[44] Amino acid analysis

Add time:09/09/2019    Source:sciencedirect.com

Publisher SummaryAmino acid analysis provides an important quantitative parameter in the characterization of isolated protein or peptide samples. Because of the availability of highly sensitive instruments for determining amino acids in the picomole range, the preparation of the sample for analysis is of prime importance in obtaining meaningful results. The first step in the determination of the amino acid composition of proteins is releasing the amino acids quantitatively without concomitant degradation. Because amino acid analyzers are expensive and require considerable expertise to operate, investigators in need of an amino acid analysis frequently depend on a facility equipped to perform such analyses. Concentration and removal of residual salts and various enzyme stabilizers without contamination is the most difficult aspect in obtaining a meaningful amino acid composition from small amounts of protein. The methods used for concentrating proteins from dilute solutions include precipitation by organic solvents or by compounds, such as trichloroacetic acid (TCA). If the sample originally contains quantities of salts, the precipitate is suspended in water and again centrifuged. Membrane proteins do not redissolve in water after acid–acetone precipitation, but some polar proteins do.

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