高效液相色谱法测定何首乌中二苯乙烯苷及大黄素-8-O-β-D-葡萄糖苷的含量

目的：探讨高效液相色谱法（HPLC法）对何首乌中有效成分二苯乙烯苷及大黄素-8-O-β-D-葡萄糖苷含量进行测定。方法：采用Phenomenex C18（250mm×4.6mm,5μm）色谱柱,流动相选择乙腈∶水=25∶75,流速为1.0mL/min,检测波长为322nm,柱温为25℃。结果：二苯乙烯苷的线性回归方程为：Y=3.247×105X+1.951×104（r=0.9999）,在4.021～40.12μg/mL范围内线性关系良好;大黄素-8-O-β-D-葡萄糖苷线性回归方程为：Y=1.501×105X+0.352×104（r=0.9998）,在0.012～25.00μg/mL范围内线性关系良好。二苯乙烯苷的平均加样回收率为99.79%,RSD=1.23%,大黄素-8-O-β-D-葡萄糖苷平均加样回收率为99.51%,RSD=1.52%。结论：高效液相色谱法测定何首乌中的两种有效成分,操作简单快速,稳定性好,精密度高,重现性好,为何首乌中有效成分的含量测定提供了有效的方法。     

益肾乌发口服液中二苯乙烯苷、大黄素、大黄素甲醚的含量测定

目的 建立益肾乌发口服液中2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷、大黄素、大黄素甲醚含量的测定方法,从指标成分角度来探讨益肾乌发口服液毒性与何首乌的关系,并为益肾乌发口服液的质量控制提供方法.方法 采用HPLC法,C18柱(4.6mm×250mm,5μm);测定二苯乙烯苷的流动相为乙腈-水(25:75),检测波长为320hm,流速为1.0 mL·min-1,柱温:25℃;测定大黄素大黄素甲醚的流动相为甲醇-0.1%磷酸溶液(80:20),检测波长为254nm,流速为1.0 mL·min-1,柱温:25℃.结果 二苯乙烯苷在0.1μg～10μg、大黄素在0.0168μg～1.68μg、大黄素甲醚在0.0208～1.04μg范围内呈良好的线性关系,r分别为0.9989(n=8)、0.9984(n=8)、0.9966(n=8);平均回收率:二苯乙烯苷为99.33%,大黄素为99.998%,大黄素甲醚为99.576%;RSD:二苯乙烯苷为0.68%,大黄素为1.11%,大黄素为1.29%.结论 高效液相色谱法测定益肾乌发口服液中二苯乙烯苷、大黄素、大黄素甲醚的含量方法操作简单、灵敏度高、干扰少、重现性好、回收率高,可用于益肾乌发口服液的含量测定.初步证实益肾乌发口服液的毒性可能与君药制何首乌中二苯乙烯苷、大黄素、大黄素甲醚的含量有关,应控制其剂量范围,避免不良反应的发生.     

炮制工艺对何首乌成分的影响

目的 分析不同炮制工艺对何首乌中的二苯乙烯苷、蒽醌、鞣质含量的影响。方法 统计何首乌经不同工艺(黑豆汁蒸片、黑豆汁炖片、黑清蒸片、黑豆汁高压蒸片、黑豆汁屉上蒸片等)炮制12 h、32 h后的二苯乙烯苷含量,不同批次何首乌生片在黑豆汁炖32 h下的二苯乙烯苷与蒽醌含量,不同炮制工艺(生片、豆汁蒸32 h、豆汁炖32 h、清蒸32 h、豆汁高压蒸12 h、豆汁屉上蒸32 h)下炮制的何首乌中蒽醌的含量以及豆汁炖不同炮制时间(0 h、12 h、24 h、36 h、48 h)二苯乙烯苷、蒽醌和鞣质含量。结果 黑豆汁高压蒸片工艺下炮制12 h的二苯乙烯苷含量最高,黑豆汁炖片共一下炮制32 h二苯乙烯苷含量最高;豆汁炖在0 h下二苯乙烯苷含量最高;在生片状态下,来源于广东的三种何首乌中的二苯乙烯苷含量最高,其次为云南,四川和贵州的二苯乙烯苷含量大致相同。在黑豆汁炖32 h下,来自于云南的何首乌片的二苯乙烯苷含量最高,其次为来自于广东的何首生片,然后为来自于贵州的何首乌片,最低的为来自于四川的何首乌片;豆汁高压蒸12 g下游离大黄素含量最高、豆汁蒸32 h下游离大黄素甲醚含量最高、生片状态下,总大黄...     

