EU,USP及 ChP对干热灭菌/除热原条件对照表 1 / 7
EU,USP及 ChP对干热灭菌/除热原条件对照表
EP 6.0 USP32-NF27 中国药典 2010
Dry heat sterilisation.
(附件 1-2)
2.6.14. BACTERIAL
ENDOTOXINS
(附件1-2)
1211 STERILIZATION
AND STERILITY
ASSURANCE OF
COMPENDIAL ARTICLES
附件2-1
85
BACTERIAL
ENDOTOXINS
TEST
附件 2-2
灭菌方法
内毒素检测方
法
For this method of terminal
sterilisation the reference
conditions are a minimum
of 160 °C for at least 2 h.
Other combinations of time
and temperature may be
used provided that it has
been
satisfactorily demonstrated
that the process chosen
delivers
an adequate and
reproducible level of lethality
when
operated routinely within the
established tolerances. The
Apparatus
Depyrogenate all
glassware and other
heat-stable apparatus
in a hot-air oven using a
validated process. A
commonly used
minimum time and
temperature is 30
minutes at 250 °C. If
employing plastic
apparatus, such as
microtitre plates and
pipette tips for
automatic pipetters, use
A typical acceptable range
in temperature in the empty
chamber is ±15 when the
unit is operating at not less
than 250 .
A microbial survival
probability of 10–12 is
considered achievable for
heat-stable articles or
components. An example
of a biological indicator for
validating and monitoring
dry-heat sterilization is a
preparation of Bacillus
APPARATUS
AND
GLASSWARE
Commonly
used minimum
time and
temperature
settings are 30
minutes at
250 .
干热灭菌条件
一般为 160~
170℃×120min
以上、170~
180℃×60min
以上或 250℃
×45min以上,
也可采用其它
温度和时间参
数。
检测用玻璃器
皿去热原温度
在 250℃,30
分钟以上。
总之,应
保证灭菌后的
产品其
SAL≤10-6。干
热过度杀灭后
产品的SAL应
≤10-12,
EU,USP及 ChP对干热灭菌/除热原条件对照表 2 / 7
procedures and precautions
employed are such as to
give an
SAL of 10− 6 or better.
Dry heat at temperatures
greater than 220 °C is
frequently
used for sterilisation and
depyrogenation of
glassware. In
this case demonstration of a
3-log reduction in heat
resistant endotoxin can be
used as a replacement for
biological
indicators (5.1.2).
apparatus shown
to be free of detectable
endotoxin and of
interfering effects
for the test.
NOTE: In this chapter,
the term ‘tube’ includes
all types of
receptacles, for
example microtitre plate
wells.
subtilis spores. Since dry
heat is frequently employed
to render glassware or
containers free from
pyrogens as well as viable
microbes, a pyrogen
challenge, where
necessary, should be an
integral part of the
validation program, e.g., by
inoculating one or more of
the articles to be treated
with 1000 or more USP
Units of bacterial
endotoxin. The test with
Limulus lysate could be
used to demonstrate that
the endotoxic substance
has been inactivated to not
more than 1/1000 of the
original amount (3 log cycle
reduction).
250℃ 45min
的干热灭菌也
可除去无菌产
品包装容器及
有关生产灌装
用具中的热原
物质。
细菌内毒
素灭活验证试
验是证明除热
原过程有效性
的试验。一般将
不小于 1000单
位的细菌内毒
素加入待去热
原的物品中,证
明该去热原工
艺能使内毒素
至少下降 3个
对数单位。细菌
内毒素灭活验
证试验所用的
细菌内毒素一
般为大肠杆菌
内毒素
EU,USP及 ChP对干热灭菌/除热原条件对照表 3 / 7
( Escherichia
coli
endoxin )。
结论
对去热原温度和时间组合没有明
确要求,对温度下限有要求。
对温度和时间有要求,同
USP,ChP.
对去热原温度和时间组合没有明
确要求,对温度下限有要求
对温度和时间有
要求,同EP,ChP.
对去热原温度
和时间组合有
建议要求,没
有强制要求。
对温度和时间
有 要 求 , 同
EP,USP.
