HPLC-MS法测定人血浆中溴己新浓度及生物等效性研究

目的:建立人血浆中溴己新的高效液相色谱-质谱测定方法.研究受试制剂克洛己新片中溴己新在人体内的药代动力学,评价其相对生物利用度和生物等效性.方法:20 名健康男性受试者随机分成2组,采用自身对照交叉给药的方式,分别单剂量口服受试或参比制剂,剂量均为溴己新 16 mg (相当于盐酸溴己新 17.5 mg )/头孢克洛 500 mg ,采用高效液相色谱-质谱法测定血药浓度.结果:志愿者口服受试和参比制剂后,溴己新的药动学参数如下:Cmax分别为(34.30 ± 7.02)ngmL-1和(35.06 ± 8.39)ngmL-1,tmax分别为(1.4 ± 0.6) h 和(1.0 ± 0.5)h,t1/2分别为(5.75 ± 0.82) h和(6.12 ± 0.62)h,CL分别为(123.9 ± 13.3)Lh-1和(129.2 ± 16.4)Lh-1 ,AUC0-24分别为(125.2 ± 16.1)nghmL-1和(119.8 ± 16.1)nghmL-1;溴己新的相对生物利用度为(106.0 ± 15.3)%.结论:试验建立的分析方法准确、灵敏、快速,统计结果表明受试制剂与参比制剂中溴己新生物等效.     

HPLC法测定盐酸溴己新片的含量及溶出度

盐酸溴己新为临床常用祛痰药，疗效确切。目前盐酸溴己新的含量测定方法有紫外分光光度法、HPLC法和毛细管电泳法。《中华人民共和国药典》2005年版收载的盐酸溴己新片的含量测定方法为紫外分光光度法，在进行溶出度实验时以0．1mol／L盐酸溶液为溶媒，该法测含量专属性不强，易受辅料干扰。盐酸溴己新在水中极微溶解，而在目前的质量标准中还没有溶出度检查，仅以崩解时限作为生物利用度的评定标准。为系统评价其药品质量，确保临床疗效，本建立了盐酸溴己新片的HPLC法，可用于该片的含量及溶出度测定，为考察盐酸溴己新片生物利用度提供了更为客观的标准。本方法简便、准确、专属性强，经献检索，尚未发现有关盐酸溴己新片溶出度实验的报道。     

愈创甘油醚片在健康人体中的药动学和生物等效性

目的研究愈创甘油醚片在健康中国人体内药动学和生物等效性。方法采用双周期交叉试验设计,20名健康男性受试者随机交叉单剂量口服愈创甘油醚片试验制剂和参比制剂0．2g,以高效液相色谱-质谱联用法测定人血浆中愈创甘油醚经时血药浓度,用DAS Ver2．0软件计算药动学参数,评价两制剂的生物等效性。结果愈创甘油醚片试验制剂和参比制剂的主要药动学参数ρmax分别为（909．3±314．7）和（874．1±301．5）μg·L^-1;tmax分别为（0．50±0．28）和（0．56±0．25）h;t1/2,分别为（1．54±0．92）和（1．53±1．08）h;AUC0→1分别为（1441-8±411．2）和（1486．9±482．8）μg·h·L^-1,AUC0→∞分别为（1456．5±418．2）和（1505．1±490．1）μg·h·L^-1;试验制剂的AUC0→1、AUC0→∞、ρmax的90％置信区间分别为参比制剂相应参数的91％-118％、91％-118％和94％-116％。以AUC0→t计算试验制剂中愈创甘油醚对参比制剂的相对生物利用度F为（99．7±15．1）％。结论经方差分析及双单侧t检验结果显示,试验制剂和参比制剂具有生物等效性。     

盐酸左氧氟沙星片在健康受试者体内的相对生物利用度与生物等效性研究

目的:研究国产盐酸左氧氟沙星片的人体相对生物利用度与生物等效性。方法:20名男性健康志愿者随机交叉单剂量口服国产盐酸左氧氟沙星受试和参比制剂（彼妥）200 mg,采用反相高效液相色谱法测定其血药浓度,计算其药动学参数和相对生物利用度,评价2种制剂的生物等效性。结果:国产盐酸左氧氟沙星受试和参比制剂的主要药动学参数:t_1/2分别为（7.37±0.88）,（7.39±0.96）h;t_max分别为（1.13±0.30）,（1.06±0.24）h;C_max分别为（2.08±0.42）,（2.03±0.35）mg·L^-1;AUC_0～24分别为（13.17±2.32）,（13.73±2.89）mg·h·L^-1。国产盐酸左氧氟沙星受试制剂的相对生物利用度为（97.7±16.7）%。结论:2制剂具有生物等效性。     

反相离子对色谱法测定溴吡斯的明的血药浓度及生物等效性研究

目的研究溴吡斯的明片的生物等效性。方法18名健康男性志愿者随机交叉单剂量口服60mg溴吡斯的明受试和参比制剂,考察两种制剂的生物等效性。血浆样本采用Sep-pak C18柱进行固相萃取后,用反相离子对色谱法测定溴吡斯的明的浓度,以计算2种制剂的药动学参数及相对生物利用度,并进行等效性评价。结果2种制剂的各主要药动学参数AUC0～18,Cmax和tmax无显著性差异,受试制剂的相对生物利用度为（102．9±11．6）％。结论双单侧t检验结果表明两制剂具有生物等效性。     

