I want to measure the amino acid contents in biological samples. I have run the standard amino acid once and got different peak for each amino acid. What should I do to prepare the standard curve for all the amino acid?
I bought the amino acid standard solution from Sigma #AA-S-18, as #1 amino acids (AA) stock and individually Asparagine, glutamine and tryptophan and made #2 extended amino acids (EAA) stock solution of 1800pmol/mL of the last three and when creating my calibration standard curve I mixed the two #1 (AA) and #2 (EAA) and made serial dilutions. I had to inject most of them individually to find their exact retention time according to my HPLC method before i started assigning the peaks to my calibration standard chromatogram. I hope this helps.