甘草酸二胺注射液细菌内毒素检查方法的研究

目的建立甘草酸二胺注射液细菌内毒素的检查方法。方法采用中国药典2005年版二部附录收载的细菌内毒素检查法。结果甘草酸二胺注射液稀释40倍时，用灵敏度为0．125EU·mL^-1的鲎试剂检测细菌内毒素，无干扰作用。结论甘草酸二胺注射液细菌内毒素限值确定为1EU·mL^-1，可用细菌内毒素检查法控制其热原。     

硫酸软骨素注射液细菌内毒素检查方法研究

目的建立检查硫酸软骨素注射液细菌内毒素的方法。方法用中国药典2005年版附录细菌内毒素检查法。结果硫酸软骨素注射液浓度0.025mg/mL,用灵敏度0.125EU/mL的鲎试剂检测细菌内毒素无干扰因素的影响。结论硫酸软骨素注射液可用鲎试剂检查细菌内毒素。     

硫酸软骨素A钠注射液细菌内毒素检查方法研究

目的建立检查硫酸软骨素A钠注射液细菌内毒素的方法。方法采用中国药典2005年版附录细菌内毒素检查法。结果硫酸软骨素A钠注射液浓度为0.15 mg.mL-1,用灵敏度为0.25 EU.mL-1的鲎试剂检测细菌内毒素,无干扰因素的影响。结论硫酸软骨素A钠注射液可用鲎试剂检查细菌内毒素。     

硫酸软骨素A钠注射液细菌内毒素检查方法研究

目的建立检查硫酸软骨素A钠注射液细菌内毒素的方法。方法采用中国药典2005年版附录细菌内毒素检查法。结果硫酸软骨素A钠注射液浓度为0．15mg·mL^-1,用灵敏度为0．25EU·mL^-1的鲎试剂检测细菌内毒素,无干扰因素的影响。结论硫酸软骨素A钠注射液可用鲎试剂检查细菌内毒素。     

硫酸软骨素注射液细菌内毒素检查方法研究

目的 建立检查硫酸软骨素注射液细菌内毒素的方法.方法 用中国药典2005年版附录细菌内毒素检查法.结果 硫酸软骨素注射液浓度0.025 mg/mL,用灵敏度0.125 EU/mL的鲎试剂检测细菌内毒素无干扰因素的影响.结论 硫酸软骨素注射液可用鲎试剂检查细菌内毒素.     

马来酸桂哌齐特注射液细菌内毒素检查方法学研究

目的建立马来酸桂哌齐特注射液的细菌内毒素检查方法。方法按照《中国药典》2010年版(二部)附录细菌内毒素检查法,用2个不同生产厂家的鲎试剂(TAL)对3批马来酸桂哌齐特注射液进行干扰试验和细菌内毒素检查。结果样品的细菌内毒素限值确定为1.0 EU/mg,供试品溶液低于或等于0.25 mg/mL时,对TAL与细菌内毒素之间的凝集反应无干扰抑制作用。结论建立的细菌内毒素检查方法可行,可用于马来酸桂哌齐特注射液的检查。     

门冬氨酸细菌内毒素检查法的研究

为建立注射用L-门冬氨酸细菌内毒素检查法,按中国药典2005年版附录86"细菌内毒素检查"进行实验和结果判断。通过加入适量1mol/L NaOH溶液和1mol/L HCl溶液调节L-门冬氨酸溶液pH值的方法,L-门冬氨酸(C=1.25%)对东方鲎试剂无干扰,检测细菌内毒素的东方鲎试剂灵敏度为0.25EU/ml。结论为注射用L-门冬氨酸可用灵敏度为0.25EU/ml的东方鲎试剂检查其细菌内毒素用以代替家兔法检查热原。     

