近日，EDQM OMCL发布了《分析用色谱柱的确认》指南，该文件概述了分析色谱柱确认的一般原则。色谱柱被认为是设备，作为色谱系统的重要组成部分，它们会影响分析结果。因此，OMCL 认为必要时应进行色谱柱的确认。

 

文件介绍了液相色谱（LC、HPLC、UHPLC）和气相色谱（GC） 程序中使用的分析色谱柱的确认要求和一般标准。

 

OMCL表示在计划、执行和记录确认过程时，在收到色谱柱时以及在色谱柱的生命周期中，都将其用作指导。

 

该文件现已翻译完成，如下：

 

QUALIFICATION OF ANALYTICAL COLUMNS

 

分析用色谱柱的确认

 

Note: Mandatory requirements in this guideline and its annexes are defined using the terms “shall” or “must”. The use of “should” indicates a recommendation. For these parts of the text other appropriately justified approaches are acceptable. The term “can” indicates a possibility or an example w ith non-binding character.

 

注：本指南及其附件中的强制性要求使用术语“应”或“必须”进行定义。使用“应该”表示推荐。对于本文件中的要求，也可以接受其他适当合理的方法。术语 “可以” 表示具有非约束性特征的可能性或示例。

 

Contents 

 

目录

 

1. INTRODUCTION

 

介绍

 

2. SCOPE

 

范围

 

3. GLOSSARY AND DEFINITIONS

 

术语和定义

 

4. CONSIDERATIONS FOR QUALIFICATION UPON PURCHASING

 

采购的确认考虑

 

5. INITIAL QUALIFICATION

 

首次确认

 

5.1 Qualification parameters

 

确认参数

 

6. PERIODICAL QUALIFICATION

 

定期确认

 

 6.1 Qualification procedure

 

确认程序

 

 6.2 Frequency of qualification

 

确认频率

 

 6.3 Qualification records

 

确认记录

 

7. IN-USE QUALIFICATION

 

使用过程中的确认

 

8. PROCEDURES FOR QUALIFICATION OF ANALYTICAL COLUMNS

 

色谱柱的确认程序

 

9. REFERENCES

 

参考文献

 

APPENDIX 1: EXAMPLES FOR QUALIFICATION OF ANALYTICAL COLUMNS FOR LC

 

附录1：液相色谱柱确认示例

 

APPENDIX 2 EXAMPLES FOR THE QUALIFICATION OF ANALYTICAL COLUMNS FOR GAS CHROMATOGRAPHY

 

附录 2 ：气相色谱柱确认示例

 

1.INTRODUCTION

 

介绍

 

The present document outlines the general principles for the qualification of analytical columns used within OMCLs. Analytical columns are considered to be equipment, and as an essential part of the chromatographic system (Ph. Eur. Chapters 2.2.28, 2.2.29), they can influence the results obtained (ISO/IEC 17025:2017, Chapter 6.4 Equipment, point 6.4.1). For this reason, the qualification of analytical columns should be carried out whenever considered necessary by the OMCL.

 

本文件概述了 OMCL 中使用的分析用色谱柱确认的一般原则。色谱柱被认为是设备，作为色谱系统的重要组成部分，它们会影响分析结果（ISO/IEC 17025：2017，第 6.4 章设备，第 6.4.1 点）。因此，OMCL 认为必要时应进行色谱柱的确认。

 

The OMCL is responsible for defining a suitable qualification scheme for use during the life cycle of the column, depending, for example, on the intended purpose and the frequency of use.

 

OMCL 负责确定在色谱柱生命周期内使用的合适确认方案，例如，根据预期目的和使用频率。

 

2.SCOPE

 

范围

 

This document describes the requirements and general criteria for the qualification of analytical columns used in Liquid Chromatography (LC, UHPLC) and Gas Chromato-graphy (GC) procedures.

 

本文件介绍了液相色谱（LC、UHPLC）和气相色谱（GC）程序中使用的分析用色谱柱的确认要求和一般标准。

 

It can be used as guidance by OMCLs when planning, performing and documenting the qualification process, upon receipt and during the life cycle of analytical columns.

 

OMCL 在规划、执行和记录确认过程时，在收到色谱柱时以及在色谱柱的生命周期中，都可以将其用作指导。

 

Exemplary procedures provided in Annexes 1 and 2 have non-binding character. They can be helpful when carrying out the required qualification. Nevertheless, it is left to the professional judgment and background experience of each OMCL to apply the most suitable procedure, in order to prove that the analytical columns are suitable for their intended use.

 

附录 1 和 2 中提供的示例程序具有非约束性。在执行所需的确认时，他们会有所帮助。然而，每个 OMCL 的专业判断和背景经验都有责任应用最合适的程序，以证明分析用色谱柱适合其预期用途。

 

3. GLOSSARY AND DEFINITIONS

 

术语

 

The terminology used in this document is consistent with that used in Ph. Eur. Chapter 2.2.46 “Chromatographic separation techniques”.

