Pre-column Derivatization RP-HPLC Determination of Amino Acids in Asparagi Radix before and after Heating Process

A pre-column derivatization RP-HPLC assay was established for the simultaneous determination of 12 kinds of amino acids in Asparagi Radix before and after heating process. PITC was used as pre-column derivatization reagent. The obtained amino acid derivatives were separated on C-18 column (250 x 4.6 mm i. d., 5 mu m) by using gradient elution with 0.1 mol/L pH6.5 acetate buffer solution-acetonitrile (93:7, v/v) as mobile phase A and acetonitrile-water (4:1, v/v) as mobile phase B. 12 kinds of amino acids could be detected at 254nm in 40min with a flow rate of 0.9mL/min. Linear range was between 0.5-500 mu g/mL and average recovery was between 75.4% -100.1 %. Experimental results showed that the amino acid content in Asparagi Radix was decreased after heating process. (C) 2013 Published by Elsevier B.V.