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| Determination of effective components in the fermentation broth of Flos Sophorae Immaturus by HPLC |
| CHEN Caiyun1, YIN Shuying2, LIN Ying1, REN Yan1, LI Gaotian1, CAO Sen1 |
1 School of Pharmacy,Binzhou Medical University,Yantai 264003,P.R.China;
2 School of Public Health and Management,Binzhou Medical University |
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Abstract Objective To establish a high performance liquid chromatography (HPLC) method to determine the content of rutin and quercetin in Flos sophorae immaturus fermentation broth.Methods Flos sophorae immaturus was fermented by Natto fungus and the broth was separated on a Phenomenex Gemini 5μ C18 110A (250 mm×4.6 mm I.D.5μm) eluted with methanol-water (0.05% formic acid) in linear gradient condition [0min→15min→20min→25min→26min→ 36min,elute rate of methanol-water (0.05% formic acid) was 35:65→50:50→55:45→ 60:40→35:65→35:65].The detection wavelength was 257nm for rutin and 254nm for quercetin.The flow rate was 1.0 mL/min;the column temperature was set at 25℃.Results There was a good linear relationship between the peak area and concentration of rutin and quercetin.The linear range was 0.031 ~ 1 mg/mL (R12= 0.9997,R22=0.9991).The precision RSD of rutin was 0.997% and quercetin was 0.392%.The stability RSD of rutin was 0% and quercetin was 0.05%.The repeatability RSD of rutin was 1.405% and quercetin was 1.649%.The average recovery rate of rutin was 93.90%,with RSD 1.55% and quercetin was 98.55%,with RSD 1.65%.The contents of rutin and quercetin were 19.9199±0.1436 mg/g and 0.6501±0.0089 mg/g, respectively.Conclusion The method can accurately and efficiently determine the content of effective components of the fermentation broth of Flos sophorae immaturus.
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Received: 03 January 2018
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