Han Jun, Danell Ryan M, Patel Jayanti R, Gumerov Dmitry R, Scarlett Cameron O, Speir J Paul, Parker Carol E, Rusyn Ivan, Zeisel Steven, Borchers Christoph H
J. Han, C. H. Borchers, University of Victoria - Genome BC Proteomics Centre, Vancouver Island Technology Park, #3101-4464 Markham St., Victoria, BC, Canada V8Z 7X8.
Metabolomics. 2008 Jun;4(2):128-140. doi: 10.1007/s11306-008-0104-8.
With unmatched mass resolution, mass accuracy, and exceptional detection sensitivity, Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FTICR-MS) has the potential to be a powerful new technique for high-throughput metabolomic analysis. In this study, we examine the properties of an ultrahigh-field 12-Tesla (12T) FTICR-MS for the identification and absolute quantitation of human plasma metabolites, and for the untargeted metabolic fingerprinting of inbred-strain mouse serum by direct infusion (DI). Using internal mass calibration (mass error ≤1 ppm), we determined the rational elemental compositions (incorporating unlimited C, H, N and O, and a maximum of two S, three P, two Na, and one K per formula) of approximately 250 out of 570 metabolite features detected in a 3-min infusion analysis of aqueous extract of human plasma, and were able to identify more than 100 metabolites. Using isotopically-labeled internal standards, we were able to obtain excellent calibration curves for the absolute quantitation of choline with sub-pmol sensitivity, using 500 times less sample than previous LC/MS analyses. Under optimized serum dilution conditions, chemical compounds spiked into mouse serum as metabolite mimics showed a linear response over a 600-fold concentration range. DI/FTICR-MS analysis of serum from 26 mice from 2 inbred strains, with and without acute trichloroethylene (TCE) treatment, gave a relative standard deviation (RSD) of 4.5%. Finally, we extended this method to the metabolomic fingerprinting of serum samples from 49 mice from 5 inbred strains involved in an acute alcohol toxicity study, using both positive and negative electrospray ionization (ESI). Using these samples, we demonstrated the utility of this method for high-throughput metabolomics, with more than 400 metabolites profiled in only 24 h. Our experiments demonstrate that DI/FTICR-MS is well-suited for high-throughput metabolomic analysis.
傅里叶变换离子回旋共振质谱仪（FTICR-MS）具有无与伦比的质量分辨率、质量精度和卓越的检测灵敏度，有潜力成为高通量代谢组学分析的强大新技术。在本研究中，我们研究了超高场12特斯拉（12T）FTICR-MS用于鉴定和绝对定量人血浆代谢物，以及通过直接进样（DI）对近交系小鼠血清进行非靶向代谢指纹分析的特性。使用内部质量校准（质量误差≤1 ppm），我们确定了在人血浆水提取物的3分钟进样分析中检测到的570个代谢物特征中约250个的合理元素组成（每个分子式包含不限量的C、H、N和O，最多两个S、三个P、两个Na和一个K），并能够鉴定出100多种代谢物。使用同位素标记的内标，我们能够获得用于胆碱绝对定量的出色校准曲线，灵敏度达到亚皮摩尔级，所用样品量比之前的液相色谱/质谱分析少500倍。在优化的血清稀释条件下，作为代谢物模拟物添加到小鼠血清中的化合物在600倍浓度范围内呈线性响应。对来自2个近交系的26只小鼠（有或没有急性三氯乙烯（TCE）处理）的血清进行DI/FTICR-MS分析，相对标准偏差（RSD）为4.5%。最后，我们将该方法扩展到对参与急性酒精毒性研究的5个近交系的49只小鼠的血清样品进行代谢组指纹分析，同时使用正电喷雾电离（ESI）和负电喷雾电离。使用这些样品，我们证明了该方法在高通量代谢组学中的实用性，仅在24小时内就对400多种代谢物进行了分析。我们的实验表明，DI/FTICR-MS非常适合高通量代谢组学分析。