EU,USP 及 ChP 对 干 热 灭 菌 / 除 热 原 条 件 对 照 表
4 / 7
附件 1 -1
EP6.0 5.1.1. METHODS OF PREPARATION
OF STERILE PRODUCTS)
Dry heat sterilisation. For this method of terminal sterilisation the reference conditions
are a minimum
of 160 °C for at least 2 h. Other combinations of time
and temperature may be used provided that it has been
satisfactorily demonstrated that the process chosen delivers
an adequate and reproducible level of lethality when
operated routinely within the established tolerances. The
procedures and precautions employed are such as to give an
SAL of 10− 6 or better.
Dry heat sterilisation is carried out in an oven equipped with
forced air circulation or other equipment specially designed
for the purpose. The steriliser is loaded in such a way that
a uniform temperature is achieved throughout the load.
Knowledge of the temperature within the steriliser during
the sterilisation procedure is usually obtained by means of
temperature-sensing elements inserted into representative
containers together with additional elements at the
previously established coolest part of the loaded steriliser.
The temperature throughout each cycle is suitably recorded.
Where a biological assessment is carried out, this is obtained
using a suitable biological indicator (5.1.2).
Dry heat at temperatures greater than 220 °C is frequently
used for sterilisation and depyrogenation of glassware. In
this case demonstration of a 3-log reduction in heat resistant
endotoxin can be used as a replacement for biological
indicators (5.1.2)
附件 1-2 2.6.14. BACTERIAL ENDOTOXINS
The test for bacterial endotoxins is used to detect or
quantify endotoxins of gram-negative bacterial origin
using amoebocyte lysate from horseshoe crab (Limulus
polyphemus or Tachypleus tridentatus). There are
3 techniques for this test : the gel-clot technique, which
is based on gel formation ; the turbidimetric technique,
based on the development of turbidity after cleavage of an
endogenous substrate ; and the chromogenic technique,
based on the development of colour after cleavage of a
synthetic peptide-chromogen complex.
The following 6 methods are described in the present
EU,USP 及 ChP 对 干 热 灭 菌 / 除 热 原 条 件 对 照 表
5 / 7
chapter :
Method A. Gel-clot method: limit test
Method B. Gel-clot method: semi-quantitative test
Method C. Turbidimetric kinetic method
182 See the information section on general monographs (cover pages)
EUROPEAN PHARMACOPOEIA 6.0 2.6.14. Bacterial endotoxins
Method D. Chromogenic kinetic method
Method E. Chromogenic end-point method
Method F. Turbidimetric end-point method
Proceed by any of the 6 methods for the test. In the event
of doubt or dispute, the final decision is made based upon
method A unless otherwise indicated in the monograph.
The test is carried out in a manner that avoids endotoxin
contamination.
Apparatus
Depyrogenate all glassware and other heat-stable apparatus
in a hot-air oven using a validated process. A commonly used
minimum time and temperature is 30 minutes at 250 °C. If
employing plastic apparatus, such as microtitre plates and
pipette tips for automatic pipetters, use apparatus shown
to be free of detectable endotoxin and of interfering effects
for the test.
NOTE: In this chapter, the term ‘tube’ includes all types of
receptacles, for example microtitre plate wells.
indicators (5.1.2).
附件 2-1
1211 STERILIZATION AND STERILITY ASSURANCE OF
COMPENDIAL ARTICLES
This informational chapter provides a general description of the concepts and
principles involved in the quality control of articles that must be sterile. Any
modifications of or variations in sterility test procedures from those described
under Sterility Tests 71 should be validated in the context of the entire
sterility assurance program and are not intended to be methods alternative to
those described in that chapter.
Within the strictest definition of sterility, a specimen would be deemed sterile
only when there is complete absence of viable microorganisms from it.
However, this absolute definition cannot currently be applied to an entire lot of
finished compendial articles because of limitations in testing. Absolute sterility
cannot be practically demonstrated without complete destruction of every
finished article. The sterility of a lot purported to be sterile is therefore defined
EU,USP 及 ChP 对 干 热 灭 菌 / 除 热 原 条 件 对 照 表
6 / 7
in probabilistic terms, where the likelihood of a contaminated unit or article is
acceptably remote. Such a state of sterility assurance can be established only
through the use of adequate sterilization cycles and subsequent aseptic
processing, if any, under appropriate current good manufacturing practice, and
not by reliance solely on sterility testing. The basic principles for validation and
certification of a sterilizing process are enumerated as follows:
1. Establish that the process equipment has capability of operating within the
required parameters.
