漂浮型和定生型铜藻中色素的液质联用分析
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摘要
为探究由气候变化、浅海海水污染加剧等因素造成的铜藻爆发性漂浮增殖现象,明确铜藻的漂浮生长机制,采用高效液相色谱-三重四级杆串联质谱(HPLC-QqQ-MS)技术建立11种色素(岩藻黄素、角黄素、玉米黄质、叶黄素、叶绿素a、叶绿素b、α-胡萝卜素、β-胡萝卜素、ε-胡萝卜素、γ-胡萝卜素和ζ-胡萝卜素)的定性定量分析方法,分析不同地区、不同生态型及不同部位的铜藻中色素的种类和含量。结果表明,共检测到10种色素,其中东极岛漂浮型铜藻中的叶绿素a、叶绿素b和β-胡萝卜素含量显著高于其他地区,而叶黄素、玉米黄质和岩藻黄素含量最低;定生型铜藻的色素总量是漂浮型铜藻的1.6倍;铜藻气囊中含有大量的叶黄素、岩藻黄素、玉米黄质、叶绿素a和叶绿素b等5种色素。本研究建立了快速、灵敏的液相色谱质谱联用定量分析方法,比较分析了不同铜藻中的10种色素种类和含量分布规律,为铜藻的健康、可控利用研究提供了基础数据支撑。
In order to study the floating growth mechanism of S. horneri, the qualitative and quantitative analyses of eleven pigments including fucoxanthin, canthaxanthin, zeaxanthin, lutein, chlorophyll a, chlorophyll b, α-carotene, β-carotene, ε-carotene, γ-carotene, ζ-caroteney were established by high performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry(HPLC-QqQ-MS), to investigate the pigment types and contents of S. horneri, which covered different regions, ecotypes and parts. The results showed that total 10 pigments except γ-carotene were detected. Among them, the contents of chlorophyll a, chlorophyll b and β-carotene in the floating S. horneri in the Dongji Island had highest levels compared to those of the other regions; while the contents of lutein, zeaxanthin and fucoxanthin were the lowest levels. The total pigment contents of the fixed-type S. horneri was 1.6 times that of the floating-type one. Moreover, the air bags of S. horneri contained a large amounts of pigments such as lutein, fucoxanthin, zeaxanthin, chlorophyll a and chlorophyll b. In summary, a rapid and sensitive quantitative methods was developed based on HPLC-QqQ-MS to investigated fluctuant of 10 pigments in different kinds of S. horneri, which will have great significance for the healthy and controllable utilization of S. horneri.
In order to study the floating growth mechanism of S. horneri, the qualitative and quantitative analyses of eleven pigments including fucoxanthin, canthaxanthin, zeaxanthin, lutein, chlorophyll a, chlorophyll b, α-carotene, β-carotene, ε-carotene, γ-carotene, ζ-caroteney were established by high performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry(HPLC-QqQ-MS), to investigate the pigment types and contents of S. horneri, which covered different regions, ecotypes and parts. The results showed that total 10 pigments except γ-carotene were detected. Among them, the contents of chlorophyll a, chlorophyll b and β-carotene in the floating S. horneri in the Dongji Island had highest levels compared to those of the other regions; while the contents of lutein, zeaxanthin and fucoxanthin were the lowest levels. The total pigment contents of the fixed-type S. horneri was 1.6 times that of the floating-type one. Moreover, the air bags of S. horneri contained a large amounts of pigments such as lutein, fucoxanthin, zeaxanthin, chlorophyll a and chlorophyll b. In summary, a rapid and sensitive quantitative methods was developed based on HPLC-QqQ-MS to investigated fluctuant of 10 pigments in different kinds of S. horneri, which will have great significance for the healthy and controllable utilization of S. horneri.
