摘要
建立了超声提取-毛细管气相色谱法分析植物细胞膜脂脂肪酸组成与含量的快速检测方法,以用于植物生理抗逆性研究。采用氯仿-甲醇-水体系超声提取植物总类脂,改变KOH浓度为0.05～0.075mol/L,利用叶绿素的皂化反应产物溶于水的性质除去色素干扰,膜脂经氢氧化钾-三氟化硼法甲酯化后上机分析。采用100∶1分流比进样,220～240℃程序升温操作,BPX70毛细管柱分离,FID检测。主要脂肪酸在12min内出峰,分离度>2,利用线性程序升温等当碳长度(ECLT)值结合标准品对照的方法对脂肪酸组分包括存在双键异位的不饱和脂肪酸进行定性。面积归一法或标准系数法定量,在31.25～5000ng的实验浓度范围内线性良好。实测了经乙烯利和多效唑处理后狗牙根叶片膜脂脂肪酸中10种脂肪酸组成与含量。该方法准确、简便、实用,可应用于植物叶片膜脂脂肪酸分析。
A rapid analytical method was developed for the determination of fatty acids in the membrane lipids of plant cell by ultrasonic extraction-capillary gas chromatography.The membrane lipids were extracted from fresh leaves using ultrasonic with Chloroform-Methanol-Water solvent.Chlorophyll could be separated according to the characteristic that the production of saponified chlorophyll could be dissolved in water when KOH concentration is 0.05～0.075 mol/L.The membrane lipids were esterifies with KOH-BF_3 method followed by gas chromatography.A fused-silica BPX70 capillary column (60 m×0.25 mm ID) and FID detector were employed in gas chromatography.The inlet mode was split with split rate 100∶1.Temperature program settings were:initial temperature 220℃,increase at 1℃/min to final temperature 240℃.Fatty acids, more importantly,the positional isomers,were identified by fatty acid methyl ester standards or applying linear programmed temperature equivalent chain length (ECL^T) values based on even-chain saturated fatty acid methyl ester(FAME) standards.Area normalization method or external standard coefficient method could be use to quantified the content of fatty acids.This method was simple,fast and practical.It has been successfully applied to the analysis of fatty acids in the membrane lipids of Cynodon dactylon treated with paclobutrazol(PP333) or ethphon.
出处
《广西农业生物科学》
CAS
CSCD
2004年第2期154-158,共5页
Journal of Guangxi Agricultural and Biological Science
基金
国家自然科学基金资助项目(30170033)
关键词
膜脂
脂肪酸
毛细管气相色谱法
超声提取
叶绿素
membrane lipid
fatty acid
capillary gas chromatography
ultrasonic extraction
chlorophyll