HPLC同时测定不同何首乌中的两种蒽醌类成分

目的 比较生何首乌、制何首乌及其发酵产品中游离型和结合型蒽醌类成分的含量.方法 采用HPLC法测定了不同何首乌蒽醌中两者的含量.色谱柱为Diamonsil C18(150 mm×4.6 mm,5 μm)柱,流动相为乙腈-0.1%磷酸(70:30),检测波长为254 nm,流速为1.0 ml·min-1.结果 大黄素、大黄素甲醚的线性范围分别为0.02～0.40 μg (r=0.9994)、0.01～0.20μg(r=0.9994),游离大黄素与大黄素甲醚测定的平均回收率分别为99.9%(RSD=1.9%)、97.9%(RSD=1.5%),总大黄素与大黄素甲醚的平均回收率分别为95.8%(RSD=3.7%)、105.9%(RSD=2.2%).结论 何首乌经炮制和发酵后,结合型蒽醌的含量明显低于生何首乌,文中方法准确可靠,可用于何首乌的质量控制.     

HPLC法测定养血生发胶囊中二苯乙烯苷、大黄素、大黄素甲醚的含量

目的建立高效液相色谱法测定养血生发胶囊中二苯乙烯苷、大黄素、大黄素甲醚含量的检测方法,从指标成分角度来探讨养血生发胶囊毒性与何首乌的关系,并且为养血生发胶囊的质量研究提供依据。方法采用高效液相色谱法,Hypersil C18柱(4.6mm×250mm,5μm);测定二苯乙烯苷采用流动相:乙腈-水(19 81),检测波长:320nm;测定大黄素、大黄素甲醚采用流动相:甲醇-0.1%磷酸溶液(80:20),检测波长:254nm。结果二苯乙烯苷在0.005～0.5mg/mL,大黄素在0.00168～0.084mg/mL,大黄素甲醚在0.00104～0.052 mg/mL,范围内呈良好线性关系;平均回收率:二苯乙烯苷98.66%,大黄素为95.80%,大黄素甲醚为97.70%;RSD:二苯乙烯苷为1.23%,大黄素为1.63%,大黄素甲醚为1.89%。结论本方法检测快速,定量准确,线性关系、重现性、稳定性较好、回收率较高,可用于养血生发胶囊的含量测定。初步证实养血生发胶囊的毒性可能与何首乌有关,蒽醌类物质可能是养血生发胶囊的毒性成分。     

何首乌九蒸九晒炮制工艺的优化研究

目的:建立测定何首乌中二苯乙烯苷、大黄素、大黄素甲醚和总多糖含量的方法,并优化何首乌传统九蒸九晒的炮制工艺。方法:采用高效液相色谱法检测二苯乙烯苷、大黄素、大黄素甲醚的含量;采用紫外分光光度法检测总多糖的含量。以蒸制时间、烘制时间、烘制温度为自变量,二苯乙烯苷、大黄素、大黄素甲醚和总多糖含量为指标,结合加权综合评分法和星点设计-效应面法优化何首乌九蒸九晒的炮制工艺并验证。结果:二苯乙烯苷、大黄素、大黄素甲醚检测进样量的线性范围分别为0.27～2.7μg(r=0.997 1)、0.063～0.63μg(r=0.999 9)、0.038～0.38μg(r=0.990 9),总多糖检测质量浓度的线性范围为2.07～12.42μg/mL(r=0.999 6);精密度、稳定性、重复性试验的RSD均小于2%;加样回收率分别为99.43%～101.06%(RSD=0.63%,n=6)、98.74%～120.33%(RSD=1.34%,n=6)、98.39%～102.44%(RSD=1.49%,n=6)、99.51%～101.98%(RSD=0.87%,n=6)。最优炮制工艺为蒸制时间4.5 h、烘制...     