2. Demonstrate that the critical control equipment and instrumentation are
capable of operating within the prescribed parameters for the process
equipment.
3. Perform replicate cycles representing the required operational range of the
equipment and employing actual or simulated product. Demonstrate that the
processes have been carried out within the prescribed protocol limits and
finally that the probability of microbial survival in the replicate processes
completed is not greater than the prescribed limits.
4. Monitor the validated process during routine operation. Periodically as needed,
requalify and recertify the equipment.
5. Complete the protocols, and document steps (1) through (4) above.
METHODS OF STERILIZATION
In this informational chapter, five methods of terminal sterilization, including
removal of microorganisms by filtration and guidelines for aseptic processing,
are described. Modern technological developments, however, have led to the
use of additional procedures. These include blow-molding (at high
temperatures), forms of moist heat other than saturated steam and UV
irradiation, as well as on-line continuous filling in aseptic processing. The
choice of the appropriate process for a given dosage form or component
requires a high level of knowledge of sterilization techniques and information
concerning any effects of the process on the material being sterilized.1
Dry-Heat Sterilization
The process of thermal sterilization of Pharmacopeial articles by dry heat is
usually carried out by a batch process in an oven designed expressly for that
purpose. A modern oven is supplied with heated, filtered air, distributed
uniformly throughout the chamber by convection or radiation and employing a
blower system with devices for sensing, monitoring, and controlling the critical
parameters. The validation of a dry-heat sterilization facility is carried out in a
manner similar to that for a steam sterilizer described earlier. Where the unit is
employed for sterilizing components such as containers intended for
intravenous solutions, care should be taken to avoid accumulation of
EU,USP 及 ChP 对 干 热 灭 菌 / 除 热 原 条 件 对 照 表
7 / 7
particulate matter in the chamber. A typical acceptable range in temperature in
the empty chamber is ±15 when the unit is operating at not less than 250 .
In addition to the batch process described above, a continuous process is
frequently employed to sterilize and depyrogenate glassware as part of an
integrated continuous aseptic filling and sealing system. Heat distribution may
be by convection or by direct transfer of heat from an open flame. The
continuous system usually requires a much higher temperature than cited
above for the batch process because of a much shorter dwell time. However,
the total temperature input during the passage of the product should be
equivalent to that achieved during the chamber process. The continuous
process also usually necessitates a rapid cooling stage prior to the aseptic
filling operation. In the qualification and validation program, in view of the short
dwell time, parameters for uniformity of the temperature, and particularly the
dwell time, should be established.
A microbial survival probability of 10–12 is considered achievable for
heat-stable articles or components. An example of a biological indicator for
validating and monitoring dry-heat sterilization is a preparation of Bacillus
subtilis spores. Since dry heat is frequently employed to render glassware or
containers free from pyrogens as well as viable microbes, a pyrogen challenge,
where necessary, should be an integral part of the validation program, e.g., by
inoculating one or more of the articles to be treated with 1000 or more USP
Units of bacterial endotoxin. The test with Limulus lysate could be used to
demonstrate that the endotoxic substance has been inactivated to not more
than 1/1000 of the original amount (3 log cycle reduction). For the test to be
valid, both the original amount and, after acceptable inactivation, the remaining
amount of endotoxin should be measured. For additional information on the
endotoxin assay, see Bacterial Endotoxins Test 85 .
附件 2-2 85 BACTERIAL ENDOTOXINS TEST
APPARATUS AND GLASSWARE
Depyrogenate all glassware and other heat-stable materials in a hot-air oven
using a validated process. 2 Commonly used minimum time and
temperature settings are 30 minutes at 250 . If employing plastic apparatus,
such as microplates and pipet tips for automatic pipetters, use only that which
has been shown to be free of detectable endotoxin and not to interfere with the
test. [note—In this chapter, the term “tube” includes any other receptacle such
as a micro-titer well.]