HPLC法测定脑通颗粒中丹酚酸B

目的 建立HPLC法测定脑通颗粒中丹酚酸B.方法 采用迪马色谱柱(250 mm×4.6 mm,5 μm);以甲醇-乙腈-甲酸-水溶液(30:10:0.5:59.5)为流动相;检测波长286 nm;体积流量1.0 mL/min:柱温40℃;进样量10μL.结果 丹酚酸B在0.441 6～2.208 0 μg线性关系良好(...     

HPLC波长切换法同时测定补肾活血方中丹参素、葛根素、大豆苷和丹酚酸B的含量北大核心CSCD

目的:建立HPLC波长切换法同时测定补肾活血方中丹参素、葛根素、大豆苷、丹酚酸B含量。方法:采用Inertsil ODS-SP C_(18)(4.6 mm×250 mm,5μm)色谱柱,以乙腈(A)-0.2%磷酸水溶液(B)为流动相梯度洗脱,柱温30℃,波长切换(0~12 min,282 nm,检测丹参素;13~22 min,250 nm,检测葛根素、大豆苷;23~36 min,286 nm,检测丹酚酸B)。结果:丹参素、葛根素、大豆苷、丹酚酸B线性范围分别为0.205 6~2.056μg(r=0.999 7)、0.1~1.0μg(r=0.999 6)、0.050 1~0.501μg(r=0.999 4)、1.0~10.0μg(r=0.999 5);平均回收率(n=6)分别为99.3%、100.7%、99.3%和98.8%,RSD分别为1.1%、1.6%、1.6%和1.6%。含量测定结果(n=3)分别为3.328 4~4.707 0、0.919 6~1.631 4、0.455 1~0.610 8、29.478 4~31.116 9 mg·g^(-1),RSD分别为1.6%~2.2%、0.4%~1.1%、1.2%~2.0%、0.8%~1.3%。结论:方法学验证结果表明,本法能为补肾活血方的质量标准研究提供参考。     

高效液相色谱法测定白术挥发油中苍术酮β-桉叶醇榄香烯的含量

目的建立高效液相色谱法测定白术挥发油中苍术酮、β-桉叶醇、β-榄香烯含量分析方法。方法采用HypersilODS2(4.6mm×250mm,5μm)色谱柱;流动相为乙腈-水(A)-水(B)60∶40,线性梯度洗脱(0~30min,40%B→10%B;30~40min,10%B→10%B);流速为1.0ml/min,柱温室温,检测波长200nm。结果苍术酮浓度在0.10~1.00mg/ml,β-桉叶醇浓度在2.00~600.00μg/ml,β-榄香烯浓度在0.70~42.00μg/ml范围内呈良好线性关系(r≥0.9998);3个组分低、中、高浓度的平均加样回收率在99.7%~101.2%,RSD(n=3)均<2.3%;重复性良好,RSD(n=6)均<1.6%。结论本方法简便、准确可靠,可用于白术挥发油中苍术酮、β-桉叶醇、β-榄香烯的含量测定。     

HPLC法同时测定心舒乐片中丹酚酸B、葛根素和羟基红花黄色素A的含量

目的:建立同时测定心舒乐片中丹酚酸B、葛根素和羟基红花黄色素A含量的方法。方法:采用高效液相色谱法。色谱柱为Welchrom C18,流动相为乙腈-0.1%磷酸(梯度洗脱),流速为1.0 ml/min,检测波长为280 nm,柱温为35℃,进样量为10μl。结果:丹酚酸B、葛根素、羟基红花黄色素A检测质量浓度线性范围分别为31.54~189.22(r=0.999 1)、17.13~102.76(r=0.999 7)、4.27~25.60μg/ml(r=0.999 5);精密度、稳定性、重复性试验的RSD<2%;加样回收率分别为96.29%~100.88%(RSD=1.73%,n=6)、97.12%~100.57%(RSD=1.15%,n=6)、97.09%~99.95%(RSD=1.01%,n=6)。结论:该方法简便实用、重现性好,专属性强,可有效控制该制剂质量。     

HPLC法同时测定复方丹参口服液中羟基红花黄色素A和丹酚酸B的含量

目的:建立同时测定复方丹参口服液中羟基红花黄色素A和丹酚酸B含量的方法。方法:采用高效液相色谱法。色谱柱为SHIMADZU ODS-C18,流动相为甲醇-0.5%磷酸(35∶65,V/V),流速为1.0 ml/min,检测波长为403 nm(羟基红花黄色素A)、286nm(丹酚酸B),柱温为35℃,进样量为20μl。结果:羟基红花黄色素A、丹酚酸B检测质量浓度线性范围分别为2.01~20.10、39.80~398.00μg/ml(r均为0.999 9);精密度、稳定性、重复性试验的RSD<2%;加样回收率分别为98.26%~101.25%、98.70%~101.35%,RSD分别为0.94%、0.71%(n=9)。结论:该方法简便可行、重复性好,可用于复方丹参口服液中羟基红花黄色素A和丹酚酸B的含量测定。     

