鲎试验测定注射用头孢雷特中细菌内毒素的方法学研究

目的：检测注射用头孢雷特中的细菌内毒素。方法：供试品干扰试验后,采用凝胶鲎试验对样品中细菌内毒素进行检测。结果：三批注射用头孢雷特浓度为2mg·ml^-1对细菌内毒素的检测无干扰作用,所测样品的头孢雷特细菌内毒素含量均小于0.5EU.mg-1。结论：所建立方法检查注射用头孢雷特中的细菌内毒素可行。     

鲎试验测定复方氨基酸(15)双肽(2)注射液中的细菌内毒素

目的:检测复方氨基酸(15)双肽(2)注射液中的细菌内毒素。方法:供试品干扰试验后,采用凝胶鲎试验对样品中的细菌内毒素进行检测。结果:三批复方氨基酸(15)双肽(2)注射液稀释至28倍时对细菌内毒素的检测无干扰作用,所测样品的细菌内毒素含量均小于7 EU.ml-1。结论:所建立方法检查复方氨基酸(15)双肽(2)注射液中的细菌内毒素可行。     

动态比浊法鲎试验定量分析催化剂的细菌内毒素

目的通过定量检测催化剂的细菌内毒素含量,为控制药品质量及筛选最佳生产工艺提供参考。方法应用动态比浊法及凝胶法鲎试验对催化剂的细菌内毒素进行定量和半定量检测。结果动态比浊法鲎试验定量检测了4批催化剂,其原液(原瓶加5.0mL水)细菌内毒素含量依次为20EumL-1-50EumL-1不等,样品在256倍稀释时其添加内毒素(2.0EumL-1)的回收率在75%-125%之间,此时对鲎试验反应无干扰作用,与凝胶法的检查结果一致。结论动态比浊法鲎试验可用来定量检测催化剂的细菌内毒素含量。     

两种方法测定灯盏细辛注射液中细菌内毒素含量

目的：建立灯盏细辛注射液细菌内毒素的定性和定量检测方法。方法：按照《中国药典》2010年版一部附录细菌内毒素检查法项下凝胶法和动态浊度法对灯盏细辛注射液进行试验。结果：灯盏细辛注射液注射液稀释25倍（凝胶法）和200倍（动态浊度法）后对鲎试剂及内毒素反应无干扰作用。结论：用凝胶法和动态浊度法检测灯盏细辛注射液中细菌内毒素的含量是可行的。     

不同提纯方法对多肽体外中和内毒素作用的影响

目的：体外检测不同方法纯化的粗肽对内毒素中和作用的强弱，为提纯粗肽的方法的筛选提供实验依据。方法：采用鲎试验反应的动态浊度法，对不同方法提纯出的粗肽在体外中和内毒素作用进行检测。结果：3种实验方法中，方法2、3的结果重复性好；6种多肽提纯方法提纯的多肽对细菌内毒素都有不同程度的中和作用，但B组样品2中和内毒素的作用最强。结论：用乙嗪加上TFA处理提纯的多肽对LPS的中和作用最强，适宜在肽提纯生产上采用。     

肾康注射液细菌内毒素检查法的研究

目的建立肾康注射液细菌内毒素检查方法,以期能够替代家兔热原法控制产品的热原,达到快速尧高效尧可靠尧简便的 目的遥方法按叶中国药典2015 年版四部曳通则1143 细菌内毒素检查法进行试验遥采用两个不同厂家的的鲎试剂,对7 个不同 批次的样品开展干扰试验,确定最小不干扰稀释倍数,并对样品进行了细菌内毒素检查遥结果供试品4倍稀释液无干扰作用, 细菌内毒素限值定为1.0 Eu/ml遥结论用细菌内毒素检查法代替热原法是可行的遥     

丹参川芎嗪注射液细菌内毒素检查法研究

目的 建立丹参川芎嗪注射液细菌内毒素检查法。方法 依据《中国药典》2015年版四部（通则1143）细菌内毒素检查法,通过预干扰预试验和干扰试验,确定丹参川芎嗪注射液最大无干扰的浓度,并进行方法学验证。结果 将丹参川芎嗪注射液稀释20倍,对细菌内毒素检测法无干扰作用。结论 丹参川芎嗪注射液内毒素检查法可用于检查丹参川芎嗪注射液。     

双黄连注射液细菌内毒素检查法的应用

目的:建立双黄连注射液的细菌内毒素检查方法.方法:按照《中国药典》2005年版附录细菌内毒素检查法进行试验.结果:双黄连注射液对鲎试剂与内毒素反应无干扰作用.结论:双黄连注射液采用细菌内毒素检查方法可靠,简单可行,可以考虑代替热原检查.     

