4. Biological Tests/Biochemical Tests/Microbial Tests
4.微生物检测
4.01 Bacterial Endotoxins Test
4.01 细菌内毒素检测
This test is harmonized with the European Pharmacopoeia and the U. S. Pharmacopeia.
本测试的EP和USP一致。 防臭地漏
Bacterial Endotoxins Test is a test to detect or quantify bacterial endotoxins of gram黄小蕾电视剧
negative bacterial origin using an amoebocyte lysate prepared from blood corpuscle
extracts of horshoe crab (Limulus polyphemus or Tachypleus tridentatus). There are
two types of techniques for this test: the gel-clot techniques, which are bad on gel
formation by the reaction of the lysate TS with endotoxins, and the photometric
techniques, which are bad on endotoxin-induced optical changes of the lysate TS. The
latter include turbidimetric techniques, which are bad on the change in lysate TS
turbidity during gel formation, and chromogenic techniques, which are bad on the 有那么一棵树
development of color after cleavage of a synthetic peptide-chromogen complex.
细菌内毒素测试是使用鲎试剂对阴性菌所产生的内毒素进行检测或者定量的测试。此种检测有两种方法，一种是凝胶法，即试剂和内毒素发生反应，会使胶体的结构发生改变凝结。另一种是光度测量法，即加入内毒素的试剂会发生光学上的变化。后面一种可以采用浊度或者颜色的的测量方法，即内毒素试剂反应会改变胶体结构发生浊度改变，内毒素的反应会导致颜色上的改变。
Proceed by any one of the techniques for the test. In the event of doubt or dispute,
the final decision is made bad on the gel-clot techniques, unless otherwi indicated.
The test is carried out in a manner that avoids endotoxin contamination.
可以用这两种方法的任意一种进行检测，但是当结果有所怀疑或争议时，除另有规定，结果以凝胶法为准。检测时应避免内毒素的污染。
1. Apparatus
Depyrogenate all glassware and other heat-stable materials in a hot-air oven using a
validated process. Commonly ud minimum time and temperature ttings are 30
minutes at 250°C. If employing plastic apparatus, such as multi-well plates and tips for
micropipettes, u only that which has been shown to be free of detectable endotoxin and
which does not interfere with the test.
1.器皿
使用验证过的规程对玻璃器皿和其他热稳定性的材料用干热法进行去热源处理。通常处理
最短时间为30分钟，最低温度是250℃。对于一些塑料制品，如多孔板和微量进样器的吸头等器具，应使用那些无热源且不会干扰检测的器具。
2. Preparation of Solutions
信息集2.溶液的制备
2.1 Standard Endotoxin Stock Solution
Prepare Standard Endotoxin Stock Solution by dissolving Japane Pharmacopoeia
Reference Standard Endotoxin in water for bacterial endotoxins test (BET). Endotoxin is
expresd in Endotoxin Units (EU). One EU is equal to one International Unit (IU) of
endotoxin.
2.1 内毒素标准储备液
将日本药典对照内毒素标准品溶于水制成内毒素标准储备液。内毒素用内毒素单位（EU）
表示。1EU相当于1IU（国际单位）。
爬楼2.2 Standard Endotoxin Solution
After mixing Standard Endotoxin Stock Solution thoroughly, prepare appropriate rial
dilutions of Standard Endotoxin Solution, using water for BET. U dilutions as soon as
possible to avoid loss of activity by adsorption.
2.2 内毒素标准溶液
将内毒素标准储备液充分混匀后，使用内毒素检测用水进行稀释，制成一系列的内毒素标准溶液的稀释液。稀释后的溶液应尽快使用，以避免因吸附作用失活。
2.3 Sample Solutions
Unless otherwi specified, prepare sample solutions by dissolving or diluting drugs, 危险品运输企业
using water for BET. If necessary, adjust the pH of the solution to be examined so that
the pH of the mixture of the lysate TS and sample solution falls within the specified pH 杭州西湖景点介绍
range for the lysate to be ud. This usually applies to a sample solution with a pH in the
range of 6.0 to 8.0. TSs or solutions ud for adjustment of pH may be prepared using
water for BET, and then stored in containers free of detectable endotoxin. The TSs or
清平乐村居诗意
solutions must be validated to be free of detectable endotoxin and interfering factors.
2.3 样品溶液
除另有规定外，用BET检测用水稀释或溶解样品来制备样品溶液。如有必要，调节待测液的PH使样品液和和使用的检测试剂的混合液与试剂制定的PH一致。通常使用的PH的范围是6.0-8.0，或者用BET用水配制调节样品PH的缓冲溶液，此缓冲溶液应在远离内毒素污染的条件下储存。所用试液或溶液需避免内毒素或其他干扰因子的污染。
3. Determination of Maximum Valid Dilution
The Maximum Valid Dilution (MVD) is the maximum allowable dilution of a sample
solution at which the endotoxin limit can be determined.
3.最大稀释倍数的测定
最大稀释倍数（MDV）是在内毒素能够被检测到的情况下所允许的最大稀释倍数。
Determine the MVD from the following equation:
MVD ＝（Endotoxin limit × Concentration of sample solution）/λ
最大稀释倍数可以通过以下公式计算：
MVD=（内毒素限度x样品液浓度）/λ
Endotoxin limit:
The endotoxin limit for injections, defined on the basis of do, equals K/M, where
K is a minimum pyrogenic do of endotoxin per kg body mass (EU/kg), and M is
equal to the maximum bolus do of product per kg body mass. When the product is
to be injected at frequent intervals or infud continuously, M is the maximum total
do administered in a single hour period.
内毒素限度：注射液的内毒素限度等于K/M，K为每公斤体重最小的内毒素致病量，M指每公斤体重中最大用药量。当产品是连续注射或连续用药时，M是指每小时的最大用药量。
