-WAXetr毛细管柱（25 m×0.53 mm×2μm）,载气为氦气,流速5.0 mL·min-1,检测器为火焰离子化检测器,检测器温度为250℃,进样温度为200℃。结果在选定的条件下布洛芬和杂质完全分离,杂质F的检测限为2.5 mg·L-1,检测方法的精密度、准确度、灵敏度均达到了预定的分析要求。结论建立的方法准确、灵敏,适用于检测布洛芬注射液中的杂质F。     

GC-MS测定坎地沙坦酯中微量的N-亚硝基二甲胺和N-亚硝基二乙胺

目的：建立GC/MS直接进样法测定坎地沙坦酯中微量的基因毒性杂质N-亚硝基二甲胺（NDMA）和N-亚硝基二乙胺（NDEA）的含量。方法：采用单四极杆GC/MS,色谱柱Agilent VF-WAX ms（长度30m,内径0.25mm,液膜厚度0.25μm）,程序升温,进样口温度200℃,流速1.0 ml?min-1;质谱采用EI源,电压为70eV,离子源温度230℃、四极杆温度150℃,5-7min检测离子为74(m/z) (NDMA),7-10min检测离子为102(m/z) (NDEA);以0.2mol?L-1氢氧化钠溶液溶解样品,以乙酸乙酯进行萃取。结果：在上述GC/MS条件下,NDMA、 NDEA及相邻色谱峰之间分离效果良好,NDMA和 NDEA分别在2.97～148.5 ng?mL-1和3.00～150.20ng?mL-1浓度范围内线性关系良好,定量限分别为0.06ppm、0.03ppm,检出限分别为0.018ppm、0.009ppm;NDMA低、中、高浓度平均回收率分别为86.3%、88.3%、91.4%,RSD分别为0.9%、0.7%、0.4%,NDEA低、中、高浓度平均回收率分别为88.1%、87.9%、91.7%,RSD分别为0.9%、1.1%、0.2%。结论：建立的GC/MS测定坎地沙坦酯中NDMA和NDEA的方法,准确度好,灵敏度高,简便可靠,对仪器污染小,可用于坎地沙坦酯中这两个基因毒性杂质的质量控制。     

气相色谱法测定头孢妥仑匹酯中N,N-二甲基甲酰胺残留量

目的 建立头孢妥仑匹酯原料药中N,N-二甲基甲酰胺残留量的检测方法。 方法 采用气相色谱法，FID检测器，色谱柱为DB-624(30m×0.53mm, 3.0μm)，柱温采用程序升温，初始柱温为110℃，保持5min，再以30℃/min的速率升至200℃，保持5min，进样口温度为230℃，检测器温度250℃，以氮气为载气，流速为3.0mL/min，进样体积1.0μL，分流比2:1。 结果 DMF能够有效分离，在4.44~66.60μg/mL范围内线性关系良好，r=0.9993；检测限为0.777μg/mL，定量限为2.664μg/mL；平均回收率为100.72%(RSD=1.76%, n=9)。结论 方法简便灵敏、结果准确、重复性好，可用于头孢妥仑匹酯中DMF残留量的检测。     

气相色谱法测定麝香草酚中的有机溶剂残留量

目的 建立了麝香草酚中有机残留溶剂异丙醇的气相色谱测定法.方法 采用DB-1毛细管色谱柱(30 m×0.25 mm×1μm),以氮气为载气,FID检测器,进样口温度为250℃,检测器温度为300℃.采用程序升温,柱温:40℃,保持8 min.然后以30℃/min的速率升至200℃,测定残留溶剂异丙醇的含量.结果 线性范...     

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.