细菌内毒素定量检测的影响因素分析及对策

细菌内毒素检查法(简称鲎试验)是利用鲎血变形细胞制成的鲎试剂与微量内毒素产生凝集反应的现象,作为判断样品中细菌内毒素含量是否符合规定的一种方法。其基本原理是利用微量内毒素激活、活化鲎试剂中的C因子和B因子,使凝固酶原转化为凝固酶,成为凝胶样的凝固蛋白,具有简便、迅速、特异性高及灵敏度高等优点。中国药典1995年板已收载,并对鲎试剂、内毒素标准、试验程序、判断标准等作了规定。 我科引进的EDS-99细菌内毒素检测系统可进行动态比浊法和显色基质法检测内毒素,分别利用细菌内毒素与鲎试剂形成凝胶过程中的浊度变化以及反应过程中产生的凝固酶使特     

医用透明质酸钠细菌内毒素检查方法的研究

本实验是为研究透明质酸钠的细菌内毒素检查方法。方法：实验中使用鲎试剂的灵敏度 为０．０３ＥＵ／ｍｌ,进行干扰试验和抗干扰试验,并对其进行了热原检查。结果：在干扰试验中,样品 表现为增强性干扰,增大样品稀释倍数不能排除干扰。添加抗增液则可有效排除干扰。根据参考 文献和热原检查的兔剂量,我们认为透明质酸钠的细菌内毒素限值应定为５ＥＵ／ｍｌ以内。     

Hemostatic safety of the bolus intravenous injection of a novel medium-chain triglyceride:fish oil emulsion

The bolus intravenous injection of a novel 8:2 medium-chain triglyceride:fish oil emulsion was recently found to increase within 60 min and for the subsequent 24-48 h the long-chain polyunsaturated omega3 fatty acid content of both leucocyte and platelet phospholipids in 12 normal subjects. The aim of the present report is to document the hemostatic safety of such a procedure in the same 12 subjects. No adverse effect was found when comparing the results obtained after administration of either the fish oil-containing emulsion or a control 5:5 medium-chain triglyceride:soybean triglyceride emulsion, whether in terms of the occlusion time in either an ADP or epinephrine test or in terms of the [CD]42b, [CD]62p, fibrinogen and PAC-1 response to ADP, collagen or thrombin receptor analog peptide 6 in platelets examined by fluorescence activated cell sorting. In conclusion, this novel procedure for the rapid enrichment of cell phospholipid in long-chain polyunsaturated omega3 fatty acids presents the required safety in a hemostatic perspective.     

Rapid changes in liver lipid composition and pancreatic islet K+ handling and secretory behaviour provoked by the intravenous administration of a medium-chain triglyceride: fish oil emulsion to long-chain polyunsaturated omega3 fatty acid-depleted rats

Second generation rats depleted in long-chain polyunsaturated omega3 fatty acids are currently used as an animal model for the insufficient dietary supply of such fatty acids often prevailing in Western populations. The present study deals mainly with the effects of a novel medium-chain triglyceride: fish oil emulsion (MCT:FO), as compared to a control medium-chain triglyceride:olive oil emulsion (MCT: OO), administered as an intravenous bolus to the omega3-depleted rats 60-120 min before sacrifice upon selected biochemical and biophysical variables. The major findings consisted of a severe decrease of the omega3 fatty acid content of liver lipids in non-injected omega3-depleted rats and its partial correction after injection of the MCT:FO emulsion. The omega3-depleted rats also displayed liver steatosis, increased incorporation of long-chain polyunsaturated omega6 fatty acids in liver phospholipids and increased activity of liver Delta9-desaturase. As judged from the effects of ouabain upon 86Rb net uptake by isolated pancreatic islets, the activity of Na+,K+-ATPase was virtually abolished in the omega3-depleted rats. The latter defect was corrected by prior intravenous injection of the MCT:FO emulsion, this coinciding with suppression of the excessive secretory response to a number of insulin secretagogues otherwise observed in the islets of omega3-depleted rats injected or not with the MCT:OO emulsion.     

Preclinical investigations of a medium-chain triglyceride:fish oil emulsion. I. Effects of bovine milk lipoprotein lipase on lipid composition

The bolus intravenous injection of a novel medium-chain triglyceride:fish oil emulsion (MCT:FO, 8:2, w:w) was recently found to increase within 60 min the leucocyte and platelet phospholipid content of long-chain polyunsaturated omega3 fatty acids. The present report deals with the effects of bovine milk lipoprotein lipase on the lipid composition of this emulsion. The results are compared to those obtained with either a pure fish oil emulsion or a medium-chain triglyceride: long-chain triglyceride:fish oil emulsion (MLF, 5:4:1). Emphasis is placed on i) differences in the fate of distinct fatty acids initially present in the triglycerides, di glycerides and phospholipids, ii) the generation of unesterified fatty acids relative to their initial content in each emulsion, and iii) the time course for these various events. The comparison between the three emulsions under consideration also provides information relevant to their respective sensitivity to lipoprotein lipase and suitability in terms of the generation of distinct unesterified fatty acids, including long-chain polyunsaturated omega3 fatty acids. Furthermore, attention is drawn to the greater efficiency for the hydrolysis of fatty acids from diglycerides as compared to triglycerides and a transient increase in the paired C8:0/C10:0 ratio in the diglycerides generated from the MCT:FO or MLF emulsion. The present study thus affords novel information relevant to the possible use of the MCT:FO emulsion in human subjects.     