注射用硫酸萘替米星细菌内毒素检查法的研究

目的　建立注射用硫酸萘替米星细菌内毒素检查法。方法　对浓度为1500、750、375u ml的硫酸萘替米星溶液进行了干扰试验。结果　浓度为375u ml的硫酸萘替米星溶液对细菌内毒素检查法无干扰作用。结论　注射用硫酸萘替米星细菌内毒素检查法是可行的。     

缬氨酸细菌内毒素检测使用LAL和TAL结果差异分析

分析了不同鲎试剂对缬氨酸细菌内毒素检测结果的差异以及找出它们差异的根本原因,得出结论:缬氨酸样品确实存在对不同鲎试剂反应差异的现象。     

活性炭吸附脱除大豆磷脂细菌内毒素的研究

采用活性炭吸附去除大豆磷脂中的内毒素.以磷脂残留的内毒素浓度和磷脂回收率为指标,考察了活性炭用量、吸附温度、吸附时间对内毒素脱除效果的影响.在单因素试验的基础上,通过正交试验,得到最佳工艺参数:活性炭用量0.6％,吸附时间40 min,吸附温度50℃.在最佳条件下,测得磷脂中细菌内毒素的浓度为11.02×10-3 EU/mL,回收率为89.65％.     

中国药典2005年版二部与2000年版二部细菌内毒素检查法的比较

对比中国药典2005年版二部与2000年版二部中有关细菌内毒素检查法的不同,归纳出新增和修改的内容。     

Detection of bacterial endotoxin in food: New planar interdigital sensors based approach

Food poisoning caused by endotoxins or Lipopolysaccharide (LPS) are associated with Gram-negative bacteria. Two major food-borne pathogens, Escherichia coli and Salmonella are examples of Gram-negative bacteria which cause a large number of outbreaks of food poisoning. New types of planar interdigital sensors have been fabricated with different coating materials to assess their response to endotoxins. A carboxyl-functional polymer, APTES (3-Aminopropyltriethoxysilane) and Thionine were chosen to be coated onto FR4 interdigital sensors. The chosen coating materials have carboxylic or amine functional groups, which were optimized to be stable in water. All coated sensors were immobilized with PmB (Polymyxin B) which has specific binding properties to LPS. The sensors were tested with different concentrations of LPS O111:B4, ranging from 0.1 to 1000 μg/ml. Analyses of sensors’ performance were based on the impedance spectroscopy method. The impedance spectra were modeled using a constant phase-element (CPE) equivalent circuit, and a principal component analysis (PCA) was used for data classification. Sensor coated with APTES has shown better selectivity for LPS detection. The experiments were repeated by coating APTES and immobilizing PmB to a new improve designed of novel interdigital sensors (thin film silicon based sensors). These sensors were observed to have better sensitivity and selectivity to the target biomolecules of LPS. Further experiments were conducted to study the effect of different coating thickness on sensor sensitivity, selectivity and stability. Different food samples contaminated with endotoxin were also tested to verify that the interdigital sensing approach is able to be used for endotoxin detection.     

Development of anti-bovine TNF-alpha mAb and ELISA for quantitating TNF-alpha in milk after intramammary injection of endotoxin

Murine mAb reactive with recombinant bovine tumor necrosis factor-alpha (r-boTNF-alpha) were produced. An ELISA using murine mAb and rabbit polyclonal antibodies, each reactive with r-boTNF-alpha to sandwich bovine TNF-alpha was developed. Secretion of TNF-alpha in quarter milk increased 1 h after injection of 0.1 mg (four cows) or 0.5 mg (four cows) Escherichia coli lipopolysaccharide (LPS) into a mammary quarter, peaked 1 to 5 h later, and returned to control levels in 24 h. There were no differences in body temperature, SCC, TNF-alpha, and blood leukocyte responses between 0.1 and 0.5 mg of LPS. To determine effects of repeated injections of LPS into the same udder, a second injection of 0.1 mg of LPS into the same quarter (two cows) 24 h after the first injection produced a strongly attenuated TNF-alpha response. However, a normal TNF-alpha response was observed when LPS was injected into a contralateral quarter (two cows) 24 h after the first LPS injection. Leukocyte counts in blood decreased and body temperature increased substantially after each injection of LPS. Quarter milk SCC increased 200-fold 8 to 12 h after the LPS injections. It would appear that these changes were not regulated by TNF-alpha secretion because the changes were also similar after the second injection of LPS into the same mammary quarter.     

