Session TOD. There are 6 abstracts in this session.

Session: Metabolomics, time: 4:00-4:25
Metabonomics: contributions to host and micro/macro-parasites interactions
Yulan Wang
Wuhan Inst of Phys and Math, Wuhan, China
We live in a wormy world; humans inevitably harbor various parasites. Therefore infection with multiple micro/macro-parasites is a common phenomenon in humans and animals. Infection with these organisms will cause immune responses as well as metabolic responses. Infection responses can be a complex issue when multiple organisms present. Metabonomics is a robust tool for studying the metabolic responses to multiple stimuli. I will present metabonomics investigation of interaction of host and multiple organisms such as Schistosoma japonicum and Salmonella typhimurium. In addition, I will show how metabonomics could generate credible hypothesis for identifying drug targets for HBV infection.

Session: Metabolomics, time: 4:25-4:50

Developments toward the detection of oscillating metabolites in the fungus Neurospora crassa

Michael Judge; Sicong Zhang; Yue Wu; John Glushka; Jonathan Arnold; Arthur Edison
University of Georgia, Athens, GA

N. crassa has a circadian clock. We are attempting to measure the metabolome of N. crassa to identify candidates for circadian synchronization. We initially grew large cultures over 2 days and extracted samples. This resulted in a rich, but noisy, dataset with the usual problems of alignment and normalization seen in NMR metabolomics experiments. Although we have detected possible oscillating features from these data, the results have been ambiguous because of high variance. Recently, we used an HR-MAS CMP probe to record the growth of a culture of N. crassa with 10 min resolution over nearly 2 days. The data are rich in detail and require no alignment or normalization with an 80 uL culture and no sample preparation.

Session: Metabolomics, time: 4:50-5:15
High-resolution NMR Spectroscopy from field Inhomogeneity and Spectral Crowding
Zhong Chen; Yuqing Huang
Xiamen University, Xiamen, China
In general NMR applications, such as metabolomics studies, spectral resolution is the key index determining the availability of resulting spectra. Due to limited chemical shift ranges and extensive J couplings, conventional proton (1H) NMR spectra are subject to spectral crowding in complex samples. In addition, there exist adverse experimental conditions in which magnetic fields suffer from spatial or temporal inhomogeneity, which constitutes the second factor degrading spectral resolution in 1H NMR applications. Therefore, an NMR method available for high-resolution NMR measurements under the condition of field inhomogeneity and spectral crowding is greatly demanded. In this abstract, a series of NMR methods based on intermolecular multiple-quantum coherences and pure-shift mechanism for high-resolution applications is introduced.

Session: Metabolomics, time: 5:15-5:30
Unravelling NMR visible and invisible components of urine by a powerful analysis tool
Panteleimon Takis1; Hartmut Schäfer2; Manfred Spraul2; Claudio Luchinat3, 4
1GIOTTO BIOTECH S.r.l., Sesto Fiorentino Florence, Italy; 2Bruker BioSpin, Rheinstetten, GERMANY; 3Magnetic Resonance Center (CERM), Sesto Fiorentino (FI), Italy; 4University of Florence, Sesto Fiorentino (FI), Italy

Urine consists of >2000 metabolites, approximately 250 of which potentially detectable by NMR.  However, chemical shift variability hampers automated assignment – and subsequent quantitation - of urine metabolites.  To overcome this problem, we considered that chemical shift variability should correlate with the variable concentrations of the metabolites. We mathematically unraveled this interrelationship by constructing >4000 mixtures, varying the concentrations of the most abundant metabolites and inorganic ions in urine, and assigning >90 signals from >60 metabolites in each mixture. We then created the ‘Urine shift predictor’ (Takis et al, Nat. Commun., 8:1662, 2017), which automatically and accurately predicts metabolite chemical shifts and concentrations of the “invisible” inorganic ions and of other urine components with analytical accuracy, for real urine samples. 

Session: Metabolomics, time: 5:30-5:45
Fast 2D 1H NMR lipidomics workflow to tackle chemical food safety issues 
Jérémy Marchand1, 2; Estelle Martineau1, 3; Yann Guitton2; Bruno Le Bizec2; Gaud Dervilly-Pinel2; Patrick Giraudeau1, 4
1CEISAM, Nantes, France; 2Laberca, Nantes, France; 3SpectroMaitrise, Nantes, France; 4Institut Universitaire de France, Paris, France
Proton Nuclear Magnetic Resonance (NMR) is an appealing solution for untargeted lipidomics. However, its applicability is hampered by the considerable overlap of lipid signals on 1D spectra and the need for dedicated sample preparation protocols. To overcome such issues, we developed a complete NMR lipidomics workflow for serum fingerprinting applications including fast 2D NMR strategies –non-uniform sampling and ultrafast schemes– enabling the automated acquisition of high-throughput and well-resolved lipid spectra. The workflow was applied to a challenging chemical food safety issue: the characterization of lipid profiles disruption in serum from β-­agonists diet-fed pigs. The results confirm the ability of fast 2D NMR to discriminate efficiently sample groups in an automated acquisition framework.

Session: Metabolomics, time: 5:45-6:00
NMR Guided Mass Spectrometry for Absolute Quantitation of Human Blood Metabolites
G. A. Nagana Gowda1; Danijel Djukovic1; Lisa Fan Bettcher1; Haiwei Gu1; Daniel Raftery1, 2
1University of Washington, Seattle, WA, USA; 2Fred Hutchinson Cancer Research Center, Seattle, WA, USA
While absolute quantitation using NMR is reasonably straightforward, this is not the case for mass spectrometry (MS) owing to numerous factors including the need for isotope labeled internal standards, their prohibitively high cost, the need to maintain their concentrations close to the target metabolites and/or the requirement of calibration curve for each target metabolite. Here we provide a new method, in which metabolites from a single serum specimen quantitated based on the recently developed NMR method [Anal. Chem. 2015, 87, 706-15] were used for absolute metabolite quantitation using MS. This approach is simple, easy to implement and, by obviating the need for internal standard for each metabolite, offers a new avenue for quantitation of blood metabolites using MS, routinely.