Equivalent GC systems performance for regulatory method compliance and validation

在更换诸如气相色谱(GC)或液相色谱(LC)系统等技术时，实验室之间或从一个分析系统迁移到另一个分析系统，无论是由于更新分析设备或从一个供应商更换到另一个供应商，都可能是一项耗时且困难的任务。分析实验室使用的仪器种类繁多，可以属于各种品牌。不同厂商生产的仪器通常采用相同的分析方法，期望其性能是相同的。

As part of the method transfer and validation, federal and governmental agencies, such as the United States Food and Drug Administration (US-FDA), and the European Medicines Agency (EMA) released specific guidelines [1,2]. Moreover, the USP Chapter 621 of the current United States Pharmacopeia has suitability procedures to test analytical methods and demonstrate equivalency when transferring them from one system to another [3]. This is also applicable for GC methods where strict chromatographic separation criteria are defined.

In this paper, two examples of how the Thermo Scientific™ TRACE™ 1300 and 1600 Series Gas Chromatograph systems perform with typical, well-known GC methods for pharmaceutical and food industry are detailed, demonstrating the compatibility with common consumables such as liners and capillary columns, simplifying the method portability assuring equivalency of the analytical performance. The instrument conditions are not included herein. Please refer to the full paper (WP-74062) for information on instrument conditions.

Residual solvent analysis in pharmaceutical products according to USP <467> method

Introduction
溶剂广泛应用于制药产品、物质和辅料的合成。为了确保患者的安全，国际协调会议(ICH)[4]和美国药典(USP)[5]发布了一些指南，设定了可接受的限度，并支持对生产和纯化过程中使用的残留溶剂的评估。残留溶剂(RS)具有低沸点和热稳定性，因此可以使用顶空-气相色谱(HS-GC)耦合火焰电离检测来确定。

残留溶剂评估的工作流程使用图1中的简化示意图进行了报告。当已知可能存在的残留溶剂时，可以使用限度试验(如程序a或程序B)或定量试验(如程序c)来确定它们。当残留溶剂未知时，必须使用程序a进行筛选试验。如果产品不符合程序A的验收标准，则必须使用程序B来证明其符合性。如果产品使用程序A和程序B不符合标准，则必须使用程序C来量化产品中存在的残留溶剂。

Results and discussion

Procedure A - Screening of unknown residual solvents
原液、标准液和测试液按照USP <467>方法制备。在当地购买了一种非处方乙酰水杨酸(分散阿司匹林，75mg)，并根据图1中的USP <467>流程进行分析。

System suitability criteria for sensitivity (peak-to-peak (PtP) signal-to-noise ratio (S/N)) and chromatographic resolution (Rs) were met with:

• S/N > 5:1 for 1,1,1-trichloroethane in Class 1 standard solution
• S/N >3:1 for all peaks in Class 1 system suitability solution (Figure 2)
• Rs between acetonitrile/dichloromethane >1 in Class 2A standard solution (Figure 2).

The innovative system design with direct connection between the gas chromatograph and the autosampler combined with the high inertness and the precise temperature and flow controls of the TRACE 1310 Gas Chromatograph allowed for an efficient chromatographic process ensuring Gaussian peak shapes with average asymmetry factor (As) of 1.2. Peak responses obtained for the un-spiked sample were lower than the corresponding peaks in Class 1 and Class 2 standard injections. According to the regulation, the test solution met the requirements for residual solvent content with no other actions required.
通过采用阀-回路顶空技术，保证了方法参数的一致性，从而从不同的HS-GC系统中简化了方法转移。报告方法参数的名称在不同的品牌可能不同，特别是对顶空自动采样器。参数的等价性在之前发表的白皮书[6]中有明确的解释。

Summary

本研究的结果表明，TRACE 1310 GC-FID符合USP <467>要求，符合规范的c-GMP制药实验室所需的色谱分离适宜标准。方法参数的等效性确保了不同HS-GC品牌的方法使用阀-回路顶空技术的安全可移植性。
Separation of 37 fatty acid methyl esters according to AOAC method 996.06 by GC-FID

Introduction

Food fat mainly consists of triglycerides and assessing the fat (trans and saturated) composition of food products as part of the nutritional information is a fundamental test for the food industry. The AOAC method 996.06 describes the determination of total, saturated and unsaturated fat in foods using capillary GC-FID by a multiple steps procedure: hydrolytic extraction followed by the derivatisation (methylation) of fatty acids to produce fatty acids methyl esters (FAMEs) which are the derivatives suitable for GC analysis [7].

Experimental
A TRACE 1610 Gas Chromatograph configured with an Instant Connect split/splitless SSL Injector and an Instant Connect Flame Ionisation Detector (FID) was coupled with an AI/AS 1610 Autosampler and used to assess the chromatographic separation performance according to AOAC method 996.06.
A standard solution was prepared by diluting Restek Food Industry FAMEs mix (30 mg/mL in dichloromethane) (P/N 35077) to 1000 μg/mL in dichloromethane/hexane.

Results and discussion

Chromatographic resolution (Rs) is fundamental for FAMEs separation, identification and quantitation and specific resolution requirements for critical peaks pair are included in AOAC method 996.06: (Rs) ≥ 1.0 for FAMEs pair of adjacent peaks (C18:3 - C20:1 and C22:1 – C23:3 – C20:4).
The chromatographic profile of 37 FAMEs separation obtained with TRACE 1610 Gas Chromatograph (equipped with Restek Rt-2560 column) is shown in Figure 2; critical pair peaks are highlighted, and the achieved resolution meets and exceeds the requirements. Peak identification and retention times are reported in Table 1.

Summary

The TRACE 1610 Gas Chromatograph equipped with Restek RT-2560 100 m, 0.25 mm, 0.2 μm capillary column is suitable for FAMEs separation in food samples according to AOAC method 996.06, meeting or exceeding resolution requirements and providing reliable peaks integration and quantification.

Conclusion

本白皮书中所考虑的例子表明，TRACE系列气相色谱仪系统具有等效的色谱性能，确保满足特定法规方法的适用性要求。在气相色谱系统的标准工作范围内应用方法参数，以及使用标准消耗品，允许顺利转移，保持所需的分析性能。

References
1. EMEA/CHMP/EWP/192217/2009 Rev. 1 Corr. 2** Committee for Medicinal Products for Human Use (CHMP).
2. FOOD AND DRUG ADMINISTRATION OFFICE OF REGULATORY AFFAIRS ORA Laboratory Manual Volume II, ORA-LAB.5.4.5 Methods, Method Verification and Validation Revision #: 02 Revision June 2020.
3. USP Chapter 621 of the current United States Pharmacopeia.
4. Impurities: Guideline for Residual Solvents Q3C(R6), ICH Harmonised Guidelines, International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human use, 2016.
5. General Chapter USP <467> Organic Volatile impurities, Chemical Tests, United States Pharmacopeia, 2012 and Interim Revision Announcement Official November 1, 2019; Official December 1, 2020 <467> Residual Solvents.
6. Thermo Scientific White Paper 10705- Investigation of key parameters for a smooth method transfer to the new Thermo Scientific TriPlus 500 Headspace Autosampler
7. AOAC 996.06-1996 (2010) - Fat (Total, Saturated, and Unsaturated)

Please refer to WP-74062 for the full paper which will include:
- Determination of gasoline range organics (GRO) in water by static headspace gas chromatography
- Separation of US EPA 16 priority polycylic aromatic hydrocarbons by GC-FID

For the full paper (WP-74062), please contact CMD.APACMarketing@thermofisher.com