延胡索HPLC指纹图谱评价及含量测定研究

目的 建立延胡索药材的高效液相色谱(HPLC)指纹图谱,对不同产地6批延胡索药材进行比较。同时对其中的6种生物碱进行色谱研究并测定其在延胡索药材中的含量。方法 使用Inertsil ODS-SP(4.6 mm×150 mm, 5μm)色谱柱,30℃柱温,流速1.0 mL·min^-1,检测波长280 nm,以乙腈(A)-0.1%磷酸溶液(B)(三乙胺调pH值至6.1)为流动相进行梯度洗脱。结果 建立了延胡索药材的指纹图谱并通过单个生物碱对照品确认了其中的6个生物碱,分别为原阿片碱、盐酸巴马汀、盐酸小檗碱、脱氢紫堇碱、延胡索乙素、延胡索甲素,这6种生物碱在对应线性范围内与其峰面积均呈现较好的线性关系(r≥0.9),并且分别测定出了其在延胡索药材中的含量。结论 此种方法操作简便、方法准确可行、耗时较短、稳定、重复性好,能够为延胡索药材的遴选提供一定的评判依据。     

UPLC-MS/MS法测定不同产地延胡索中原阿片碱、盐酸巴马汀、脱氢紫堇碱、延胡索乙素和延胡索甲素

目的 建立UPLC-MS/MS法同时测定不同产地延胡索药材中原阿片碱、盐酸巴马汀、脱氢紫堇碱、延胡索乙素和延胡索甲素。方法 采用Waters UPLC HSS T3 C18色谱柱（100 mm×2.1 mm,1.7μm）,以乙腈–0.1%醋酸铵为流动相,梯度洗脱,体积流量0.3 mL/min,柱温40℃,进样量5μL。ESI正离子模式采集,采集模式：MRM（多反应检测）,离子源温度150℃,毛细管电压3.0 kV,锥孔电压50 V,脱溶剂气流量800 L/h,脱溶剂气温度400℃。结果 原阿片碱、盐酸巴马汀、脱氢紫堇碱、延胡索乙素和延胡索甲素分别在0.20～10.12、0.25～12.58、1.02～51.20、0.85～42.64、0.39～19.44 ng线性关系良好;平均回收率分别为97.48%、100.16%、101.61%、96.05%、104.17%,RSD值分别为1.70%、2.05%、2.55%、0.74%、3.24%。不同产地的延胡索药材中5种生物碱的含量有明显差异,浙江产地延胡索药材中5种生物碱的含量相对其他产地较高,5种生物碱中脱氢紫堇碱的含量最高。结论 UPLC...     

延胡索酸碱水提物中生物碱含量比较

目的比较不同浓度酸碱水溶液对延胡索中原阿片碱、盐酸巴马汀、盐酸小檗碱、去氢紫堇碱、延胡索乙素和紫堇碱6个生物碱提取的影响。方法用氨水、冰醋酸配制不同酸碱浓度水溶液,供试品加热回流提取;采用Agilent Eclipse XDB-C_(18)(4.6 mm×250 mm,5μm),乙腈-0.6%醋酸溶液(pH 5.0)为流动相进行梯度洗脱,检测波长280 nm,柱温25℃,流速1 mL/min,测定6个生物碱及总生物碱含量。结果原阿片碱、盐酸巴马汀、盐酸小檗碱、去氢紫堇碱、延胡索乙素和紫堇碱分别在14.34~143.36、12.61~126.07、12.80~128.01、12.22~122.16、15.31~153.14、11.41~114.09μg/mL内呈良好线性关系,R2≥0.999 0;平均加样回收率分别为99.31%、97.64%、101.41%、96.29%、99.63%、98.74%,RSD值均小于3.0%。结论延胡索不同酸碱提取物生物碱含量有较大差异,酸性提取物生物碱含量均高于碱性提取物。     

HPLC测定广西产钩藤药材中的钩藤碱

目的评价广西不同产地钩藤Uncaria rhynchophylla的药材质量,确定药材的最佳采收期。方法采用HPLC测定28批不同产地和1个产地不同时期钩藤药材钩藤碱的含量,采用Gemini C18色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇-水(含0.002 mol.L-1三乙胺,用冰乙酸调pH7.5)(64∶36),流速1.0 mL.min-1,检测波长254 nm。结果 28批不同产地钩藤药材中有11批药材能检出钩藤碱,但含量差异较大;有17批药材未能检出钩藤碱成分;不同时期钩藤中钩藤碱的动态积累差异较大。结论广西不同产地钩藤药材的质量有一定差异;钩藤药材于秋、冬二季采收似乎较合理。     

一测多评结合内标加入法测定吐根及其制剂中生物碱类成分的含量

目的建立以盐酸小檗碱为加入内标,测定吐根药材及其制剂中盐酸吐根碱、盐酸吐根酚碱的一测多评测定方法。方法采用高效液相色谱法,以Capcell pak C_(18) MGⅡ(250 mm×4.6 mm,5μm)为色谱柱,乙腈(A)-体积分数0.1%磷酸溶液(B)为流动相,梯度洗脱,流速为1.0 mL·min~(-1),柱温30℃,检测波长为205 nm。以外加的盐酸小檗碱为内参物,分别建立盐酸吐根碱、盐酸吐根酚碱的相对校正因子,利用该相对校正因子计算药材及其制剂中该两种成分的量;同时利用外标法测定盐酸吐根碱、盐酸吐根酚碱的量,比较两种测定方法的差异,采用方差分析比较不同产地吐根药材的两种成分的含量差异。结果在线性范围内,盐酸小檗碱与盐酸吐根碱、盐酸吐根酚碱的相对校正因子分别为0.499 5、0.439 2,在不同实验条件下相对校正因子重现性良好,两种生物碱类成分的计算值与实测值无显著性差异。结论采用一测多评法测定吐根药材及其制剂中生物碱类成分准确且可行,可用于吐根药材及其制剂的质量控制。     

