流动注射抑制化学发光法测定痕量儿茶酚

目的:建立快速测定痕量儿茶酚的新方法。方法:实验发现在碱性介质中,儿茶酚能强烈抑制luminol与K_3Fe(CN)_6的化学发光反应。本文以儿茶酚在一定浓度范围内与luminol-K_3Fe(CN)_6化学发光强度降低值的呈线性关系为基础,采用流动注射技术,测定了痕量的儿茶酚。结果:测定的线性范围0.1 ng·mL~(-1)～0.1 μg·mL~(-1),检出限为40pg·mL~(-1)(3σ,3倍的标准偏差),RSD小于3.5％(n=9)。本法成功地用于对合儿茶酚水样和人体尿样的测定。结论:该法简便快速,灵敏度高,选择性好。     

高效液相色谱法测定血浆和组织中纳曲酮浓度

目的:建立紫外检测高效液相色谱方法测定动物样本中纳曲酮浓度。方法:采用YWG-C18色谱柱（4.6mm×150mm,5μm）,UV1000紫外检测器,甲醇-乙腈-磷酸盐缓冲液（pH4.0）（15:10:75）为流动相,流速:1.0mL·min-1;以乙酰苯胺为内标,样本经有机相提取纯化后进样,检侧波长230nm。结果:测定血浆中药物浓度时的线性范围 5～40ng·mL-1,回收率91.4％～94.5％,日内误差（RSD）<12％,日间误差（RSD）<7.5％;肝组织测定时线性范围5～200ng·mL-1,回收率78.8％～82.3％,日内误差（RSD）<17％,日间误差（RSD）<4.5％;肾组织测定时线性范围5～200ng·mL-1,回收率79.4％～83.4％,日内误差（RSD）<8.1％,日间误差（HSD）<8.2％。结论:本方法快速、准确、重复性好,可以满足临床和动物试验样品测定的要求。     

反相高效液相色谱法测定人体血浆中扑尔敏的浓度

目的:建立一种测定人体血浆中扑尔敏浓度的高效液相色谱法。方法:取人体血浆1.5 mL,加内标右美沙芬后,用三氯甲烷提取,取其有机相于60℃水浴挥干,剩余物加流动相复溶,并用乙酸乙酯洗涤,取水相20μL进样。流动相为0.025mol·L~(-1)磷酸二氢铵(用磷酸调pH为3.5)-乙腈(70:30),色谱柱为Diamonsil ODS C_(18)柱(5μm,4.6mm×150mm),检测波长为200 nm,流速为1.0mL·min~(-1),柱温为室温。结果:本方法线性范围为0.5～16ng·mL~(-1),,r=0.998 7,最低检测限为0.25ng·mL~(-1),方法回收率为96.6％～108.0％,日内RSD为4.1％～5.9％,日间RSD为4.6％～5.9％。结论:本方法灵敏度高,特异性强,重现性好,可用于人体血浆中扑尔敏浓度的测定。     

UPLC-MS/MS法同时测定如意金黄散中12种化学成分的含量

摘要：目的:建立超高效液相色谱-串联质谱（UPLC-MS/MS）法同时测定中成药如意金黄散中12种化学成分的含量。方法:采用C18色谱柱（100 mm×2.1 mm, 1.7μm）,流动相A为水（含0.1%甲酸）,B为甲醇（含0.1%甲酸）,梯度洗脱,流速为0.3 ml·min-1,柱温为45℃;采用电喷雾离子源（ESI）,多反应监测（MRM）模式,正负离子检测。结果:大黄素、大黄酸、大黄酚、大黄素甲醚、芦荟大黄素、姜黄素、小檗碱、黄柏碱、厚朴酚、橙皮苷、甘草苷和甘草酸的线性范围分别为8.64～864 ng·ml-1、11.02～1 102 ng·ml-1、10.68～1 068 ng·ml-1、10.78～1 078 ng·ml-1、11.12～1 112 ng·ml-1、12.42～1 242 ng·ml-1、11.88～1 188 ng·ml-1、9.47～947 ng·ml-1、9.32～932 ng·ml-1、13.50～1 350 ng·ml-1、11.72～1 172 ng·ml-1和10.52～1 052 ng·ml-1,各成分在线性范围内具有良好的线性关系（r>0.99）;加样回收率均在96.5%～104.3%范围内,RSD小于4.4%（n=9）。3批如意金黄散中大黄素、大黄酸、大黄酚、大黄素甲醚、芦荟大黄素、姜黄素、小檗碱、黄柏碱、厚朴酚、橙皮苷、甘草酸和甘草苷的含量测定均值分别为0.763 0,0.466 0,0.875 0,0.329 0,0.234 0,1.814 0,4.396 0,1.700 0,3.516 0,2.085 0,0.763 0,0.948 0 mg·g-1。结论:研究结果表明,该方法简单、快速、灵敏、准确,可以用于如意金黄散中多种成分的含量测定。     

