SE168:/S01/M1

Sample Set Information

ID	TSE1330
Title	Automation of chemical assignment for identifying molecular formula of S-containing metabolites by combining metabolomics and chemoinformatics with 34S labeling
Description	Introduction Sulfur-containing metabolites (S-metabolites) in organisms including plants have unique benefits to humans. So far, few analytical methods have explored such metabolites. Objectives We aimed to develop an automatic chemically assigning platform by metabolomics and chemoinformatics with 34S labeling to identify the molecular formula of S-metabolites. Results We identified 35 molecular formulae for known S-metabolites and characterized 72 for unknown. Chemoinformatics required around 1.5 min to analyze a pair of the non-labeled and 34S-labeled data of the organ. Conclusion In this study, we developed an automation platform for automatically identifying the presence of S-metabolites. We identified the molecular formula of known S-metabolites, which are accessible in free databases, together with that of unknown. This analytical method did not focus on identifying the structure of S-metabolites, but on the automatic identification of their molecular formula.
Authors	Nakabayashi, R., Tsugawa, H.,Mori, T. and Saito, K.
Reference	Metabolomics, November 2016, 12:168, DOI: 10.1007/s11306-016-1115-5
Comment

http://link.springer.com/article/10.1007/s11306-016-1115-5

Sample Information

ID	S01
Title	Arabidopsis thaliana (ecotype Columbia-0)
Organism - Scientific Name	Arabidopsis thaliana
Organism - ID	NCBI taxonomy:3702
Compound - ID
Compound - Source
Preparation	Arabidopsis thaliana (ecotype Columbia-0) was used in this study. Non-labeled and 34S-labeled Arabidopsis plants were prepared by SI Science CO., LTD (Saitama, Japan). The plants were individually grown in pots filled with vermiculite, which were set in a plant growth room at 25 °C under a 16 h light/8 h dark cycle for 8 weeks. A non-labeled and an 34S-labeled liquid fertilizer (Table S1) was added to the plants daily. Different parts of the flower, including the silique, leaf, stem, and root, in 8-week-old Arabidopsis plants were harvested and immediately lyophilized at −55 °C. The lyophilized materials were stored at room temperature with silica gel.
Sample Preparation Details ID
Comment

Analytical Method Information

ID	M1
Title	FTICR-MS (Direct infusion)
Method Details ID	MS01
Sample Amount	100 μL
Comment

Analytical Method Details Information

ID	MS01
Title	FTICR-MS (Direct infusion)
Instrument	FTICR-MS solariX 7.0 T (Bruker Daltonics)
Instrument Type
Ionization	ESI
Ion Mode	negative
Description	Extraction of metabolites The freeze-dried samples were extracted with 50 μL of 80 % MeOH containing 2.5 μM 10-camphorsulfonic acid, 1.25 μM reserpine, 2.5 μM ampicillin, and 2.5 μM CHAPS as internal standards for calibration per mg dry weight using a mixer mill (MM300, Retsch) with zirconia beads for 7 min at 18 Hz and 4 °C. After 10 min of centrifugation, the supernatant was filtered with an HLB μElution plate (Waters). Extracts were routinely diluted with 80 % MeOH at a ratio of 1:1000 (extract: 80 % MeOH). Direct infusion analysis Ultra-high-resolution metabolome data were acquired using an FTICR-MS solariX 7.0 T (Bruker Daltonics) with an ESI source. The following settings were applied: MS Detection; Acquisition Mass Control (mass range, m/z 100–800; estimated resolution power, 530,000 at m/z 400; transient length, 3.9147 s); Data Storage (save full profile spectrum, on; save FID file, on); Accumulation (scan time, 100; source accumulation, 0.01 s; ion accumulation time, 0.1 s; ion cooling time, 0 s; time of flight, 0.45 s); API Source (source type, ESI; capillary, 4500 V; end plate offset, 1000 V); Source Gas Tune (nebulizer 1.0 bar; dry gas, 2.0 L/min; dry temperature, 200 °C); Source Optics (capillary exit, −220 V; deflector plate, −200 V; funnel 1, −150 V; skimmer 1, −25 V; funnel RF amplitude, 180 Vpp); Octopole (frequency, 5 MHz; RF amplitude, 350 Vpp); Collision Cell (collision voltage, 0.8 V; DC extract bias, −0.9 V; RF frequency, 2 MHz; collision RF amplitude, 850 Vpp); Transfer Optics (time of flight, 0.45 ms; frequency, 6 MHz; RF amplitude, 400 Vpp); Infinity Cell (transient exit lens, 20 V; analyzer entrance, 10 V; side kick, 0 V; side kick offset, 1.5 V; front trap plate, −0.5 V; back trap plate, −0.5 V; sweep excitation power, 20 %); Multiple Cell Accumulations (number of ICR cell fills, 1); Gated Trapping (gated trapping mode, off); Polarity, negative. MS spectra were internally calibrated using the MS spectra of internal standards.
Comment_of_details