基于UPLC-MS/MS的特征多肽识别技术鉴别中成药中的阿胶

目的 建立基于超高效液相色谱-三重四级杆质谱(UPLC-MS/MS)的特征多肽识别技术检测中成药中阿胶成分的专属鉴别方法.方法 将中成药供试品进行提取及胰蛋白酶酶解,以阿胶特有的特征多肽离子对m/z 539.8(双电荷)→612.4和m/z 539.8(双电荷)→923.8作为鉴别离子对,采用UPLC-MS/MS液质联...     

特征肽技术用于龟甲(浙龟甲)中掺巴西龟的检查研究

目的 找出巴西龟特征肽段,建立龟甲（浙龟甲）中掺入巴西龟的专属性检查方法。方法 样品以水提法提取,胰蛋白酶酶解,利用超高效液相色谱-四级杆飞行时间质谱和数据处理软件查找巴西龟特征肽,并利用蛋白质搜库技术初步测序;利用找到的特征肽段,建立超高效液相色谱-串联四级杆质谱的多反应监测的巴西龟掺伪专属性检查方法。结果 找出巴西龟特征性肽段m/z 400.23（双电荷）,并初步测定序列,通过分析该特征肽段二级质谱图,确定了专属性检测离子对m/z 400.23（双电荷）→374.05,142.90,并建立了龟甲（浙龟甲）中掺入巴西龟的专属性检查方法。20批样品中,11批样品检出巴西龟成分。结论 该方法的专属性强,符合分析检测的方法技术要求,可用于龟甲（浙龟甲）掺入巴西龟的检查。     

超高效液相色谱-三重四极杆质谱法检测孕康颗粒中阿胶的特征肽及掺杂猪皮源成分

摘要：目的:建立孕康颗粒中阿胶的特征肽成分及掺杂猪皮源成分的专属性检测方法。方法:采用胰蛋白酶对孕康颗粒进行酶解,利用超高效液相色谱-三重四极杆质谱（UPLC-QQQ MS）对阿胶的专属性特征肽段及猪皮源成分进行检测。色谱柱:Agilent Zorbax Eclipse-C18柱（2.1 mm×100 mm,1.5μm）;流动相:乙腈（A）-0.1%甲酸溶液（B）,梯度洗脱;流速:0.3 ml·min-1;柱温:40℃。选择阿胶特征分子离子峰m/z 539.8（双电荷）→612.4,923.8及猪皮源特征离子峰m/z 774.5（双电荷）→977.8,1 034.6作为检测离子对,离子化模式为ESI+,进行多反应监测。结果:8批市售孕康颗粒样品中均可检出阿胶的特征肽段,均未检出猪皮源成分。结论:建立的方法专属性强、准确可靠,可用于孕康颗粒中阿胶特征肽成分及掺杂猪皮源成分的检测。     

超高效液相色谱-质谱法检测妇宁颗粒中阿胶及牛皮源成分

摘要：目的:建立妇宁颗粒中阿胶及掺杂牛皮源成分的专属性检查方法。方法:利用胰蛋白酶对妇宁颗粒中所含的阿胶进行酶解,采用超高效液相色谱-质谱,以阿胶特征分子离子峰m/z 539.8（双电荷）→612.4、m/z 539.8（双电荷）→923.8及牛皮源特征离子对m/z 641.3 （双电荷）→726.2、m/z 641.3 （双电荷）→783.3作为检测离子对,离子化模式为ESI+,以多反应监测模式检测。结果:6批妇宁颗粒样品均检出了阿胶,且均未检出牛皮源成分。结论:该检测方法准确、可靠、专属性强,可用于妇宁颗粒中阿胶及牛皮源成分的检测。     

超高效液相色谱-三重四极杆质谱法检测复方阿胶浆中阿胶

目的:建立复方阿胶浆中阿胶的专属性检测方法。方法:采用胰蛋白酶对复方阿胶浆中阿胶成分进行酶解,利用超高效液相色谱-三重四极杆质谱(RRLC-QQQ/MS)对阿胶的专属性特征分子离子峰进行检测。采用Agilent SB-C18(2.1 mm×100 mm,1.8μm)色谱柱,以0.1%甲酸水溶液为流动相A,以乙腈为流动相B,梯度洗脱(0~25 min,95%A→80%A,5%B→20%B;25~40 min,80%A→50%A,20%B→50%B)。流速0.3 m L·min-1,柱温40℃,进样量5μL;选择阿胶特征分子离子峰m/z 539.8(双电荷)→612.4和m/z 539.8(双电荷)→923.8作为检测离子对,离子化模式为ESI+,进行多反应监测。结果:3批市售样品中均可检出阿胶的特征分子离子峰,即同时检出m/z 539.8(双电荷)→612.4和m/z 539.8(双电荷)→923.8离子对。结论:所建立的方法经方法学验证,阴性样品及其他胶类如黄明胶、龟甲胶和鹿角胶等对复方阿胶浆样品测定无干扰,样品检出浓度为含5%阿胶的复方阿胶浆样品。方法专属性强,可用于复方阿胶浆中阿胶的检测。     

