Sensitive Analysis of Underivatized Amino Acids Using UHPLC with Charged Aerosol Detection
نویسندگان
چکیده
Overview Purpose: To develop fast and sensitive approaches for the analysis of underivatized free amino acids and peptides using the latest UHPLC column technology and charged aerosol detection. Methods: Presented here are two distinct methods using the same UHPLC analytical column. The first method uses large amounts of ion-pairing agents to resolve 18 free amino acids. The second method uses less ion-pairing agent and an inverse gradient approach that enables a direct comparison of charged aerosol detection to traditional UV techniques. Protein digestion is monitored on the autosampler tray. Results: The first method enabled the analysis of 18 amino acids in under 9 minutes with low nanogram sensitivity for all analytes. The tryptic digest method not only allowed the separation of amino acid residues and peptides, but the time course of digestion could be followed using this automated system. The results showed that charged aerosol detection is a complementary technique to the well established low wavelength UV with added sensitivity for small peptides and many amino acids.
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