Simultaneous Determination of 10 Flavonoids in Astragalus membranaceus by HPLC / 中国药房

ABSTRACT

OBJECTIVE:To establish a method for simultaneous determination of 10 flavonoids in Astragalus membranaceus. METHODS:HPLC method was adopted. The determination was performed on Agilent SB-C18 column with mobile phase consisted of acetonitrile-0.3% formic acid (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 254 nm, and the column temperature was 35 ℃. The sample size was 10 μL. RESULTS:The linear ranges of calycosin-7-O-glucoside,iso-quercitrin,genistin,ononin,calycosin,quercetin,genistein,kaempferol,isorhamnetin and formononetion were 0.03029-1.5145μg (r=0.9994),0.01500-0.7500 μg(r=0.9995),0.00739-0.3695 μg(r=0.9991),0.12011-6.0055 μg(r=0.9998),0.03836-1.918 μg (r=0.9999),0.02989-1.4945 μg(r=0.9995),0.00704-0.352 μg(r=0.9994),0.01683-0.8415 μg(r=0.9995),0.00454-0.227μg(r=0.9999),0.01336-0.668 μg(r=0.9999),respectively. RSDs of precision,stability and reproducibility tests were all lower than 2.0% . The recoveries were 99.55% -100.45%(RSD=0.36% ,n=6) ,99.34% -101.00%(RSD=0.59% ,n=6) , 98.05%-100.36%(RSD=1.27%,n=6),99.73%-100.13%(RSD=0.18%,n=6),99.70%-100.30%(RSD=0.22%,n=6), 99.67%-103.27%(RSD=1.37%,n=6),98.13%-104.41%(RSD=2.37%,n=6),96.35%-100.06%(RSD=1.46%,n=6), 99.47%-101.13%(RSD=0.60%,n=6),99.70%-100.06%(RSD=0.15%,n=6),respectively. CONCLUSIONS:This method is convenient,sensitive,stable and reproducible,can be used for simultaneous determination of 10 flavonoids in A. membranaceus.