-
PA/PH/OMCL (11) 04 - OMCL Guideline on
qualification of HPLC equipment (Annex
1)
OMCL Network of
the Council of Europe
QUALITY MANAGEMENT DOCUMENT
欧洲委员会
OMCL
网络
质量管理文件
PA/PH/OMCL (11) 04
QUALIFICATION OF EQUIPMENT
设备的确认
ANNEX 1:
QUALIFICATION OF HPLC EQUIPMENT
附件
1
：
HPLC
(
高效液相色谱仪
)
设备的确认
Full
document title and reference
文件全名和索引号
Qualification of Equipment
设备的确认
Annex 1:
Qualification of HPLC equipment
PA/PH/OMCL (11) 04
< br>附件
1
：
HPLC
设备的确
认
PA/PH/OMCL (11) 04
Guideline
指南
The present
document was also accepted by EA
as
recommendation document to be used in the
context of Quality Management System
audits of
OMCLs
本文件也被
EA
认可并推荐在
OMCL
的质量管理体系审计中使用。
May 2005
2005
年
5
月
June 2005
200
5
年
6
月
1st July 2011
<
/p>
2011
年
7
月
1
号
This document replaces document
PA/PH/OMCL
(07) 17 DEF
本文件替代文件
PA/PH/OMCL (07) 17
DEF
Document type
文件类型
Legislative
basis
立法基础
Date
of first adoption
首次采用日期
Date of
original entry into force
首次生效日期
Date of
entry into force of revised
document
修订版生效日期
Previous
titles/other references
原文件
名
/
其它索引号
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PA/PH/OMCL (11) 04 - OMCL Guideline on
qualification of HPLC equipment (Annex
1)
Custodian Organisation
监管组织
The present document was elaborated by
the
OMCL Network/ EDQM of the Council
of
Europe
本文件由
OMCL
网络
/
欧
洲
EDQM
制
定。
GEON
Concerned Network
相关网络
ANNEX 1 OF THE OMCL NETWORK
GUIDELINE
“QUALIFICATION OF
EQUIPMENT”
OMCL<
/p>
网络指南“设备的确认”之附件
1
QUALIFICATION OF HPLC
EQUIPMENT
HPLC
设备的确认
Introduction
概述
The present document is the first Annex
of the core document “Qualification of Equipment”,
and it
should be used in combination
with it when planning, performing and documenting
the HPLC
equipment qualification
process.
本文件是核心文件“设备的确认”第
1
个附件，在计划、实施和记录
HPLC<
/p>
仪器的确认过程时，
应将本文件与核心文件一起使用。
The core document
contains the general introduction and the Level I
and II of qualification, common to
all
type of instruments, and the present annex
contains HPLC instrument-related recommendations
on
parameters to be checked and the
corresponding typical acceptance Limits, as well
as practical
examples on the
Methodology that can be used to carry out these
checks.
核心文件包括了简介和第一阶段和第二阶段
的确认，它们适用于所有类型的仪器。
本附件包括
了
HPLC
仪器相关的需要检查的参数和相应典型
的可接受标准的推荐，以及可用于进行这此检查
的实用方法学举例。
When qualifying HPLC
equipment, it should be noted that it is
acceptable to check at Level III and IV
several
of
the
mentioned
parameters
at
the
same
time
in
a
combined
test
procedure
(e.g.
“overall”
system performance
test giving information on peak area precision,
retention time precision, gradient
reproducibility, etc).
在进行
HPLC
设备确认时，应注意也可以对第三阶段和第四阶段几个提到
的参数采用联合检测
程序同时进行检查（例如“全面”系统性能检查，同时给出峰面积精
密度、保留时间精密度、
梯度重复性等的信息）
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PA/PH/OMCL (11) 04 - OMCL Guideline on
qualification of HPLC equipment (Annex
1)
TABLE III
表三
Level III.
Periodic and motivated instrument checks
第三阶段
定期和专门的仪器检查
Examples of requirements for HPLC
instruments and detectors
HPLC
仪器和检测器要求举例
Instrument module
Parameter to be checked
Typical tolerance Limits
仪器模块
需检查的参数
典型误差限度
Solvent delivery system
●
Flow rate
流速
●
±
5 %
●
Proportioning
accuracy and
溶剂输送系统
●
±
2
precision (gradient test)
配比准
确度和精密度（梯度测试）
●
≤
0.2 %
●
Proportioning ripple
配比波纹
Injector
?
Volume precision
进样量精密
?
RS
D
≤
1.0 %
进样器
度
?
Carry-over
残留
?
see Annex I
见附件
1
Autosampler
?
Thermostatting accuracy and
?
±
3
°
C
自动进样器
precision
恒温准确度和精密
度
Oven or cooling device
?
