吐温80体外溶血研究

目的 考察吐温80的体外溶血作用,为制剂生产和临床安全用药提供依据.方法在570 nm波长下,采用紫外分光光度法评价吐温80对家兔红细胞的体外溶血作用.结果吐温80可引起家兔红细胞体外溶血和红细胞形态改变.结论 吐温80具有一定的体外溶血性.     

注射用多西他赛致溶血反应的实验研究

目的探讨注射用多西他赛引起急性溶血反应的机制。方法采用紫外分光光度法对市售制剂进行体外溶血作用的测定。结果多西他赛注射液各浓度条件均存在不同程度的溶血情况,其溶血程度与药物浓度呈剂量依赖性,与空白辅料组相比其溶血作用无统计学意义（P〉0.05）。结论注射用多西他赛引起急性溶血反应与其制剂中使用的高浓度吐温80直接相关。     

重楼总皂苷溶血作用实验研究

目的:研究重楼总皂苷的溶血作用。方法:通过常规体外试管法(肉眼观察法)及改进的体外溶血性试验法(分光光度法)来观察重楼总皂苷的溶血作用。结果:重楼总皂苷浓度≤0.01mg·mL~(-1)时无溶血现象发生;而当其>0.01mg·mL~(-1)时则出现溶血,并且随着重楼总皂苷浓度的增加溶血率也增加,直至接近100%。肉眼观察法和分光光度法结果一致。结论:重楼总皂苷低浓度时无溶血作用,而大于一定浓度时则具有溶血作用,且溶血强度与皂苷浓度呈剂量依赖性。     

油茶皂苷的体外溶血试验研究

目的观察油茶皂苷（sasanquasaponin,SQS）的溶血作用和抗氧化剂对SQS溶血的影响。方法通过常规体外试管法（肉眼观察法）及改进的体外溶血性试验法（分光光度法）来观察溶血作用。结果SQS试液浓度≤0．39×10^-4mg·mL^-1时溶血率〈5％,无溶血现象发生,SQS＋VitC组各管溶血率也明显低于SQS组同浓度各管溶血率;肉眼观察法和分光光度法结果一致。结论SQS低浓度时无溶血作用,而大于一定浓度时则具有溶血作用,且溶血强度与浓度呈剂量依赖性。加入抗氧化剂VitC后再加入SQS。SQS的溶血百分率明显下降,不同程度地抑制SQS的溶血作用。     

复方茵陈注射液体外溶血试验观察

目的 评价复方茵陈注射液的体外溶血作用.方法 通过常规体外试管法(肉眼观察法)及改进的体外溶血性试验法(分光光度法)来观察3个批次的复方茵陈注射液对兔红细胞的溶血作用.结果 复方茵陈注射液在试验浓度内均无溶血现象发生.肉眼观察法和分光光度法结果一致.结论 复方茵陈注射液对兔红细胞无溶血作用.     

香丹注射液溶血性实验及药物安全性检测问题探讨

目的：建立体外溶血试验方法,评价香丹注射液的体外溶血作用,为药物安全性检测提供参考。方法：用常规体外试管法、分光光度法和氰化高铁血红蛋白法3种方法探讨3批香丹注射液对兔红细胞的溶血作用。结果：香丹注射液按常规体外试管法有部分溶血发生,与其他两种方法结果不一致,说明肉眼观察影响结果判断,分光光度法更准确可靠。结论：香丹注射液可能有溶血作用,故需要对药品进行溶血性检测;采用抗凝兔血紫外分光光度法更能准确检查该制剂的潜在溶血倾向,保证临床用药安全。     

中药注射剂溶血检查的两种方法比较

目的:常规体外试管法与分光光度法在中药注射剂溶血检查中的应用比较。方法:通过吸收波长、离心转速及温育时间的研究,建立了分光光度法测定溶血率的方法,并与常规体外试管法进行了3种中药注射剂溶血试验的结果对比。结果:分光光度法可以定量地测定中药注射剂的溶血程度,较常规体外试管法更能准确地判断药品的质量。结论:建议对于中药注射剂应使用分光光度法评价其溶血程度。     

葛根素注射液对不同物种红细胞体外溶血性比较

目的比较葛根素注射液对不同物种红细胞体外溶血的差异。方法将不同浓度葛根注射液与Beagle犬、人和家兔红细胞在温度为(37±0.5)℃的生化孵箱内孵育6 h,用分光光度计在波长540 nm处测定各管的吸光率,计算溶血率。结果葛根素注射液对Beagle犬、人及家兔红细胞体外最大溶血率分别为65.35%、81.89%和135.05%;对3种红细胞体外溶血的半效溶血浓度分别为2.79、3.96和5.47 mg/ml;对3种红细胞的毒-效反应关系的斜率分别为1.45、1.96和0.49。此外,溶剂丙二醇在0.25和0.5 ml对3种红细胞均无溶血性(溶血率5%)。结论葛根素注射液对犬、人和家兔红细胞均能产生体外溶血作用,并有物种差异(敏感性:犬人家兔);溶剂丙二醇对葛根素注射液致红细胞溶血作用无影响。     

