摘要
目的优化桦树皮中桦木酸的提取纯化工艺,并研究其对斑马鱼(Danio rerio)的毒性效应。方法采用超声辅助乙醇提取桦木酸,通过单因素实验及响应面分析优化工艺参数,再经重结晶法进行纯化,探究其对模式生物斑马鱼的毒性效应,观察并计算斑马鱼的半数致死浓度(half lethal concentration,LC_(50))、生存率、畸形率和主要毒性靶器官。结果经优化验证后,最佳工艺条件为乙醇浓度75%、料液比1:25(g/mL)、超声温度45℃、超声时间30 min,桦木酸得率达(9.15±0.01)mg/g。以异丙醇为溶剂进行桦木酸重结晶,纯化的最佳工艺参数为料液比1:25(mg/mL)、重结晶温度0℃,重结晶时间24 h,此时桦木酸纯度达到96.49%。桦木酸对斑马鱼的LC_(50)为0.905mg/mL,致畸的主要表现为心包水肿、卵黄囊水肿、肾性水肿和胃肠道发育延迟,质量浓度为0.2~0.6mg/mL的桦木酸没有显示出急性毒性作用。结论优化后的纯化工艺可有效提取桦树皮中的桦木酸,且桦木酸在受试剂量下对斑马鱼胚胎及幼鱼无明显的急性毒性及致畸作用。
Objective To optimize the extraction and purification process of betulinic acid from birch bark,and investigate its toxic effects on zebrafish(Danio rerio).Methods Betulinic acid was extracted by ultrasonic-assisted ethanol,the process parameters were optimized by single-factor experiments and response surface analysis,and then purified by recrystallization to investigate the toxic effects on the model organism zebrafish,and the half lethal concentration(LC_(50)),survival rate,deformation rate and main toxic target organs were observed and calculated.Results After optimization and verification,the optimum process conditions were 75% ethanol concentration,1:25(g/mL)material-to-liquid ratio,45℃ sonication temperature and 30 min sonication time,and the extraction rate of betulinic acid reached(9.15±0.01)mg/g.Betulinic acid recrystallization was performed with isopropyl alcohol as solvent,the optimal process parameters for the purification were the material-liquid ratio of 1:25(mg/mL),recrystallization temperature of 0℃ and recrystallization time of 24 h.The purity of betulinic acid was 96.49%.The LC_(50)of betulinic acid for zebrafish was 0.905 mg/mL,the main teratogenic manifestations were pericardial edema,yolk sac edema,nephrogenic edema and delayed gastrointestinal development,and birchic acid at concentrations of 0.2-0.6 mg/mL did not show acute toxic effects.Conclusion The optimized purification process is effective in extracting betulinic acid from birch bark,which has no significant acute toxicity or teratogenic effects on zebrafish embryos and larvae at reagent levels.
作者
李镕基
陈海燕
赵峰
栾立
聂海民
LI Rong-Ji;CHEN Hai-Yan;ZHAO Feng;LUAN Li;NIE Hai-Min(College of Food Science and Engineering,Jilin Agricultural University,Changchun 130118,China;College of Life Science,Changchun Institute of Science and Technology,Changchun 130118,China;College of Pharmacy,Changchun University of Chinese Medicine,Changchun 130117,China;Horticultural Specialties Workstation,Deyhui City,Changchun 130300,China)
出处
《食品安全质量检测学报》
CAS
北大核心
2023年第3期68-77,共10页
Journal of Food Safety and Quality
基金
省部共建食品营养与安全国家重点实验室开放课题项目(TJKD20200812)
长春科技学院青年启动基金项目(CCKJ20221101)。
关键词
桦树皮
桦木酸
超声辅助提取
重结晶
斑马鱼
急性毒性
birch bark
betulinic acid
ultrasonic-assisted extraction
recrystallization
Danio rerio
acute toxicity
作者简介
李镕基,硕士,主要研究方向为营养与代谢机制研究。E-mail:lirongji6@126.com;通信作者:陈海燕,硕士,教授,主要研究方向为营养与代谢机制研究。E-mail:100361@cstu.edu.cn。
