LC-MS/MS测定酒石酸溴莫尼定中苯胺类基因毒性杂质

目的 建立LC-MS/MS方法测定酒石酸溴莫尼定中苯胺类基因毒性杂质（6-氨基喹喔啉、4-硝基邻苯二胺、6-氨基-5-溴喹喔啉）的含量。方法 采用色谱柱ZORBAX Eclipse Plus C₁₈（2.1 mm×50 mm，1.8 μm），流动相为0.1%乙酸水溶液（A）-乙腈（B），梯度洗脱，流速0.35 mL·min^-1，柱温为35℃；采用Shimadzu LC30 UPLC-AB Science 4000+三重四极杆液质联用仪和多反应离子监测模式二级质谱MS/MS（ESI）源检测，正离子模式采集数据。结果 6-氨基喹喔啉、4-硝基邻苯二胺、6-氨基-5-溴喹喔啉分别在9.98~199.50，9.94~198.74，10.03~200.55 ng·mL^-1内线性关系良好；低、中、高浓度（50，100，150 ng·mL^-1）回收率为94.8%~110.8%，RSD均<4.2%；溶液在12 h内稳定性良好。结论 本方法操作简便，结果可靠，可以用于酒石酸溴莫尼定中苯胺类基因毒性杂质（6-氨基喹喔啉、4-硝基邻苯二胺、6-氨基-5-溴喹喔啉）的控制。     

HPLC测定奥美沙坦酯中的基因毒性杂质

目的 建立HPLC测定奥美沙坦酯中潜在的基因毒性杂质[杂质1 ：N-（三苯基甲基）-5-（4''-溴甲基联苯-2-基）四氮唑,杂质2 ：N-三苯甲基-5-（4'',4''-二溴甲基联苯-2-基）四氮唑]的含量和限度。方法 采用Phenomenex C₁₈柱（250 mm×4.6 mm,5 μm）;流动相：0.1%冰乙酸水溶液-0.1%冰乙酸乙腈溶液（15：85）;检测波长：254 nm;流速：1.5 mL·min^-1;柱温：25℃。结果 杂质1 和杂质2 均在0.030 97~0.247 7 μg·mL^-1内线性良好（r分别为0.999 6和0.998 7）,平均回收率分别为94.37%和94.43%,RSD分别为2.38%和2.72%（n=9）。结论 该方法专属性强,准确、灵敏,可以作为奥美沙坦酯中基因毒性杂质1 和杂质2 的液相分析方法。     

气相色谱法测定异环磷酰胺原料药中3种有关物质

目的 建立气相色谱（GC）法测定异环磷酰胺原料药中3种有关物质（起始物料3-氨基丙醇、杂质D和杂质F）。方法 采用HP-55% Phenyl Methyl Siloxane色谱柱（30 m×320 μm，0.25 μm）；采用程序升温的方式，起始柱温50℃，保持2 min后以每分钟5℃的速率升温至180℃，再以每分钟20℃的速率升温至260℃；进样口温度为380℃；检测器温度为380℃；分流比5∶1。结果 起始物料峰、杂质D、杂质F与周围峰分离度分别为2.6、3.0、1.6；异环磷酰胺起始物料3-氨基丙醇在39.80～199.0 μg·mL^-1线性关系良好（r=0.999 1），杂质D在39.48～197.4 μg·mL^-1线性关系良好（r=0.999 1），杂质F在2.000～20.000 μg·mL^-1线性关系良好（r=0.999 9）； 3-氨基丙醇和杂质D的检测限分别为9.95、9.87 μg·mL^-1，杂质F因色谱柱柱效降低且进样量过大未能进行检测限测定；3-氨基丙醇、杂质D、杂质F的定量限分别为19.90、19.74、2.000 μg·mL^-1；起始物料、杂质D和杂质F回收率均在90.09%～115.90%，回收率RSD分别为4.8%、7.2%、2.3%。结论 该方法的专属性、精密度、线性关系良好，准确可靠，符合异环磷酰胺中3种有关物质的检测要求。     

