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Measurement of BCAA in Milks and Supplements With an Enzyme Assay: Confirmation of Results with HPLC
Author(s) -
Keenan Kelly,
Do Daniel,
Ngo Kathleen
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.752.1
Subject(s) - isoleucine , leucine , valine , amino acid , high performance liquid chromatography , chromatography , ingredient , enzyme , hydrolysis , chemistry , hydrolyzed protein , biochemistry , food science
Branched chain amino acids (BCAA) are comprised of three types of amino acids (valine, leucine, and isoleucine) and are a common type of supplement or a key ingredient in various types of protein supplements. Given their popularity, a laboratory experiment was devised to measure the level of BCAA in various types of milks or supplements. To make this experiment accessible to many, an enzyme assay was adapted to measure the level of BCAAs. It is expected that this project can be done in advanced high school Biology laboratories or college Biochemistry laboratories. The samples are hydrolyzed to break all bonds releasing the free amino acids and the BCAAs are measured using an enzyme assay that detects only these amino acids. In order to validate this method, the samples were also measured for BCAA using reverse phase‐high performance liquid chromatography (RP‐HPLC). It was demonstrated that all three BCAA survived the hydrolysis process with high recoveries. When these amino acids were added to samples, the recoveries were also very high. The results from RP‐HPLC confirm the results from the enzyme assay and demonstrate that this method makes for a viable, accurate and easily accessible laboratory experiment.