引文
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[6] 关万春, 陈亨, 王铁杆, 陈少波, 徐军田. 阳光紫外辐射对铜藻的生长及荧光参数的影响[J]. 水产学报, 2016, 40(1): 83-91
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[9] Karadas F, Pappas A C, Surai P F, Speake B K. Embryonic development within carotenoid-enriched eggs influences the post-hatch carotenoid status of the chicken[J]. Comparative Biochemistry & Physiology Part B Biochemistry & Molecular Biology, 2005, 141(2): 244-251
[10] Rocchi L, Rustioni L, Failla O. Chlorophyll and carotenoid quantifications in white grape (Vitisvinifera L.) skins by reflectance spectroscopy[J]. Vitis, 2016, 55(1): 11-16
[11] Damm A, Guanter L, Verhoef W, Schl?pfer D, Garbari S, Schaepman M E. Impact of varying irradiance on vegetation indices and chlorophyll fluorescence derived from spectroscopy data[J]. Remote Sensing of Environment, 2015, 156: 202-215
[12] 雷琼, 王芝林, 白雪. 胶束毛细管电泳法测定龙胆属藏药中环烯醚萜类成分[J]. 中药材, 2018, 41(8): 1930-1933
[13] Wei W L, Guo B Y, Lin J M. Ultra-high concentration of amylose for chiral separations in capillary electrophoresis[J]. Journal of Chromatography A, 2009, 1216(9): 1484-1489
[14] Gupa Ⅴ, Kumar M, Brahmbhatt H, Reddy C R K, Seth A, JhaB. Simultaneous determination of different endogenetic plant growth regulators in common green seaweeds using dispersive liquid-liquid microextraction method[J]. Plant Physiology and Biochemistry, 2011, 49(11): 1259-1263
[15] Kai J, Feng X M, Liu K. Development of a subcritical fluid extraction and GC-MS validation method for polychlorinated biphenyls (PCBs) in marine samples[J]. Journal of Chromatography B, 2013, 923/924: 37-42
[16] Daood H G, Czinkotal B, ágoston H, Biacs P. High-performance liquid chromatography of chlorophylls and carotenoids from vegetables[J]. Journal of Chromatography A, 2015, 472(1): 296-302
[17] Suzuki K, Kamimura A, Hooker S B. Rapid and highly sensitive analysis of chlorophylls and carotenoids from marine phytoplankton using ultra-high performance liquid chromatography (UHPLC) with the first derivative spectrum chromatogram (FDSC) technique[J]. Marine Chemistry, 2015, 176(20): 96-109
[18] Moreau R A, Powell M J, Agnew J, Young D H. Chlorophyll-derived porphyrins co-chromatograph with phospholipids in high performance liquid chromatographic separations of plant lipid classes[J]. Phytochemical Analysis, 2015, 9(1): 1-4
[19] Christ B, Hauenstin M, H?rtensteiner S. A liquid chromatography-mass spectrometry platform for the analysis of phyllobilins, the major degradation products of chlorophyll in Arabidopsis thaliana[J]. Plant Journal for Cell & Molecular Biology, 2016, 88(3): 505-518
[20] Zhai X J, Chen F, Zhu C R, Lu Y N. A simple LC-MS/MS method for the determination of cortisol, cortisone and tetrahydro-metabolites in human urine: Assay development, validation and application in depression patients[J]. Journal of Pharmaceutical and Biomedical Analysis, 2015, 107: 450-455
[21] Zhao H M, Wang Y F, Yuan B, Liu S, Man S, Xu H Y, Lu X M. A novel LC-MS/MS assay for the simultaneous determination of melatonin and its two major metabolites, 6-hydroxymelatonin and 6-sulfatoxymelatonin in dog plasma: Application to a pharmacokinetic study[J]. Journal of Pharmaceutical and Biomedical Analysis, 2016, 117: 390-397