HPLC测定奥格列汀中基因毒性杂质残留量

目的 建立奥格列汀原料药中基因毒性杂质的HPLC测定方法。方法 采用Zorbax SB-C₁₈色谱柱（250 mm×4.6 mm,5μm）,进样量：20 μl;流动相为0.1%乙酸水-乙腈（65：35）;流速为1mL·min^-1;紫外检测器,检测波长为220 nm;柱温为25℃,色谱乙腈为溶剂。结果 该方法专属性良好,测得苯磺酸异丙酯在4~60 μg·mL^-1内线性关系良好,平均回收率为98.56%（n=9,RSD=3.78%）,溶液在8 h内稳定。结论 该法操作简便,重复性好,结果准确可靠,可用于奥格列汀原料药中基因毒性杂质的测定。     

UHPLC-HRMS定量检测莫西沙星中5种亚硝胺类化合物

目的 基于超高效液相色谱和高分辨质谱建立同时检测莫西沙星中5种亚硝胺类基因毒性杂质的定量分析方法。方法 采用ACE Excel 1.7 C₁₈-PFP(100 mm×2.1 mm，1.7 μm)色谱柱，0.1%甲酸水-甲醇为流动相，0.3 mL·min^-1的流速进行梯度洗脱。采用四极杆高分辨静电场轨道阱质谱在电喷雾正离子条件下运用平行反应监测模式对5种亚硝胺类化合物进行定量检测。结果 5种亚硝胺类基因毒性杂质的定量限均≤0.5 ng·mL^-1。NPYP、NEMA和NDEA的标准曲线在1.0~100 ng·mL^-1(NPIP和NDBA为0.1~100 ng·mL^-1)呈良好线性关系(R²≥0.998)。该方法在盐酸莫西沙星药物中的回收率为93.1%~114.1%，RSD为0.7%~4.6%。结论 本方法结果准确、灵敏度高、重复性好，适用于盐酸莫西沙星中亚硝胺类基因毒性杂质的含量检测。     

依帕司他原料药有关物质的HPLC测定方法改进研究

目的 采用改进的高效液相色谱法测定依帕司他原料药有关物质。方法 采用依利特Hypersil BDS C₁₈色谱柱（250 mm×4.6 mm,5 μm）,以乙腈-pH6.5磷酸盐缓冲液（0.025 mol·L^-1 KH₂PO₄+0.025 mol·L^-1 Na₂HPO₄,用H₃PO₄调节pH至6.5）为流动相,梯度洗脱,流速为1.0 mL·min^-1,检测波长为396 nm和280 nm,柱温为30℃,进样量为20 μL。结果 依帕司他与相邻杂质能较好地分离。立体异构体杂质在浓度0.010~30 μg·mL^-1内线性关系良好,检测限和定量限分别为0.80,2.4 ng·mL^-1。精密度、稳定性、准确度、耐用性均符合要求。结论 该方法可用于依帕司他原料药有关物质的检查。     

HPLC法测定阿奇霉素颗粒中的基因毒性杂质对甲苯磺酸乙酯

目的 建立阿奇霉素颗粒中基因毒性杂质对甲苯磺酸乙酯的高效液相色谱（HPLC）测定方法。方法 采用Waters XBridge C₁₈（250 mm×4.6 mm,5 μm）色谱柱,以磷酸盐缓冲液（2.2 g·L^-1无水磷酸氢二钾溶液,用稀磷酸调节pH值至8.9）为流动相A;甲醇-乙腈（25∶75）为流动相B,进行梯度洗脱,体积流量为1.0 mL·min^-1,检测波长为225 nm,柱温为60℃,进样量为50 μL。进行专属性、线性关系、定量限和检测限、精密度、重复性、回收率、稳定性、耐用性试验。结果 空白溶剂、空白辅料、混合溶液中邻近对甲苯磺酸乙酯的杂质L均不干扰对甲苯磺酸乙酯检测;对甲苯磺酸乙酯质量浓度在0.05～0.20 μg·mL^-1（R²＝0.999 9）与峰面积呈良好线性关系;检测限为0.016 7 μg·mL^-1,定量限为0.050 0 μg·mL^-1;平均回收率为95.76%～96.48%,RSD为0.42%～1.53%;精密度、重复性、稳定性、耐用性均符合要求。3批样品中均未检出对甲苯磺酸乙酯。结论 建立的方法专属性强、灵敏度高、特异性强、准确度高,可用于阿奇霉素颗粒中基因毒性杂质对甲苯磺酸乙酯的限度控制与检测。     

