Hosaka K, Kikuchi T, Mitsuhida N, Kawaguchi A
J Biochem. 1981 Jun;89(6):1799-803. doi: 10.1093/oxfordjournals.jbchem.a133380.
A rapid and sensitive spectrophotometric assay for free fatty acids using acyl-CoA synthetase and acyl-CoA oxidase is described. It is sensitive to as low as 5 nmol of free fatty acids, and the standard curve is linear up to 100 nmol. The assay consists of the measurement of H2O2 produced from free fatty acids by acyl-CoA synthetase and acyl-CoA oxidase. The quantity of H2O2 is determined by the absorbance at 550 nm in the presence of catalase and 4-amino-3-hydrazino-5-mercapto-1,2,4-triazole (AHMT). This method shows a broad specificity to long-chain fatty acids and the recoveries of added fatty acids (C12-C18) are more than 90%. The presence or absence of serum components or Escherichia coli cell-free extracts has no significant effect on the recovery of added palmitic acid.
本文描述了一种使用酰基辅酶A合成酶和酰基辅酶A氧化酶的快速灵敏的游离脂肪酸分光光度测定法。该方法对低至5 nmol的游离脂肪酸敏感，标准曲线在100 nmol范围内呈线性。该测定法包括测量酰基辅酶A合成酶和酰基辅酶A氧化酶将游离脂肪酸转化产生的H2O2。H2O2的量通过在过氧化氢酶和4-氨基-3-肼基-5-巯基-1,2,4-三唑（AHMT）存在下550 nm处的吸光度来测定。该方法对长链脂肪酸具有广泛的特异性，添加脂肪酸（C12 - C18）的回收率超过90%。血清成分或大肠杆菌无细胞提取物的存在与否对添加的棕榈酸的回收率没有显著影响。