延胡索中5种生物碱的含量测定及稳定性研究

目的:建立延胡索中5种主要生物碱的含量测定方法,并比较产地初加工方法和炮制方法对延胡索中生物碱含量的影响。方法:采用超声提取法提取样品,以高效液相色谱法测定收集自浙江磐安的多个不同延胡索样品及其炮制品中5种主要生物碱的含量。结果:延胡索丙素、巴马汀、小檗碱、去氢延胡索甲素与延胡索乙素检测浓度的线性范围分别为0.05～0.50、0.05～0.50、0.02～0.20、0.20～2.00、0.20～2.00μg.mL-1,平均加样回收率在96.7%～100.2%之间。延胡索生品根据药典规定的原药材处理方式处理后,其生物碱含量降低一半左右,在此基础上的进一步炮制过程对原药材中5种生物碱的含量影响不大。结论:详备的延胡索原药材制备、炮制与生物碱含量检测方法的制定对延胡索药材的质量稳定将起到重要的作用。     

不同产地黄连中6个生物碱含量测定

目的测定不同产地黄连药材加工品中盐酸药根碱、盐酸非洲防己碱、盐酸表小檗碱、盐酸黄连碱、盐酸巴马汀、盐酸小檗碱的含量,为黄连药材的真伪鉴别提供依据。方法色谱条件为Phenomenex Gemini-NX C18色谱柱（250 mm×4.6 mm,5μm）;流动相为乙腈-30 mmol.L-1碳酸氢铵溶液（每1 000 mL碳酸氢铵溶液含7 mL氨水,1 mL三乙胺）梯度洗脱;流速1.0 mL.min-1;检测波长270 nm;柱温30℃。结果黄连药材中6个生物碱含量均较高,且其中盐酸药根碱、盐酸防己碱、盐酸表小檗碱、盐酸黄连碱、盐酸巴马汀和盐酸小檗碱的含量分别不低于0.30%,0.40%,1.10%,2.02%,1.45%,6.58%。结论方法准确、可行,可作为黄连药材质量的定量依据。     

宽筋藤总生物碱含量测定方法研究

目的建立宽筋藤药材中总生物碱的含量测定方法。方法以盐酸药根碱为对照品,采用紫外-可见分光光度法测定宽筋藤总生物碱的含量,测定波长为430nm。结果盐酸药根碱在0.001 1~0.006 5mg·mL-1范围内与吸光度呈良好线性关系,回归方程:Y=14.747 X-0.010 7(r=0.999 5)。盐酸药根碱平均回收率为100.4%,RSD=2.0%(n=6)。结论 16批宽筋藤药材总生物碱含量测定结果表明,所建立的方法简便、准确,可用于宽筋藤药材总生物碱的含量测定。     

中药附子质量控制研究

目的HPLC法同时测定附子饮片（生品和炮制品）中3种双酯型生物碱含量,达到控制附子质量及指导临床安全用药的目的。方法以附子饮片（生品和炮制品）为研究对象,采用Agilent Extend—C18（4．6mm×150ram,5lxm）色谱柱,以乙腈-40mmol·L^-1乙酸铵（氨水调pH10．0）为流动相,建立乌头碱、新乌头碱和次乌头碱的含量测定方法,采用外标法测定药材中3种生物碱的含量,并比较各样品之间的差异。结果3种生物碱分离度良好,回收率均在95％－105％。黑顺片中乌头碱、新乌头碱及次乌头碱的含量与其余五批生附片及另一批白附片相比,差别较大。白附片与生附片之间差异较小。同一产地的5批生附片中各生物碱的含量亦存在差异。结论通过同时控制乌头碱、新乌头碱和次乌头碱的含量来控制附子饮片（生品和炮制品）的质量较为合理有效,且简单易行。     

不同地区市售麻黄药材中盐酸麻黄碱、盐酸伪麻黄碱和总生物碱的含量测定

目的:对全国不同地区市售麻黄药材中盐酸麻黄碱、盐酸伪麻黄碱和总生物碱的含量进行测定,从化学成分的角度对其质量进行评估。方法:收集了28个批次来源于全国7个不同产地、21个不同地区的麻黄药材,按照2010版《中国药典》的方法测定其中盐酸麻黄碱、盐酸伪麻黄碱的含量;建立酸性染料比色法测定药材中的总生物碱。结果:有10个批次的麻黄药材(占全部批次的35.7%)达不到2010版《中国药典》的要求(盐酸麻黄碱和盐酸伪麻黄碱的总质量分数不得少于0.80%),且28个批次的样品中两种生物碱的总质量分数最高与最低相差45倍,总生物碱的质量分数最高与最低相差33倍。结论:所建立的酸性染料比色法测定麻黄总生物碱的含量方法简便、重复性好。市售麻黄药材以草麻黄为主,有效成分生物碱的含量差异较大,劣质情况较为严重。有必要加强麻黄药材的市场监管,保证其临床应用的安全、有效。     

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.