Analysis of monoamine oxidase enzymatic activity by reversed-phase high performance liquid chromatography and inhibition by β-carboline alkaloids occurring in foods and plants
Analysis of monoamine oxidase enzymatic activity by reversed-phase high performance liquid chromatography and inhibition by β-carboline alkaloids occurring in foods and plants
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DOI：
10.1016/j.chroma.2005.12.009
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发表时间：
2006-07-07
影响因子：
4.1
通讯作者：
Chaparro, Carolina
中科院分区：
文献类型：
--
作者：
Herraiz, Tomas;Chaparro, Carolina
单胺氧化酶（MAO）是一种位于线粒体外膜的黄素腺嘌呤二核苷酸（FAD）酶，催化生物胺和异生物胺的氧化脱氨。我们已经使用色谱法通过使用犬尿胺作为非选择性底物来测量MAO-酶活性，随后通过RP-HPLC-DAD和HPLC-质谱法（MS）分析其MAO-氧化产物。应用该方法研究了食品、植物和哺乳动物中四氢-β-咔啉和β-咔啉生物碱对MAO重组酶的动力学参数、抑制作用和反应产物。HPLC分析表明，四氢-β-咔啉或β-咔啉未被MAO修饰。几种β-咔啉类化合物，如1-甲基-1，2，3，4-四氢-β-咔啉和1-甲基-1，2，3，4-四氢-β-咔啉，是MAO-A的抑制剂;而它们相应的6-羟基衍生物（6-羟基-1-甲基-6-羟基-咔啉和6-羟基-1-甲基-咔啉）则没有这种活性。四氢-β-咔啉-3-羧酸不能抑制MAO酶。相反，它们的氧化产物，即全芳族β-咔啉（norharman和harman），作为良好的MAO抑制剂。采用固相萃取（SPE）和反相高效液相色谱法（RP-HPLC）从香肠样品中分离得到两种四氢-β-咔啉类化合物（色托林和1-甲基色托林），其提取物对单胺氧化酶A（MAO-A）具有良好的抑制作用。这些结果表明，β-咔啉从食物，植物和哺乳动物可以发挥抑制作用的单胺氧化酶。(c)2005 Elsevier B. V.保留所有权利。
Monoamine oxidase (MAO) is a flavin adenine dinucleotide (FAD)-containing enzyme located at the outer membranes of mitochondria that catalyzes the oxidative deamination of biogenic and xenobiotic amines. We have used a chromatographic method to measure MAO-enzymatic activity by using kynuramine as a non-selective substrate with its MAO-oxidation product subsequently analyzed by RP-HPLC-DAD and HPLC-mass spectrometry (MS). This method was applied to study the kinetic parameters, inhibition and reaction products of MAO recombinant enzymes in presence of tetrahydro-beta-carboline and P-carboline alkaloids occurring in foods, plants and mammals. Analysis by HPLC showed that tetrahydro-beta-carbolines or beta-carbolines were not modified by MAO. Several beta-carbolines such as tryptoline (1,2,3,4-tetrahydro-beta-carboline) and 1-methyltryptoline (1-methyl-1,2,3,4-tetrahydro-beta-carboline) were inhibitors of MAO-A; instead their corresponding 6-hydroxy-derivatives (6-hydroxytryptoline and 6-hydroxy-1-methyltryptoline) lacked this activity. Tetrahydro-beta-carboline-3-carboxylic acids were unable to inhibit MAO enzymes. In contrast, their oxidation products, i.e. the fully aromatic beta-carbolines (norharman and harman), acted as good inhibitors of MAO. Two tetrahydro-beta-carbolines (i.e. tryptoline and 1-methyltryptoline) occurring in foods were isolated by solid-phase extraction (SPE) and RP-HPLC from selected samples of sausages and the corresponding extracts exhibited good inhibition properties over MAO-A. These results suggest that beta-carbolines from foods, plants, and mammals may exert inhibitory actions on MAO enzymes. (c) 2005 Elsevier B.V. All rights reserved.