卡泊芬净及其杂质的LC-QTOF-MS定性分析

目的 采用LC-QTOF-MS分析醋酸卡泊芬净原料药及其强降解产物的杂质，研究卡泊芬净及其相关杂质(杂质A、B、C、D和E)的质谱特征。方法 采用Waters CORTECS^® C₁₈⁺(4.6 mm×150 mm，2.7 μm)色谱柱，流动相A为0.1%甲酸-水溶液，流动相B为0.1%甲酸-乙腈，流速为0.6 mL·min^-1，梯度洗脱；ESI-QTOF-MS正离子扫描检测。结果 在一级质谱中，除杂质D主要显示单电荷准分子离子峰外，卡泊芬净及其余4个杂质均以多电荷准分子离子峰的响应较高；在二级质谱中，卡泊芬净及其他含乙二胺结构的杂质均会丢失乙二胺及其所连基团产生碎片离子m/z 103 3；卡泊芬净及其相关杂质主要通过肽键的断裂生成一系列碎片离子，也可以通过丢失氨基酸残基中的羟基、酰基或氨基生成碎片离子；杂质A和杂质C分别显示高丰度的特征碎片离子m/z 137.070 8和m/z 77.071 1，可以与卡泊芬净及其他杂质区分。结论 卡泊芬净及其5个杂质均具有明显的质谱特征，研究结果可为卡泊芬净生产过程中可能出现的未知杂质的结构鉴定提供参考，以便及早发现生产工艺中的潜在问题，降低产品质量风险。     

液质联用技术用于醋酸阿托西班注射液中杂质结构鉴定

目的:采用高效液相色谱(HPLC)、超高效液相色谱-四极杆串联静电场轨道阱高分辨质谱(UPLC-Q Orbitrap MS)技术对醋酸阿托西班注射液中杂质结构进行鉴定。方法:HPLC法采用Inertsil ODS-2 C₁₈色谱柱(250 mm×4.6 mm, 5μm),流动相A为乙腈-甲醇-三氟乙酸溶液(pH 3.2)(15∶10∶75),流动相B为乙腈-甲醇(60∶40),梯度洗脱，流速1.2 mL·min^-1,检测波长220 nm。UPLC-Q Orbitrap MS法采用BEH300 C₁₈色谱柱(150 mm×2.1 mm, 1.7μm),流动相为0.1%甲酸水溶液(A)-0.1%甲酸乙腈溶液(B),梯度洗脱，流速0.2 mL·min^-1;采用电喷雾离子源，选择正离子模式进行Full MS/dd-MS²扫描。结果:对5家企业各1批醋酸阿托西班注射液进行了有关物质测定，以面积归一化计算，含量>0.1%的杂质峰个数分别为9、10、13、9和6,总杂含量分别为0.35...     

UPLC-MS/MS测定酒石酸伐尼克兰中基因毒性杂质N-亚硝基伐尼克兰

目的 建立酒石酸伐尼克兰原料药和片剂中基因毒性杂质N-亚硝基伐尼克兰超高效液相色谱-串联三重四级杆质谱(UPLC-MS/MS)的检测方法。方法 采用ACQUITY UPLC^? CSH^TM Phenyl-Hexyl(150 mm×3.0 mm,1.7μm)色谱柱;0.1%甲酸水溶液为流动相A,0.1%甲酸的甲醇溶液为流动相B,梯度洗脱;流速为0.45 mL·min^–1,柱温为50℃;采用ESI离子源正离子扫描,多反应监测(MRM)模式下,对基因毒性杂质进行定量检测。结果 杂质在0.10～10.04ng·mL^–1具有良好的线性关系;原料药的低、中、高3个浓度的加样回收率(n=3)分别为103.58%(RSD=3.30%),98.65%(RSD=2.73%),92.00%(RSD=1.98%);片剂的低、中、高3个浓度的加样回收率(n=3)为91.53%(RSD=0.78%),96.76%(RSD=3.12%),93.01%(RSD=2.21%);检测限与定量限分别为0.014 ng·mL^–1<...     