银杏达莫注射液溶血试验方法研究及结果评价

目的:采用溶血检测方法对供试品在试验条件下的溶血和凝聚潜在性做出评价,为其临床安全使用提供参考.方法:根据《中药、天然药物刺激性和溶血性研究的技术指导原则》的要求,采用常规体外试管溶血法(观察法)和改进的体外溶血性试验法(分光光度法),对银杏达莫注射液进行了溶血性研究.结果:常规体外试管溶血法对94批次银杏达莫注射液进行溶血检查,结果:为全部合格;采用改进的体外溶血性试验法(分光光度法),根据溶血率判定标准,检测的64批银杏达莫注射液的溶血率均小于5％.结论:通过对分光光度法与常规方法的比较可以看出,分光光度法操作较复杂,但检测准确,数据详实,可以更直接准确地反应制剂对血细胞的影响,有助于及时发现安全风险.可以作为常规方法的有效补充.     

吐温-80不同浓度对依普黄酮胶囊溶出度的影响

目的考察吐温-80不同浓度对依普黄酮胶囊溶出度的影响。方法分别配制含不同浓度吐温-80的依普黄酮胶囊,采用紫外分光光度法等测定依普黄酮的含量及溶出度,计算参数Td、T50、m,并对T50、Td值进行两两比较。结果吐温-80浓度在1%~3%范围内,随其浓度增加,T50、Td值均减小,其最佳浓度为2.5%。结论制剂时适量加入吐温-80可明显增加依普黄酮的溶出度(P<0.01)。     

不同有机溶剂对人精子体外运动参数的影响比较

目的研究常用有机溶剂对正常男性精子体外运动活力的影响。方法取正常男性精子,采用上游优化法处理,制备成精子悬液并分成4组,前3组分别加入二甲亚砜,使二甲亚砜终浓度分别为0.5%,1%和2%,pH值均为7.5,渗透压为290 mosm.(kg.H2O-1),D组加入等量Ham’s F10培养液作对照。分别于孵育15,30,45和60 m in后采用CASA进行分析。参照上述方法,考察有机溶剂甘油、聚乙二醇、吐温80对精子体外运动活力的影响。甘油终浓度分别调至1%,5%和10%,聚乙二醇终浓度分别调至1%,5%和10%,吐温80终浓度分别调至1%,5%和10%。结果2%二甲亚砜作用于精子60 m in能导致精子全部死亡,10%甘油作用60 m in后对精子运动参数抑制率达73.8%,10%聚乙二醇400作用于人精子60 m in可致其全部死亡,10%吐温80作用60 m in后对精子运动参数抑制率达95.5%。加入不同浓度二甲亚砜、甘油、聚乙二醇400和吐温80的各组前向运动精子百分率、曲线运动速度、直线运动速度和平均路径速度均较空白对照组显著降低(均P<0.05)。结论高浓度有机溶剂体外可抑制正常男性精子存活率和运动活力。     

聚山梨酯80刺激肥大细胞RBL-2H3脱颗粒作用的评价

目的：研究不同类别及厂家的聚山梨酯80直接刺激肥大细胞RBL-2H3脱颗粒的作用,为建立完善的注射用辅料引发类过敏反应的筛选评价体系提供依据。方法：以不同剂量的聚山梨酯80处理RBL-2H3细胞,测定β-氨基己糖苷酶释放量,并通过MTT法进一步研究有脱颗粒作用的受试物对RBL-2H3细胞的毒性作用。结果：8种聚山梨酯80样品具有不同程度的直接刺激RBL-2H3细胞脱颗粒的作用,且呈浓度依赖性,其中上海试剂采购供应站提供的受试物作用极强,威尔制药口服级受试物作用较强,威尔制药A、威尔制药B、威尔制药注射级、威尔制药4个受试物作用相近,2个进口品种的作用略弱。在产生直接刺激RBL-2H3细胞脱颗粒作用的浓度下,上海试剂采购供应站的聚山梨酯80有明显的细胞毒性,并呈浓度依赖性。结论：聚山梨酯80能够直接刺激RBL-2H3细胞脱颗粒,其作用强度不同反映了产品的质量差异;上海试剂采购供应站提供的聚山梨酯80直接刺激RBL-2H3细胞脱颗粒的作用可能由其细胞毒性引起。     

