Determination of 13 amino acids in Asini Corii Colla by RP-HPLC with precolumn derivatization / 中草药

ABSTRACT

Objective: To establish a RP-HPLC method for the simultaneous determination of 13 amino acids in Asini Corii Colla. Methods: Asini Corii Colla was hydrolyzed for 3 h at 120°C using 6 mol/L hydrochloric acid, and the amino acids were determined by RP-HPLC after being derived with phenyl isothiocyanate. HPLC was performed on an Eclipse AAA column (75 mm × 4.6 mm, 3.5 μm) with acetonitrile-110 mmol/L ammonium acetate solution (199) as mobile phase A and acetic acid-acetonitrile-water (0.058020) as mobile phase B in the gradient mode at a flow rate of 0.6 mL/min. The column temperature was 42°C, and the detection wavelength was set at 254 nm. Results: The 13 amino acids were separated within 36 min with a good linearity (r > 0.9977) in the range of the test concentration. The average recoveries (n = 3) were 95.2%-104.4% and the RSD values were less than 5.0%. Conclusion: The method is simple, accurate, and repeatable, which could be available for the quality control of Asini Corii Colla. It may also serve as a good reference for the determination of amino acids in other pharmaceuticals.