HPLC同时测定乐脉颗粒中的丹酚酸B和丹参素钠

目的建立同时测定乐脉颗粒中丹酚酸B和丹参素钠含量的方法。方法采用HPLC法,色谱柱为Shim-pack C18柱(150 mm×4.6 mm,5μm),流动相A为甲醇,B为5%冰醋酸溶液,梯度洗脱。流速1.0 ml.min-1;检测波长286 nm;柱温35℃。结果丹酚酸B和丹参素钠分别在0.1933~2.8995、0.0807~1.2105μg线性关系良好,r均为0.9999,丹酚酸B和丹参素钠的平均回收率分别为98.54%、98.36%,RSD分别为0.69%、0.57%(n=6)。结论所建方法操作简便、结果准确,重复性好,可用于该制剂的质量控制。     

藤丹胶囊质量标准方法的改进

目的完善藤丹胶囊的质量标准。方法修订川芎、黄芪、三七的TLC方法、增加丹参、防己的TLC方法,建立HPLC法测定三七中人参皂苷Rg_1、人参皂苷Rb_1、三七皂苷R1的含量和丹参中丹酚酸B的含量测定。三七的含量测定:以Agilent 5 TC-C_(18)(250mm×4.6mm,5μm)为色谱柱;以乙腈为流动相A,以0.1%磷酸溶液为流动相B进行梯度洗脱;流速1.0m L/min;柱温:35℃;丹参的含量测定:以Agilent Extend C_(18)(250mm×4.6mm,5μm)为色谱柱;流动相:乙腈-1%甲酸溶液(20︰80);流速:1.0m L/min;柱温:35℃。结果三七皂苷R1进样量在1.1545~4.1562μg、人参皂苷Rg_1进样量在5.833~20.9988μg、人参皂苷Rb_1进样量在3.849~13.8564μg、丹酚酸B进样量在1.1~6.6μg范围内,与峰面积呈良好的线性关系,r=0.9998、r=0.9999、r=0.9996、r=0.9999;加样回收率分别为:平均回收率分别为98.39%、101.11%、101.11%,104.05%。结论完善后的质量标准可行,能有效控制藤丹胶囊的质量。     

UPLC法同时测定丹参片中6种酚酸类成分的含量

目的采用超高效液相色谱法(UPLC)同时测定8个厂家74批次丹参片中6种酚酸类成分(丹参素钠、原儿茶醛、迷迭香酸、紫草酸、丹酚酸B和丹酚酸A)的含量。方法采用UPLC法,色谱柱为Waters BEH C_(18)(100 mm×2.1 mm,1.7μm)。以乙腈(A)-1 mL·L~(-1)磷酸溶液(B)为流动相,梯度洗脱(0～14 min,6%B→23%B;14～20 min,23%B→39%B);柱温:25℃;样品管理器温度:4℃;流速:0.3 mL·min~(-1);检测波长:280 nm。结果丹参素钠、原儿茶醛、迷迭香酸、紫草酸、丹酚酸B和丹酚酸A的进样量分别在0.203 1～8.125 0,0.020 8～0.832 0,0.087 9～3.516 0,0.085 3～3.412 0,0.125 8～5.034 0和0.125 4～5.015 0μg范围内与色谱峰峰面积呈良好的线性关系;平均回收率(n=6)分别为99.2%,99.6%,99.5%,98.7%,99.3%和98.9%,RSD值分别为0.49%,0.79%,0.48%,0.92%,0.40%和0.88%。结论所建立的方法经方法学验证简便、快捷、准确、重复性好,可作为丹参片的质量控制方法。     

HPLC法测定丹参颗粒中丹酚酸B的含量

目的建立丹参养心颗粒中丹酚酸B含量的测定方法。方法采用HPLC法测定丹酚酸B的含量,色谱柱：OSD C18反相色谱柱（5μm,4.6×250 mm）;流动相：乙腈-2.5%甲酸（20：80）;流速：1.0 ml/min;检测波长：286 nm;柱温：35℃。结果平均回收率为99.55%,RSD%=0.42%。结论方法简便可行,重现性好,可很好地控制丹参养心颗粒的内在质量。     

益气强心胶囊的质量标准研究

目的:建立益气强心胶囊的质量标准。方法:采用薄层色谱(TLC)法对益气强心胶囊中的黄芪、红景天、丹参、葶苈子进行定性鉴别;采用高效液相色谱法测定制剂中丹酚酸B的含量:色谱柱为VP-ODS柱(150 mm×4.6 mm,5μm),流动相为甲醇-乙腈-2%甲酸溶液(24∶9∶67,V/V/V),检测波长为286 nm,柱温为35℃,流速为1 ml/min。结果:黄芪、红景天、丹参、葶苈子的TLC斑点清晰、分离度好,且阴性对照无干扰。丹酚酸B的质量浓度在0.027 8⁰.139 0 mg/ml范围内与其峰面积积分值呈良好的线性关系(r=0.999 9);精密度、稳定性、重复性试验的RSD<2%;平均加样回收率为100.29%,RSD=1.91%(n=6)。结论:所建标准可用于益气强心胶囊的质量控制。     

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.     

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.