Perturbation and age-related changes in the fatty acid pattern of soleus muscle phospholipids and triglycerides in rats depleted in long-chain polyunsaturated omega3 fatty acids

Altered D-glucose metabolism prevails in the soleus muscle of rats depleted in long-chain polyunsaturated omega3 fatty acids (omega3). In these animals, the prior intravenous injection of an omega3-rich medium-chain triglyceride:fish oil emulsion (omega3-FO rats), as compared to that of an omega3-poor medium-chain triglyceride:olive oil emulsion (omega3-OO rats), may either correct or aggravate selected metabolic variables. This study deals with the fatty acid pattern of soleus phospholipids and triglycerides in control animals versus omega3-depleted rats not injected with any lipid emulsion (omega3-NI rats) and in omega3-OO versus omega3-FO rats. In each group of omega3-depleted rats, age-related changes were also monitored. The omega3-depleted rats displayed low long-chain polyunsaturated omega3 fatty acid content, facilitated metabolism of long-chain polyunsaturated omega6 fatty acids, and increased Delta9-desaturase activity. Both the age-related changes in lipid variables and those attributable to the prior intravenous injection of the omega3-rich lipid emulsion consisted either in a move towards normalization or in the opposite direction, i.e. towards aggravation of the defect found in the omega3-depleted rats. Emphasis is placed, therefore, on the unusual situation found in the soleus muscle of omega3-depleted rats, in which both lipid and metabolic variables may be either favourably or adversely affected by the same environmental factor(s).     

Sustained enrichment of liver phospholipids and triglycerides in eicosapentaenoate after a bolus intravenous injection of a medium-chain triglycerides:fish oil emulsion to streptozotocin (Type 1) and Goto-Kakizaki (Type 2) diabetic rats

This study deals with the sustained enrichment of liver phospholipids and triglycerides in long-chain polyunsaturated omega3 fatty acids (omega3) found after the bolus intravenous injection of a novel medium-chain triglyceride:fish oil emulsion (MCT:FO) to streptozotocin (Type 1) and Goto-Kakizaki (Type 2) diabetic rats. Twenty hours after injection of the MCT:FO emulsion, the relative concentration of omega3 was indeed higher in liver phospholipids and triglycerides than that found in rats injected with either saline or a control medium-chain triglyceride:long-chain triglyceride emulsion. This coincided with a decrease in the ponderal percentage of C18:3omega3, C20:4omega6 and/or C22:4omega6 in liver triglycerides. The present study further documents differences between streptozotocin-induced and Goto-Kakizaki diabetic rats in terms of body weight, glycemia, liver triglyceride content and the fatty acid pattern of both liver phospholipids and triglycerides, as well as a close correlation in the latter animals between liver and plasma phospholipids or triglycerides as far as the ratio in the relative concentration of selected fatty acids representative of desaturase and elongase activities is concerned. In light of these and previous findings, it is proposed that the beneficial metabolic and functional events of the MCT:FO emulsion may display not solely a rapid but also sustained time course.     

Protection of aortic endothelial function in both normal and diabetic rats by intravenous bolus injection of a medium-chain triglyceride: fish oil emulsion

The bolus intravenous injection of a novel medium-chain triglyceride:fish oil emulsion (MCT:FO) was recently proposed as a tool to provoke a rapid and sustained increase of cell phospholipid content in long-chain polyunsaturated omega3 fatty acids, for instance in selected subjects prior to anesthesia and surgery. In this study, therefore, the possible protective effect of MCT:FO upon aortic endothelial function was investigated in both normal and diabetic rats. The animals were injected intravenously 20 h before sacrifice with 1.0 ml of either saline, MCT:FO or a control medium-chain triglyceride:long-chain triglyceride emulsion. The vasomotor response of isolated aortic rings was then explored by assessing the relaxation provoked by increasing concentrations of acetylcholine in rings contracted with phenylephrine. Such measurements were performed before and after exposure of the aortic rings to suitable concentrations of oxidized LDL. In both normal and diabetic rats, the prior injection of the MCT:FO emulsion protected the aortic rings against the deleterious effect of oxidized LDL. In the diabetic rats, a beneficial effect of the MCT:FO emulsion was even observed prior to exposure of the aortic rings to oxidized LDL. These findings support the view that this novel procedure is indeed appropriate to protect endothelial function against oxidative stress.     

Preclinical investigations of a medium-chain triglyceride:fish oil emulsion III. Experiments in cultured endothelial cells

The bolus intravenous injection of a novel medium-chain triglyceride:fish oil emulsion was recently reported to provoke a rapid and sustained increase in the cell phospholipid content of long-chain polyunsaturated omega3 fatty acids in both rats and human subjects. This report deals mainly with a comparison between this and other emulsions, as well as albumin-bound omega3 fatty acids, in terms of the time course, reversibility and concentration dependency for the incorporation of the omega3 fatty acids in the phospholipids of cultured endothelial cells. The results document that the new emulsion is quite efficient for a rapid and sustained enrichment of phospholipids in long-chain polyunsaturated omega3 fatty acids. The potential beneficial effects of such an enrichment in terms of aortic endothelial and cardiac function are emphasized.