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NEaar: North East Amino Acid Racemization

Amino acid enantiomers are separated by reverse phase HPLC (following a modified method of Kaufman & Manley, 1998). Samples are derivitised with o-phthaldialdehyde and thiol N-isobutyryl-L-cysteine automatically prior to injection. The resulting diastereomeric derivatives are then separated on Hypersil C18 BDS column (sphere d. 5 µm; 250 x 3mm) using a linear gradient of a sodium acetate buffer, methanol and acetonitrile on an Agilent 1100 or 1200 Rapid Resolution liquid chromatograph. The fluorescence intensity of derivitised amino acids are measured (Ex=230nm, Em=445nm) and normalised to the internal standard, L-homo-arginine. All samples are run in duplicate. Amino acid standards and chemical blanks are also run daily.  

Example HPLC trace (click to enlarge)

Image:

Contact

  • Kirsty Penkman
    Department of Chemistry
    University of York
    Heslington
    York YO10 5DD
    United Kingdom
    (+44) 01904 322574
    e-mail: kirsty.penkman@york.ac.uk

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