This document specifies a method for the separation of fatty acid methyl esters (FAME) from middle distillates by liquid chromatography (LC) and for the determination of the pattern of the fatty acid methyl esters by gas chromatography (GC) according to EN 14103. The pattern of the fatty acid methyl esters might be used for calculation of the average molecular mass of FAME according to EN 14078 [1]. Independently from the origin of the middle distillate, this method is applicable to FAME of vegetable or animal origin that contain fatty acid methyl esters between C6:0 and C24:1. The method is suitable for the separation and determination of FAME from middle distillates with FAME contents of at least 2 % (V/V). NOTE For the purpose of this document, the terms % (V/V) and % (m/m) are used to express volume fractions in % or mass fractions in %.