高效液相色谱法测定复方利福平片中利福平、异烟肼及吡嗪酰胺的含量

用HPLC法测定复方利福平片中利福平、异烟肼及吡嗪酰胺的含量。方法 :采用SpherisorbCN(2 50mm× 4 6mm ,5μm)色谱柱 ,流动相采用 0 0 1mol/L庚烷磺酸钠溶液 (pH2 2 )—乙腈 (44∶56,v/v) ,流速为 2 0ml/min,检测波长为 2 54nm。结果 :利福平在 71 76～ 1 66 40 μg/ml、异烟肼在 47 64～ 1 1 1 1 6μg/ml、吡嗪酰胺在 1 37 76～ 32 1 44μg/ml范围内线性良好 ,r分别为 0 9999、 0 9991、 0 9999(n =9) ,平均回收率分别为 :99 57%、 99 51 %、 1 0 0 1 0 % ,RSD分别为 :0 71 %、 0 51 %、0 62 % (n =7)。结论 :本法简便、准确、灵敏度高     

高效液相色谱法测定利福肼片的含量

目的建立高效液相色谱(HPLC)法测定利福肼片含量的方法.方法HPLC法,以岛津C18(4.6 mm×150mm,5μm)色谱柱,流动相为0.02 mol·L-1磷酸氢二钠溶液(磷酸调节pH 4.5)-甲醇(25∶75,v/v),检测波长为254 nm.结果线性范围及回归方程利福平10～250 mg·L-1,Y=6.121X-1.756 2(r=0.999 0);异烟肼5～125 mg·L-1,Y=4.261 24X-2.382 6(r=0.999 7);平均回收率利福平99.08%,RSD为1.02%(n=6);异烟肼99.75%,RSD为1.17%(n=6).结论本法专属性强,准确,灵敏,可作为利福肼片含量测定的方法.     

HPLC法测定乙胺吡嗪利福异烟片中4组分含量

目的：建立高效液相色谱法，测定乙胺吡嗪利福异烟片中利福平、异烟肼、吡嗪酰胺和盐酸乙胺丁醇含量。方法：利福平、异烟肼和吡嗪酰胺用C18色谱柱（150mm×4．6mm，5μm），以-0．01M磷酸二氢钾溶液-乙腈为流动相进行梯度洗脱，流速1．0ml·min^-1，检测波长254nm。盐酸乙胺丁醇选用C18色谱柱（150mm×4．6mm，5μm），四氢呋喃-0．4％庚烷磺酸钠（含有0．016％硫酸铜，用磷酸调pH至4．5）（25：75）为流动相，流速1．0ml·min^-1，检测波长258nm。结果：利福平、异烟肼、吡嗪酰胺及盐酸乙胺丁醇线性范围分别为16～160μg·ml^-1（r=0．9999），16—160μg·ml^-1（r=0．9999），53—532μg·ml^-1（r=0．9998），80～320μg·ml^-1（r=0．9998），平均回收率为99．3％～99．5％。结论：方法简便，准确，可用于乙胺吡嗪利福异烟片中各组分的质量控制。     

毛细管区带电泳法测定头孢他啶粉针剂的含量

目的　建立测定头孢他啶粉针剂含量的毛细管区带电泳法。方法　未涂层毛细管柱 (75 μm× 48.5 cm,有效长度 40 cm) ,电压 3 0 k V,检测波长 2 3 0 nm,温度 2 0℃ ,进样 5 0 m bar× 3 s。运行缓冲液为 2 5 mmol·L-1硼砂缓冲液 (用磷酸调 p H 8.0 )。结果　头孢他啶在 1 9.3 6～ 3 0 9.76 mg· L-1范围内呈良好的线性关系 ,r=0 .9999;最低检测限 0 .5 mg·L-1;加样回收率为 96 .8% ,RSD为 0 .6 5 % ;日内精密度 (RSD <3 .0 % ) ,日间精密度 (RSD <3 .0 % )。结论　本法简便快速、专属性强、准确、可靠     

高效液相色谱法测定头孢替唑钠的含量

目的　采用高效液相色谱法测定头孢替唑钠的含量。 方法 　采用 Agilent Hypersil ODS C1 8色谱柱 ( 4 .0× 12 5 mm,5μm) ,p H3 .6磷酸盐 /柠檬酸缓冲液 -乙腈 ( 88∶ 12 ) ,流速为 1.0 ml·min- 1 ;检测波长为 2 5 4nm。结果 　头孢替唑的浓度在 0 .0 45 2 2～ 0 .45 2 2 mg· m L- 1范围内线性关系良好 ,回归方程为 :Y=3 0 160 .44 83 X+67.6860 19,r=0 .9999。精密度实验 ( n=6)的 RSD为 0 .0 4%。平均回收率 99.0 %( RSD=0 .3 7%n=9)。 结论 　本方法简单 ,快速 ,结果准确 ,重现性好     

