用HPLC法和微生物法测定人血浆中头孢丙烯浓度的比较

目的:比较用HPLC法和微生物法测定人血浆中头孢丙烯浓度的差异。方法:10名健康志愿者单剂量口服头孢丙烯片500 mg,分别用HPLC法和微生物法测定血浆中药物浓度。结果:HPLC法线性范围为0．143～14．300μg／mL,低、中、高浓度(0．357 5、1．430 0、7．150 0μg／mL)的回收率分别为(101．75±7．71)％、(96．89±2．55)％和(98．70±1．67)％(n= 6);3种浓度的日内、日间RSD分别为7．58％0、2．63％、1．69％和7．11％、3．77％、2．01％(n=6)。微生物法线性范围为0．25～3．00μg／mL,低、中、高浓度(0．50、1．75、3．00μg／mL)的回收率分别为(96．37±5．99)％、(108．26±8．39)％和(105．12±10．35)％(n=5);3种浓度的日内、日间RSD分别为9．84％、7．75％、6．22％和10．05％、8．31％、7．87％(n=5)。结论:两种方法回收率和精密度均符合要求,受试者各时间点平均血药浓度测定值无显著性差异。     

测定泰妥拉唑含量的3种方法研究

目的:建立测定泰妥拉唑含量的方法。方法:采用银量法、紫外分光光度法和高效液相色谱法分别对泰妥拉唑进行含量测定。结果:银量法平均回收率为99．9％,RSD为0．13％(n=5),方法精密度为O．09％(n=5); HPLC法在泰妥拉唑浓度为5．020～50．20μg·mL-1的范围内,峰面积与浓度线性关系良好(r=0．999 7),精密度为0．20％(n=6),回收率在99．5％～100．0％;UV法在2．0～12μg·mL-1的范围内线性关系良好(r=0．999 9),精密度为0．81％(n=5),回收率在99．4％～100．3％。结论:3种方法均适用于泰妥拉唑的含量测定,且都有较高的准确度和精密度。银量法结果准确,无需对照品;UV法快速、简便;HPLC法专属性好,可测定主药含量,同时还可进行杂质限度检查。     

盐酸金刚烷胺片的含量测定

目的建立盐酸金刚烷胺片的含量测定方法。方法采用紫外-可见分光光度法测定,测定波长为(412±1)nm。结果盐酸金刚烷胺浓度在2．5-15μg／mL范围内与吸收度呈良好的线性关系(r=0．999 5,n=6),平均回收率为99．88％,RSD=0．69％(n=6)。结论紫外-可见分光光度法灵敏,准确可靠,重现性好,可用于盐酸金刚烷胺片的含量测定。     

RP-HPLC法测定氢溴酸加兰他敏口服液含量及有关物质

目的:采用反相高效液相色谱法测定氢溴酸加兰他敏的含量及其有关物质。方法:采用Kromasil C_(18)色谱柱(5 μm,4.6mm×150 mm);以乙腈-水相(20:80,每 800 mL水相中含有2.67 mL 二丁胺,用磷酸调节pH为 9.0±0.05)为流动相;流速1.0 mL·min~(-1);检测波长:280 nm;柱温:室温;进样量:20 μL。结果:反相高效液相色谱法测定的线性范围为30～210 μg·mL~(-1);r=0.999 9;日内精密度:RSD为0.73％(n=5);日间精密度:RSD为0.72％(n=5),氢溴酸加兰他敏含量测定高、中、低3种浓度的回收率分别为98.89％、99、84％和99.56％,RSD分别为0.53％、0.40％和0.49％。结论:本法简便快速、准确、专属性好。     

紫外分光光度法测定醋酸洗必泰含漱剂中醋酸洗必泰含量

目的:建立醋酸洗必泰含漱剂中醋酸洗必泰的含量测定方法。方法:采用紫外分光光度法,以乙醇为溶剂,测定波长为(259±1)nm。结果:醋酸洗必泰浓度在3．0-15．0 μg/mL范围内和吸收度有良好线性关系,回归方程为A=0．060 78 C-0．001 21,r=0．999 8(n=7), 平均回收率为99．78％,RSD=0．43％(n=5)。结论:该方法灵敏度高、操作简便、专属性好、重现性好,可用于该制剂的质量控制。     

