干旱区生态环境知识资源中心(Arid): Optimization of extraction solvents, solid phase extraction and decoupling for quantitation of free isoprenoid diphosphates in Haematococcus pluvialis by liquid chromatography with tandem mass spectrometry

Arid

DOI	10.1016/j.chroma.2019.03.051
	Optimization of extraction solvents, solid phase extraction and decoupling for quantitation of free isoprenoid diphosphates in Haematococcus pluvialis by liquid chromatography with tandem mass spectrometry
	Jin, Hui 1; Lao, Yong Min 1,2; Zhou, Jin 1; Zhang, Huai Jin 3; Cai, Zhong Hua 1
通讯作者	Cai, Zhong Hua
来源期刊	JOURNAL OF CHROMATOGRAPHY A
ISSN	0021-9673
EISSN	1873-3778
出版年	2019
卷号	1598 页码:30-38
英文摘要	Isoprenoid diphosphates are important precursors actively participating in many downstream metabolisms; they are often in modified forms, e.g., protein-coupled or esterified form. Therefore, in vivo level of free isoprenoid diphosphates is quite low, (0) over tilde .07 nmol/g fresh weight in plants. In order to directly measure the isoprenoid diphosphate pool during stress-induced accumulation of astaxanthin in Haematococcus pluvialis, the present study optimized several pretreatment procedures to enrich free isoprenoid diphosphates for high-pressure liquid chromatography, with tandem mass spectrometry (HPLC-MS/MS) detection. Specifically, different extraction solvents, e.g., water, methanol, chloroform, and mixture of water, methanol, and chloroform (1:1:1, V/V/V), and solid phase extraction (SPE) columns (OASIS@ WAX and HLB Cartridges) were compared; and gentle decoupling by NaOH or trifluoroacetic acid (TFA) was introduced to release free isoprenoid diphosphates. Results found that solvent mixture of water, methanol and chloroform (1:1:1, V/V/V) showed the highest extraction efficiency (R-E) for five isoprenoid diphosphates, ranging from 76.83% to 92.43%; HLB column showed the balanced recoveries ranging from 75.29% to 87.54%; and incubation with low NaOH ((4) over tilde .7 mmol/L) at 4 degrees C significantly increased detectable isoprenoid diphosphates in algal cells, some of which were undetectable or in trace level before NaOH decoupling. The method was applied to H. pluvialis cells under various stresses. Low levels of isoprenoid diphosphates were determined in most of the stresses used, e.g., 0.19 +/- 0.09 to 0.98 +/- 0.06 mg/g fresh weight (FW) for IPP/DMAPP, 0.35 +/- 0.07 mg/g FW for GGPP and undetectable for FPP and GPP; while isoprenoid diphosphates were significantly accumulated in the dark to 3.27 +/- 0.05, 0.17 +/- 0.09, 1.81 +/- 0.16 and 0.58 +/- 0.07 mg/g FW for IPP/DMAPP, GPP, FPP and GGPP, respectively. These results implied that isoprenoid diphosphates were exhausted by downstream carotenogenesis under stress. Our work emphasizes NaOH decoupling for exact quantitation of in vivo isoprenoid diphosphates. (C) 2019 Elsevier B.V. All rights reserved.
英文关键词	Isoprenoid diphosphates Haematococcus pluvialis High-pressure liquid chromatography with tandem mass spectrometry (HPLC-MS/MS) Extraction solvent Solid phase extraction (SPE) Decoupling
类型	Article
语种	英语
国家	Peoples R China
收录类别	SCI-E
WOS记录号	WOS:000472241600003
WOS关键词	DIMETHYLALLYL DIPHOSPHATE ; FARNESYL DIPHOSPHATE ; NONRADIOACTIVE ASSAY ; BIOSYNTHESIS ; ACCUMULATION ; PRODUCTS ; TRANSFERASE ; SYNTHASE ; TISSUE ; PLANTS
WOS类目	Biochemical Research Methods ; Chemistry, Analytical
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
来源机构	清华大学
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/216837
作者单位	1.Tsinghua Univ, Grad Sch Shenzhen, Shenzhen Publ Platform Screening & Applicat Marin, Shenzhen 518055, Peoples R China; 2.Shenzhen Univ, Inst Adv Study, Shenzhen 518060, Peoples R China; 3.Tsinghua Univ, Sch Life Sci, Beijing 100084, Peoples R China
推荐引用方式 GB/T 7714	Jin, Hui,Lao, Yong Min,Zhou, Jin,et al. Optimization of extraction solvents, solid phase extraction and decoupling for quantitation of free isoprenoid diphosphates in Haematococcus pluvialis by liquid chromatography with tandem mass spectrometry[J]. 清华大学,2019,1598:30-38.
APA	Jin, Hui,Lao, Yong Min,Zhou, Jin,Zhang, Huai Jin,&Cai, Zhong Hua.(2019).Optimization of extraction solvents, solid phase extraction and decoupling for quantitation of free isoprenoid diphosphates in Haematococcus pluvialis by liquid chromatography with tandem mass spectrometry.JOURNAL OF CHROMATOGRAPHY A,1598,30-38.
MLA	Jin, Hui,et al."Optimization of extraction solvents, solid phase extraction and decoupling for quantitation of free isoprenoid diphosphates in Haematococcus pluvialis by liquid chromatography with tandem mass spectrometry".JOURNAL OF CHROMATOGRAPHY A 1598(2019):30-38.