高效液相色谱-质谱联用法测定人血浆中兰索拉唑的浓度

目的：建立高效液相色谱-质谱联用法测定人血浆中兰索拉唑的浓度。方法：血浆样品经处理后。采用WatersAdan—tis C18色谱柱（100mm×2．1mm,3μm）,流动相为甲醇-0．01％甲酸溶液（70：30）,以电喷雾电离源（ESI）正离子检测,奥美拉唑为内标。结果：兰索拉唑血浆质量浓度的线性范围为5．0～1500．0ng·ml-1（r=0．9998）,,提取回收率为78．2％。88．9％（n=5）,方法回收率为90．3％～101.9％（n=5）,日内和日间RSD均小于10％。结论：本法专属性强,样品处理方便,灵敏度高,适用于兰索拉唑药动学研究。     

高效液相色谱法测定人血浆中奥美拉唑与兰索拉唑

目的：建立高效液相色谱法测定人血浆中奥关拉唑和兰索拉唑血药浓度的方法。方法：在血浆中同时加入奥美拉唑、兰索拉唑及内标物泮托拉唑,以叔丁基甲醚提取。流动相为甲醇-乙腈-水（2：29：69）,流速为1．0ml·min^-1;检测波长为285nm,柱温为30℃。结果：奥美拉唑、兰索拉唑与内标物泮托拉唑分离良好,在0．02-4．0μg·ml^-1浓度范围为奥美拉唑线性方程为Y=6．889X-5．787×10^-4,r=0．9999;兰索拉唑线性方程为Y=10．758X＋6．667×10^-3,r=0．9998,奥美拉唑与兰索拉唑日内、日问精密度RSD均小于10％。结论：该法简单、准确、灵敏,适用于测定人血浆中奥美拉唑及兰索拉唑浓度。     

HPLC-MS／MS法测定大鼠血浆中兰索拉唑及其代谢产物的浓度

目的：建立大鼠血浆中兰索拉唑及其代谢产物5-羟基兰索拉唑、兰索拉唑砜的HPLC-MS／MS测定方法。方法：色谱条件：色谱柱：Diamonsil C18柱（150mm×2．1mm,5um）;流动相：乙腈-水（合0．01％甲酸及2mmol·L-1的醋酸铵（43：57,V／V）;流速：0．3ml·min-1;柱温：40℃;进样量10ul。质谱条件：电喷雾离子源（ESI）,以多反应监测离子方式测定兰索拉唑及其代谢产物,选择性监测质荷比（m／z）为368．0／163．9（兰索拉唑）,384．1／179．9（5-羟基兰索拉唑）,383．9／115．9（兰索拉唑砜）,326．0／280．1（内标奥美拉唑）。样品用乙腈沉淀蛋白处理。结果：兰索拉唑、5-羟基兰索拉唑、兰索拉唑砜的线性范围分别为11．40～4560．00,1．26～504．00,1．24～496．00ng·ml-1;定量下限分别为11．40,1．26,1．24ng·ml-1;批内、批间精密度RSD均〈15％。结论：该方法灵敏、准确、快速、专属性好,适用于兰索拉唑及其代谢产物在大鼠体内的药代动力学研究。     

HPLC法测定复方异丙安替比林片中异丙安替比林和氯唑沙宗的含量

目的:建立复方异丙安替比林片含量测定的HPLC方法。方法:色谱柱为Zorbax SB-C18(4．6×150 mm,5μm),流动相为乙腈-水-冰醋酸(68:32:1．0),检测波长为280 nm,流速为1．0 ml·min-1。结果:异丙安替比林和氯唑沙宗线性范围分别为5．4-27．1μg·ml-1(r=0．999 8),5．3-26．7μg·ml-1(r=0．999 8),检测限分别为0．20 ng和0．22 ng,平均回收率分别为99．6％,RSD0．4％和99．9％RSD0．6％(n=9)。结论:该方法简便、结果准确,可同时测定复方制剂中2种成分。     

HPLC法测定兰索拉唑胶囊中兰索拉唑的含量

目的建立并优化测定兰索拉唑胶囊中兰索拉唑含量的高效液相色谱法。方法色谱柱为依利特C18(4.6 mm×250mm,5μm),流动相为乙腈-0.1 mol.L-1磷酸盐缓冲液(pH7.2)(75∶25),检测波长为285 nm。结果兰索拉唑在浓度为线性范围为8～160 mg.L-1,相关系数为0.999 9,平均加样回收率为100.1%,RSD=1.19%。结论该方法操作简单、快速、准确,可用于兰索拉唑胶囊中兰索拉唑的含量测定。     