LC-MS/MS法测定瑞戈非尼中4-(4-氨基-3-氟苯氧基)-N-甲基吡啶-2-甲酰胺

摘 要 目的：建立抗癌新药瑞戈非尼中遗传毒性杂质4-(4-氨基-3-氟苯氧基)-N-甲基吡啶-2-甲酰胺（AFP-PMA）的含量测定方法。方法: 采用LC MS/MS法测定;利用Waters XBridge Shield RP18色谱柱（150 mm×3.0 mm,3.5 μm）进行分离,以5 mmol·L^-1乙酸铵溶液（A）-乙腈（B）为流动相,采用梯度洗脱（0~9 min,5% B→90% B）,流速为1.0 ml·min^-1,柱温50 ℃;ESI离子源,正离子扫描方式,多反应监测（MRM）模式扫描,检测离子通道为m/z262.2→244.1。结果: AFP-PMA 浓度在2.41～980.90 ng·ml^-1范围内与峰面积呈良好线性关系（r=0.999 8）;定量限为8.02 ng·ml^-1;检出限为2.41 ng·ml^-1,约为供试品溶液浓度的0.000 241%;平均加样回收率为100.95%,RSD为2.37%（n=9）。结论:本方法专属性强、灵敏度高、准确度好,可用于瑞戈非尼中4-(4-氨基3-氟苯氧基)N-甲基吡啶-2甲酰胺的含量测定。     

HPLC测定赛利司他有关物质及降解产物

目的 建立HPLC测定赛利司他有关物质的方法。方法 采用Agilent Phenyl色谱柱（250 mm×4.6 mm,5 μm）,以5 mmol·L^-1磷酸二氢钾（用10%磷酸调pH值至4.0）-乙腈为流动相进行梯度洗脱,流速为1.0 mL·min^-1,检测波长为226 nm。结果 各杂质与主峰之间的分离度良好。开环杂质（杂质A）、2-十七烷基氧-6-甲基-4氢-3,1-苯并噁嗪-4-酮（杂质B）、2-十八烷基氧-6-甲基-4氢-3,1-苯并噁嗪-4-酮（杂质C）浓度分别在0.049 5~1.981 μg·mL^-1,0.059 9~0.399 1 μg·mL^-1,0.059 6~0.397 5 μg·mL^-1内与峰面积呈良好的线性关系,r均为0.999 9;杂质A、B、C加样回收率的平均值分别为104.2%,107.0%和107.8%,RSD分别为2.09%,1.77%和2.18%。结论 本方法简便、准确可靠,适用于赛利司他中有关物质的控制。     

HPLC测定尼美舒利颗粒中的有关物质

摘 要 目的： 建立尼美舒利颗粒有关物质的HPLC测定方法。 方法: 采用Agilent HC C₁₈（250 mm×4.6 mm,5 μm）色谱柱;流动相：乙腈 0.01 mol·L^-1磷酸二氢铵溶液（氨水调pH至7.0）（4060）;检测波长：230 nm;流速：1.0 ml·min^-1;柱温：30℃;进样量：20 μl。结果: 尼美舒利在0.10～0.30 mg·mL^-1（r=0.999 9）浓度范围内线性关系良好,各杂质峰与主峰分离度均大于2.0,检测限为0.3 ng,定量限为1.0 ng。结论:本方法专属性强,准确度高,重现性良好,可用于尼美舒利颗粒的质量控制。     

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.

---

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.     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.     

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.