ISO 12966-2:2017 PDF

Animal and vegetable fats and oils — Gas chromatography of fatty acid methyl esters — Part 2: Preparation of methyl esters of fatty acids

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Title (English):

ISO 12966-2:2017

Animal and vegetable fats and oils — Gas chromatography of fatty acid methyl esters — Part 2: Preparation of methyl esters of fatty acids

Document status:: Active
Format:: Electronic (PDF)
Number of pages:: 15
Publication date:: March 31, 2017
Edition:: ISO IS 12966 edition 2 version 1
ICS:: 67.200.10

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Description (English):

ISO 12966-2:2017 specifies methods of preparing the methyl esters of fatty acids. It includes methods for preparing fatty acid methyl esters from animal and vegetable fats and oils, fatty acids and soaps. To cover different requirements, four methylation methods are specified, namely: a) a "rapid" transmethylation procedure under alkaline conditions; b) a "general" transmethylation/methylation procedure under sequential alkaline and acid conditions; c) a BF3 transmethylation procedure; d) an alternative procedure using acid-catalysed transmethylation of glycerides. Methyl esters so produced are used in various analytical procedures requiring such derivatives, e.g. gas-liquid chromatography (GLC), thin-layer chromatography (TLC) and infrared spectrometry (IR). Milk and milk products (or fat coming from milk and milk products) are excluded from the scope of this document.

Technical details

Technical committee: ISO/TC 34/SC 11 - Animal and vegetable fats and oils
SKU: ISO 12966-2:2017

Abstract

Overview

ISO 12966-2:2017 specifies validated methods for preparing fatty acid methyl esters (FAMEs) from animal and vegetable fats, oils, free fatty acids and soaps for subsequent analysis. The standard defines four methylation/transmethylation procedures to suit different sample types and analytical needs, and provides practical guidance on reagents, apparatus, sample preparation and checks for completeness of derivatization. Milk and milk products (and fats from these sources) are explicitly excluded from the scope.

Key Topics

Four prescribed methylation methods
- Rapid transmethylation under alkaline conditions (fast, routine for many edible fats).
- General sequential alkaline then acid transmethylation/methylation (broad applicability).
- Boron trifluoride (BF3) transmethylation/methylation (powerful but toxic; use with caution).
- Alternative acid-catalysed transmethylation of glycerides.
Reagents and catalysts - includes alkali catalysts (e.g., KOH/NaOH, sodium methoxide) and methanolic BF3 complexes; emphasizes excess methanol and low water content.
Analytical considerations
- Prevention of incomplete conversion and formation of artefacts.
- Sensitivity of labile functional groups (keto, epoxy, hydroxyl, hydroperoxy), cyclopropyl/cyclopropenyl and acetylenic fatty acids to some methods.
- Volatility and recovery issues for short-chain fatty acid methyl esters.
Quality control and verification
- Procedures include apparatus, stepwise protocols and an informative TLC method (annex) to verify completeness of derivatization.
Safety and practical limitations
- Toxicity and shelf-life concerns for BF3 reagents; water and sample matrix effects; impact on GC column life when injecting acidic/basic reaction media.

Applications

ISO 12966-2:2017 is intended for laboratories and organizations involved in lipid analysis and routine quality control:

Food and ingredient manufacturers assessing fatty acid composition of oils and fats.
Analytical chemistry and food testing laboratories preparing FAMEs for gas chromatography (GC/GLC), thin-layer chromatography (TLC) and infrared spectrometry (IR).
Research groups in nutrition, lipidomics and oleochemistry profiling fatty acid distributions.
Regulatory and compliance testing where standardized, reproducible FAME preparation is required.

Practical outcomes include reliable FAME preparation for accurate GC-based fatty acid profiling, informed method selection depending on sample matrix, and reduced risk of artefacts or losses of short-chain esters.

Related Standards

Part of the ISO 12966 series (gas chromatography of fatty acid methyl esters). Other parts cover chromatographic conditions and identification; consult the ISO catalogue for related documents and cross-references.

Related standards

Mentioned in the description and other ISO standards

Abstract

Overview

Key Topics

Four prescribed methylation methods
- Rapid transmethylation under alkaline conditions (fast, routine for many edible fats).
- General sequential alkaline then acid transmethylation/methylation (broad applicability).
- Boron trifluoride (BF3) transmethylation/methylation (powerful but toxic; use with caution).
- Alternative acid-catalysed transmethylation of glycerides.
Reagents and catalysts - includes alkali catalysts (e.g., KOH/NaOH, sodium methoxide) and methanolic BF3 complexes; emphasizes excess methanol and low water content.
Analytical considerations
- Prevention of incomplete conversion and formation of artefacts.
- Sensitivity of labile functional groups (keto, epoxy, hydroxyl, hydroperoxy), cyclopropyl/cyclopropenyl and acetylenic fatty acids to some methods.
- Volatility and recovery issues for short-chain fatty acid methyl esters.
Quality control and verification
- Procedures include apparatus, stepwise protocols and an informative TLC method (annex) to verify completeness of derivatization.
Safety and practical limitations
- Toxicity and shelf-life concerns for BF3 reagents; water and sample matrix effects; impact on GC column life when injecting acidic/basic reaction media.

Applications

ISO 12966-2:2017 is intended for laboratories and organizations involved in lipid analysis and routine quality control:

Food and ingredient manufacturers assessing fatty acid composition of oils and fats.
Analytical chemistry and food testing laboratories preparing FAMEs for gas chromatography (GC/GLC), thin-layer chromatography (TLC) and infrared spectrometry (IR).
Research groups in nutrition, lipidomics and oleochemistry profiling fatty acid distributions.
Regulatory and compliance testing where standardized, reproducible FAME preparation is required.

Related Standards

Part of the ISO 12966 series (gas chromatography of fatty acid methyl esters). Other parts cover chromatographic conditions and identification; consult the ISO catalogue for related documents and cross-references.

ISO 12966-2:2017 PDF

Technical details

Abstract

Overview

Key Topics

Applications

Related Standards

Related standards

Abstract

Overview

Key Topics

Applications

Related Standards

Related standards

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