Thermostatting
accuracy
?
±
2 °
C
柱温箱或冷却装置
恒温准确度
UV/DAD detector
?
Linearity
线性
?
r
2
≥
0.999
UV/DAD
检测器
?
Wavelength
accuracy
波长准
?
±
2 nm
确度
Fluorescence
detector
?
Wavelength accuracy
excitation
?
±
3 nm
荧光检测器
波长准确度激发
?
Wavelength
accuracy emission
?
±
3 nm
波长精密度发射
?
Sensitivity
灵敏度
?
±
see Annex I
Electrochemical
?
Accuracy of the signal
信号精
?
see Annex I
见附件
1
detector
电化学检测
密度
器
?
Stability of the signal
信号稳定
?
see Annex I
见附件
1
性
RID detector
?
Signal/Noise ratio
信噪比
?
see Annex I
见附件
1
?
0.1 mV/min
RID
检测器
?
Drift over time
飘移
CD detector
?
Signal/Noise ratio
信噪比
?
see Annex I
见附件
1
CD
检测器
?
Sensitivity
灵敏度
?
see Annex I
见附件
1
?
Drift over time
飘移
?
see Annex I
见附件
1
?
r > 0.999
?
Linearity
线性
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PA/PH/OMCL (11) 04 - OMCL Guideline on
qualification of HPLC equipment (Annex
1)
TABLE IV
表四
Level IV. In-
use instrument checks
第四阶段
在用仪器的检查
Examples
of requirements for HPLC instruments with UV or
DAD detectors
配备
UV
或
DAD
检测器的
HPLC
仪器的要求举例
Typical
tolerance Limit
典型误差限度
Parameter
to be checked
需检查参数
?
System
suitability check for the
According to
Ph. Eur. or MAH dossier or
validated
in-house Method
Method
方法系统适用性检查
根据欧洲药典或
MAH
文件或公司内部
经过验证的方法
?
RSD ≤ 1.5 %
the
main peaks in the test solution)
(unless otherwise prescribed in the
system
峰面积准确度（适用于供试液中主峰
suitability
of the Method, e.g. specific
requirements from Ph. Eur. 2.2.46, API
面积）
monographs or
MA dossiers)
除非在方法
的系统适用性里有描述
，例如
EP2.2.46
特定要求，原料药各论或
MA
文件
?
Retention time
precision
RSD ±
5 %
保留时间精密度
?
Carry-over (by
comparing
≤ 0.2 %
consecutive standard (of the substance
being quantified) and blank
injections)
残留
（通过
比较连续
（进
行定量检测的物质）标准和空白进
样）
?
Signal/Noise ratio (to be applied for
According to Ph. Eur.
related substances test)
根据欧洲药典
信噪比（用于有关物质检测）
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PA/PH/OMCL (11) 04 - OMCL Guideline on
qualification of HPLC equipment (Annex
1)
ANNEX I
附件
1
Level III. Periodic and motivated
instrument checks
第三阶段
定期和专门的仪器检查
This
Annex contains practical examples of tests and
their associated tolerance Limits for several
parameters related to the performance
of the different modules of a HPLC.
本附件包含的是检测
HPLC
仪器的不同模
块性能的一些参数的实际例子及其参数的误差限度。
These examples can be considered by the
OMCLs as possible approaches to perform the Level
III of
the equipment qualification
process: “Periodic and motivated instrument
checks”.
OMCLs
认
为这些例子可以用来执行设备确认过程的阶段
III
“定期和专
门的仪器检测”。
HPLC
SOLVENT DELIVERY SYSTEM
HPLC
溶剂传输系统
The following tests are
proposed for the periodic and motivated check of
the HPLC solvent delivery
system: flow
rate and gradient test.
以下测
试建议用于对
HPLC
溶剂传输系统的周期性主动检查：流速和
梯度检测
FLOW RATE
流速
Materials:
材料
Volumetric flask of 5 or 10 ml
5ml
或
1
0ml
的容量瓶
Calibrated chronometer
校正过的气压计
Settings:
设置
Mobile phase: degassed water
流动相：脱过气的水
No column (open end)*
无柱（开放式）
*
Flow
rate: adjusted between 0.5 and 3.0 ml/min
流速：调整到
0.5
至
3.0ml/min
之间
If high-pressure mixing systems are
installed, this test has to be done on each
solvent channel.
如果有安装高压混合系统，本测试应在每一溶剂通道进行。
* For certain equipment,
e.g. in the case of low flow rates, the check
would be performed by using a
column or
a backpressure regulator.
