High throughput phospholipid fatty acid analysis of soils
High throughput phospholipid fatty acid analysis of soils
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DOI：
10.1016/j.apsoil.2012.06.005
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发表时间：
2012-10-01
影响因子：
4.8
通讯作者：
Sasser, Myron
中科院分区：
文献类型：
--
作者：
Buyer, Jeffrey S.;Sasser, Myron
磷脂脂肪酸（PLFA）分析被广泛用于土壤和其他类型环境样品中微生物生物量和群落组成的测量。通常，根据每个实验室使用的确切设备，分析涉及许多步骤，需要1.5-3天准备小批量（即20-24个样品和空白）。然后使用气相色谱（GC）或气相色谱-质谱（GC- ms）分析样品，需要更多的时间来获得数据。我们开发了一种在1.5天内制备96个土壤样品和空白的方法，吞吐量提高了4到5倍。所有干燥和离心步骤都在离心蒸发器中进行。将试管中的土壤样品干燥过夜，然后进行Bligh-Dyer脂质提取。将萃取物干燥，溶解在氯仿中，并装在96孔固相萃取板上。磷脂被洗脱到96孔格式的玻璃小瓶中，干燥，并进行酯交换。所得脂肪酸甲酯用气相色谱分析，并相对于内标定量。高通量方案比传统方案使用更小的溶剂体积，结合96孔格式的使用，使样品制备速度更快。10种不同土壤的生物标志物PLFA浓度在两种方案之间高度相关，尽管不相同。PLFA生物标志物的多变量分析表明，两种方案对不同土壤产生相似的模式。高通量协议可能是有用的实验室进行大量的PLFA分析。Elsevier B.V.出版
Phospholipid fatty acid (PLFA) analysis is widely used to measure microbial biomass and community composition in soil and other types of environmental samples. As typically performed, the analysis involves many steps and 1.5-3 days are required to prepare a small batch (i.e. 20-24 samples and blanks), depending on the exact equipment employed in each laboratory. Gas chromatography (GC) or gas chromatography-mass spectrometry (GC-MS) is then used to analyze the samples, requiring further time to obtain the data. We have developed a method for preparing 96 soil samples and blanks in 1.5 days, a 4- to 5-fold increase in throughput. All drying and centrifuging steps take place in a centrifugal evaporator. Soil samples in test tubes are dried overnight and then a Bligh-Dyer lipid extraction is performed. The extract is dried, dissolved in chloroform, and loaded onto a 96-well solid phase extraction plate. Phospholipids are eluted into glass vials in a 96-well format, dried, and transesterified. The resulting fatty acid methyl esters are analyzed by GC and quantified relative to an internal standard. The high throughput protocol uses much smaller solvent volumes than the traditional protocol, which combined with the use of the 96-well format leads to much faster sample preparation. Biomarker PLFA concentrations for 10 different soils were highly correlated, although not identical, between the two protocols. Multivariate analysis of the PLFA biomarkers indicated that the two protocols produced similar patterns for the different soils. The high throughput protocol may be useful to laboratories performing large numbers of PLFA analyses. Published by Elsevier B.V.