HPLC法测定复方利血平片中维生素B1、B6、泛酸钙和氢氯噻嗪的含量

目的：建立复方利血平片中维生素B1、维生素B6、泛酸钙和氢氯噻嗪含量测定的HPLC法。方法：采用phenomenex的Synergi Hydro—RP色谱柱（150mm×4．6mm，4μm），以乙腈、0．02mol·L^-1磷酸氢二钾溶液和0．005mol·L^-1庚烷磺酸钠（用磷酸调节pH至3．0±0．05）为流动相，采用梯度洗脱，流速为1．0mL·min^-1，用二极管阵列检测器检测，检测波长210nm，柱温36℃。结果：维生素B1的线性范围为7．752～77．52μg mL^-1（r=0．9992），精密度RSD为0．99％（n=7），重现性RSD为1．62％（n=7），平均回收率为98．63％，RSD为1．59％（n=9）；维生素B6的线性范围为7．580～75．80μg·mL^-1（r=0．9995），精密度RSD为0．62％（n=7），重现性RSD为1．17％（n=7），平均回收率为99．17％，RSD为1．26％（n=9）；泛酸钙的线性范围为7．904—79．04μg·mL^-1（r=0．9997），精密度RSD为0．58％（n=7），重现性RSD为0．95％（n=7），平均回收率为98．37％，RSD为0．92％（n=9）；氢氯噻嗪的线性范围为25．62～256．2μg·mL^-1（r=0．9995），精密度RSD为0．19％（n=7），重现性RSD为1．36％（n=7），平均回收率为99．12％，RSD为1．06％（n=9）。结论：本方法操作简便，结果准确、可靠，可同时测定复方利血平片中维生素B1、维生素B6、泛酸钙和氢氯噻嗪的含量。     

HPLC测定混合糖电解质注射液中果糖、葡萄糖和木糖醇的含量

目的建立混合糖电解质注射液中果糖、葡萄糖和木糖醇的含量测定方法。方法色谱柱：Waters SugarPakI（300mm×6．5mm）;流动相：水;流速：0．5mL.min^-1’;示差折光检测器。结果果糖、葡萄糖和木糖醇峰分离度良好;果糖的线性范围为20～100μg（r=0．9999）;平均回收率为99．53％,RSD=O．81％（n=9）;葡萄糖的线性范围为40-200μg（r=0．9997）;平均回收率为99．75％,RSD=0．47％（n=9）;木糖醇的线性范围为10-50ug（r=0．9997）;平均回收率为99．78％,RSD=1．02％（n=9）。结论本方法简便、准确,可用于混合糖电解质注射液中果糖、葡萄糖和木糖醇的含量测定。     

梯度洗脱高效液相色谱法测定辛伐他汀分散片的含量和有关物质

目的建立辛伐他汀分散片含量测定及有关物质的检查方法。方法采用Supeleo C18柱（33mm×4．6mm,3μm）,以乙腈-0．1％磷酸（50：50）为流动相A、0．1％磷酸乙腈溶液为流动相B,梯度洗脱,流速为3．0mL／min,检测波长为238nrn。结果辛伐他汀与辛伐他汀酸、洛伐他汀及强制破坏产生的降解产物均分离良好,辛伐他汀质量浓度在20．02-180．2μg／mL范围内与峰面积呈良好的线性关系。回归方程A：12493C＋2199,r=0．9999（n=7）;日内精密度RSD为O．59％（n=6）;日间精密度RSD为0．69％（n=6）;平均回收率为99．9％．RSD=0．5l％（n=9）;供试品溶液在6h内基本稳定;检测限为17．26ng／mL。结论该方法专属性强、灵敏度高,可用于辛伐他汀分散片的含量测定和有关物质检查。     

用HPLC法测定复方盐酸阿替卡因注射液中两种组分的含量

目的:建立用HPLC法测定复方盐酸阿替卡因注射液中盐酸阿替卡因和肾上腺素两组分含量的方法。方法:采用C_8柱和C_(18)柱分别建立两组分的色谱分离检测系统,其中测定盐酸阿替卡因采用的流动相为含庚烷磺酸钠的乙腈与磷酸盐缓冲液,检测波长为276 nm;测定肾上腺素时采用了水和甲醇双流动相切换洗脱装置,检测波长为220 nm。结果:盐酸阿替卡因的浓度线性范围为4～400μg/mL(r=0．9999),低、中、高3个浓度溶液的加样回收率分别为(103．83±0．85)％、(101．26±0．44)％、(99．15±0．31)％(n=5),日内RSD分别为1．69％、0．60％、0．21％(n=5),日间RSD分别为1．73％、0．67％、0．22％(n=5)。肾上腺素的浓度线性范围为1～100μg/mL(r=0．999 9)。低、中、高3个浓度溶液的加样回收率分别为(102．89±0．85)％、(99．30±0．43)％、(100．39±0．82)％(n=5),日内RSD分别为0．94％、0．71％、0．34％(n=5),日间RSD分别为1．37％、0．96％、0．55％(n=5)。结论:该方法精密、简便、准确,可用于复方盐酸阿替卡因注射液的含量测定。     

反相离子对-高效液相色谱法测定河豚毒素

目的建立了反相离子对-高效液相色谱法测定河豚毒素含量的方法。方法选用SHIMADZUODS色谱柱(150×6mm5μm),以0．2％(v／v)醋酸液为流动相,流速为1．2mL／min,检测波长为230nm。结果河豚毒素线性范围20～100μg／mL,r=0．9960(n=5);回收率为93．32％,RSD=5．41％(n=5);日内精密度为6．10％,日间精密度为7．42％(n=5)最低检测限50ng。结论方法准确、快速、简便,可作为迅速确定河豚毒素含量的测定方法。     

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.