*
对于某些设备，例如，流速较低时，检测时需要用到一根色谱柱或背压调节器进行检查。
Method
方法
:
Set the flow rate at an
appropriate level and measure the time needed to
fill the volumetric flask up to
the
mark. Record the time needed.
< br>将流速设定在一个合适的水平，测量将容量瓶装满至刻度的时间。记录所用的时间。
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PA/PH/OMCL (11) 04 - OMCL Guideline on
qualification of HPLC equipment (Annex
1)
＝
＝
(
V
×
60)/t
f
................. measured flow rate
[ml/min]
测得的流速
t
................. elapsed
time to fill up to mark [s]
充满至刻度所需的时间
V
............... volume of the
volumetric flask [ml]
容量瓶的体积
D
＝
10<
/p>
0
×
(f
－
F)/F
D
............... deviation [%]
偏差
F
............... adjusted flow rate
[ml/min]
调节后的流速
f
................. measured flow rate
[ml/min]
测量的流速
Li
mits
限度
:
±
5%
GRADIENT COMPOSITION AND RIPPLE
梯度组成和波动
Settings:
设置
Stainless steel capillary e.g. 2000 x
0.12 mm installed instead of a column
不锈钢毛细管，例如
2000X0.12mm
< br>，代替色谱柱安装于仪器
Detection:
UV-Detector adjusted to 265 nm
检测器：紫外检测器调节至
265nm
Mobile phase A: degassed water
流动相
A
：
脱气水
Mobile phase B: degassed
water containing 0.5% acetone
< br>流动相
B
：含
0.5%
的脱气水
Flow rate: 1.0
ml/min
流速：
1.0ml/
分钟
Method
方法
:
The test is carried out in
the following way by using a gradient program
depending on the number of
solvent
channels and the configuration of the system:
根据溶剂通道的数量和系统的配置，使用下列不同系统参数和
梯度进行测试：
A-B
A-B and A-C
A-C,
A-B and B-D
% mobile phase B
time [min]
% mobile phase A
(water)
%
流动相
B
(water-acetone mixture)
时间【分钟】
%
流动相
A
（水）
（水
-
丙酮混合物）
0.0
100
0
0.1
90
10
10
90
10
10.1
50
50
20
50
50
20.1
10
90
30
10
90
30.1
0
100
40
0
100
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PA/PH/OMCL (11) 04 - OMCL Guideline on
qualification of HPLC equipment (Annex
1)
40.1
100
0
Start the test by pumping
water for at least 10 min to equilibrate the
system.
开始测试前，先用水平衡系统至少
10
分钟。
The
zero % value at the start of the test is the
baseline. All steps are measured at the beginning
of the
horizontal part of the line
either by software or manually on the paper print
using a liner. The height of
the 100%
water/acetone mixture is used as the 100% value in
the following calculation.
测试开始时的
0%
值是基线值。所有步骤均在基线的水平部分开
始时由软件或人工测量，并打印
在纸上。
100%
水
/
丙酮混合物的高度在以下计算中作为
100%
值。
＝
100* h/H
%H
............ calculated
composition
计算组成
h
................ height of
the measured line
测量线的高度
H
............... height of
the 100% water/acetone mixture line (mobile phase
B)
100%
水
< br>/
丙酮混合物线的高度（流动相
B
）
＝
%H
－
G
d
................ deviation
偏差
G
............... gradient composition
adjusted [% acetone/water solution mixture =
mobile phase B]
梯度组成调节【
%
丙酮
/
水溶液混合物
=
流动相
B
】
Limits
限度
:
Absolute
deviation:
±
2 of
the adjusted value
绝对偏差：
调整值的±
2
The ripple
of the gradient composition is the percentage of
noise of the 50% line from the gradient
program.
梯度组分的波动为梯度程序噪声基线的
50%
＝
100* N
/h
50
%R
............ ripple
波纹
h
50
............. height of the 50% line
50%
线的高度
N
............... height of
the noise line, measured during 1 minute in the
linear region
噪声线高度，在线
性区域
1
分钟期间测量的结果
Limits
限度
:
≤ 0.2 %
HPLC INJECTOR
HPLC
进样器
Volume precision and carry-
over are the tests proposed for the periodic and
motivated check of the
HPLC injector.
建议在
HPLC
进样器的定期和专门检查中进行容量
(
体积
)
精密度和残留检查。
VOLUME PRECISION AND CARRY-OVER
体积精密度和残留
Solutions:
溶液
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PA/PH/OMCL (11) 04 - OMCL Guideline on
qualification of HPLC equipment (Annex
1)
Solvent A: methanol :
water R, 60 : 40.
溶液
A
：乙醇：水（试剂）
= 60
：
40
Reference solution (a): dissolve 15.0
mg methyl-, ethyl-, and propylparabene in 100.0 ml
of solvent A.
