HPLC法同时测定天山雪莲中紫丁香苷、绿原酸和芦丁的含量

目的:采用液相色谱法同时测定天山雪莲中紫丁香苷、绿原酸和芦丁的含量。方法:Kromasil ODS-1色谱柱(250 mm×4.6 mm,5μm),柱温为35℃;以乙腈(A)-0.4%磷酸(B)为流动相,梯度洗脱(0 min,13%A;15 min,13%A;16 min,15%A;40 min11,15%A),流速1 mL·min~(-1);双波长检测:257 nm(检测紫丁香苷、芦丁),327 nm(检测绿原酸)。结果:紫丁香苷存5.104～51.04 μg·mL~(-1)(r=0.9998)、绿原酸在38.82～258.8μg·mL~(-1)(r=0.9991)、芦丁在39.42～262.8μg·mL~(-1)(r=0.9997)范围内都呈良好线性关系;紫丁香苷、绿原酸和芦丁的回收率分别为99.7%(RSD=1.0%),102.9%(RSD=1.5%),101.7%(RSD=1.2%);供试品溶液在12 h 内稳定。结论:该方法快速简单,线性关系良好。     

美妮达搽剂的定量方法的研究

目的制定美妮达搽剂的质量标准,采用反相高效液相色谱法测定美妮达搽剂中的有效成分甲硝唑、氯霉素的含量,同时对氯霉素降解的二醇物进行含量控制。方法选用Inertsustain C_(18)柱(4.6mm×250 mm,5μm),以甲醇(A)-0.08%冰醋酸(B)系统梯度洗脱,洗脱条件为:0~13 min(25%A),13~18 min(25%A→80%A),18~25 min(80%A),25~30 min(80%A→25%A);检测波长为271nm,柱温35℃,流速:1.0 mL·min~(-1)。结果美妮达搽剂中甲硝唑、氯霉素以及氯霉素二醇物3种成分分离良好,甲硝唑、氯霉素检测浓度分别在38.1~304.8μg·mL~(-1)(r=1)、88.05~704.4μg·mL~(-1)(r=1)与峰面积呈良好的线性关系,甲硝唑低﹑中﹑高浓度的加样回收率在101.1%~102.4%(RSD<2.0%),氯霉素低、中﹑高浓度的加样回收率在98.6%~100.9%(RSD<2.0%);氯霉素二醇物浓度在3.065~24.52μg·mL~(-1)(r=0.9999)与峰面积呈良好的线性关系,低﹑中﹑高浓度的加样回收率在96.0%~96.8%(RSD<1.0%),检测限为0.25 ng。结论该法同时检测美妮达搽剂中甲硝唑、氯霉素以及氯霉素二醇物的含量,操作简单,准确可靠,可用于该制剂的质量控制。     

HPLC法同时测定复方土茯苓胶囊中落新妇苷和芦丁的含量

目的:建立同时测定复方土茯苓胶囊中落新妇苷和芦丁含量的方法。方法:采用高效液相色谱法。色谱柱为WatersSymmetry-C1(8250mm×4.6mm,5μm),流动相为乙腈-0.1%磷酸(15∶85,V/V),检测波长为291nm,流速为1.0mL·min-1,柱温为35℃。结果:落新妇苷和芦丁的检测浓度分别在2.00～20.00、47.96～239.80mg·L-1范围内与各自峰面积积分值呈良好线性关系(r均为0.9999);二者平均加样回收率分别为100.40%和101.51%,RSD分别为1.61%和0.66%(n均为9)。结论:本方法简便、准确、稳定,可用于复方土茯苓胶囊的质量控制。     