[22] Yao J F, Zhou N, Bai L, Xu P X, Liu K L, Xue M. Simultaneous determination of five novel luteinizing hormone-releasing hormone antagonists by LC-MS and pharmacokinetics in rats following cassette dosing[J]. Journal of Chromatography B, 2014, 962: 94-101
[23] 戴超, 郑鹭飞, 刘佳萌, 王凤忠, 孙玉凤, 卢嘉, 郑旭, 王淼, 贺妍, 郭静, 范蓓. 液质联用法分析贮藏因素对马铃薯中α-茄碱含量的影响[J]. 核农学报, 2017, 31(11): 2200-2205
[24] Patil R B, Kore B A. Phytoconstituents, pigments, gas chromatography mass spectrometry analysis, and allelopathy effect of Alternanthera ficoidea L. P. Beauv [J]. Asian Journal of Pharmaceutical & Clinical Research, 2017, 10(2):103-108
[25] Stanislaw W, Dariusz D, Roman K. Two-step dispersive-solid phase extraction strategy for pesticide multiresidue analysis in a chlorophyll-containing matrix by gas chromatography–tandem mass spectrometry[J]. Journal of Chromatography A, 2015, 1412: 22-32
[26] 郑立洋, 陈娟娟, 徐继林, 周成旭, 严小军. 高效液相色谱-三重四极杆质谱同时测定与筛查海洋浮游藻类中的5种色素[J]. 色谱, 2014, 32(9): 992-998
[27] Luo Q J, Yue S, Zheng L Y, Chen J J, Xu J L, He S, Yan X J. Characterization of phytoplankton communities in response to macronutrient treatments determined by high-performance liquid chromatography mass spectrometry[J]. Analytical Letters, 2016, 49(11): 1758-1771
[28] Li N, Hu J J, Wang S, Cheng J, Hu X L, Lu Z Y, Lin Z J, Zhu W M, Bao Z M. Isolation and identification of the main carotenoid pigment from the rare orange muscle of the Yesso scallop[J]. Food chemistry, 2010, 118(3): 616-619
[29] Takaaichi S. Carotenoids in algae: Distributions, biosyntheses and functions[J]. Marine Drugs, 2011, 9(6): 1101-1118
[30] Canfifld L M, Krinsky N Ⅰ, Olson J A. Carotenoids in Human Health[M]. USA: Annals of the New York Academy of Sciences, 1993: 156-166
[31] 张义浩, 王志铮, 吴常文, 赵盛龙, 严世强, 侯伟芬. 舟山群岛定生海藻种类组成、生态分布及区系特征研究[J]. 浙江海洋学院学报(自然科学版), 2002, 21(2): 98-105
[32] Le M, LinX M, Zou Z Y,Xu X R, Li Y, Xu R. A 12-week lutein supplementation improves visual function in Chinese people with long-term computer display light exposure [J]. British Journal of Nutrition, 2009, 102(2): 186-190
[33] Osuka A, Kume T. Fucoxanthin and peridininpheophorbide-α molecules as a new light-harvesting model[J]. Tetrahedron Letters, 1998, 39(7): 655-658
[34] Osuka A, Kume T, Haggquist G W, JávorfiT, Lima J C, Melo E, NaqviK R. Photophysical characteristics of two model antenna systems: a fucoxanthin–pyropheoporbide dyad and its peridinin analogue[J]. Chemical Physics Letters, 1999, 313(3/4): 499-504
[35] Katoh T, Nagashima U, Mimuro M. Fluorescence properties of the allenic carotenoid fucoxanthin: Implication for energy transfer in photosynthetic pigment systems[J]. Photosynthesis Research, 1991, 27(3): 221-226
[2] 张鹏, 蔡一凡, 王铁杆, 钟晨辉, 谢起浪, 陈少波, 王宁. 浙江沿海不同地理群体铜藻Sargassum horneri的AFLP分析[J]. 浙江农业学报, 2015, 27(9): 1586-1592
[3] 孙建璋, 陈万东, 庄定根, 郑海羽, 林力, 逢少军. 中国南麂列岛铜藻Sargassum horneri实地生态学的初步研究[J]. 南方水产, 2008, 4(3): 58-63
[4] 孙建璋, 庄定根, 王铁干, 陈万东, 杨加波. 南麂列岛铜藻的研究[J]. 现代渔业信息, 2009, 24(5): 19-21
