纤维素酶联合微波提取石崖茶中黄酮类成分的工艺优化及其成分表征

本实验以黄酮得率为指标,在单因素实验的基础上,采用Box-Behnken响应面法优化纤维素酶联合微波提取石崖茶中黄酮类成分的工艺条件,采用扫描电镜(scanning electron microscope,SEM)对提取后的物料进行微观形态观察,利用高效液相色谱-超高分辨四级杆飞行时间串联质谱(high performance liquid chromatography quadrupole time-of-flight mass spectrometry,HPLC-Q-TOF-MS)对工艺优化后的黄酮提取物进行成分表征。结果表明,以pH为4的50%乙醇为溶剂,料液比1:40 g/mL,加入0.08%纤维素酶,51℃酶解60 min之后,再采用450 W微波处理3 min,该条件下石崖茶中黄酮类成分的得率可达35.01%±0.75%,比单独采用纤维素酶、微波处理分别提高了66.87%和18.12%。相对于单独采用纤维素酶或微波处理,二者联合作用时,石崖茶细胞壁破碎更为彻底,利于成分溶出。HPLC-Q-TOF-MS初步鉴定工艺优化后的提取物中含13种成分,分别为儿茶素、原花青素、表儿茶素、木犀草苷、野漆树苷、山茶苷B、槲皮苷、山茶苷A、芹菜素、金圣草黄素、野蔷薇苷、棕榈酸、熊果酸。本实验优化的工艺稳定、可行,可用于石崖茶中黄酮类成分的提取。

Optimization for Cellulase-microwave Extraction Process and Its Chemical Composition of Flavonoids Substance from Adinandra nitida Merr.ex H.L.Li

The extraction yield of flavonoids was taken as the index,based on the single factor experiments,Box-Behnken response surface methodology was used to optimize the cellulase-microwave extraction process of flavonoids from Adinandra nitida Merr.ex H.L.Li with the extraction yield of flavonoids as the index. The cell wall microstructure of the extraction were observed by scanning electron microscope(SEM)and the composition of flavonoids from Adinandra nitida Merr.ex H.L.Li were analyzed by high performance liquid chromatography quadrupole time-of-flight mass spectrometry(HPLC-Q-TOF-MS). Results showed that the optimal extraction were:Extraction with 50% ethanol at a material/liquid ratio of 1:40 g/mL,pH4.0 with an enzyme dose of 0.08%,cellulase hydrolysis for 60 min at 51℃,followed by the assistance of microwave treatment for 3 min at 450 W power. Under these conditions,the average yield of flavonoids from Adinandra nitida Merr.ex H.L.Li. was 35.01%±0.75%,which was 66.87% and 18.12% higher than that of cellulase and microwave treatment alone. The cell wall of Adinandra nitida Merr.ex H.L.Li was broken more thoroughly than that of cellulase or microwave treatment alone,which was favorable for dissolution of components. 13 components obtained by optimized extraction were detected through HPLC-Q-TOF-MS analysis,including catechin,procyanidin,epicatechin,luteoloside,rhoifolin,camellianin B,quercitroside,camellianin A,apigenin,chrysoeriol,rosamultin,palmitic acid and ursolic acid. This study would provide a preliminary basis for the development and utilization of Adinandra nitida Merr.ex H.L.Li.