UHPLC-MS/MS法测定盐酸普萘洛尔缓释片中基因毒性杂质N-亚硝基普萘洛尔

目的 建立盐酸普萘洛尔缓释片中基因毒性杂质N-亚硝基普萘洛尔的超高效液相色谱-串联质谱(UHPLC-MS/MS)检测方法。方法 Waters ACQUITY UPLC CSH^TM C₁₈色谱柱(3.0 mm×150 mm, 1.7μm),10 mmol·L^-1甲酸铵的水溶液(含0.1%甲酸)作为流动相A,乙腈溶液(含0.1%甲酸)作为流动相B,梯度洗脱,流速为0.5 mL·min^-1,柱温为50℃,进样器温度为5℃,进样体积为10μL,采用多反应监测(MRM)模式,对盐酸普萘洛尔缓释片中的N-亚硝基普萘洛尔进行定量检测。结果 N-亚硝基普萘洛尔在1～20 ng·mL^-1范围内具有良好的线性关系。低、中、高3个浓度的加样回收率(n=3)在98.4%～103.2%之间,RSD≤2.7%。检测限和定量限分别为0.09 ng·mL^-1和0.3 ng·mL^-1。检出盐酸普萘洛尔缓释片中基因毒性杂质N-亚硝基普萘洛尔含量为1.8μg·g^...     

UPLC测定注射用醋酸西曲瑞克中有关物质的含量

目的建立UPLC测定注射用醋酸西曲瑞克的有关物质含量的方法。方法采用色谱柱ACQUITY UPLC CSHTM C₁₈ (2.1 mm×150 mm,1.7μm),流动相A为0.05 mol·L^-1高氯酸钠溶液(磷酸调节pH值至2.0),流动相B为0.05 mol·L^-1高氯酸钠溶液-乙腈(30∶70)(磷酸调节pH值至2.0),梯度洗脱,流速为0.2 mL·min^-1,检测波长为226 nm,进样量为1μL。结果杂质A、B、C、D、E、F的检测浓度范围分别为0.127 8～6.390 4,0.124 4～6.221 3,0.126 8～6.340 8,0.120 5～6.025 6,0.120 9～6.047 3,0.127 7～6.389 4μg·mL^-1,各杂质检出限约为0.04 ng,杂质B、D、E定量限约为0.12 ng,杂质A、C、F定量限约为0.13 ng,平均加样回收率分别为97.4%,95.2%,101.5%,105.3%,100.7%,97.4%,RSD分别...     

高效液相色谱-电喷雾检测器测定熊去氧胆酸的有关物质

目的:建立高效液相色谱-电喷雾检测器法测定熊去氧胆酸中的10个有关物质。方法:采用SHISEIDO Capcell PAK C₁₈ MGII(150 mm×4.6 mm, 5μm)色谱柱,以0.1%甲酸-甲醇-乙腈(30∶45∶25)为流动相A,乙腈为流动相B,梯度洗脱,流速1.0 mL·min^-1,柱温40℃,检测器温度35℃,采集频率5 Hz,过滤常数3.6 s。采用主成分自身对照法计算已知杂质和其它杂质的含量,并对建立的方法进行方法学验证。结果:熊去氧胆酸与各杂质分离度良好;熊去氧胆酸、杂质A、杂质B、杂质C、杂质E的质量浓度均在1.0～100.0μg·mL^-1范围内与峰面积呈良好的线性关系,定量限均为1.0μg·mL^-1,检测限均为0.5μg·mL^-1,杂质的平均回收率在99.3%～100.1%,RSD(n=9)不高于2.0%;供试品溶液在10℃条件下放置24 h内稳定;微调液相色谱参数后,对有关物质的检测结果无影响。3批样品有关物质结果显示,杂质A的含量均小于1.0%...     

