Determination of Amino Acids in CHO Cells by Reversed-phase High Performance Liquid Chromatography with o-Phthaldialdelhyde Pre-column Derivatization

Nineteen amino acids in Chinese hamster ovary (CHO) cells were determinated by reversed-phase high performance liquid chromatography with o-phthaldialdelhyde pre-column derivatization. The separation of amino acid derivatives was carried out on a Hypersil AA-ODS column by gradient elution, and was detected with an ultraviolet detector. The detection limits of the ultraviolet detector ranged from (0.10) to (0.23 μmol/L); the retention time precision was below (0.3%) and area precision was below 3% over 12 consecutive runs for a (25μmol/L) amino acid standard solution. A good linear relationship exists between the peak area ratio of amino acids and internal standard in certain range of concentrations, and the correlation coefficients ranged from (0.997 9) to (0.999 9). Concentrations of intracellular amino acids were quantitatively analyzed with standarded curves method based on internal standardization, and the recovery of amino acids ranged from 87% to 106%.