黑豆、半成品和成品淡豆豉中6种异黄酮含量变化比较

目的 比较发酵前的黑豆及半成品和成品淡豆豉中大豆苷、黄豆黄苷、染料木苷、大豆苷元、黄豆黄素和染料木素6种异黄酮的含量变化。方法 采用高效液相色谱法测定异黄酮的含量,色谱条件为DiamonsilC₁₈色谱柱(4.6×250 mm,5μm),柱温30℃,检测波长为260 nm,流动相为0.2%醋酸水(A)-甲醇(B),梯度洗脱,流速为1.0 ml/min。结果 该方法测定6种异黄酮成分在测定范围内线性良好(r≥0.999 3),且回收率符合要求,黑豆经发酵后其中的6种异黄酮含量明显提升,且成品淡豆豉比半成品淡豆豉中含量更多。结论 本研究建立的高效液相色谱法能够准确测定淡豆豉中6种异黄酮的含量。发酵可提升黑豆中的异黄酮的含量,且在发酵过程中结合型异黄酮向游离型异黄酮苷元转化。     

HPLC法同时测定中国、泰国葛根中异黄酮的含量

目的:建立HPLC法同时测定中国、泰国葛根中异黄酮大豆苷、大豆苷元和染料木素的含量,为其质量控制提供方法学参考.方法:色谱柱为AT.LICHROM ODS-C18柱(250 mm ×4.6 mm,5μm);流动相为甲醇和水,梯度洗脱;流速:0.8 ml·min-1;检测波长250 nm;柱温为25℃.结果:大豆苷、大豆苷元和染料木素分别在0.15 ～0.88 μg,0.10 ～0.63 μg,0.15 ～0.89 μg范围内呈良好线性关系,r分别为0.9997,0.9995,0.9996.其平均回收率在99.5％～100.4％范围内,RSD均小于3％.结论:建立的方法简单、准确,分离效果好,首次用于中国、泰国葛根的定量分析和比较分析,为葛根的资源研究提供参考.     

薄层扫描法测定染料木素和大豆苷原的含量

目的建立染料木素和大豆苷原的薄层扫描测定方法。方法薄层色谱条件是:吸附剂:聚酰胺-6薄膜,展开剂:苯-丁酮-甲醇(3∶1∶0.5),结果染料木素和大豆苷原均在250nm处有最大吸收,本法线性关系良好(染料木素:r=0.9993;大豆苷原:r=0.9995)。染料木素的平均回收率98.52%(n=5);大豆苷原的平均回收率99.45%(n=5)。结论方法简便,准确,重现性好,可用于测定染料木素和大豆苷原的含量。     

HPLC法同时测定葛根中5种异黄酮的含量

目的建立能同时测定葛根中5种异黄酮含量的HPLC法。方法采用Phenomenex C 18色谱柱(250 mm×4.6 mm,5μm);柱温:40℃;流动相A为甲醇,B为水,进行梯度洗脱;流速为1.0 mL·min^-1;检测波长为260 nm。结果葛根素、大豆苷、染料木苷、大豆苷元和染料木素的回归方程分别为y 1=35462.65 x 1+899.4(r 1=0.9999),y2=89343.6 x2-720.2(r2=0.9999),y 3=139651.625 x3+1351.7(r 3=0.9998),y4=129138.175 x4-1339.5(r 4=0.9999)和y5=216079.6 x5-3679.8(r5=0.9999),分别在0.015~0.150,0.048~0.480,0.049~0.490,0.049~0.490和0.052~0.520μg范围内线性关系良好,平均回收率分别为98.3%,99.6%,97.5%,100.1%和96.0%。结论该方法同时测定了葛根中5种异黄酮的含量,结果准确、可靠,为葛根质量评价奠定了基础。     

HPLC法同时测定保健食品中6种大豆异黄酮类成分的含量

目的:建立同时测定大豆类保健食品中大豆苷、黄豆黄苷、染料木苷、大豆苷元、黄豆黄素、染料木素6种成分含量的高效液相色谱法。方法:色谱柱为 Agilent Zorbax SB-C_(18)(4.6 mm×150 mm,5 μm),采用流动相乙腈-0.2%甲酸进行梯度洗脱,流速为1 mL·min~(-1),检测波长为255 nm。结果:大豆苷、黄豆黄苷、染料木苷、大豆苷元、黄豆黄素、染料木素的线性范围分别为0.42～3.15,0.20～2.25,0.41～3.06,0.15～1.14,0.051～0.38,0.21～1.55 μg;加样回收率(n=5)分别为95.86%,96.87%,96.85%,96.63%,96.48%,96.67%。结论:本方法简便、灵敏、准确,可用于含大豆异黄酮类保健食品中以上6种成分的含量测定和产品质量控制。     

HPLC-MS/MS法同时测定大鼠血浆中槐角苷及染料木素

目的:建立快速、灵敏的液相色谱-串联质谱方法测定大鼠血浆中槐角苷及其苷元染料木素。方法:血浆经过乙酸乙酯提取,以大豆苷元为内标,采用Zorbax SB-C18(2.1 mm×30 mm,3.5μm)色谱柱,流动相为甲醇-0.001%甲酸铵溶液(加氨水调节pH至7.5)(45∶55),流速0.2 mL·min-1;质谱条件为电喷雾离子源(ESI源),负离子模式检测,扫描方式为多反应监测(MRM),定量离子对为m/z 252.9→223.7(大豆苷元)、m/z 431.1→267.6(槐角苷)、m/z 268.8→132.8(染料木素)。结果:槐角苷浓度在1.072～536 ng·mL-1,染料木素浓度在1.068～534 ng·mL-1的范围内线性关系良好(r>0.995);日间和日内精密度RSD均小于10%,低、中、高3个浓度的提取回收率在85%～97%之间。结论:本法可用于大鼠血浆中槐角苷和染料木素的测定及其药代动力学研究。     

