Study On The Synchronous Extraction Process And Hypoglycemic Activity In Vitro Of Chlorogenic Acid And Total Flavonoids From Honeysuckle

Honeysuckle is not only a traditional Chinese medicine in my country,but also one of the homologous substances of medicine and food.Its main active components are chlorogenic acid and flavonoids.Honeysuckle has antibacterial,antiviral,anti-inflammatory,hypoglycemic,hypolipidemic and other biological activities,and has high medicinal value and broad application prospects.At present,most of the research on honeysuckle is focused on chlorogenic acid,and flavonoids are less studied.The utilization of honeysuckle resources is mainly reflected in the extraction of chlorogenic acid,resulting in a waste of honeysuckle resources.In this paper,taking honeysuckle as the research object,the simultaneous extraction process and hypoglycemic activity in vitro of chlorogenic acid and total flavonoids were studied,in order to provide reference for the development of chlorogenic acid and total flavonoids hypoglycemic health care products of honeysuckle.The main findings are as follows:（1）A high-performance liquid chromatography method was established for simultaneous determination of chlorogenic acid and total flavonoids in honeysuckle.The liquid phase conditions are as follows:chromatographic column kromasil 100-5-C18（4.6 mm×250 mm,5μm）;mobile phase A phase is 0.4%phosphoric acid aqueous solution and phase B is methanol.Chromatographic gradient elution.The flow rate was 0.8 m L/min,the injection volume was 4μL,the detection wavelength was 327 nm,and the column temperature was 25℃.The results of methodological investigation showed that chlorogenic acid and total flavonoids had a good linear correlation within the determination range,the linear correlation coefficients were all above 0.99994,and the recovery rate was above 92%.The method had high precision and good stability.（2）The synchronous extraction process of chlorogenic acid and total flavonoids of honeysuckle was studied by heating extraction method in water bath.Taking the"normalized value"of the overall evaluation of chlorogenic acid and total flavonoids as the index,on the basis of single factor experiment,the Box-Behnken response surface method was used to optimize the ratio of liquid to material,extraction time,extraction temperature and ethanol concentration The optimal extraction process was as follows:the ratio of liquid to material was25:1（m L/g）,the extraction time was 82 min,the extraction temperature was 57℃,and the volume fraction of ethanol was 66%.Under the process conditions,the yields of chlorogenic acid and total flavonoids in honeysuckle were 3.08±1.41%and 6.34±1.48%,respectively.（3）The flavonoids in honeysuckle were qualitatively analyzed by UPLC-QTOF-MS technology.Through the comparison of standard products,four compounds of rutin,hypericin,quercetin and luteolin in honeysuckle were identified.Combined with academic literature data and database search results,15 compounds including quercetin-3-O-glucoside in honeysuckle were inferred.（4）The in vitro hypoglycemic activity of honeysuckle extract was analyzed.The test results showed that the inhibition rate of honeysuckle extract toα-amylase reached 53.50%,and the IC₅₀ was 8.79 mg/m L.The type of inhibition ofα-amylase by honeysuckle extract is reversible competitive inhibition.The inhibition rate of honeysuckle extract toα-glucosidase reached 70.95%,and the IC₅₀ was 0.034 mg/m L.The type of inhibition ofα-glucosidase by honeysuckle extract is reversible anticompetitive inhibition.