The conflict between in vitro release studies in human biorelevant media and the in vivo exposure in rats of the lipophilic compound fenofibrate

The performance of four different lipid-based (Tween 80-Captex 200P, Tween 80-Capmul MCM, Tween 80-Caprol 3GO and Tween 80-soybean oil) and one commercially available micronized formulation (Lipanthyl Micronized(?)) of the lipophilic compound fenofibrate was compared in vitro in various biorelevant media and in vivo in rats. In simulated gastric fluid without pepsin (SGF(sp)) and fasted state simulated intestinal fluid (FaSSIF), only Tween 80-Captex 200P system resulted in a stable fenofibrate concentration, but no supersaturation was obtained. The other three lipid based systems created fenofibrate supersaturation; however they did not maintain it. In fed state simulated intestinal fluid (FeSSIF), all lipid-based formulations resulted in complete dissolution of fenofibrate during the experiment, which represented a supersaturated state for Tween 80-Capmul MCM and Tween 80-Caprol 3GO systems. In both FaSSIF and FeSSIF, all lipid-based formulations yielded a higher fenofibrate concentration than the micronized formulation. Contrary to the in vitro results, no significant difference in the in vivo performance was observed among the four tested lipid-based formulations both in the fasted and the fed states. The in vivo performance of all lipid-based formulations was better than that of Lipanthyl Micronized(?), in the fasted as well as in the fed state. The fact that for the lipid based systems the in vitro differences in pharmaceutical performance were not translated into in vivo differences can be attributed to the continuous excretion of bile in the gastrointestinal tract of rats, causing enhanced solubilizing capacity for lipophilic drugs. This study clearly points to the conflicting situation that might arise during the preclinical phase of the development of lipid based formulations of lipophilic drugs as the performance of such systems is very often evaluated by both in vitro release studies in human biorelevant media as well as in vivo studies in rats. Care must be taken to select a relevant animal model.     

注射用七叶皂苷钠体外溶血作用试验研究

目的：探讨不同种类实验动物血细胞对七叶皂苷钠体外溶血活性的影响以及比较国内外同类品种体外溶血活性。方法：参照《中国药典》2005年版一部“溶血与凝聚”检查法。结果：用兔血细胞考核的9批产品,其中2批出现红细胞微溶无凝聚,采用羊血细胞则均未出现红细胞溶血与凝聚;用兔、羊、牛血细胞分别测得本公司2批产品的溶血指数（完全溶血浓度）为1／20．8万、1／15．6万、1／15．6万以上;用成年黄牛血细胞测得七叶皂苷钠国家对照品溶血指数为1／10万,略高于德国Reparil（1／20万）。结论：应采用同一来源的一种常用动物--家兔血细胞检测药品体外溶血活性以及在相同条件下检测国内同类品种体外溶血活性不高于国外。     

Optimization of the in vitro release of zomepirac from suspensions and suppositories

In an in vitro study the release rates of zomepirac acid (I) and zomepirac sodium dihydrate (II) from suspensions in liquid paraffin towards an aqueous phase were determined. When calcium was added to the aqueous phase a cake of zomepiraecalcium was formed at the interface paraffin/buffer and the release rates of I and/or II decreased extensively. The influence of additives, i.e. Tween 80, PVP and lecithin, in the suspension on this caking process was determined. It was found that Tween 80 was the most effective additive. The results obtained were ‘transformed’ to the release of II from suppositories (in vitro studies in a modified Paddle set-up). Again Tween 80 was the most effective additive; the influence of lecithin and PVP was negligible.     

Class III antiarrhythmic action linked with positive inotropy: acute electrophysiological and inotropic effects of amiodarone in vitro

Negative inotropy is an adverse feature of most antiarrhythmic drugs. Positive inotropy, however, has been demonstrated for some drugs with class III antiarrhythmic action. Although amiodarone exerts its antiarrhythmic effect by an interplay of different actions on cardiac cells, it has been regarded to be the prototype class III drug due to its prolongation of action potential duration. The present study was designed to test the hypothesis that class III antiarrhythmic action and positive inotropy may be linked. We compared the effects of amiodarone in Cordarone and its solvent Tween 80 on automaticity, refractoriness and inotropy. Two series of experiments were done; one with spontaneously beating rat atria to study the effects on sinus node function, and one with electrically stimulated left atria to study the effects on excitability, refractoriness and inotropy. Amiodarone 1 x 10(-4) M decreased spontaneous heart rate by 13% and prolonged sinus node recovery time by 105%. Without affecting the excitability amiodarone prolonged the effective refractory period by 12%. At the same time contractile force increased by 12%. Lower concentration of amiodarone (5 x 10(-6) M) or Tween 80 had no significant effects. In conclusion, amiodarone exerts acute electrophysiological and inotropic effects in vitro. The class III antiarrhythmic action of amiodarone is linked with positive inotropy.     

