Assay of Amino Acids in Ganoderma Lucidum Spore Powder by Pre-column Derivatization HPLC
YIN Jiao-yang,YAN Shu,TIAN Shu-xia
School of Pharmacy, Tianjin Medical University, Tianjin (300070), China
Abstract:
Objective To establish a pre-column derivatization-high performance liquid chromatography method for the determination of amino acids in ganoderma lucidum spore powder. Methods The amino acids in ganoderma lucidum spore powder were derivatized with o-phthalaldehyde (OPA) and 9-fluorenyl methoxycarbonyl chloride(FMOC). The chromatographic conditions were Advance Bio AAA (100 mm×4.6 mm, 2.7μm) column, and Mobile phase A was 0.01 mol/L disodium hydrogen phosphate and 0.01 mol/L sodium borate solution (pH: 8.2±0.1). Mobile phase B was methanol: acetonitrile: water (45:45:10), flow rate was 0.8 mL/min, the column temperature was 45 ℃, the injection volume was 10 μL and the detection wavelength was fixed at 338 nm and 262 nm. Results 17 amino acids in Ganoderma lucidum spore powder were separated satisfactorily in 60 minutes. All the components exhibited good linear correlation in the given concentration range (r ≥0.999, n=6). The recovery rate of the sample was between 95.42% and 99.38%. The RSD of the precision, repeatability and stability of the experimental method were all less than 3.0%. Conclusion The methodological verification shows that the established experimental method is suitable for the determination of 17 amino acids in ganoderma spore powder.