超高效液相色谱-质谱法同时检测妇科止带胶囊中阿胶、龟甲胶成分

摘要：目的:建立妇科止带胶囊中阿胶、龟甲胶成分的专属性检测方法。方法:利用胰蛋白酶对妇科止带胶囊中阿胶、龟甲胶进行酶解,采用超高效液相色谱-质谱,以阿胶特征分子离子峰m/z 539.8 （双电荷）→612.4、m/z 539.8 （双电荷）→923.8及龟甲胶特征分子离子峰m/z 631.3（双电荷）→546.4、m/z 631.3（双电荷）→921.4作为检测离子对,离子化模式为ESI+,进行多反应监测。结果:10批妇科止带胶囊样品均检出了阿胶及龟甲胶成分。结论:建立的检查方法准确、可靠、专属性强,可用于妇科止带胶囊中阿胶及龟甲胶成分的检测。     

高效液相色谱-质谱联用技术测定妇科止带片中阿胶

目的 建立妇科止带片中阿胶的检测方法.方法 利用超高效液相色谱-三重四极杆串联质谱(UPLC-QQQ/MS)对妇科止带片中阿胶的特征分子离子峰进行检测,选择阿胶特征分子离子峰m/z 539.8(双电荷)→612.4和m/z 539.8(双电荷)→923.8作为检测离子对,离子化模式为ESI+,进行多反应监测.结果 16批市售样品中均可检出阿胶的特征分子离子峰,即同时检出m/z 539.8(双电荷)→612.4和m/z 539.8(双电荷)→923.8离子对.结论 所建立的方法经方法学验证,该方法专属性强,可用于妇科止带片中阿胶的检测,为妇科止带片的质量控制提供参考.     

固相萃取-液质联用法用于乌鸡白凤丸中动物药的质量控制初探

摘要：目的:建立乌鸡白凤丸中鹿角胶、鹿角霜和鳖甲的鉴别和掺伪检查方法。方法:采用胰蛋白酶对乌鸡白凤丸样品酶解处理,利用固相萃取-超高效液相色谱-三重四极杆质谱（SPE-UPLC-QQQ）对样品中的鹿、鳖、猪、牛、马、驴、角鳖、佛罗里达鳖源性成分的专属性特征离子进行检测。结果:方法的专属性、精密度和灵敏度均符合分析检测的技术要求。12批样品检测到了处方中规定的鹿、鳖源性成分,部分样品也检出了处方中未规定的猪、牛源性成分。结论:所建立的方法经方法学验证,专属性强,为乌鸡白凤丸的质量控制及标准研究提供了参考。     

超高效液相色谱-三重四级杆质谱法检测阿归养血糖浆中阿胶及牛皮源成分

目的：建立阿归养血糖浆中阿胶及掺杂牛皮源成分的专属性检测方法。方法：用胰蛋白酶对阿归养血糖浆处方中胶类成分进行酶解，采用超高效液相色谱-三重四极杆质谱（UPLC-QQQ/MS），选择阿胶特征分子离子峰m/z 540.0（双电荷）→612.2、m/z 540.0（双电荷）→924.4及牛皮源特征离子对m/z 641.3（双电荷）→726.2、m/z 641.3（双电荷）→783.3作为检测离子对，离子化模式为ESI⁺，进行多反应监测。结果：26批阿归养血糖浆中未检出阿胶的有13批，其中3批检出牛皮源成分，1批检出牛皮源成分但未超出限度，9批均未检出阿胶及牛皮源成分；26批样品检出阿胶的有13批，其中同时检出牛皮源成分的有1批，未检出牛皮源成分的为12批。结论：该检测方法专属性强，可用于阿归养血糖浆中阿胶及牛皮源的检测。     

UPLC-QTOF-MS结合主成分分析法用于龟甲胶、鹿角胶中添加牛皮源成分的检测研究

目的:建立龟甲胶、鹿角胶中非法掺入牛皮源成分的检测方法。方法:采用胰蛋白酶对龟甲胶、鹿角胶、牛皮源成分进行酶解,利用超高效液相色谱-四极杆飞行时间质谱(UPLC-QTOF-MS)进行测定,采用Markerlynx软件对3种胶类的液质数据进行主成分分析,找出鉴别牛皮源成分的专属性特征肽段,并对特征肽的序列进行确认。结果:对19批样品进行检测发现,其中8批龟甲胶及8批鹿角胶中可检出牛皮源成分的特征肽段。结论:所建立的方法经验证,专属性较强,可用于龟甲胶、鹿角胶中非法掺入牛皮源成分的检测。     

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.     

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.