 

本文件中使用的术语与欧洲药典第2.2.46 章“色谱分离技术”中使用的术语一致。

 

4.CONSIDERATIONS FOR QUALIFICATION UPON PURCHASING

 

采购确认的注意事项

 

Incoming columns shall be verified for physical damage due to shipping, for compliance with the purchase order and for completeness of the documentation.

 

应确认到货色谱柱是否因运输而造成物理损坏、是否符合采购订单以及文件的完整性。

 

The OMCL shall retain records related to the management of columns for the applicable requirements described in ISO/IEC 17025:2017 clause 6.4.13.

 

应根据 ISO/IEC 17025：2017第 6.4.13 条中描述的适用要求保留与色谱柱的管理相关的记录。

 

5.INITIAL QUALIFICATION

 

初始确认

 

The analytical column shall be verified upon receipt or before the first use by checking the documentation (e.g. certificate of analysis) or by performing qualification of the column according to a pre-defined procedure.

 

分析用色谱柱应在收到时或首次使用前通过检查文件（例如COA）或根据预定的程序对色谱柱进行确认。

 

Verification/qualification shall be documented (including, e.g. date of verification/qualification, reference to the procedure for qualification, reference materials or reagents used, results, chromatographic equipment used, pass/fail conclusion, etc.). This record can be used to make a complaint in cases where the column does not comply with the manufacturer’s specification.

 

验证/确认应有记录（包括验证/确认日期、确认程序引用、使用的标准物质或试剂、结果、使用的色谱设备、通过/失败结论等）。此记录可用于在色谱柱不符合制造商标准时进行投诉。

 

5.1 Qualification parameters

 

确认参数

 

It is the responsibility of the OMCL to choose the most appropriate procedure, parameters and acceptance criteria. These should be appropriate to the intended use of the column. These parameters can include resolution, symmetry factor, theoretical plates, etc. Some typical limits are given as examples in Annexes 1 and 2.

 

OMCL 有责任选择最合适的程序、参数和接受标准。它们应适合色谱柱的预期用途。这些参数可以包括分离度、对称因子、理论踏板等。附件1 和附件 2 中给出了一些典型的限度标准作为示例。

 

6. PERIODICAL QUALIFICATION

 

定期确认

 

The qualification of the columns can be performed periodically using the same test compounds/procedure. A systematic record of the qualification results should be maintained in the laboratory.

 

可以使用相同的测试化合物/程序定期进行色谱柱的确认。确认结果的系统记录应在实验室中保存。

 

6.1 Qualification procedure

 

确认程序

 

At least one of the procedures chosen for initial qualification can be applied for periodical qualification. The first chromatogram obtained with a new column in the laboratory can serve as a reference of column performance at the initial time point and used as a reference for subsequent verifications. Slight variations can be observed on chromatograms due to different LC or GC configuration of the equipment (e.g. dead volumes), operating environment, system electronics, etc. These can be taken into consideration whenever the acceptance criteria are set in-house.

 

至少可用初始确认的程序进行定期确认。在实验室中使用新色谱柱获得的第一张色谱图可以作为该色谱柱初始性能的参考，并作为后续验证的参考。由于不同 LC 或GC 的设备配置（例如死体积）、运行环境、系统电子元件等，色谱图上可能会观察到细微的不同。当设定内控接受标准时，可以考虑这些因素。

 

6.2 Frequency of qualification

 

确认频率

 

The laboratory can define and document the frequency of qualification whenever periodic qualification is considered appropriate and useful. The frequency depends on the use of the column (e.g. routine or “spot” analysis). For instance, a qualification according to section 5 (above) could be performed at fixed time intervals (e.g. once a year) or depending on the extent of use, e.g. after a defined number of injections.

 

当认为定期确认适当和有用时，实验室可以定义和记录确认频率。频率取决于色谱柱的使用情况（例如日常或“点”分析）。例如，根据第5 节（上文）的确认可以按固定的时间间隔（例如每年一次）进行，也可以根据使用程度进行，例如在规定的进样次数后进行。

 

6.3 Qualification records

 

确认记录

 

A record of the qualification results can help the user to monitor the column performance over time and inform the decision to discard it. A column is considered suitable for the intended use when the acceptance/system suitability criteria are met.

 

确认结果的记录可以帮助用户监测色谱柱性能随时间的变化，并告知是否需要丢弃该色谱柱。当符合接受/系统适用性标准时，色谱柱被认为适合预期用途。

 

7. IN-USE QUALIFICATION

 

使用过程中的确认

 

In-use qualification corresponds to the system suitability test for the method used. It shall be performed on a regular basis before each test and/or during the sequence of injections. In cases where the system suitability criteria are not met, the qualification of the column can be performed, to verify the consistency with the initial performance.