对照液（
a
）：取
15.0mg
尼泊金
M
、尼泊金乙酯和尼泊金丙酯溶于
100.
0ml
的溶剂
A
中
Reference solution (b): Dilute 1.0
ml of reference solution (a) to 10.0 ml of solvent
A.
对照液（
b
< br>）：取
1.0ml
对照液（
a<
/p>
）用溶于
10.0ml
溶剂
A
中
Reference
solution (c): Dilute 1.0 ml of reference solution
(b) to 100.0 ml of solvent A.
< br>对照液（
c
）：取
1.0ml<
/p>
对照液（
b
）用溶于
100.0ml
溶剂
A
中
Settings:
设置
Column: Lichrospher 100 RP8, 5um, 125 x
4 mm, without precolumn
色谱柱：
Lichrospher 100 RP8, 5
um, 125 x 4 mm,
无衬管
Mobile phase: methanol : water = 60 :
40
流动相：乙醇：水
= 60
：
40
Flow rate: 1.0 ml/min
流速：
1.0ml/
分钟
Detection: 254 nm
检测器：
254nm
Injection volume: 20
μ
l
进样体积：
20ul
Method
方法
:
Injection scheme:
进样序列
—
6x reference solution (b)
对照液（
b
）
6
针
—
1x
reference solution (a)
对照液（
a
）
1
针
—
1x solvent A (blank injection 1)
溶剂
A
（空
白
1
）
1
针<
/p>
—
1x reference solution (b)
<
/p>
对照液（
b
）
1
针
—
1x solvent A (blank
injection 2)
溶剂
A
（空白
2
）
1
针
—
1x reference solution (c)
对照液（
c
）
1
针
Limit
限度
s:
Repeatability
of
peak
areas:
The
relative
standard
deviation
of
the
peak
areas
of
all
peaks
in
the
chromatogram obtained
with the reference solution (b) should
be
≤
1.0 %.
峰面积重复性：对照液（
b
）色谱图中所有峰的峰面
积相对标准偏差应≤
1.0 %
Carry-over: The percentage of the peak
area corresponding to propylparabene in the blank
injection 1
does not exceed 0.5% of 10
times the peak area of the propylparabene peak in
the chromatogram
obtained with the
reference solution (b) injected after the blank
injection.
残留：尼泊金丙酯在空白图谱中的峰
面积不超过在空白后进样的对照液（
b
）中尼泊金丙酯的对
应峰面积的
10
倍的
0.5%
The percentage of the
peak area corresponding to propylparabene in the
reference solution (c) is 0.9
–
1.1% of the
peak area of the propylparabene peak in the
chromatogram obtained with the reference
1
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PA/PH/OMCL (11) 04 - OMCL Guideline on
qualification of HPLC equipment (Annex
1)
solution (b) injected
after the blank injection.
在对照液（
c
）中尼泊金丙酯对应的峰面积为空白后进样的对照
液（
b
）中尼泊金丙酯峰面积的
0.9
-1.1%
。
HPLC AUTOSAMPLER
HPLC
自动进样器
Thermostatting accuracy and
precision can be tested in the frame of the
periodic and motivated check
of the
HPLC Autosampler.
对
< br>HPLC
自动进样器的定期和专门检查可以检测温度的准确度和精确度。
THERMOSTATTING
ACCURACY
温度准确度
Materials:
材料
Calibrated temperature probe.
校正过的温度探头
Method
方法
:
Select a temperature along
the operational or required temperature range of
the equipment. Wait until
the system is
equilibrated.
在可操作范围或要求的温度范
围内选择并设定一个温度，等待系统平衡
By means
of the calibrated probe, measure the actual
temperature in the autosampler and compare it to
the selected temperature.
<
/p>
采用校正过的温度探头，测量自动进样器的实际温度，与设定的温度进行比较
Repeat the same procedure at
different pre-selected points covering the
temperature range.
在选定的温度范围内的不同温度点重复相同的步骤
Limit
限度
s:
The actual temperature may
not differ more than
±
3°
C with respect
to the selected temperature.
实际温度与设定的温度相比差值不超过±
3°
C
THERMOSTATTING PRECISION
温度精确度
Materials:
材料
Calibrated temperature probe.
校正过的温度探头。
Method
方法
:
Select a temperature along
the operational or required temperature range of
the equipment. Wait until
the system is
equilibrated.
选择一个操作温度或要求的仪器温度范围，等待系统平衡。
By means of the calibrated probe, make
“n” measurements over a p
re-established
period of time.
Compare the mean of the
“n” measurements to the selected temperature.
采用校正过的温度探头，在预设的时间内进行
n
次测量，比较
n
次测量值的
平均值与设定的温度。
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