高效液相色谱法测定异福酰胺片的含量

为控制异福酰胺片剂质量,采用HPLC法测定异福酰胺片中异烟肼、利福平和吡嗪酰胺的含量。利福平选择极性较小的流动相,乙腈:0.05mol&#183;L~(-1)磷酸二氢铵(40:65);异烟肼和吡嗪酰胺选择极性较大的流动相,乙腈:0.05 mol&#183;L~(-1)磷酸二氢铵(2:98);均采用C_(18)柱,紫外检测器,检测波长分别为334nm和263nm。利福平、异烟肼和吡嗪酰胺的回收率分别为99.9%、99.8%和99.5%,RSD分别为1.33%、0.5%和1.1%。异烟肼、利福平及吡嗪酰胺日内RSD分别为2.78%、1.62%及1.31%,日间RSD分别为3.84%、2.12%及1.98%,其最低检出量分别为35μg、50μg及25μg。结果表明,采用HPLC法测定异福酰胺片的含量具有各种成分互不干扰,结果准确等特点。     

高效液相色谱法测定鼻腔洗剂Ⅱ号中氯霉素含量

目的 :建立高效液相色谱法测定鼻腔洗剂Ⅱ号中氯霉素含量的方法。方法 :采用高效液相色谱法。IntersilC1 8分析色谱柱 (4.6mm× 2 50mm ,5μm ) ,流动相为甲醇 磷酸盐缓冲液 (含 0 .0 5mol·L- 1 三乙胺 ,磷酸调 pH7.5 ,50∶50 ) ,流速 1mL·min- 1 ,检测波长 2 75nm。结果 :氯霉素与其相邻峰的分离度大于 1 .5 ,理论板数为 750 0 ,线性回归方程Y =-2 .738+0 .0 0 0 0 3776X ,r =0 .9999,线性范围 2 5 .98～ 2 59.8mg·L- 1 ,平均回收率为 1 0 1 .2 % ,RSD 2 .6 %。结论 :用高效液相色谱法测定鼻腔洗剂Ⅱ号中氯霉素含量 ,操作简便 ,结果准确     

牛黄上清片质量标准的研究

目的　对牛黄上清片的质量标准进行研究。方法　采用薄层色谱法对牛黄上清片中牛黄、冰片、黄连进行定性鉴别 ;采用高效液相色谱法测定大黄的有效成分大黄素、大黄酚的含量。结果　薄层色谱鉴别明显 ;大黄素线性范围为 2 5～ 2 0mg·L-1 ,回归方程 ^Y =4 4 2 8 X +2 2 9 5 6 (r =0 9999) ,精密度RSD为 0 74 % ;大黄酚线性范围为 1 5～ 4 5mg·L-1 ,回归方程 ^Y =[7 1 0 5× 1 0 6] X - 2 36 4 2 94 (r =0 9998) ,精密度RSD为 0 78%。结论　该方法简便 ,专属性强 ,重现性好 ,可作为牛黄上清片质量控制标准     

RP-HPLC测定替拉扎明及其有关物质

目的　建立测定替拉扎明含量及其有关物质检查的高效液相色谱法。方法　采用HypersilC18色谱柱(2 5 0mm× 4.6mm ,id 10 μm) ,甲醇 - 5 0mmol·L-1磷酸盐缓冲液 (10∶90 ,氢氧化钠调pH =6 .5± 0 .1)为流动相 ,在检测波长 2 6 6nm处进行含量测定 ;在相同条件下 ,以甲醇 - 5 0mmol·L-1磷酸盐缓冲液 (2 5∶75 ,氢氧化钠调pH =6 .5±0 .1)为流动相 ,对有关物质进行检查。结果　替拉扎明在 1.2 7～ 5 0 .8μg·ml-1范围内 ,峰面积与其浓度线性关系良好 (r =0 .9999) ,最低检测限为 2 .5× 10 -2 ng (S/N =3) ,重复进样RSD =0 .6 9% (n =5 ) ,日内RSD =0 .83%(n =5 ) ,日间RSD =0 .39% (n =5 ) ,重复性试验RSD =0 .2 1% (n =6 ) ,加样回收率为 99.6 2 %。结论　本法准确、简便、快速 ,可用于替拉扎明原料药及其制剂的质量控制     

异福酰胺片(Ⅱ)人体生物等效性评价

目的研究20名健康志愿者单剂量经口给予异福酰胺片(Ⅱ)受试制剂和参比制剂后的人体生物等效性。方法采用反相高效液相色谱法分别测定给药后受试者血浆中利福平、异烟肼、吡嗪酰胺3种有效成分的质量浓度,用DAS软件求其药动学参数及相对生物利用度。结果利福平、异烟肼、吡嗪酰胺的线性范围分别为0.40～25.0 mg·L-1、0.10～10.0 mg·L-1、0.30～60.0 mg·L-1,相关系数均大于0.99,日内和日间精密度均小于15%,提取回收率均大于70%,受试制剂中利福平、异烟肼、吡嗪酰胺的相对生物利用度分别为(97.5±25.5)%、(108.7±24.0)%、(109.0±17.4)%。结论受试制剂与参比制剂生物等效。     

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.