UHPLC-MS/MS法测定盐酸二甲双胍制剂中2种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(UHPLC-MS/MS)法同时测定盐酸二甲双胍制剂中2种基因毒性杂质N-亚硝基二甲胺(N-nitrosodimethylaminen, NDMA)和N-亚硝基二乙胺(N-nitrosodiethylamine, NDEA)的含量的方法。方法 采用多反应监测(MRM)模式检测，离子源为APCI(+),色谱柱为ACE Excel 3 C₁₈-AR柱(100 mm×4.6 mm, 3μm),以体积分数0.1%甲酸水溶液(A)-质量分数0.1%甲酸甲醇溶液(B)为流动相梯度洗脱，流速0.5 mL·min^-1,进样体积5μL。结果 NDMA和NDEA的线性范围分别为0.99～99μg·L^-1(r>0.999 9)和0.53～53μg·L^-1(r>0.999 9),平均回收率为90.9%～98.9%,定量限分别为0.99和0.53μg·L^-1,检测限分别为0.3和0.2μg·L^-1。用建立的方法测定不同剂型的6批样品，2批...     

UPLC-MS/MS测定硫酸庆大霉素原料及滴眼液中的7种基因毒性杂质

目的 采用UPLC-MS/MS快速测定庆大霉素原料及滴眼液中7种基因毒性杂质的含量。方法 采用ODS-3色谱柱(100 mm×3.0 mm, 5μm),流动相A为含0.1%甲酸的甲醇溶液，流动相B为含0.1%甲酸的水溶液，梯度洗脱，流速0.3 mL·min^-1,柱温35℃,正离子(ESI+)模式下单离子检测扫描模式检测。结果 7种基因毒性杂质在实验浓度范围内线性关系良好，回收率结果准确可靠，27批样品中均未检出7种基因毒性杂质。结论 所用方法简便快捷，准确可靠，适合快速检测基因毒性杂质。     

用LC-MS/MS法同时测定大鼠血浆中多黏菌素B和替加环素的浓度

目的 建立大鼠血浆中多黏菌素B和替加环素测定的液相色谱串联质谱(LC-MS/MS)方法,并研究其在大鼠体内的药代动力学特征。方法 用3%三氯乙酸-甲醇溶液(50∶50)对大鼠血浆进行沉淀蛋白处理。色谱柱：Symmetry C₁₈(150.0 mm×4.6 mm, 3.5μm),流动相：0.1%甲酸水-0.1%甲酸乙腈,流速：0.6 mL·min^-1,柱温：40℃;离子源：电喷雾离子源,正离子检测模式,多反应检测。考察该方法的专属性、标准曲线和定量下限、精密度与回收率、稳定性以及重现性。结果 替加环素的线性范围为25～2 500 ng·mL^-1,定量下限为25 ng·mL^-1,提取回收率为95.89%～107.90%;多黏菌素B_1+1-I的线性范围为82～8 200 ng·mL^-1,定量下限为82 ng·mL^-1,提取回收率为93.84%～97.70%;多黏菌素B₂的线性范围为9～900 ng·mL^-1<...     

UHPLC-MS/MS法测定依那普利氢氯噻嗪复方制剂中潜在基因毒性杂质

目的 通过合成基因毒性杂质N-亚硝基氢氯噻嗪(NO-HZCT),建立超高效液相色谱-串联质谱法(UHPLC-MS/MS)测定依那普利氢氯噻嗪制剂中N-亚硝基氢氯噻嗪。方法 参考文献方法合成NO-HZCT,采用高分辨质谱对其相对分子量和结构进行确定;采用Agilent Eclipse Plus C₁₈ RRHD(3.0 mm×150 mm, 1.8μm)色谱柱,以10 mmol·L^-1甲酸铵-0.1%甲酸的水溶液作为流动相A,以0.1%甲酸的乙腈溶液作为流动相B,梯度洗脱,体积流量0.6 mL·min^-1;采用ESI离子源正离子扫描,多反应监测(MRM)模式下,对NO-HZCT进行定量检测。结果 NO-HZCT质量浓度在0.51～50.67 ng·mL^-1范围内具有良好的线性关系,相关系数(r)为0.999 7;低、中、高3个浓度的加样回收率(n=3)分别为93.10%(RSD 3.7%)、104.30%(RSD 1.0%)和106.48%(RSD 1.8%);检测限和定量限分别为0.08 ng·mL...     

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.     

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.     

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.