HPLC法测定何首乌中二苯乙烯苷的含量

目的：建立高效液相色谱法（HPLC）测定何首乌中二苯乙烯苷含量的方法。方法：采用高效液相色谱法（HPLC）,以十八烷基硅烷键合相硅胶为固定相（4.6mm×250mm,5μm）,色谱纯乙腈-水（25∶75）,检测波长325nm,流速为1.0ml/min,色谱柱柱温30℃,测定何首乌中主要成分二苯乙烯苷的含量。结果：何首乌中二苯乙烯苷进样量在0.200～4.000μg范围内,与峰面积呈良好的线性关系,其回归方程为Y=152.92X＋45.67（r=0.9997）;平均回收率为99.96%。结论：该方法操作简单、方便、准确、重复性好,用于测量二苯乙烯苷的含量可信度高。     

一测多评法测定决明子中蒽醌苷元类和萘并吡喃酮苷类有效成分含量

目的建立决明子中蒽醌苷元类和萘并吡喃酮苷类有效成分含量的一测多评法。方法采用Sunfire C18色谱柱(4.6mm×250 mm,5μm),柱温30℃,乙腈-0.1%磷酸水溶液梯度洗脱,检测波长278 nm和284 nm,测定橙黄决明素与红镰霉素龙胆二糖苷、大黄酚和大黄素甲醚间的相对校正因子,并考察其重现性,比较计算值与测得值的差异。结果红镰霉素龙胆二糖苷、橙黄决明素、大黄酚和大黄素甲醚分别在:0.0503~1.5078、0.3197~9.5899、0.5070~15.2108、0.4027~12.0814μg范围呈良好线性;回归方程分别为:Y=4.95X+2.01(R2=0.9998)、Y=1.03X+0.03(R2=0.9999)、Y=3.98X-0.12(R2=0.9993)、Y=4.81X+0.26(R2=0.9996);用一测多评法对6批决明子中大红镰霉素龙胆二糖苷、大黄酚和大黄素甲醚进行测定,与外标法实测值基本一致。结论该方法可靠,结果准确,可用于决明子的质量控制。     

不同炮制辅料对何首乌药效成分含量的影响

目的研究不同炮制辅料对何首乌药效成分含量的影响,为选择何首乌炮制辅料提供依据。方法分别采用5种不同炮制辅料制备制何首乌,高效液相色谱法按照2015年版中国药典中制何首乌成分含量测定方法分别测定制何首乌中二苯乙烯苷、游离蒽醌的含量,比较不同炮制辅料对何首乌中药效成分含量的影响。结果炮制辅料基本都会降低制何首乌中二苯乙烯苷和游离蒽醌的含量。不同炮制辅料制备的制何首乌中二苯乙烯苷含量从高到低依次为空白对照何首乌>米泔水制何首乌>生姜汁制何首乌>甘草汁制何首乌>熟地汁制何首乌>黑豆汁制何首乌,游离蒽醌含量从高到低依次为甘草汁制何首乌>空白对照何首乌>米泔水制何首乌>熟地汁制何首乌>黑豆汁制何首乌>生姜汁制何首乌。不同炮制辅料制备的何首乌中二苯乙烯苷含量均高于2015年版中国药典中的要求,而游离蒽醌含量均未达到2015年版中国药典中的要求。结论不同炮制辅料对制何首乌中药效成分含量的影响不同。     

益肾降糖胶囊质量控制方法的研究

目的 建立益肾降糖胶囊的质量控制方法.方法 2018年9月至2019年3月,运用薄层色谱法(TLC)对益肾降糖胶囊中的制何首乌、赤芍、玄参、黄芪、山药、黄芩进行定性鉴别;采用高效液相色谱法测定芍药苷、二苯乙烯苷和黄芩苷的含量.采用Ultimate?XB-C18(4.6×250 mm,5μm)色谱柱;以甲醇-0.2％磷酸溶液为流动相,梯度洗脱;流速为1.0 mL/min;检测波长为230 nm(芍药苷)和320 nm(二苯乙烯苷、黄芩苷),柱温为40℃.结果 制何首乌、赤芍、玄参等TLC图斑点清晰,分离度好,阴性对照无干扰.芍药苷、二苯乙烯苷、黄芩苷质量浓度分别在31.320～234.900μg/mL(r=0.9991),3.939～14.772μg/mL(r=0.9995)、36.080～270.600μg/mL(r=0.9995),范围线性关系良好;平均加样回收率分别为98.96％,99.91％,99.89％,(RSD分别为0.76％,1.16％,1.41％,n=6).结论 该方法准确简便,重现性好,可用于益肾降糖胶囊的质量控制.     

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.

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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.     

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.     

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.