Rapid Quantitative Analysis of Lipids Using a Colorimetric Method in a Microplate Format
Rapid Quantitative Analysis of Lipids Using a Colorimetric Method in a Microplate Format
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DOI：
10.1007/s11745-010-3494-0
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发表时间：
2011-01-01
期刊：
影响因子：
1.9
通讯作者：
VanderGheynst, Jean S.
中科院分区：
文献类型：
--
作者：
Cheng, Yu-Shen;Zheng, Yi;VanderGheynst, Jean S.
建立了高通量分析总脂的磺基磷酸香草醛(SPV)比色法。开发的方法使用的反应混合物在整个分析过程中一直保持在96孔微孔板中。与其他脂类测量方法相比，新的分析方法具有以下优点：(1)背景吸光度可以很容易地针对每一孔进行校正，(2)处理和转移反应混合物中含有的硫酸的风险更小，(3)显色更一致，提供更准确的吸光度测量，以及(4)该分析方法可用于从多种来源提取的脂类的定量测量。与使用荧光染料的其他分光光度方法不同，所开发的分析的最佳光谱和反应条件不随样品来源而变化。用该方法测定了4株微藻提取液中的脂质含量。当使用新方法测量脂肪含量并与使用宏观重量法获得的结果进行比较时，在脂肪含量测定中没有发现显著差异。
A colorimetric sulfo-phospho-vanillin (SPV) method was developed for high throughput analysis of total lipids. The developed method uses a reaction mixture that is maintained in a 96-well microplate throughout the entire assay. The new assay provides the following advantages over other methods of lipid measurement: (1) background absorbance can be easily corrected for each well, (2) there is less risk of handling and transferring sulfuric acid contained in reaction mixtures, (3) color develops more consistently providing more accurate measurement of absorbance, and (4) the assay can be used for quantitative measurement of lipids extracted from a wide variety of sources. Unlike other spectrophotometric approaches that use fluorescent dyes, the optimal spectra and reaction conditions for the developed assay do not vary with the sample source. The developed method was used to measure lipids in extracts from four strains of microalgae. No significant difference was found in lipid determination when lipid content was measured using the new method and compared to results obtained using a macro-gravimetric method.