贵州产菝葜属及肖菝葜属8个品种中落新妇苷分析及抗炎作用的比较

目的:采用高效液相色谱分析贵州产菝葜属及肖菝葜属8个品种中的活性成分落新妇苷,并通过对8个品种间慢性抗炎试验的对比研究,以对品种间进行药效与质量的评价。方法:RP-HPLC测定落新妇苷含量,色谱条件:依利特HypersilODS2色谱柱(5μm,4.6 mm×250 mm),流动相为乙腈-0.1%冰醋酸(21∶79);流速1 mL.min-1;柱温为30℃;检测波长291nm,理论塔板数以落新妇苷计不得低于5 000。结果:结论:落新妇苷在0.003～3.168μg范围内与峰面积呈良好线性关系,加样回收率(n=9)为100.43%,RSD%=1.5%,8个品种中落新妇苷含量为0.005%～4.493%,含量最低者是小叶菝葜与托柄菝葜,均为0.005 1%,而菝葜中落新妇苷的含量仅为0.009 0%,土茯苓中落新妇苷含量最高为4.5%,是2010年版中国药典(一部)中土茯苓药材中落新妇苷最低限量的10倍以上。通过抗炎试验研究,8个品种对小鼠棉球肉芽形成均具有显著抑制作用,其中土茯苓较其他品种抗炎作用好。本实验可为贵州产菝葜属及肖菝葜属新的药用资源开发利用及土茯苓药材的质量控制提供一定的参考依据。     

HPLC法同时测定复方氨酚苯海拉明片中4种成分的含量

目的:建立高效液相色谱法同时测定复方氨酚苯海拉明片中对乙酰氨基酚、咖啡因、盐酸苯海拉明和盐酸麻黄碱的含量。方法:采用迪马 C_(18)色谱柱(200 mm×4.6mm,5 μm);流动相为乙腈(A)-水(磷酸调节 pH 至2.1)(B),梯度洗脱[0 min(15%A)→7 min(50%A)→8 min(15%A)→18 min(15%A)];流速:1.0 mL·min~(-1);检测波长:215 nm;柱温:室温。结果:对乙酰氨基酚、咖啡因、盐酸苯海拉明和盐酸麻黄碱的线性范围分别为0.064～0.51 mg·mL~(-1)(r=0.9999)、0.067～0.54 mg·mL~(-1)(r=0.9995)、0.020～0.14 mg·mL~(-1)(r=0.9998)和0.017～0.14 mg·mL~(-1)(r=0.9999);平均回收率分别为100.1%(RSD=0.5%)、99.6%(RSD=0.8%)、101.4%(RSD=1.0%)和99.2%(RSD=0.6%)。结论:本法简便快速,结果准确可靠,可作为该复方制剂中4种成分的质量控制方法。     

复方瓜子金颗粒中秦皮乙素、蒙花苷和polygalasaponin ⅩⅪ的HPLC-DAD-ELSD法测定

目的:建立一种同时测定复方瓜子金颗粒中秦皮乙素、蒙花苷和 polygalasaponin XXI 含量的 HPLC-DAD-ELSD 方法。方法:采用 HPLC-DAD—ELSD 法,以 Diamonsil C_(18)(250 mm×4.6 mm,5 μm)为色谱柱,乙腈(A)-0.05%三氟醋酸水溶液(B)为流动相,梯度洗脱[0～10 min(15%A),10～30 min(15%A→28%A),30～40 min(28%A),40～50 min(28%A→32%A)],流速为1.0 mL·min~(-1),二极管阵列检测器的检测波长为340 nm,蒸发光散射检测器的漂移管温度为105℃、载气流速为2.6 L·min~(-1)。结果:秦皮乙素、蒙花苷和 polygalasaponin XXI 的线性范围分别为4.75～118.75μg·mL~(-1)(r=0.9998),4.02～100.50μg·mL~(-1)(r=0.9999),19.62～157.00μg·mL~(-1)(r=0.9993);加样回收率(n=9)分别为102.7%,100.9%,97.3%。结论:测定方法准确、重现,可为复方瓜子金颗粒的质量控制提供借鉴。     

HPLC双波长切换同时测定玄参药材中哈巴苷和哈巴俄苷的含量

目的:建立以双波长切换方式同时测定玄参药材中哈巴苷和哈巴俄苷的含量的方法。方法:采用Shimpack VP-ODS C_(18)柱(250 mm×4.6,5μm)色谱柱,以乙腈(A)-磷酸(0.03%)(B)为流动相进行梯度洗脱:0-10 min,3%-10%(A);10-20 min,10%-33%(A);20-25 min,33%-50%(A);25-30 min,50%-80%(A);30-35 min,80%(A);35-37min,80%-3%(A);检测波长:前10 min为278 nm,10-15 min为210 nm,15 min后为278 nm,柱温为30℃,进样量为10μL,流速为1 mL·min~(-1)。结果:哈巴苷在0.011824-0.11824 mg·mL~(-1)范围内与哈巴俄苷在0.00416-0.0416 mg﹒mL~(-1)范围内线性关系良好。结论:该检测方法稳定可行,可用于同时测定玄参药材中哈巴苷和哈巴俄苷的含量。     