野葛、粉葛与云南葛中葛根素、大豆苷、大豆苷元、染料木素的含量比较

目的：比较野葛、粉葛与云南葛中葛根素、大豆苷、大豆苷元和染料木素的含量。方法：样品以30%乙醇回流提取后采用高效液相色谱法测定含量。色谱柱为Uvis-201（250mm×4.6mm,5μm）,流动相为甲醇-水（梯度洗脱）,流速为1.0mL·min-1,检测波长为250nm,柱温为室温。结果：葛根素、大豆苷、大豆苷元、染料木素的进样量分别在0.68～5.44μg（r=0.9992）、0.143～1.144μg（r=0.9993）、0.0245～0.1960μg（r=0.9992）、0.088～0.704μg（r=0.9994）范围内与各自峰面积积分值呈良好线性关系;平均加样回收率分别为98.1%、99.5%、101.1%、99.2%,RSD分别为2.17%、2.42%、2.53%、2.65%（n均为6）。葛根素含量高低排序为河南野葛〉云南葛〉山西野葛〉大别山野葛〉广西粉葛;大豆苷含量为云南葛〉河南野葛〉大别山野葛〉山西野葛〉广西粉葛;大豆苷元含量为河南野葛〉云南葛〉山西野葛〉大别山野葛〉广西粉葛;染料木素含量为河南野葛〉云南葛〉山西野葛〉大别山野葛〉广西粉葛。结论：野葛、粉葛与云南葛中葛根素、大豆苷、大豆苷元、染料木素含量差别较大,且云南葛为野葛的伪品之一,三者不能同等应用。     

HPLC同时测定豆豉片中5种有效成分含量

目的建立高效液相色谱法同时测定豆豉片中栀子苷、大豆苷、染料木苷、大豆苷元、染料木素含量的方法。方法采用Hypersil ODS 2色谱柱(200 mm×4.6 mm,5-m);乙腈为流动相A,甲醇为流动相B,1.8%冰醋酸溶液为流动相C,洗脱程序为0 min(A:8%,B:10%,C:82%)→28 min(A:8%,B:10%,C:82%)→28.1 min(A:0%,B:38%,C:62%)→55 min(A:0%,B:38%,C:62%),流速为1 m L·min-1,检测波长为254 nm,柱温:30℃。结果栀子苷、大豆苷、染料木苷、大豆苷元、染料木素分别在12.36~396.00,3.50~112.00,3.36~116.00,3.00~96.00和1.63~52.00μg·m L-1内线性关系良好,平均回收率分别为100.0%,100.3%,99.8%,99.4%,99.9%,RSD分别为0.14%,0.97%,0.83%,0.98%,1.23%。结论本方法灵敏度高、专属性强、重复性好,可用于本制剂的质量控制。     

高效液相色谱法同时测定葛根提取物中5种异黄酮的含量

目的:建立葛根提取物中5种异黄酮含量测定的HPLC方法。方法:采用Hypersil C18色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.02 mol·L-1醋酸铵(醋酸调pH=4.6)为流动相,进行梯度洗脱,流速为1.0mL·min-1,检测波长为260 nm,柱温为35℃。结果:葛根素、大豆苷、染料木苷、大豆素、染料木素分别在0.204～510.4μg·mL-1(r=0.999 9),0.201～503.0μg·mL-1(r=0.999 9),0.233～581.8μg·mL-1(r=0.999 9),0.201～502.4μg·mL-1(r=0.999 9),0.195～487.6μg·mL-1(r=0.999 9)范围内线性关系良好。平均回收率(n=6)分别为101.52%(RSD=0.69%),102.66%(RSD=0.30%),98.90%(RSD=0.26%),98.64%(RSD=0.29%),100.16%(RSD=0.20%)。结论:本方法快速、简便、精密度和重现性好、灵敏度高,可作为葛根提取物的含量控制方法。     

RP-HPLC法测定大豆异黄酮的含量

目的：采用反相高效液相色谱法,建立大豆提取物中6种大豆异黄酮-大豆苷,黄豆黄苷,染料木苷,大豆苷元,黄豆黄素,染料木素的含量测定方法.方法：色谱柱：welchrom-C18柱（250 mm×4.6 mm;5 μm）,流动相：甲醇-水-乙腈=35∶65∶8（磷酸调节pH到2.6-3.0）;流速：0.8 ml·min-1;检测波长：254 nm,柱温：35℃.结果：线性范围：大豆苷1.48～29.54 mg·L-1（r=0.999 8）,黄豆黄苷1.58～31.64 mg·L-1（r=0.999 8）,染料木苷1.09～21.84 mg·L-1（r=0.999 8）,大豆苷元0.97～19.46 mg·L-1（r=0.999 8）,黄豆黄素1.20～23.96 mg·L-1（r=0.999 9）,染料木素1.31～26.26 mg·L-1（r=0.999 9）.大豆苷,黄豆黄苷,染料木苷,大豆苷元,黄豆黄素,染料木素平均回收率分别为101.56%,100.45%,102.75%,100.83%,100.92%,101.53%,RSD分别为1.66%,1.10%,1.89%,1.10%,1.52%,1.35%（n=6）.结论：本方法便捷、灵敏、准确、重复性好,适用于大豆产品中大豆异黄酮含量检测.     

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.     

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.