Establishment of reversed phase high performance liquid chromatography for flavonoids in chickpeas and evaluation of their in vitro efficacy
[Objective]To establish a reversed phase high performance liquid chromatography method for the effective components of chickpea flavonoids,and evaluate the in vitro pharmacological effects of chickpeas and chickpea total flavonoids through antibacterial tests.[Methods]Reversed phase high performance liquid chromatography was used for analysis,Zorbax SB-C18(4.6 mm×150.0 mm,5 μm)column was used,with mobile phase A(methanol)∶B(0.5%phosphoric acid aqueous solution)=45∶55.The flow rate was 0.8 mL/min for equal elution,and the column temperature was 30℃.The precision,stability,repeatability and sample recovery were verified by HPLC.Filter paper diffusion method,Oxford cup method and micro broth dilution method were used to investigate the antibacterial effect of chickpea(100 mg/mL)and chickpea total flavonoids(20 mg/mL).[Results]After equal elution,three flavonoids,rutin,naringin dihydrochalcone,and quercetin were well separated,and their chromatographic peaks showed good separation within 30 min,with a linear range of 0.003 75-0.060 00 mg/mL(R2=0.996-0.999).The stability and repeatability test results indicate that,the content of rutin was 0.032 mg/g,and the content of naringin dihydrochalcone was 0.052 mg/g,quercetin was not detected.In the antibacterial effect experiment,the filter paper diffusion method and Oxford cup method did not appear bacteriostatic circle,and the pore plates of the micro broth dilution method bacteriostatic experiment were muddy.[Conclusion]The reversed phase high performance liquid chromatography method used in this experiment has certain reliability and is suitable for the separation and determination of two flavonoids,rutin and naringin dihydrochalcone in chickpeas.Through the verification of three antibacterial methods,chickpea and chickpea total flavonoids did not have obvious antibacterial effect.
ChickpeasTotal flavonoidsReversed phase high performance liquid chromatographyAntibacterial effect
万方数据
维普
