ISO 13903:2005 describes the determination of free (synthetic and natural) and total (peptide-bound and free) amino acids in feeding stuffs, using an amino acid analyser or HPLC equipment. It is applicable to the following amino acids: sum of cystine and cysteine; methionine; lysine; threonine; alanine; arginine; aspartic acid; glutamic acid; glycine; histidine; isoleucine; leucine; phenylalanine; proline; serine; tyrosine; valine.
The method does not distinguish between the salts of amino acids, nor does it differentiate between D and L forms of amino acids. It is not valid for the determination of tryptophan or hydroxy analogues of amino acids.
Limits of quantification depend on the chromatographic equipment, but levels as low as: 0,3 g/kg total lysine; 0,25 g/kg total methionine; 0,35 g/kg total cystine plus cysteine; 0,2 g/kg total threonine; 0,035g/kg free lysine; 0,035g/kg free methionine; and 0,03g/kg free threonine can typically be analysed.