近日，EDQM OMCL發布了《分析用色譜柱的確認》指南，該文件概述了分析色譜柱確認的一般原則。色譜柱被認為是設備，作為色譜系統的重要組成部分，它們會影響分析結果。因此，OMCL 認為必要時應進行色譜柱的確認。

 

文件介紹了液相色譜（LC、HPLC、UHPLC）和氣相色譜（GC） 程序中使用的分析色譜柱的確認要求和一般標準。

 

OMCL表示在計劃、執行和記錄確認過程時，在收到色譜柱時以及在色譜柱的生命周期中，都將其用作指導。

 

該文件現已翻譯完成，如下：

 

QUALIFICATION OF ANALYTICAL COLUMNS

 

分析用色譜柱的確認

 

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強制性要求使用術語“應”或“必須”進行定義。使用“應該”表示推薦。對于本文件中的要求，也可以接受其他適當合理的方法。術語 “可以” 表示具有非約束性特征的可能性或示例。

 

Contents 

 

目錄

 

1. INTRODUCTION

 

介紹

 

2. SCOPE

 

范圍

 

3. GLOSSARY AND DEFINITIONS

 

術語和定義

 

4. CONSIDERATIONS FOR QUALIFICATION UPON PURCHASING

 

采購的確認考慮

 

5. INITIAL QUALIFICATION

 

首次確認

 

5.1 Qualification parameters

 

確認參數

 

6. PERIODICAL QUALIFICATION

 

定期確認

 

 6.1 Qualification procedure

 

確認程序

 

 6.2 Frequency of qualification

 

確認頻率

 

 6.3 Qualification records

 

確認記錄

 

7. IN-USE QUALIFICATION

 

使用過程中的確認

 

8. PROCEDURES FOR QUALIFICATION OF ANALYTICAL COLUMNS

 

色譜柱的確認程序

 

9. REFERENCES

 

參考文獻

 

APPENDIX 1: EXAMPLES FOR QUALIFICATION OF ANALYTICAL COLUMNS FOR LC

 

附錄1：液相色譜柱確認示例

 

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

 

附錄 2 ：氣相色譜柱確認示例

 

1.INTRODUCTION

 

介紹

 

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 中使用的分析用色譜柱確認的一般原則。色譜柱被認為是設備，作為色譜系統的重要組成部分，它們會影響分析結果（ISO/IEC 17025：2017，第 6.4 章設備，第 6.4.1 點）。因此，OMCL 認為必要時應進行色譜柱的確認。

 

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 負責確定在色譜柱生命周期內使用的合適確認方案，例如，根據預期目的和使用頻率。

 

2.SCOPE

 

范圍

 

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.

 

本文件介紹了液相色譜（LC、UHPLC）和氣相色譜（GC）程序中使用的分析用色譜柱的確認要求和一般標準。

 

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 在規劃、執行和記錄確認過程時，在收到色譜柱時以及在色譜柱的生命周期中，都可以將其用作指導。

 

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.

 

附錄 1 和 2 中提供的示例程序具有非約束性。在執行所需的確認時，他們會有所幫助。然而，每個 OMCL 的專業判斷和背景經驗都有責任應用最合適的程序，以證明分析用色譜柱適合其預期用途。

 

3. GLOSSARY AND DEFINITIONS

 

術語

 

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

 

本文件中使用的術語與歐洲藥典第2.2.46 章“色譜分離技術”中使用的術語一致。

 

4.CONSIDERATIONS FOR QUALIFICATION UPON PURCHASING

 

采購確認的注意事項

 

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

 

應確認到貨色譜柱是否因運輸而造成物理損壞、是否符合采購訂單以及文件的完整性。

 

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.

 

應根據 ISO/IEC 17025：2017第 6.4.13 條中描述的適用要求保留與色譜柱的管理相關的記錄。

 

5.INITIAL QUALIFICATION

 

初始確認

 

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.

 

分析用色譜柱應在收到時或首次使用前通過檢查文件（例如COA）或根據預定的程序對色譜柱進行確認。

 

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.

 

驗證/確認應有記錄（包括驗證/確認日期、確認程序引用、使用的標準物質或試劑、結果、使用的色譜設備、通過/失敗結論等）。此記錄可用于在色譜柱不符合制造商標準時進行投訴。

 

5.1 Qualification parameters

 

確認參數

 

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 有責任選擇最合適的程序、參數和接受標準。它們應適合色譜柱的預期用途。這些參數可以包括分離度、對稱因子、理論踏板等。附件1 和附件 2 中給出了一些典型的限度標準作為示例。

 

6. PERIODICAL QUALIFICATION

 

定期確認

 

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.

