脑蛋白水解物注射液的质量评价

目的:通过测定脑蛋白水解物注射液各种氨基酸的含量,证明所采用方法的可行性与优越性的同时,对脑蛋白水解物注射液进行质量评价。方法:各种氨基酸含量测定采用异硫氰酸苯酯柱前衍生化高效液相色谱法:色谱柱为HiQsil C18V(4.6mm×250mm,5μm);流动相A为0.1mol.L-1醋酸钠溶液(pH6.5)-乙腈(94∶6),B为乙腈-水(80∶20);二元梯度洗脱;检测波长为254nm;流速为1mL.min-1;柱温为38℃;结果分析采用内标对比法,杂质检查分析采用归一化法分析。测定了8个厂家脑蛋白水解物注射液中各种游离氨基酸、总游离氨基酸及杂质含量,并进行了初步的质量评价。结果:8家药厂所生产的脑蛋白水解物注射液的质量参差不齐,A厂家所生产的脑蛋白水解物注射液质量相对较好。结论:本实验方法适用于脑蛋白水解物注射液的质量控制。     

异硫氰酸苯酯柱前衍生法测定复方氨基酸注射液(3AA)中氨基酸含量

目的:建立采用异硫氰酸苯酯柱前衍生化高效液相色谱法测定复方氨基酸注射液(3AA)的含量。方法:色谱柱:Diamonsil C18(2)(250 mm×4.6 mm,5μm);流动相:A为0.1 mol/L醋酸钠溶液(pH 6.5)-乙腈(93∶7);B为乙腈-水(80∶20)梯度洗脱,流速:1.0 ml/min;柱温:40℃;检测波长:254 nm。结果:各个氨基酸的浓度与其相应色谱峰面积的线性相关系数均达到0.999以上;各个氨基酸的浓度与其相应色谱峰面积呈良好的线性关系,回收率在98.5%～99.6%。结论:该方法适用于测定复方氨基酸注射液(3AA)的含量,具有准确、快速和灵敏度高等优点。     

HPLC—ELSD法测定疏血通注射液中17种未衍生氨基酸含量

目的:建立高效液相色谱-蒸发光散射检测法(HPLC-ELSD),直接测定疏血通注射液中17种未衍生氨基酸含量的方法。方法:以异亮氨酸等氨基酸为标准品;色谱柱:Prevail~(TM) C_(18)(4.6mm×250mm,5μm);流动相A:乙腈,流动相B:5.0mmol·L~(-1)七氟丁酸的0.7%三氟醋酸溶液,流动相A与B比例:0min为0∶100,5min为0∶100,8min为15∶85,25min为35∶65;漂移管温度:115℃;气体流量:2.5L·min~(-1);流速:0.8mL·min~(-1);柱温:35℃;进样量:10μL。结果:在25min内即可完成对疏血通注射液中17种氨基酸的分离测定,氨基酸质量浓度0.071～2.285g·L~(-1)时,其对数值与峰面积对数值线性关系良好,17种氨基酸的加样回收率为92.9%～105.1%,信噪比为3时,异亮氨酸检出限为20mg·L~(-1)。结论:该法不需要专门的氨基酸分析仪和衍生化氨基酸,操作简便,有足够的灵敏度,结果准确可靠。     

反相离子对色谱法测定硫酸奈替米星及其注射液的含量

目的:测定硫酸奈替米星及其注射液的含量。方法:采用反相离子对色谱法。色谱柱为Shim-pack VP-ODS(150 mm×4.6 mm,5μm,日本岛津);柱温:40℃;流动相为含0.61％庚烷磺酸钠的0.1 mol·L-1磷酸二氢钾溶液(用磷酸调pH为2.5)-乙腈(76:24);流速为1.0 mL·min-1,检测波长为205 nm。结果:测得原料的平均回收率为99.8％,RSD为0.6％,注射液的平均回收率为100.4％,RSD<0.6％。结论:方法准确可靠,适合硫酸奈替米星及其注射液的含量测定。     

氨基酸分析法测定复方骨肽注射液中多肽含量

目的建立复方骨肽注射液中多肽含量的氨基酸分析法。方法氨基酸水解方法:6mol.L-1盐酸110℃水解22h;氨基酸衍生方法:异硫氰酸苯酯(PITC)法;HPLC条件:Agilent Eclipse AAA色谱柱;流动相A:0.1mol.L-1乙酸钠(用冰醋酸调至pH 6.5)-乙腈(97∶3);流动相B:乙腈-水(4∶1);流速:1.0mL.min-1;检测波长:254nm。结果 18种氨基酸的线性、精密度及稳定性良好;除水解样品中的胱氨酸与色氨酸外,其他氨基酸的回收率均达到定量要求。结论方法准确、专属性强,可作为复方骨肽注射液中多肽的定量分析方法。     

