海带中氧化脂类成分的液质联用分析研究

目的: [HTSS]利用电喷雾电离源负离子模式和选择离子模式(SRM),建立起高效液相色谱-三重四极杆质谱测定海带中氧化脂类的定量分析方法。方法: 海带样品经液氮研磨,乙酸乙酯提取后上机分析。采用Hypersil Gold C₁₈色谱柱(2.1 mm×100 mm,3 μm),柱温30℃,以乙腈(A)-0.2%乙酸水溶液(B)为流动相,梯度洗脱(0~40 min,30%A→100%A),流速0.2 mL·min^-1;采用电喷雾电离源负离子电离模式,选择反应监测(SRM)扫描模式。结果: 13种氧化脂类成分在Hypersil Gold C₁₈色谱柱上得到良好分离。方法在0.01~1.00 mg·L^-1浓度范围内线性相关系数除9,15-dioxo-11-hydroxy-prosta-5,13-dien-1-oic acid为0.9941外均大于0.9973,方法的检测限(S/N=3)为0.040~0.209 μg·L^-1,定量限(S/N=10)为1.365~6.993 μg·L^-1,基质效应在68.5%~110.3%之间,回收率为68.3%~113.8%,RSD均小于15%。另外,借助液相色谱-四极杆-飞行时间质谱系统,对海带中除有对照品的成分外可能存在的氧化脂类(15-HETE)成分进行了结构确认,并对海带中有对照品的氧化脂类成分进行了精确定量分析,对没有对照品的氧化脂类成分进行了半定量分析,结果检测到7个氧化脂类成分,分别为9,12,13-trihydroxy-10-octadecenoic acid[(109.8±10.2)ng·g^-1],12-oxo-10,15-phytodienoic acid[(229.3±12.9)ng·g^-1],13-hydroxy-9,11,15-octadecatrienoic acid[(4537.4±141.6)ng·g^-1],13-hydroxy-9,11-octadecadienoic acid[(673.4±9.9)ng·g^-1],13-hydroperoxy-9,11-octadecadienoic acid[(5591.0±103.3)ng·g^-1],13-oxo-9,11,15-octadecatrienoic acid[(6872.1±223.4)ng·g^-1],15-hydroxy-5,8,11,13-eicosatetraenoic acid[(2651.9±92.5)ng·g^-1]。结论: 本法操作简单,准确可靠,适用于藻类氧化脂类成分的含量检测。

1 Yang J,Schmelzer K,Georgi K,et al.Quantitative profiling method for oxylipin metabolome by liquid chromatography electrospray ionization tandem mass spectrometry[J].Anal Chem,2009,81(19):8085
2 Mosblech A,Feussner I,Heilmann I.Oxylipins:structurally diverse metabolites from fatty acid oxidation[J].Plant Physiol Biochem,2009,47(6):511
3 Blée E.Impact of phyto-oxylipins in plant defense[J].Trends Plant Sci,2002,7(7):315
4 Pohnert G.Diatom/copepod interactions in plankton:the indirect chemical defense of unicellular algae[J].Chembiochem,2005,6(6):946
5 Baranowski R,Pacha K.Gas chromatographic determination of prostaglandins[J].Mini-Rev Med Chem,2002,2(2):135
6 Wheelan P,Zirrolli JA,Murphy RC.Analysis of hydroxy fatty acids as pentafluorobenzyl ester,trimethylsilyl ether derivatives by electron ionization gas chromatography/mass spectrometry[J].J Am Soc Mass Spectrom,1995,6(1):40
7 Küpper FC,Gaquerel E,Boneberg EM,et al.Early events in the perception of lipopolysaccharides in the brown alga Laminaria digitata include an oxidative burst and activation of fatty acid oxidation cascades[J].J Exp Bot,2006,57(9):1991
8 Adas F,Picart D,Berthou F,et al.Liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry of ω-and (ω-1)-hydroxylated metabolites of elaidic and oleic acids in human and rat liver microsomes[J].J.Chromatogr B,1998,714(2):133
9 Ritter A,Goulitquer S,Salaün JP,et al.Copper stress induces biosynthesis of octadecanoid and eicosanoid oxygenated derivatives in the brown algal kelp Laminaria digitata[J].New Phytol,2008,180(4):809
10 Unterwurzacher I,Koal T,Bonn GK,et al.Rapid sample preparation and simultaneous quantitation of prostaglandins and lipoxygenase derived fatty acid metabolites by liquid chromatography-mass spectrometry from small sample volumes[J].Clin Chem Lab Med,2008,46(11):1589