In this Application Update, we combine into a single method individual improvements made to the standard AAA-Direct gradient program. These individual improvements are now in the AAA-Direct manual. This update shows that such combined changes to the AAA-Direct method improve the performance of our previously published method. This new method successfully determines amino acids in yeast extract-peptone-dextrose yeast culture medium (YPD Broth), and Dulbecco’s modified Eagle’s (with F-12), M199, L-15 (Leibovitz), and McCoy’s 5A media for mammalian cell culture.
Amino acids present in protein formulations serve as buffers, bulking agents, stabilizers, and antioxidants. Eight specific amino acids are commonly used as excipients in protein therapeutic formulations due to their physicochemical properties. These include Arginine, Aspartic acid, Glutamic acid, Lysine, Proline, Glycine, Histidine, and Methionine. The UHPLC column, an Acclaim PA2 column, provides excellent separation characteristics for these un-labeled amino acids. Gradient conditions are adjusted by selecting appropriate ion-pairing reagents, pH and level of organic solvents.
Therapeutic proteins (antibodies and vaccines) vary considerably due to the nature and dose of the protein molecule. Aggregation is a major degradation pathway of protein therapeutics during storage. Stabilization of these protein formulations can be enhanced through the addition of specific excipients such as surfactants, amino acids and sugars. The separation of therapeutic protein and amino acid excipients was performed using a 2D approach. An integrated UHPLC system with a UV and universal charged aerosol detection offering multi-mode detection for the simultaneous analysis of both non-chr
A gradient HPLC method was developed for the direct analysis of sodium and glutamate present in a popular food product from India called Maggi 2-Minute Noodles. Chromatographic separation was performed using the mixed mode Acclaim Trinity P1 column. One advantage in using this multi-modal column is that several ions such as sodium and chloride can be simultaneously measured along with glutamate present in the Maggi spice mixture. A simple direct method for the analysis of various amino acids is possible using the universal charged aerosol detector (CAD) as previously published.
The Thermo Scientific HPLC system is applied for the analysis of glutamate in beer. The sample preparation is automated, and the separation is performed using a Thermo Scientific Acclaim PolarAdvantage column, with UV detection at 330 nm. Chromatogram A is beer and chromatogram B is beer spiked with 250 µg/mL glutamic acid.