柱前衍生高效液相色谱法测定复方氨基酸注射液中氨基酸的含量

目的建立以2,4-二硝基氟苯为衍生化试剂,高效液相色谱法(HPLC)测定复方氨基酸注射液(18AA)中各氨基酸含量的方法。方法采用Agilent HC-C 18色谱柱,乙腈-水(60∶40)为流动相A,乙腈-磷酸盐缓冲液(9∶91)流动相B,梯度洗脱,流速1.0 mL·min-1,检测波长360 nm,柱温27℃。结果复方氨基酸注射液中各氨基酸分离度良好,线性相关系数均大于0.9989,平均回收率均在98%~102%之间,RSD%均小于2%。结论本方法可以用于复方氨基酸注射液(18AA)中氨基酸检测。     

柱前衍生RP-HPLC法测定人参中17种氨基酸的含量

目的:建立同时测定人参中17种游离氨基酸和总氨基酸含量的方法,并应用于不同生长周期、炮制方法和生长部位的人参研究。方法:采用异硫氰酸苯酯对样品进行柱前衍生,以反相高效液相色谱法测定,外标法计算含量。色谱柱为UltimateAAA氨基酸分析专用柱(250mm×4.6mm,5μm),流动相A为三水合醋酸钠缓冲溶液(pH6.5)-乙腈溶液(93:7,V/V)、B为80%乙腈-水(80:20,V/V),梯度洗脱,检测波长为254nm。结果:17种氨基酸的检测浓度线性范围为0.0025～2.5000μmoL·mL-1,r均>0.9980。17种游离氨基酸的平均加样回收率在87.30%～100.01%之间,RSD在0.59%～2.94%之间(n均为6);测定总氨基酸时的平均加样回收率在91.50%～100.07%之间,RSD在0.41%～2.00%之间(n均为6)。结论:本方法灵敏、准确、重复性好,可用于人参及其相关产品的质量控制。     

HPLC法测定盐酸硫必利注射液的含量及其有关物质

目的采用高效液相色谱法测定盐酸硫必利注射液含量及其有关物质。方法以C18柱为色谱柱(250×4.6mm,5μm);0.005mol·L-1辛烷磺酸钠溶液-甲醇-乙腈(60∶15∶25)(用磷酸调pH值至3.0)为流动相;流速1.0ml·ml-1;检测波长235nm。结果该方法测定的线性范围为9.0～150μg·ml-1(r=1.0000)。结论该方法准确、灵敏、快捷,适用于该产品的质量控制。     

DTDPA-PITC联合柱前衍生化HPLC法同时测定复方氨基酸注射液中18种氨基酸的含量

目的:建立同时测定含有半胱氨酸或胱氨酸的复方氨基酸注射液中18种氨基酸含量的方法。方法:采用高效液相色谱法,以二硫代二丙酸(DTDPA)和异硫氰酸苯酯(PITC)联合作为柱前衍生化试剂。色谱柱为Kromasil C18,流动相[A为0.1 mol/L醋酸盐缓冲液(pH 5.38)-乙腈(93∶7,V/V)、B为乙腈-水(80∶20,V/V),采用梯度洗脱],检测波长为254 nm,流速为1.0 ml/min,进样量为3μl,柱温为35Ⅴ。结果:18种氨基酸在各自测定浓度的20%¹80%的范围内与各自峰面积积分值呈良好的线性关系(r≥0.999 0);精密度、稳定性、重复性试验的RSD≤1.91%;平均回收率在95.7%180%的范围内与各自峰面积积分值呈良好的线性关系(r≥0.999 0);精密度、稳定性、重复性试验的RSD≤1.91%;平均回收率在95.7%⁹9.7%之间,RSD≤1.99%(n=9)。结论:该方法准确、高效,实现了同时对复方氨基酸注射液中半胱氨酸以及其他17种氨基酸的准确定量。     

柱前衍生化HPLC法测定齿瓣石斛中总氨基酸的含量

目的:建立测定齿瓣石斛中总氨基酸含量的方法。方法:以2,4-二硝基氟苯为衍生试剂,与齿瓣石斛中氨基酸柱前衍生,采用高效液相色谱法测定其含量。色谱柱为SymmetryC1(8250mm×4.6mm,5μm),流动相A为0.05mol·L-1醋酸钠-醋酸缓冲液(pH6.4)-1%N,N-二甲基甲酰胺,流动相B为乙腈-水(1:1,V/V),梯度洗脱,检测波长为360nm,柱温为30℃,进样量为20μL。结果:13种氨基酸的线性相关系数均>0.9990,平均加样回收率范围在99.34%～101.93%之间,RSD在0.54%～1.91%之间(n=6)。结论:本方法快速、简便、结果准确,可为齿瓣石斛的深入研究奠定基础。     

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.