High-Throughput Quantification of Bioactive Lipids by MALDI Mass Spectrometry: Application to Prostaglandins
High-Throughput Quantification of Bioactive Lipids by MALDI Mass Spectrometry: Application to Prostaglandins
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DOI：
10.1021/ac201224n
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发表时间：
2011-09-01
影响因子：
7.4
通讯作者：
Caprioli, Richard M.
中科院分区：
文献类型：
--
作者：
Manna, Joseph D.;Reyzer, Michelle L.;Caprioli, Richard M.
生物系统中分析物的分析和定量是细胞功能代谢组学研究的关键组成部分。最广泛使用的方法是色谱分离，然后是质谱分析，这需要大量的时间来制备样品和顺序色谱。我们介绍了一种基于MALDI质谱的新型高通量，无分离的方法，允许对目标代谢组进行平行分析。通过前列腺素和甘油前列腺素的分析证明了概念验证。衍生化，以纳入一个带电的部分到含酮前列腺素显著增加相对于未酸化样品的信噪比。这增加了动态范围（板上15-2000 fmol），改善了线性度（r(2) = 0.99）。该方法适用于酶学和药物发现的高通量筛选方法。应用于细胞代谢组学也被证明。
Analysis and quantification of analytes in biological systems is a critical component of metabolomic investigations of cell function. The most widely used methods employ chromatographic separation followed by mass spectrometric analysis, which requires significant time for sample preparation and sequential chromatography. We introduce a novel high-throughput, separation-free methodology based on MALDI mass spectrometry that allows for the parallel analysis of targeted metabolomes. Proof-of-concept is demonstrated by analysis of prostaglandins and glyceryl prostaglandins. Derivatization to incorporate a charged moiety into ketone-containing prostaglandins dramatically increases the signal-to-noise ratio relative to underivatized samples. This resulted in an increased dynamic range (15-2000 fmol on plate) and improved linearity (r(2) = 0.99). The method was adapted for high-throughput screening methods for enzymology and drug discovery. Application to cellular metabolomics was also demonstrated.