克林霉素磷酸酯片在健康人体内的相对生物利用度研究

目的研究克林霉素磷酸酯片和盐酸克林霉素胶囊在人体的相对生物利用度。方法采用随机、开放、双周期交叉试验设计,20名男性健康受试者分别单剂量口服试验制剂克林霉素磷酸酯片300mg或参比制剂盐酸克林霉素胶囊300mg。采用HPLC-UV法测定给药后不同时间采集的血样中克林霉素的血药浓度。结果受试者单次口服300mg克林霉素磷酸酯片和300mg盐酸克林霉素胶囊后,克林霉素的AUC0～10分别为（10.9±1.2）、（11.2±1.2）mg.h.L-1,Cmax分别为（3.7±0.6）、（3.7±0.6）mg.L-1;tmax分别为（0.83±0.09）、（0.83±0.10）h,t1/2分别为（2.0±0.1）、（2.1±0.1）h。方差分析结果表明2种制剂的参数之间没有显著性差异,克林霉素磷酸酯片与盐酸克林霉素胶囊的相对生物利用度为（97.8±5.4）%。结论克林霉素磷酸酯片和盐酸克林霉素胶囊具有生物等效性。     

愈创甘油醚双层缓释片与普通片在犬体内药动学的比较研究

目的:比较愈创甘油醚双层缓释片与普通片在犬体內药动学特征。方法:Beagle犬单剂量给予600mg愈创甘油醚双层缓释片或愈创甘油醚普通片后,用RP-HPLC法测定犬血浆中愈创甘油醚的药物浓度,计算药动学参数和相对生物利用度。结果:Beagle犬单剂量口服给药后,受试制剂和参比制剂中愈创甘油醚的AUC_0-12h分别为（15.96±6.41）μg·h·ml^-1和（19.23±4.96）μg·h·ml^-1;C_max分别为（7.31±2.86）μg·ml^-1和（14.24±5.34）μg·ml^-1;t_max分别为（0.79±0.25）h和（1.08±0.38）h;t_1/2分别为（2.50±1.67）h和（0.42±0.05）h;MRT_0-1分别为（2.37±0.61）h和（1.42±0.30）h。结论:愈创甘油醚双层缓释片的C_max较普通片显著降低,MRT_0-1和t_1/2明显延长,差异有统计学意义（P<0.05）,说明该受试制剂具备一定的缓释药动学特征。     

国产与进口头孢呋辛酯片的人体生物等效性研究

目的:评价国产头孢呋辛酯片的生物等效性.方法:24名健康志愿者随机交叉,自身对照口服单剂量国产头孢呋辛酯片及进口头孢呋辛酯片各500 mg后,进行人体相对生物利用度以及生物等效性研究.服药间隔为1周.血药浓度测定采用微生物法.结果:受试头孢呋辛酯片及参比头孢呋辛酯片的实测平均达峰时间Tmax分别为2.23±0.33 h和2.13±0.54 h;平均峰浓度Cmax分别为3.97±0.55和4.16±0.67 mg.L-1;消除半衰期T1/2ke分别为1.14±0.13及1.15±0.15 h.血药浓度-时间曲线下面积AUC0-t分别为10.57±1.91及10.60±1.97 mg.h.L-1;受试药与参比药比较,相对生物利用度F为100.6±13.1%.Tmax 、Cmax 与AUC0-t经生物等效性检验(双向单侧t检验)P<0.05.结论:国产头孢呋辛酯片与进口参比头孢呋辛酯片相比,其人体相对生物利用度为100.6±13.1%,且两制剂具生物等效性.     

HPLC测定盐酸溴己新片含量的不确定度评估

目的:建立HPLC测定盐酸溴己新片含量的不确定度评估的方法。方法:通过建立HPLC含量测定的数学模型,找出影响不确定度的因素,并对各个分量进行评估。由此计算合成不确定度,最终给出测定结果的扩展不确定度和置信水平。结果:盐酸溴己新含量的扩展不确定度为1.9%(k=2),含量测定结果为97.8%±1.9%(k=2)。结论:建立的不确定度评估方法可用于HPLC测定盐酸溴己新片含量的不确定度,使结果更加可靠。     

盐酸阿呋唑嗪片剂的人体生物等效性研究

12名健康志愿者采用交叉给药方法,分别单剂量口服5mg国产盐酸阿呋唑嗪片及进口盐酸阿呋唑嗪片,进行两种制剂的生物等效性研究.采用HPLC荧光检测法测定血浆中阿呋唑嗪的浓度, 血药浓度-时间数据经3p97生物利用度计算程序处理拟合,符合血管外给药二室开放模型, 试验结果表明:单次口服5 mg国产盐酸阿呋唑嗪片和进口盐酸阿呋唑嗪片后,达峰时间分别为1.46±0.54h和1.42±0.29 h,峰浓度分别为39.79±5.29μg·L-1和40.05±5.88μg·L-1,0～24h的药时曲线下面积分别为225.40±26.66μg·h·L-1和235.04±24.13μg·h·L-1.其主要药代动力学参数经方差分析和双单侧t检验,无显著性差异(P>0.05),两种制剂具有生物等效性.国产盐酸阿呋唑嗪片的相对生物利用度为96.9%±15.8%.     

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.     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

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.     

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.