Simultaneous determination of six compounds in the alcohol extract from Mongolian medicine Qinggan Manu-4 and the establishment of fingerprints by HPLC
Objective To establish a HPLC method and fingerprints study for the simultaneous determination of loganic acid,gentiopicroside,paeoniflorin,vitexin,liquiritin,and ammonium glycyrrhizinatein,and other components in the alcohol extract of Mongolian medicine Qinggan Manu-4.Methods The Agilent Eclipse XDB-C18 column(250 mm×4.6 mm,5 μm)was used,with the detection wavelength of 230 nm for paeoniflorin,vitexin and liquiritin,237 nm for loganic acid,250 nm for glycyrrhizic acid,and 270 nm for gentiopicroside.The mobile phase was 0.1%phosphoric acid aqueous solution-acetonitrile with gradient elution.The flow rate was 1.0 mL/min,the column temperature was 30℃,and the injection volume was 10 μL.Results The mass concentrations of loganic acid,gentiopicroside,paeoniflorin,vitexin,liquiritin and ammonium gentiopicroside in the alcohol extract of Mongolian medicine Qinggan Manu-4 showed good linear relationships with the chromatographic peak area in the range of 0.020 3-0.121 0,0.053 3-0.317 7,0.021 3-0.127 0,0.011 5-0.069 0,0.014 1-0.083 8 and 0.035 1-0.209 1 mg/mL,respectively(r≥0.999 8).The average recovery rates of the six components were 94.75%,99.49%,92.32%,95.82%,101.29%and 98.04%,with the RSDs of 1.11%,0.76%,0.99%,2.75%,1.09%and 2.43%,respectively(n=6).The HPLC fingerprint of the alcoholic extract of Mongolian medicine Qinggan Manu-4 was established,using the chromatogram of sample S1 as the reference chromatogram,the control fingerprints were generated through multi-point correction and full-spectrum peak matching.A total of ten common peaks were identified,and after comparison with the reference substance,eight components were identified.Conclusion The established method and fingerprints are accurate,reliable,reproducible and specific,which provide a basis for the quality control and subsequent development of Mongolian medicine Qinggan Manu-4.