干旱区生态环境知识资源中心(Arid): 基于高效液相色谱法的扇贝毒素-2制备方法研究

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	基于高效液相色谱法的扇贝毒素-2制备方法研究
其他题名	PREPARATION OF PECTENOTOXIN-2 BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
	王玉 1; 彭吉星 2; 吴海燕 2; 郭萌萌 2; 佟蒙蒙 3; 谭志军 2
来源期刊	海洋与湖沼
ISSN	0029-814X
出版年	2018
卷号	49 期号:3 页码:568-576
中文摘要	本文以渐尖鳍藻(Dinophysis acuminata)为实验原料,建立了基于高效液相色谱法(HPLC)的扇贝毒素-2(Pectenotoxin-2, PTX2)的制备方法。研究重点对鳍藻细胞的破碎、PTX2毒素的提取及HPLC纯化等三个步骤的方法进行筛选和优化。结果表明,反复冻融与超声波细胞破碎相结合可获得最大的细胞破碎率和毒素提取率,分别为99.2%和81.5%。采用四种有机试剂(甲醇、丙酮、乙醚、二氯甲烷)及三种固相萃取小柱(Solid Phase Extraction, SPE)(Oasis HLB SPE, Phenomenex Strata-X SPE, Sep-Pak C_(18) SPE)对PTX2毒素进行提取,其中乙醚-二氯甲烷的提取效率最高,达78.2%。在HPLC对PTX2毒素的纯化实验中发现,流动相的pH会影响毒素组分的分离,其中在碱性流动相(含6.7mmol/L氨水的水溶液和乙腈溶液, pH=10.94)条件下, PTX2的分离效果最佳,响应值最高。实验使用Phenomenex Kinetex C_(18)(4.8mm*250mm, 5mum)色谱柱,在上述碱性流动相条件下,分离得到纯度为93.2%的PTX2毒素。本研究选定的藻细胞破碎、PTX2毒素提取、及HPLC纯化方法简单快捷,高效易操作,为从大规模培养的鳍藻细胞中制备高纯度PTX2毒素提供了技术支持。
英文摘要	A protocol for preparing pectenotoxin-2 (PTX2) from a large-scale culture of Dinophsis acuminata was conducted in high-performance liquid chromatography (HPLC). Methods of cell disruption, toxin extraction, and HPLC purification were selected and optimized. The results indicate that the most efficient cell disruption and toxin extraction were achieved in the combination of freezing+ thawing+ ultrasound, resulting in 99.2% and 81.5%, respectively. PTX2 was extracted using liquid-liquid extraction (methanol, acetone, diethyl ether, and dichloromethane) and Solid Phase Extraction columns (Oasis HLB SPE, Phenomenex Strata-X SPE, Sep-Pak C_(18) SPE). PTX2 extraction efficiency reached maximum (78.19%) with diethyl ether and dichloromethane. Studies on the optimization conditions of HPLC showed that the pH value of the mobile phase had an impact on the chromatographic reaction. When mobile phase was in alkaline condition (containing 6.7mmol/L ammonium hydroxide in acetonitrile/water, pH=10.94), PTX2 could be better chromatographically separated by Phenomenex Kinetex C_(18) (4.8mm *250mm, 5mum). The final PTX2 reached 93.2% in purity. This method was rapid, effective, and comprehensive for extraction, separation, and purification of PTX2 from D. acuminata cells with HPLC.
中文关键词	扇贝毒素-2 ; 鳍藻 ; 高效液相色谱 ; 萃取 ; 制备
英文关键词	pectenotoxin-2 Dinophysis acuminata high-performance liquid chromatography extraction preparation
语种	中文
国家	中国
收录类别	CSCD
WOS类目	CHEMISTRY ANALYTICAL
WOS研究方向	Chemistry
CSCD记录号	CSCD:6267743
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/237816
作者单位	1.浙江大学海洋学院海洋生物研究所;;中国水产科学研究院黄海水产研究所, ;;, 舟山;;青岛, ;; 316021;;266071; 2.中国水产科学研究院黄海水产研究所, 青岛, 山东 266071, 中国; 3.浙江大学海洋学院海洋生物研究所;;舟山市疾病预防控制中心, ;;, 舟山;;舟山, ;; 316021;;316021
推荐引用方式 GB/T 7714	王玉,彭吉星,吴海燕,等. 基于高效液相色谱法的扇贝毒素-2制备方法研究[J],2018,49(3):568-576.
APA	王玉,彭吉星,吴海燕,郭萌萌,佟蒙蒙,&谭志军.(2018).基于高效液相色谱法的扇贝毒素-2制备方法研究.海洋与湖沼,49(3),568-576.
MLA	王玉,et al."基于高效液相色谱法的扇贝毒素-2制备方法研究".海洋与湖沼 49.3(2018):568-576.