QUANTITATIVE-DETERMINATION OF METHANOGENIC CELLS BASED ON ANALYSIS OF ETHER-LINKED GLYCEROLIPIDS BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY
QUANTITATIVE-DETERMINATION OF METHANOGENIC CELLS BASED ON ANALYSIS OF ETHER-LINKED GLYCEROLIPIDS BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY
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DOI：
10.1016/0922-338x(92)90553-7
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发表时间：
1992-01-01
期刊：
影响因子：
--
通讯作者：
KOGA, Y
中科院分区：
文献类型：
--
作者：
DEMIZU, K;OHTSUBO, S;KOGA, Y
介绍了一种在自然环境中测定产甲烷细胞的方法。该方法基于对产甲烷菌和其他古细菌特有的醚连接甘油脂的测定。以总脂为原料，经乙酰解和酸甲醇解合成古醇和卡尔达酚，并采用高效液相色谱法定量鉴定其为二硝基苯甲酰衍生物。核心脂质总量与产甲烷细胞数量成正比。该方法灵敏度高，在500 μ l的培养物或污泥样品中，产甲烷细胞的最低检出限为2.5 μ g（干重）。获得了热自养甲烷菌和亨盖特甲烷螺旋菌的相同标准曲线。利用这种方法，可以估计厌氧消化器污泥样品中的产甲烷细胞。尽管污泥中含有大量的各种掺假物，但加入污泥中的热自养分枝杆菌细胞的回收率为83%，并通过该方法进行了定量分析。
A method for the quantitative determination of methanogenic cells in natural environments is described. The method is based on the determination of ether-linked glycerolipids that is specific to methanogens and other archaebacteria. Core lipids (archaeol and caldarchaeol), prepared from total lipids by a combination of acetolysis and acid methanolysis, were quantitated as the corresponding dinitrobenzoyl derivatives by high-performance liquid chromatography. The total amount of core lipids was proportional to the amount of methanogenic cells. This method was high sensitivity, and the lowest limit of detection of methanogenic cells was 2.5-mu-g (dry weight) in a 500-mu-l sample of culture or sludge. An identical standard curve was obtained for Methanobacterium thermoautotrophicum and Methanospirillum hungatei. With this method, it was possible to estimate methanogenic cells in a sample of sludge from an anaerobic digestor. In spite of contamination by large amounts of multifarious adulterants in the sludge, 83% of M. thermoautotrophicum cells added to the sludge was recovered and quantitated by this method.