The use of bacterial alkaline phosphatase assay for rapid monitoring of bacterial counts on spinach

ABSTRACT:  This study was undertaken to evaluate the applicability of bacterial alkaline phosphatase (ALP) activity for rapid monitoring of total mesophilic bacteria counts in spinach. A set of fresh and decayed spinach mixtures were tested to rapidly (10 min) monitor spinach bacterial counts. To assay ALP activity, Lumigen APS-5 was used as a substrate. Bovine ALP was reduced after heat treatment at 75 °C for 1 min; in contrast, bacterial ALP activity increased. To differentiate bacterial ALP from other ALP, heat treatment (75 °C, 1 min) was applied before measurement. As a result, a regression equation was established between the actual mesophilic aerobic bacteria count and ALP activity of spinach mixtures ( r = 0.90). The predicted total mesophilic aerobic bacterial count calculated from the fitted regression line (predicted log₁₀ CFU/g = 0.00056 × ALP values + 1.4002) showed a high correlation with the actual observed total bacterial count ( r = 0.93). The ALP assay is a simple and rapid method to utilize for estimation of existing or contaminating microorganism levels on spinach.     

菌落总数测试卡用凝胶培养基和显色剂的优化

筛选适宜的冷水溶解凝胶,优化凝胶培养基成分和显色剂浓度,制备菌落总数测试卡并与倾注培养法进行菌落总数检测比较。结果显示:卡拉胶和CMC以2∶1的质量比混合、复合凝胶粉和培养基粉以2∶1的质量比混合、TTC实际浓度为0.025mg/mL时,制备的菌落总数测试卡具有较好的检测效果,与倾注法比较无显著差异。说明该方案优化的凝胶与显色剂配方可用于制备菌落总数测试卡。     

The triadic preference test

A triadic preference test was developed as an alternative to the paired preference test. The three stimuli in the test comprised a putatively identical placebo pair and a different stimulus. This was in contrast to the regular paired preference test that utilizes a placebo pair. Such a test requires the presentation of two pairs of stimuli: a putatively identical placebo pair and a test pair. The triadic preference test only requires one triad. With the regular test, the majority of consumers respond to the placebo pair with a preference response. It is generally assumed that these consumers are responding to extraneous factors: those factors that elicit a preference response that are different from the sensory attributes of the food under assessment. As an attempt to minimize the possibility of responses to extraneous factors when assessing the test pair, it has been suggested to only use those consumers who chose the ‘No Preference’ option for the placebo pair. However, this form of ‘screening’ is not viable because the resulting ‘screened’ sample size is greatly reduced to approximately one third. However, in the present study, with the triadic preference test, the resulting ‘screened’ sample size ranged 76.5–94% of the total. Thus, this form of ‘screening’ against consumers who demonstrated response to extraneous factors for the placebo pair, was now feasible.     

应用全自动核糖体RNA基因指纹技术快速鉴定细菌菌种

应用RiboPrinter全自动核糖体RNA基因指纹技术对4株未知菌种进行快速鉴定,并与生理生化试验结合16SrDNA序列测定方法相比较。结果表明,两种方法均成功地完成了4株未知菌种的鉴定,其结果分别为大肠杆菌、嗜热链球菌、地衣芽孢杆菌和枯草芽孢杆菌。相比之下,生理生化试验结合16SrDNA序列测定的方法经济实用,而RiboPrinter系统更为简便、快速,在8h内快速地完成了4株未知菌种的鉴定,并且为菌株的基因分型以及溯源分析奠定了基础。