The role of various surfactants on the release of salbutamol from suppositories

Salbutamol is a selective beta(2)-adrenoreceptor agonist with different pharmacological effects. In this research because of the simplicity of suppository application in elderly and its higher plasma concentration than tablets as well as its particular indication in premature labour, salbutamol suppositories were prepared. The suppositories were formulated containing 10 mg of the drug and Witepsol H15, the oleaginous soluble base using melting method. To optimize the release rate of drug, different surfactants namely, sodium lauryl sulphate (SLS) as an ionic surfactant and Tween 80 as well as Arlacel 60 as non-ionic surfactants with different HLBs were chosen. The effect of surfactant concentration on the release rate of salbutamol from suppositories were also investigated. All prepared formulations fulfilled the specifications set down in British Pharmacopoeia. The results showed that Tween 80 (2%w/w) and SLS (0.75%w/w) caused an increase in dissolution rate of salbutamol from suppositories. As anionic surfactants, such as SLS, cause greater damage on mucosa than non-ionic surfactant, such as Tween 80, this study recommended that Tween 80 could be added in suppository formulation in order to increase the dissolution rate of salbutamol. It was also shown that the release rate of salbutamol altered linearly with the amount of Tween 80 in suppository formulations.     

Interaction of non-ionic surfactants with hepatic CYP in Prochilodus scrofa

Cytochromes P450 (CYP) constitute a superfamily of hemeproteins that play a vital role in the metabolism of a wide variety of endogenous and xenobiotic compounds. Xenobiotic metabolism and the role of CYP are of particular interest in studies regarding the prevention of the damage caused by chemical pollutants. We investigated, in this study, the interaction of Triton X-100 and Tween 80 with CYP and antioxidant defenses in Curimbatá, a Brazilian fish. Aiming to clarify the effects of non-ionic surfactants in the monooxigenase system of fish through in vitro study, the effects of Triton X-100 and Tween 80 were analyzed using monooxygenases and antioxidant system as experimental model. Total CYP and EROD were strongly inhibited by Triton X-100 and Tween 80 in a concentration-dependent way; the content of CYP was reduced until zero while EROD activity was completely inhibited in the presence of Triton X-100 and more than 40% inhibited in the presence of Tween 80. Each surfactant causes a different effect on each antioxidant enzyme. No effect was detected in SOD activity in the presence of even Triton X-100 or Tween 80. Triton X-100 increase catalase activity, while Tween 80 decreases this enzyme activity. The molecular structure of the surfactants causes the alteration of this system, since they are able to interact with the microsomal protein, especially with monooxigenase's components, altering their conformation and, consequently destroying their function. Our results suggest that surfactants can interact with components of the microsomal system leading to inhibition of CYP. Therefore, CYP activity, which has been used as a biomarker of xenobiotic exposure, should be used as a marker in association with other enzymes.     

Immune hemolytic anemia caused by drugs

INTRODUCTION: Drug-induced immune hemolytic anemia (DIIHA) is a rare cytopenia; about 130 drugs have been incriminated. The antibodies causing DIIHA can be i) drug-independent (drug not needed to be present to detect antibodies in vitro)-DIIHA caused by this type of antibody presents clinically and serologically as an autoimmune hemolytic anemia (AIHA) with red cell (RBC) autoantibodies in patients' sera and in eluates from their RBCs; or (2) drug-dependent (antibodies react in vitro with RBCs only in the presence of drug, on the RBC membrane or when added to the patient's plasma and RBCs). AREAS COVERED: Literature is reviewed regarding pathophysiology of DIIHA (mechanisms; incidence of drugs involved; the clinical, hematological, and serological characteristics of the most common antibodies causing DIIHA). EXPERT OPINION: DIIHA is often poorly investigated and many reports do not provide data to support the diagnosis (i.e., no serology to support an immune etiology). The three most common drugs currently causing DIIHA are piperacillin, cefotetan, and ceftriaxone. All three (especially piperacillin) can cause in vitro and in vivo effects mimicking AIHA, and in transfused patients, hemolytic transfusion reactions. It is important to exclude DIIHA in such patients as the only treatment needed is to discontinue the drug.     

吐温-80对大鼠心血管系统作用的影响

目的 :探讨吐温 -80对心血管系统的影响。方法 :观察不同浓度吐温 -80静脉注射后对大鼠心血管系统的影响。结果 :静脉注射吐温 -80量大时即可出现心血管系统抑制现象 ,小剂量吐温 -80未见对心血管系统产生明显影响。结论 :临床输注含吐温 -80制剂时应控制速度或剂量。