[5] 陈军, 王寅初, 余秋瑢, 毕燕会, 何培民, 刘正一. 绿潮暴发期间我国青岛漂浮铜藻的分子鉴定[J]. 生物学杂志, 2016, 33(1): 39-42
[6] 关万春, 陈亨, 王铁杆, 陈少波, 徐军田. 阳光紫外辐射对铜藻的生长及荧光参数的影响[J]. 水产学报, 2016, 40(1): 83-91
[7] 包楠欧, 史定刚, 关万春, 孙敏, 张鹏, 彭欣, 王铁杆, 陈少波, 仇建标. CO2及光强对南麂列岛铜藻生长的影响[J]. 浙江农业学报, 2014, 26(3): 649-655
[8] Fromme P, Melkozernov A, Jordan P, Krauss N. Structure and function of photosystem Ⅰ: Interaction with its soluble electroncarriers and external antenna systems[J]. FEBS Letters, 2003, 555(1): 40-44
[9] Karadas F, Pappas A C, Surai P F, Speake B K. Embryonic development within carotenoid-enriched eggs influences the post-hatch carotenoid status of the chicken[J]. Comparative Biochemistry & Physiology Part B Biochemistry & Molecular Biology, 2005, 141(2): 244-251
[10] Rocchi L, Rustioni L, Failla O. Chlorophyll and carotenoid quantifications in white grape (Vitisvinifera L.) skins by reflectance spectroscopy[J]. Vitis, 2016, 55(1): 11-16
[11] Damm A, Guanter L, Verhoef W, Schl?pfer D, Garbari S, Schaepman M E. Impact of varying irradiance on vegetation indices and chlorophyll fluorescence derived from spectroscopy data[J]. Remote Sensing of Environment, 2015, 156: 202-215
[12] 雷琼, 王芝林, 白雪. 胶束毛细管电泳法测定龙胆属藏药中环烯醚萜类成分[J]. 中药材, 2018, 41(8): 1930-1933
[13] Wei W L, Guo B Y, Lin J M. Ultra-high concentration of amylose for chiral separations in capillary electrophoresis[J]. Journal of Chromatography A, 2009, 1216(9): 1484-1489
[14] Gupa Ⅴ, Kumar M, Brahmbhatt H, Reddy C R K, Seth A, JhaB. Simultaneous determination of different endogenetic plant growth regulators in common green seaweeds using dispersive liquid-liquid microextraction method[J]. Plant Physiology and Biochemistry, 2011, 49(11): 1259-1263
[15] Kai J, Feng X M, Liu K. Development of a subcritical fluid extraction and GC-MS validation method for polychlorinated biphenyls (PCBs) in marine samples[J]. Journal of Chromatography B, 2013, 923/924: 37-42
[16] Daood H G, Czinkotal B, ágoston H, Biacs P. High-performance liquid chromatography of chlorophylls and carotenoids from vegetables[J]. Journal of Chromatography A, 2015, 472(1): 296-302
[17] Suzuki K, Kamimura A, Hooker S B. Rapid and highly sensitive analysis of chlorophylls and carotenoids from marine phytoplankton using ultra-high performance liquid chromatography (UHPLC) with the first derivative spectrum chromatogram (FDSC) technique[J]. Marine Chemistry, 2015, 176(20): 96-109
[18] Moreau R A, Powell M J, Agnew J, Young D H. Chlorophyll-derived porphyrins co-chromatograph with phospholipids in high performance liquid chromatographic separations of plant lipid classes[J]. Phytochemical Analysis, 2015, 9(1): 1-4
[19] Christ B, Hauenstin M, H?rtensteiner S. A liquid chromatography-mass spectrometry platform for the analysis of phyllobilins, the major degradation products of chlorophyll in Arabidopsis thaliana[J]. Plant Journal for Cell & Molecular Biology, 2016, 88(3): 505-518
[20] Zhai X J, Chen F, Zhu C R, Lu Y N. A simple LC-MS/MS method for the determination of cortisol, cortisone and tetrahydro-metabolites in human urine: Assay development, validation and application in depression patients[J]. Journal of Pharmaceutical and Biomedical Analysis, 2015, 107: 450-455