 

使用过程中的确认对应于所用方法的系统适用性测试。应在每次测试前和/或在序列进样期间定期进行。一旦不符合系统适应性标准，可以执行色谱柱确认，以验证与初始性能的一致性。

 

The analytical procedure (e.g. mobile phase used) and the number of samples injected can contribute to the degradation of the stationary phase. The information related to the tested samples and column usage/storage conditions shall be traceable in the records (for example: samples analysed and matrix for finished products, chromatography conditions (mobile phase, temperature, pH value…).

 

分析程序（例如所使用的流动相）和进样的样品数量会导致固定相降解。与测试样品和色谱柱使用/储存条件相关的信息应在记录中可追溯（例如：分析的样品和成品基质、色谱条件（流动相、温度、pH 值......

 

8. PROCEDURES FOR QUALIFICATION OF ANALYTICAL COLUMNS

 

分析用色谱柱的确认程序

 

Qualification of column performance can be done using one or more of the following approaches:

 

可以使用以下一种或多种方法对色谱柱性能进行验证：

 

a) The column can be tested under the same conditions described in the quality control leaflet provided by the manufacturer. The same parameters should be checked and the results should be compared with the manufacturer’s results. Alternatively, other test mixtures may be used.

 

该色谱柱可以在制造商提供的质量控制手册中描述的相同条件下进行测试。应检查相同的参数，并将结果与制造商的结果进行比较。或者，可以使用其他测试混合物。

 

b)If the column is used only for specific test methods, it might be appropriate to perform the qualification with the system suitability test described for the test method.

 

如果该色谱柱仅用于特定的测试方法，则可能适合使用针对该测试方法所描述的系统适应性测试进行确认。

 

c)Columns with similar stationary phases can be qualified using a general test mixture/procedure.

 

具有相似固定相的色谱柱可以使用通用测试混合物/程序进行确认。

 

Most of the examples given below in Annexes 1 and 2 are qualification procedures provided by manufacturers.

 

下面附件 1 和 2 中给出的示例是大多数制造商提供的确认程序。

 

The parameters tested for the qualification of the column should be calculated and reported as defined by Ph. Eur. Chapter 2.2.46. and the results used for verification of the column performance.

 

应按照欧洲药典第2.2.46 章的规定计算和报告用于色谱柱确认的参数。以及用于确认色谱柱性能的结果。

 

Consider any differences that might occur between the parameters defined in the manufacturer specification or instrument default calculations and the requirements of Ph. Eur. Chapter 2.2.46. (Note: the tailing factor calculated by the instrument software is quite often USP tailing by default).

 

考虑制造商标准或仪器默认计算中定义的参数与欧洲药典第2.2.46 章的要求之间可能出现的任何差异。（注意：默认情况下，仪器软件计算的拖尾因子通常是USP 拖尾）。

 

9.REFERENCES

 

参考文献

 

1)ISO/IEC 17025:2017 General requirements for the competence of testing and calibration laboratories.

 

2)Ph. Eur. Chapter 2.2.28. “Gas chromatography”.

 

3) Ph. Eur. Chapter 2.2.29 “Liquid chromatography”.

 

4) Ph. Eur. Chapter 2.2.46 “Chromatographic separation techniques”.

 

5)Column performance test mixture for liquid chromatography - standard reference material 870. NIST.

 

6) HPLC column classification. USP Pharmacopeia Forum, Vol. 31(2).

 

APPENDIX 1

 

附录1

 

TABLE 1: EXAMPLES FOR QUALIFICATION OF ANALYTICAL COLUMNS FOR LC

 

表 1：用于 LC 的分析用色谱柱的确认示例

 

The chromatographic conditions provided below apply to a column size of 250 mm (length) × 4.6 mm (diameter). These examples are also applicable for qualification of superficially porous particle (SPP) columns. When a column with other dimensions or SPP is used, the chromatographic conditions should be adjusted according to Ph. Eur. chapter 2.2.46.

 

下面提供的色谱条件适用于250 mm（长度）× 4.6 mm（直径）的色谱柱尺寸。这些示例也适用于表面多孔颗粒（SPP）色谱柱的确认。当使用其他尺寸或SPP 的色谱柱时，应根据欧洲药典第2.2.46 章调整色谱条件。

 

 

 

 

APPENDIX 2

 

附录2

 

TABLE 1: EXAMPLES FOR THE QUALIFICATION OF ANALYTICAL COLUMNS FOR GAS CHROMATOGRAPHY

 

表 1：气相色谱柱的确认示例

 

 

 

 

Plate number

 

塔板数

 

The column performance may be calculated for a given component as the plate number (also referred to as number of theoretical plates).

 

对于给定组分，色谱柱性能可以用塔板数（也称为理论塔板数）来计算。

 

This parameter is very useful for comparisons of GC columns. Table 2 below represents the requirements for the average column performance, given in plates/metre, according to the internal diameter and polarity of the phase.

 

此参数对于比较GC 色谱柱非常有用。下表2 显示了根据固定相的内径和极性对平均色谱柱性能的要求，以板/米为单位。

 

Table 2. Comparison of average efficiency by phase polarity and internal diameter.

 

表 2.按相极性和内径比较平均效率。