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Methods Bacterial endotoxin was inspected using the method in specified the Addenda of China Pharmacopoeia( second part ,2005 edition).

方法以不同厂家的鲎试剂对3批华蟾素注射液分别进行干扰试验。

The method of the soluble peptidoglycan isolation by short dose benzylpenicillin sodium and sephadex G-100 was founded; through the SLP reagent kit, the limulus test and the improved LOWRY reagent kit detecting, the prepared soluble peptidoglycan include shorter LPS and lower protein, its purity coefficient reach 93%.

建立并完善了青霉素诱导和Sephadex G-100凝胶柱法制备金黄色葡萄球菌可溶性肽聚糖的方法;经SLP试剂、鲎试验及改良的LOWRY试剂检测,所制备的金黄色葡萄球菌可溶性肽聚糖中LPS和蛋白的污染均较少,纯度达到93%以上; 3。

Isolations of the s.aureus insoluble peptidoglycan:(1) the s.aureus insoluble peptidoglycan was gained by cold, hot and hypersound affection repetat of s.aureus(ATCC25923);(2) the s.aureus insoluble peptidoglycan was gained by 10% trichloracetic acid, 5% trichloracetic acid,NH4HCO3 buffer with trypsin digestion of s.aureus(ATCC25923) in order. 2. Isolations of the s.aureus soluble peptidoglycan and quality control:(1) the s.aureus soluble peptidoglycan was released by short dose benzylpenicillin sodium and purified by sephadex G-100.(2) The quality control in the process of isolation: the soluble peptidoglycan was detected by SLP reagent kit; the LPS was detected by kinetic turbidimetric limulus test;and the protein was detected by improved LOWRY reagent kit.

金黄色葡萄球菌可溶性肽聚糖的分离提取及其质量控制(1)采用青霉素钠诱导法和葡聚糖G-100(Sephadex G-100)凝胶柱分离提取金黄色葡萄球菌可溶性肽聚糖;(2)金黄色葡萄球菌可溶性肽聚糖提取过程中的质量控制:采用SLP试剂盒对提取物定性检测,应用动态浊度法鲎试验测定提取物中内毒素含量,采用改良LOWRY试剂检测提取物中蛋白的含量;(3)采用青霉素诱导和Sephadex G-100凝胶柱分离提取金黄色葡萄球菌可溶性肽聚糖的条件:青霉素使用量、加入青霉素后孵育时间和菌液浓度等优化选择。

The soluble peptidoglycan is gained by short dose benzylpenicillin sodium and sephadex G-100; the SLP reagent kit revealed the prepared soluble peptidoglycan is peptidoglycan; the limulus test showed that the content of LPS in the prepared soluble peptidoglycan(10ng/ml) is lower (0.01ng/ml); the improved LOWRY reagent kit showed the content of protein in the prepared soluble peptidoglycan(1.0mg/ml) was 0.06mg/ml; the optimization condition is 0.75mg/ml of the dosage of benzylpenicillin sodium,1h of the incubation time of benzylpenicillin sodium addition and 5×108CFU/ml of the bacterial concentration.

应用青霉素诱导和Sephadex G-100凝胶柱成功分离得到金葡菌可溶性PGN;所制备的可溶性PGN经SLP试剂检测结果为阳性;动态浊度鲎实验检测制备的可溶性PGN(10ng/ml)中LPS含量小于0.01ng/ml;改良LOWRY试剂检测可溶性PGN(1.0mg/ml)中蛋白含量为0.06mg/ml;青霉素诱导和Sephadex G-100凝胶柱法提取金葡菌可溶性PGN的最佳条件是:青霉素的使用浓度为0.75mg/ml,菌液浓度为5×108CFU/ml,加入青霉素后孵育时间为1h。

Methods: The interference evaluation experiment proves that IVIG exerts strong inhibitions in Bacterial Endotorxin Test. While the interfering factors can't be eliminated by treatments such as dilution and neutralisation, they are liable to be inhibited provided that the IVIG samples are to be diluted by 1:4 with the Dilution Solution Ⅰ.The BET for IVIG preparation diluted by 1:4 with the Dilution Solution Ⅰ is carried out comply with the Chinese Pharmacopoeia(2000 Edition) The lysate with sensitivity of 0.125Eu/ml is used in the test,and the results of the BET are comparable to that of Pyrogen Test carried out in rabbits.

通过干扰评价实验证明IVIG对细菌内毒素检查法有强烈的抑制作用,单纯用简单稀释和调整pH值法无法消除干扰,如果用稀释剂对IVIG作1:4以上的稀释,即可消除样品对该检查法的干扰,利用灵敏度为0.125EU/ml的鲎试剂,将样品用稀释剂稀释4倍,按中国药典(2000版)进行细菌内毒素的检查,结果与家兔热原质试验进行比较。

We collected venous blood of 4 millilitre from all the children,and quantitative determination blood LPS, LBP, mCD14 by limulus reagent kinetic turbimetric assay, flow cytometry technique and ELISA method.

所有SIRS患儿及正常儿童各取静脉血4ml,分别利用鲎试剂动态比浊定量测定法、流式细胞分析技术、酶联免疫试验进行血液内毒素、mCD14、LBP的定量测定。

The test for endotoxins can substitute for the pyrogens test on the condition of the sample using sensitivity 0.25 EU.ml-1 TAL detected and 1∶1 dilution.

甲硝唑葡萄糖注射液经1∶1稀释后,使用灵敏度为0.25 EU.ml-1鲎试剂进行内毒素检查可替代其热原检查。

We tested bacterial endotoxin in the Nao Weizhi Injection with tachypleus amebocyte lysate and compared this method with the rabbit′s method for the pyrogens test mentioned in pharmacopeia.

文摘:用鲎试剂法对脑维治注射液中的细菌内毒素进行检测,并与《中国药典》家兔法检查热原进行对比。

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