香蕉果肉类胡萝卜素提取与测定方法  

Optimization of Extraction and Determination of Carotenoids from Banana Pulp

作　　者：范迎康 李莉 詹妮 况梦宇 邓贵明[2] 吕培涛 盛鸥[2] FAN Yingkang;LI Li;ZHAN Ni;KUANG Mengyu;DENG Guiming;LYU Peitao;SHENG Ou(College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350000,China;Institute of Fruit Science,Guangdong Academy of Agricultural Sciences/Key Laboratory of Biology and Genetic Resources Utilization of South Subtropical Fruit Trees,Ministry of Agriculture and Rural Affairs/Guangdong Key Laboratory of Tropical and Subtropical Fruit Trees,Guangzhou,Guangdong 510640,China)

机构地区：[1]福建农林大学园艺学院,福建福州350000 [2]广东省农业科学院果树研究所/农业农村部南亚热带果树生物学与遗传资源利用重点实验室/广东省热带亚热带果树研究重点实验室,广东广州510640

出　　处：《热带作物学报》2022年第9期1945-1952,共8页Chinese Journal of Tropical Crops

基　　金：“十三五”国家重点研发计划重点专项(No.2019YFD1000203);广东省农科院“金颖之星”人才项目(No.R2020PY-JX002);广东省现代农业产业技术体系创新团队项目(No.2021KJ109)

摘　　要：大规模开展香蕉类胡萝卜素种质资源评价,发掘高类胡萝卜素的品种资源,提高主栽品种中类胡萝卜素的含量,对于提升香蕉产业竞争力具有重要的作用。开展此项研究需要建立高效的香蕉果肉类胡萝卜素提取和测定体系。本文拟建立基于超高效液相色谱仪(Agilent 1290),适用于香蕉果肉的类胡萝卜素提取及测定的方法。利用YMC-C30色谱柱,在450 nm检测波长和柱温20℃条件下,比较了不同的料液比(g∶mL,1∶6、1∶9、1∶12、1∶15)、定容溶液的比例(V∶V,MTBE∶甲醇=1∶0、1∶1、1∶2)、流动相(V∶V,甲醇∶乙腈=4∶0、3∶1、2∶2、1∶3、0∶4)、流速(0.6、0.8、1.2 mL/min)等对类胡萝卜素组分鉴定和含量测定的影响。结果表明,当提取试剂正己烷∶丙酮∶无水乙醇=2∶1∶1(V∶V∶V),果肉与提取试剂的料液比为1∶9,超声波破碎30 min,重复3次,定容溶液MTBE∶甲醇=1∶1时,香蕉类胡萝卜素组分分离的效果最好。当色谱条件A相以甲醇∶乙腈,B相以MTBE(100%)为流动相,进样量为10μL,流速为1mL/min进行梯度洗脱时,可以很好的鉴定并分离类胡萝卜素的主要组分。利用优化后的体系进行检测不同类胡萝卜素含量的香蕉品种,发现香蕉果肉中的主要类胡萝卜素组分为叶黄素、α-胡萝卜素、β-胡萝卜素。高类胡萝卜素含量的‘Frenchsomber’香蕉品种约含9.79μg/g,而低含量的‘中蕉8号’香蕉品种只有约0.201μg/g,这也表明不同香蕉品种中类胡萝卜素的含量差异相对较大。本文中以香蕉果肉为材料所建立的类胡萝卜素提取与检测方法,操作简单,精确度高,为科研工作者在香蕉类胡萝卜素的功能研究方面提供参考。Large-scale evaluation of banana carotenoids germplasm resources, exploration of varieties with high carotenoid resources and improvement of carotenoid contents in main varieties are of great importance to enhance the competitiveness of banana industry. It is necessary to establish an efficient extraction. Meanwhile, it is as well important to grope a determination system for carotenoids in banana fruit pulp. Therefore, this paper primarily aims to establish an optimized protocol for extraction and determination of carotenoids from banana pulp. In this paper, Ultra Performance Liquid Chromatography(UPLC, Agilent 1290) was used to determine carotenoids in banana pulp. By performing on a YMC C30 column at the time when column temperature maintaining at 20℃ and the Ultraviolet detection wavelength at 450 nm, we compared the effects of different ratios of solid-liquid(g∶m L=1∶6, 1∶9, 1∶12, 1∶15), different constant volume solution(MTBE∶methanol, V∶V=1∶0, 1∶1, 1∶2), different mobile phase(methanol∶acetonitrile, V∶V=4∶0, 3∶1, 2∶2, 1∶3, 0∶4,), and different flow rates(0.6 mL/min, 0.8 mL/min, 1 mL/min, 1.2 mL/min) on the identification and content determination of carotene in banana fruit pulp. The results showed that when the extraction solution of hexane/acetone/anhydrous ethanol was 2∶1∶1(V∶V∶V), the solid-liquid ratio of pulp to extraction reagent was 1∶9(g/mL), ultrasonic crushing was performed for 30 minutes, repeated 3 times and the ratio for constant volume solution methanol/MTBE was 1∶1(V∶V), the separation effect of carotenoids component in banana fruit pulp was the best. Moreover, the main composition of carotenoids were efficiently separated and identified when we used the binary mobile phase consisting solvent A(methanol∶acetonitrile=1∶1) and solvent B(MTBE, 100%), under a linear gradient elution procedure, the injection volume of 10 μL and at the flow rate of 1 mL/min. The optimized system was used to detect the carotenoid contents of different kinds of

关 键 词：香蕉 类胡萝卜素 组分鉴定 超高效液相色谱 体系优化 

分 类 号：S668.1[农业科学—果树学]