Study On Extraction Technology Of Proanthocyanidins From Grape Seeds

In this paper,grape seeds was used as raw material to study the extraction and purification technology of procyanidins,and the pilot-scale amplification,the pilot scale experiment was carried out,and designed the production process of grape seed proanthocyanidin.The vanillin-hydrochloric acid method was established as a quantitative method for the determination of procyanidins,and the regression equation of standard curve fitted well;in addition,the High Performance Liquid Chromatography(HPLC)was established for the qualitative analysis of proanthocyanidins.The degrading grape seed was extracted by cellulase-ultrasonic assisted method,taking the extraction rate of grape seed procyanidins as an index,the effects of pH value,cellulase concentration,ethanol concentration,ratio of materia to liquid and ultrasonic time on the extraction rate were investigated,through L9(34)orthogonal experiments and verification experiments,we determined the optimal extraction conditions of Proanthocyanidins:cellulase addition amount 120 u/g,ethanol concentration 55%,material ratio 1:40,ultrasonic time 20 min,the extraction rate of procyanidins from grape seed was 17.170%under the experimental conditions,which is 17.20%higher than that of the pure ultrasonic method under the same experimental conditions.Column chromatography was used to separate and purify crude extract of grape seed proanthocyanidin,with AB-8 macroporous adsorption resin as adsorbent,through static and dynamic adsorption experiments found:the adsorption behavior of AB-8 macroporous adsorption resin on proanthocyanidins accords with Langmuir equation;taking the resin adsorption capacity as index,the effects of the sample concentration and the adsorption velocity on the dynamic adsorption of resin were investigated;when 400 mL,8 mg/mL procyanidin solution was passed through the resin layer at a rate of 4 BV/h,the dynamic adsorption capacity of resin was 88.64 mg/mL,with 20%to 60%ethanol solution for gradient elution,the final desorption rate reached 90.80%;the deactivation and regeneration properties of AB-8 resin were studied,and proposed an effective regeneration treatment method;after purification by macroporous resin,the purity of proanthocyanidins was up to 95.48%,and the HPLC chromatograms of samples were consistent with the standard spectrum.The scale-up experiment of grape seed proanthocyanidin was carried out,through the independent design and development of ultrasonic extraction equipment and column chromatography equipment for separation and purification of grape seeds procyanidins,extraction kettle with effective volume of 30 L can realize a single treatment of grape seeds was 3 kg,get 117.9 g of proanthocyanidins products,its extraction rate and purity was 15.47%and 93.59%respectively,basically meet the market quality requirements.According to the experimental results,we designed the production process of annual production of 10 t grape seed procyanidins.