不同产地知母药材的含量测定及指纹图谱研究

目的收集不同产地的知母药材,测定其芒果苷和知母皂苷BⅡ的含量,建立知母药材的HPLC-UV-ELSD指纹图谱测定方法。方法采用HPLC-UV-ELSD法测定3个不同产地的15批知母药材。色谱条件:Agilent ZORBAX SB-C_(18)色谱柱(250 mm×4.6 mm,5μm)。乙腈(A)-1 mL·L~(-1)乙酸(B)梯度洗脱,梯度为:0～5 min,2%A→5%A;5～12 min,5%A→20%A;12～18 min,20%A→25%A;18～23 min,25%A;23～40 min,25%A→100%A;40～45 min,100%A。柱温:30℃;流速:1.0 mL·min~(-1);波长:258 nm。ELSD参数:漂移管温度:105℃;氮气流速:2.6 L·min~(-1)。结果 15批知母药材中芒果苷的含量为0.803%～1.687%,知母皂苷BⅡ的含量为3.253%～7.332%。建立了知母药材的指纹图谱,包含11个共有峰,通过对照品指认了芒果苷和知母皂苷BⅡ2个峰,对15批知母药材指纹图谱的相似度进行评价,其相似度均大于0.9。结论该方法操作简便、快捷,可用于知母药材的质量研究。     

基于双波长融合HPLC图谱的银屑病外用方中7种活性成分定量分析

目的:测定银屑病外用方中7个主要活性成分(新落新妇苷、落新妇苷、新异落新妇苷、异落新妇苷、黄杞苷、异黄杞苷、柠檬苦素)含量。方法:采用Waters Xbrige BEH C₁₈色谱柱(150 mm×4.6 mm, 3.5μm),以水(A)-乙腈(B)为流动相,梯度洗脱(0～5 min, 17%B;5～10 min, 17%B→20%B;10～15 min, 20%B;15～25 min, 20%B→100%B),流速1.0 mL·min^-1,柱温30℃,进样量5μL,检测波长为292 nm(0～19 min,新落新妇苷、落新妇苷、新异落新妇苷、异落新妇苷、黄杞苷、异黄杞苷)、210 nm(19～25 min,柠檬苦素),使用MATLAB软件编程,对CSV格式数据进行双波长融合。结果:新落新妇苷、落新妇苷、新异落新妇苷、异落新妇苷、黄杞苷、异黄杞苷和柠檬苦素线性范围分别为39.06～1 250μg·mL^-1(r=0.999 9)、26.25～840μg·mL^-1(r=0.999 7)、18.64～59...     

高效液相色谱-二极管阵列-电化学联用技术测定八角莲中3种成分的含量

目的:建立高效液相色谱-二极管阵列-电化学联用技术同时测定八角莲中鬼臼毒素、山萘酚和槲皮素3种成分含量的新方法。方法:采用 Zorbax SB-C_(18)(150 mm×4.6 mm,5.0 μm)色谱柱;以甲醇(A)-0.2%醋酸(B)为流动相,采用梯度洗脱:0 min 时 A-B(50:50),12 min 时 A-B(70:30);流速为0.8 mL·min~(-1);柱温为25℃;二极管阵列检测波长为290 nm;单安培检测器的工作电位为 0.4 V。结果:鬼臼毒素、山萘酚及槲皮素的线性范围分别为46.9～469μg·mL~(-1)(r=0.9992),17.2～172μg·mL~(-1)(r=0.9991),8.44～84.4μg·mL~(-1)(r=0.9994);回收率分别为98.04%～98.51%(RSD<1.0%),96.30%～97.28%(RSD<1.5%),95.81%～97.72%(RSD<2.2%)。结论:本法简便,重复性好,适用于八角莲中鬼臼毒素、山萘酚和槲皮素3种成分的同时测定。     

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.     

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.