HPLC-ESI^＋-MS测定人血浆中氯氮平、去甲氯氮平及N—氧化氯氮平

目的:建立HPLC-MS法同时测定氯氮平及其代谢产物去甲氯氮平、N-氧化氯氮平血药浓度。方法:0.5 mL血浆用C18固相萃取小柱进行萃取。色谱柱为C18柱(5μm,3.9 mm×150 mm),柱温40℃;流动相为乙腈-水(36:64,含10 mmol·L-1醋酸铵与0.01％甲酸,pH=5.96),流速0.95 mL·min-1,进入MS检测器的分流比3:1。质谱条件采用电喷雾电离源(ESI+),电离毛细管电压3.90 kV,第一级锥孔电压38 V,用选择离子检测,定量分析检测的离子分别为m/z 326.9(氯氮平)、312.6(去甲氯氮平)、342.6(N-氧化氯氮平)。结果:氯氮平,去甲氯氮平、N-氧化氯氮平最低检测限分别为0.2,0.1,3 ng·mL-1;线性范围分别为20～2000 ng·mL-1(r=0.9994),10～1600 ng·mL-1(r=0.9990),10～800 ng·mL-1(r=0.9969)。三者的萃取回收率大于75％,方法回收率均大于95.0％。日内、日间变异(RSD)均小于10％。结论:该方法灵敏度高、快速,简便,无杂质干扰,适合于氯氮平、去甲氯氮平、N-氧化氯氮平血药浓度监测及药代动力学研究。     

流动注射化学发光法测定阿莫西林的研究

目的:建立快速测定痕量阿莫西林的新方法。方法:实验发现,在盐酸介质中,高锰酸钾能氧化阿莫西林产生较强的化学发光,据此,采用流动注射技术,建立了一种测定痕量阿莫西林的化学发光分析法。结果:该法的线性范围为1.0×1017 -5.0×10-5g·mL-1;检出限为6.0×10-8 g·mL-1;对1.0×10-5g·mL-1的阿莫西林进行连续11次平行测定,相对标准偏差为1.56％。结论:本方法快捷、简便且具有很高的灵敏度,可用于阿莫西林胶囊中阿莫西林含量的测定,并与药典规定的方法进行对照,结果满意。     

液相色谱-电喷雾质谱联用法测定人血浆中托吡酯

目的:建立测定人血浆中托吡酯的液相色谱-电喷雾质谱联用(LC/ESI-MS)法。方法:待测血浆0.2 mL经甲醇沉淀除去蛋白,离心,取上清液10μL在Diamonsil C18柱上分离,流动相为甲醇-2.5 mmol·L-1醋酸铵(80:20),流速0.4 mL·min-1,LC/ESI-MS选择离子检测,负离子模式,用于定量分析的离子分别为m/z338(托吡酯)和m/z 356(吲哚美辛,内标)。结果:血浆中无干扰测定的内源性物质,每个样品分析时间小于7 min;本法线性范围为40-2560 ng·mL-1,最低定量浓度为40 ng·mL-1;日内、日间RSD分别小于3.8％和13.4％,相对偏差小于5.9％。结论:该法操作简便、快速、准确、灵敏度高,适用于临床药物动力学研究。     

液相色谱-质谱联用法测定人参皂苷Re在健康人血浆的浓度

目的建立液相色谱-质谱联用法测定人参皂苷Re在健康人血浆中浓 度的方法。方法血浆样品用固相萃取法处理。用电喷雾离子化和正离子多 离子反应监测方式检测人参皂苷Re。结果该方法人参皂苷Re线性范围为 1．05～1 050 ng·mL-1;定量下限为1．05 ng·mL-1;方法回收率在99．3％～ 104．3％;日内、日间变异系数(RSD)均<15％。结论该法准确、灵敏、特异, 适用于健康人血浆人参皂苷Re浓度测定。     

高锰酸钾-甲醛-盐酸米托蒽醌化学发光体系的研究

目的:建立盐酸米托蒽醌含量测定的化学发光分析法。方法:基于在酸性介质中,盐酸米托蒽醌可以显著增强高锰酸钾-甲醛体系的化学发光强度,结合流动注射技术而建立起的一种测定盐酸米托蒽醌的流动注射化学发光新方法。结果:该法的线性范围为0．01-1.0 mg·L-1(r=0.999 2),平均回收率为97．9％-101．9％。对0．5 mg·L-1的盐酸米托蒽醌连续平行测定11次,RSD为2．8％。根据IUPAC建议,计算其检出限为6．4μg·L-1 (3σ)。结论:该方法灵敏度高、线性范围宽、简单、快速,可用于针剂中盐酸米托蒽醌含量的测定。     

UPLC-MS/MS法测定大补阴丸中小檗碱等4种成分含量

目的：建立同时测定大补阴丸中小檗碱、黄柏碱、毛蕊花糖苷、芒果苷的含量方法。方法：采用液质联用技术,以Agilent ZORBAX SB-C18为色谱柱,以甲醇-甲酸铵溶液为流动相,等度洗脱。结果：小檗碱、黄柏碱、毛蕊花糖苷、芒果苷的线性范围分别为5～500 ng·mL-1（r=0.9991）、0.5～50 ng·mL-1（r=0.9994）、1～100 ng·mL-1（r=0.9987）、1.5～150 ng·mL-1（r=0.9990）;加样回收率为98.35%～102.55%,RSD均＜1.60%;平均含量分别为3.55 、0.322、0.511、0.886 mg·g-1。结论：该法专属性强、快速灵敏,可用于大补阴丸中上述4种成分的含量测定。     

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.     

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.