摘要
通过优化梯度洗脱条件和类胡萝卜素提取条件,探索建立基于超高效液相色谱(ultraperformanceliquid chromatography,UPLC)检测卤虫体内类胡萝卜素组成及含量的有效方法.结果表明:类胡萝卜素提取选用丙酮-甲醇(体积比为2:1)作为提取溶剂,在超声波处理30 min、65℃浸提20 min的条件下提取效果最佳.UPLC选用ACQUITYH-ClassBEHC18色谱柱(1.7μm,2.1 mm×150 mm)以及二极管阵列(PDA)检测器.梯度洗脱条件:10%超纯水+90%甲醇-乙腈溶液(体积比为3:7)16 min,20%甲基叔丁基醚+80%甲醇-乙腈溶液(体积比为3:7)14 min,流量0.3 mL/min,进样量10μL,柱温25℃,检测波长475 nm.在此条件下可以在30 min内有效检测出卤虫初孵无节幼体中的角黄素和海胆酮的含量,分别为(572.60±11.60)μg/g和(18.02±0.41)μg/g(以干质量计),且验证了无节幼体体内不含虾青素、玉米黄质和β-胡萝卜素.该方法快速、准确、灵敏、有效,为后续卤虫类胡萝卜素营养强化及类胡萝卜素代谢模型的建立提供了有效的技术手段.
In our study an effective method based on ultra performance liquid chromatography(UPLC) that can detect the composition and content of carotenoids in Artemia sp.was established via optimizing the conditions of gradient elution and carotenoid extraction.The results showed that the carotenoid extraction procedure for Artemia samples was better to use acetone-methanol (volume ratio 2∶1) solvent with ultrasonic treatment 30 min,then extraction at65℃f or 20 min.UPLC selected ACQUITY H-Class BEH C18(1.7μm,2.1 mm×150 mm) column and photo-diode array (PDA) detector.Gradient elution conditions were 10%ultrapure water+90%methanol-acetonitrile solution(volume ratio 3∶7) for 16 min,20%methyl tert-butyl ether+80%methanol-acetonitrile solution (volume ratio 3∶7) for 14min,flow rate 0.30 m L/min,injection volume 10μL,column temperature 25℃and detection wavelength475 nm.Under these conditions,the contents of canthaxanthin and echinenone in newly hatched Artemia nauplii can be effectively detected in 30 min,which were(572.60±11.60)μg/g and (18.02±0.41)μg/g (dry weight),respectively.However,no astaxanthin,zeaxanthin,and β-carotene were detected.The optimized UPLC method in the current study has proven fast,accurate,sensitive and effective.Therefore,this method provides an effective technical means for future carotenoids nutrition enhancement in Artemia as well as the establishment of carotenoid metabolism model.
作者
陈慧聪
焦烨磊
王伟隆
黄旭雄
CHEN Huicong;JIAO Yelei;WANG Weilong;HUANG Xuxiong(Centre for Research on Environmental Ecology and Fish Nutrition of the Ministry of Agriculture,Shanghai Ocean University,Shanghai 201306,China;National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University,Shanghai 201306,China;Shanghai Collaborative Innovation Center for Cultivating Elite Breeds and Green-culture of Aquaculture Animals,Shanghai 201306,China)
出处
《天津科技大学学报》
CAS
2022年第6期9-17,共9页
Journal of Tianjin University of Science & Technology
基金
沪农科推字(2019)第2-5号
国家自然科学基金青年科学基金项目(31902385)。