HPLC法同时测定复方氨酚苯海拉明片中对乙酰氨基酚和咖啡因含量

目的:建立同时测定复方氨酚苯海拉明片中对乙酰氨基酚和咖啡因含量的高效液相色谱法。方法:采用Hypersil ODS2色谱柱(200 mm×4．6 mm,5μm),流动相:0．01 mol·L-1磷酸二氢钾溶液(用磷酸调节pH至2．6-甲醇(7∶3);检测波长:273 nm。结果:对乙酰氨基酚的线性范围为39．1～117．0μg·ml-1(r=0．999 9)、咖啡因的线性范围为4．0-12．0μg·ml-1 (r=0．999 8);对乙酰氨基酚平均回收率为99．3％(n=9),RSD=1．5％;咖啡因平均回收率为100．1％(n=9),RSD=1．9％。结论:本法简便,准确,重现性好,可用于该制剂的质量控制及评价。     

HPLC法测定麻仁丸中大黄素和大黄酚的含量

目的:建立麻仁丸中大黄素和大黄酚含量测定的方法。方法:色谱柱为Thermo ODS C18柱(250 mm×4．6 mm,5μm),流动相为甲醇-0．1％磷酸溶液(85:15),流速为1．0 ml·min-1,检测波长为254 nm。结果:大黄素在0．01-0．29μg(r =0.999 5);大黄酚在0．03-0．52μg(r=0．999 4)范围内线性关系良好。大黄素及大黄酚的平均回收率分别为100.4％(RSD =0．9,n=5),100．1％(RSD=0．3％,n=5)。结论:方法准确可靠。     

川乌和制川乌中单酯及双酯型生物碱成分的含量测定

目的:建立川乌和制川乌中单酯及双酯型生物碱成分的高效液相色谱含量测定方法。方法:采用DiamonsilTM C18 5μm, 4．6 mm×250 mm色谱柱,流动相A[乙腈-四氢呋喃(25:15)]-流动相B[0．1 mol·L-1醋酸铵(1000 mL含冰醋酸 0．5 mL)]梯度洗脱,流速:0．8 mL·min-1,检测波长:235 nm。结果:乌头碱、次乌头碱、新乌头碱、苯甲酰新乌头碱和苯甲酰次乌头碱分离良好,各成分浓度与HPLC峰面积呈良好的线性关系,线性范围分别是:乌头碱0．01-0．36(r=0．9996),0．32 -5．16μg(r=0．9999);次乌头碱0．01-1．06(r=0．9999),0．05-1．32 μg(r=0．9999),1．29-20．60μg(r=0．9999);新乌头碱0．22-3．59μg(r=0．9999);苯甲酰新乌头碱0．02-0．10(r=0．9997),0．13-2．04μg(r=0．9998);苯甲酰次乌头碱 0．02-0．12(r=0．9996),0．15-2．41μg(r=0．9999)。平均回收率分别为105．0％(RSD=4．4％),104．9％(RSD=1．8％), 100．4％(RSD=3．0％);107．6％(RSD=3．3％),104．2％(RSD=1．5％),102．9％(RSD=1．9％);112．2％(RSD=3．2％), 106．3％(RSD=1．5％),103．5％(RSD=2．3％);104．1％(RSD=2．2％);100．8％(RSD=2．9％)。结论:建立的HPLC法专属,准确,耐用性良好。采用本法测定了23批不同产地及不同炮制方法得到的川乌和制川乌药材中新乌头碱、次乌头碱、乌头碱、苯甲酰新乌头碱和苯甲酰次乌头碱的含量,为川乌及饮片质量标准的建立提供了依据。     

兰索拉唑片在人体内的药动学研究

目的 采用RP-HPLC法进行兰索拉唑的血药浓度检测和药动学研究.方法 采用Agilent C18色谱柱(250 mm×5mm,5μm),流动相为乙腈-1‰三乙胺水溶液(pH7) (30∶70),流速1.0 mL·min-1,进样量20 μL,内标为奥美拉唑.血浆样品经乙酸乙酯提取后,于285 nm处检测.结果 0.05～1.60 μg· mL-1兰索拉唑与峰面积比值的线性关系良好(r ＝0.9997),最低定量限为20 ng·mL-1(S/N ＞3),日内RSD＜ 7.94％(n＝5),日间RSD ＜9.41％(n＝5),提取回收率＞95.79％.结论 所用方法可用于临床上兰索拉唑片血药浓度的检测及药动学研究.     

RP-HPLC测定兰索拉唑制剂中主药及有关物质的含量

目的 建立测定注射用兰索拉唑及兰索拉唑肠溶片中主药及有关物质含量的反相高效液相色谱法.方法 以Shim-pack C18(5 μm,250 mm×4.6 mm)为色谱柱;流动相为甲醇-水-三乙胺-磷酸(620∶380∶5∶1.5);检测波长为UV 284 nm;流速为1 mL·min-1.结果 兰索拉唑在16～320 μg·mL-1(r=0.999 9)内呈良好线性关系;注射用兰索拉唑和兰索拉唑肠溶片平均回收率分别为100.0%和99.63%, RSD为分别为1.08%和1.03%(n=9).结论 该方法简便、快速、准确、重现性好,可同时适用于分离和测定注射用兰索拉唑及肠溶片中有关物质及主药含量.     

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.