A micromethod using gas-liquid chromatography for measuring individual fatty acids liberated during interaction of triglyceride-rich lipoproteins and lipoprotein lipase.
A micromethod using gas-liquid chromatography for measuring individual fatty acids liberated during interaction of triglyceride-rich lipoproteins and lipoprotein lipase.
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一种使用气液色谱法测量富含甘油三酯的脂蛋白和脂蛋白脂肪酶相互作用过程中释放的单个脂肪酸的微方法。
DOI：
10.1016/0003-2697(80)90405-4
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发表时间：
1980
影响因子：
2.9
通讯作者：
Robinson,K
中科院分区：
文献类型：
--
作者：
Kashyap,ML;Mellies,MJ;Brady,D;Hynd,BA;Robinson,K
富含甘油三酯的脂蛋白（TRL）被脂蛋白脂肪酶（LPL）分解代谢，同时释放脂肪酸（FA）。本文详细介绍了一种用于测定LPL和TRL在体外相互作用过程中释放的脂肪酸的灵敏和特异性微方法。采用制备式超离心分离TRL。从牛乳中分离得到LPL。TRL和LPL在含白蛋白的缓冲液中孵育。用异丙醇和己烷萃取游离脂肪酸，再用三甲基-（α,α，α-三氟-间甲苯基）氢氧化铵进行二次萃取。采用气液色谱法测定甲酯类FA衍生物。空腹人血浆中正常空腹极低密度脂蛋白（VLDL）产生6个峰，分别对应于脂肪酸14:0、16:0、16:1、18:0、18:1和18:2。主要的脂肪酸峰包括16:0,18:1和18:2。该方法使用非常小（小于1毫克）量的VLDL TG检测微克（纳摩尔）范围内的脂肪酸。游离FA与酶浓度和时间呈线性关系，并表现出典型的底物饱和动力学。这种方法的发展将允许更直接的方法与调节LPL-TRL相互作用中特定FA释放的因素有关。
Triglyceride-rich lipoproteins (TRL) are catabolized by lipoprotein lipase (LPL) with liberation of fatty acids (FA). This communication details a sensitive and specific micromethod for the measurement of fatty acids liberated during the in vitro interaction of LPL and TRL. TRL was isolated by preparative ultracentrifugation. LPL was isolated from bovine milk. TRL and LPL were incubated in an albumin-containing buffer. Liberated FA were extracted using isopropanol and hexane followed by a second extraction using trimethyl-(α,α,α-trifluoro-m-tolyl)ammonium hydroxide. Methyl ester FA derivatives were measured by gas-liquid chromatography. Normal fasting very-low-density lipoproteins (VLDL) from fasting human plasma yielded six peaks corresponding to fatty acids 14:0, 16:0, 16:1, 18:0, 18:1, and 18:2. The major peaks comprised fatty acids 16:0, 18:1, and 18:2. The method detected fatty acids in the microgram (nanomolar) range using very small (less than 1 mg) amounts of VLDL TG. FA liberation was linear with enzyme concentration and time and showed typical substrate saturation kinetics. Development of this method will allow more direct approaches relating to factors regulating the liberation of specific FA in the LPL-TRL interaction.