[21] Zhao H M, Wang Y F, Yuan B, Liu S, Man S, Xu H Y, Lu X M. A novel LC-MS/MS assay for the simultaneous determination of melatonin and its two major metabolites, 6-hydroxymelatonin and 6-sulfatoxymelatonin in dog plasma: Application to a pharmacokinetic study[J]. Journal of Pharmaceutical and Biomedical Analysis, 2016, 117: 390-397
[22] Yao J F, Zhou N, Bai L, Xu P X, Liu K L, Xue M. Simultaneous determination of five novel luteinizing hormone-releasing hormone antagonists by LC-MS and pharmacokinetics in rats following cassette dosing[J]. Journal of Chromatography B, 2014, 962: 94-101
[23] 戴超, 郑鹭飞, 刘佳萌, 王凤忠, 孙玉凤, 卢嘉, 郑旭, 王淼, 贺妍, 郭静, 范蓓. 液质联用法分析贮藏因素对马铃薯中α-茄碱含量的影响[J]. 核农学报, 2017, 31(11): 2200-2205
[24] Patil R B, Kore B A. Phytoconstituents, pigments, gas chromatography mass spectrometry analysis, and allelopathy effect of Alternanthera ficoidea L. P. Beauv [J]. Asian Journal of Pharmaceutical & Clinical Research, 2017, 10(2):103-108
[25] Stanislaw W, Dariusz D, Roman K. Two-step dispersive-solid phase extraction strategy for pesticide multiresidue analysis in a chlorophyll-containing matrix by gas chromatography–tandem mass spectrometry[J]. Journal of Chromatography A, 2015, 1412: 22-32
[26] 郑立洋, 陈娟娟, 徐继林, 周成旭, 严小军. 高效液相色谱-三重四极杆质谱同时测定与筛查海洋浮游藻类中的5种色素[J]. 色谱, 2014, 32(9): 992-998
[27] Luo Q J, Yue S, Zheng L Y, Chen J J, Xu J L, He S, Yan X J. Characterization of phytoplankton communities in response to macronutrient treatments determined by high-performance liquid chromatography mass spectrometry[J]. Analytical Letters, 2016, 49(11): 1758-1771
[28] Li N, Hu J J, Wang S, Cheng J, Hu X L, Lu Z Y, Lin Z J, Zhu W M, Bao Z M. Isolation and identification of the main carotenoid pigment from the rare orange muscle of the Yesso scallop[J]. Food chemistry, 2010, 118(3): 616-619
[29] Takaaichi S. Carotenoids in algae: Distributions, biosyntheses and functions[J]. Marine Drugs, 2011, 9(6): 1101-1118
[30] Canfifld L M, Krinsky N Ⅰ, Olson J A. Carotenoids in Human Health[M]. USA: Annals of the New York Academy of Sciences, 1993: 156-166
[31] 张义浩, 王志铮, 吴常文, 赵盛龙, 严世强, 侯伟芬. 舟山群岛定生海藻种类组成、生态分布及区系特征研究[J]. 浙江海洋学院学报(自然科学版), 2002, 21(2): 98-105
[32] Le M, LinX M, Zou Z Y,Xu X R, Li Y, Xu R. A 12-week lutein supplementation improves visual function in Chinese people with long-term computer display light exposure [J]. British Journal of Nutrition, 2009, 102(2): 186-190
[33] Osuka A, Kume T. Fucoxanthin and peridininpheophorbide-α molecules as a new light-harvesting model[J]. Tetrahedron Letters, 1998, 39(7): 655-658
[34] Osuka A, Kume T, Haggquist G W, JávorfiT, Lima J C, Melo E, NaqviK R. Photophysical characteristics of two model antenna systems: a fucoxanthin–pyropheoporbide dyad and its peridinin analogue[J]. Chemical Physics Letters, 1999, 313(3/4): 499-504
[35] Katoh T, Nagashima U, Mimuro M. Fluorescence properties of the allenic carotenoid fucoxanthin: Implication for energy transfer in photosynthetic pigment systems[J]. Photosynthesis Research, 1991, 27(3): 221-226