 

可以使用相同的測試化合物/程序定期進行色譜柱的確認。確認結果的系統記錄應在實驗室中保存。

 

6.1 Qualification procedure

 

確認程序

 

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.

 

至少可用初始確認的程序進行定期確認。在實驗室中使用新色譜柱獲得的第一張色譜圖可以作為該色譜柱初始性能的參考，并作為后續驗證的參考。由于不同 LC 或GC 的設備配置（例如死體積）、運行環境、系統電子元件等，色譜圖上可能會觀察到細微的不同。當設定內控接受標準時，可以考慮這些因素。

 

6.2 Frequency of qualification

 

確認頻率

 

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.

 

當認為定期確認適當和有用時，實驗室可以定義和記錄確認頻率。頻率取決于色譜柱的使用情況（例如日常或“點”分析）。例如，根據第5 節（上文）的確認可以按固定的時間間隔（例如每年一次）進行，也可以根據使用程度進行，例如在規定的進樣次數后進行。

 

6.3 Qualification records

 

確認記錄

 

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.

 

確認結果的記錄可以幫助用戶監測色譜柱性能隨時間的變化，并告知是否需要丟棄該色譜柱。當符合接受/系統適用性標準時，色譜柱被認為適合預期用途。

 

7. IN-USE QUALIFICATION

 

使用過程中的確認

 

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.

 

使用過程中的確認對應于所用方法的系統適用性測試。應在每次測試前和/或在序列進樣期間定期進行。一旦不符合系統適應性標準，可以執行色譜柱確認，以驗證與初始性能的一致性。

 

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（例如所使用的流動相）和進樣的樣品數量會導致固定相降解。與測試樣品和色譜柱使用/儲存條件相關的信息應在記錄中可追溯（例如：分析的樣品和成品基質、色譜條件（流動相、溫度、pH 值......

 

8. PROCEDURES FOR QUALIFICATION OF ANALYTICAL COLUMNS

 

分析用色譜柱的確認程序

 

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

 

可以使用以下一種或多種方法對色譜柱性能進行驗證：

 

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.

 

該色譜柱可以在制造商提供的質量控制手冊中描述的相同條件下進行測試。應檢查相同的參數，并將結果與制造商的結果進行比較。或者，可以使用其他測試混合物。

 

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.

 

如果該色譜柱僅用于特定的測試方法，則可能適合使用針對該測試方法所描述的系統適應性測試進行確認。

 

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

 

具有相似固定相的色譜柱可以使用通用測試混合物/程序進行確認。

 

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

 

下面附件 1 和 2 中給出的示例是大多數制造商提供的確認程序。

 

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.

 

應按照歐洲藥典第2.2.46 章的規定計算和報告用于色譜柱確認的參數。以及用于確認色譜柱性能的結果。

 

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).

 

考慮制造商標準或儀器默認計算中定義的參數與歐洲藥典第2.2.46 章的要求之間可能出現的任何差異。（注意：默認情況下，儀器軟件計算的拖尾因子通常是USP 拖尾）。

 

9.REFERENCES

 

參考文獻

 

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

 

附錄1

 

TABLE 1: EXAMPLES FOR QUALIFICATION OF ANALYTICAL COLUMNS FOR LC

 

表 1：用于 LC 的分析用色譜柱的確認示例

 

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.

 

下面提供的色譜條件適用于250 mm（長度）× 4.6 mm（直徑）的色譜柱尺寸。這些示例也適用于表面多孔顆粒（SPP）色譜柱的確認。當使用其他尺寸或SPP 的色譜柱時，應根據歐洲藥典第2.2.46 章調整色譜條件。

 

 

 

 

APPENDIX 2

 

附錄2

 

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

 

表 1：氣相色譜柱的確認示例

 

 

 

 

Plate number

 

塔板數

 

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

 

對于給定組分，色譜柱性能可以用塔板數（也稱為理論塔板數）來計算。

 

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.

 

此參數對于比較GC 色譜柱非常有用。下表2 顯示了根據固定相的內徑和極性對平均色譜柱性能的要求，以板/米為單位。

 

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

 

表 2.按相極性和內徑比較平均效率。