SE158:/S1/M1

Sample Set Information

ID	TSE1314
Title	Boosting Sensitivity in Liquid Chromatography-Fourier Transform Ion Cyclotron Resonance-Tandem Mass Spectrometry for Product Ion Analysis of Monoterpene Indole Alkaloids.
Description	In metabolomics, the analysis of product ions in tandem mass spectrometry (MS/MS) is noteworthy to chemically assign structural information. However, the development of relevant analytical methods are less advanced. Here, we developed a method to boost sensitivity in liquid chromatography-Fourier transform ion cyclotron resonance-tandem mass spectrometry analysis (MS/MS boost analysis). To verify the MS/MS boost analysis, both quercetin and uniformly labeled (13)C quercetin were analyzed, revealing that the origin of the product ions is not the instrument, but the analyzed compounds resulting in sensitive product ions. Next, we applied this method to the analysis of monoterpene indole alkaloids (MIAs). The comparative analyses of MIAs having indole basic skeleton (ajmalicine, catharanthine, hirsuteine, and hirsutine) and oxindole skeleton (formosanine, isoformosanine, pteropodine, isopteropodine, rhynchophylline, isorhynchophylline, and mitraphylline) identified 86 and 73 common monoisotopic ions, respectively. The comparative analyses of the three pairs of stereoisomers showed more than 170 common monoisotopic ions in each pair. This method was also applied to the targeted analysis of MIAs in Catharanthus roseus and Uncaria rhynchophylla to profile indole and oxindole compounds using the product ions. This analysis is suitable for chemically assigning features of the metabolite groups, which contributes to targeted metabolome analysis.
Authors	Nakabayashi R, Tsugawa H, Kitajima M, Takayama H, Saito K.
Reference	Front Plant Sci. 2015 Dec 17;6:1127. doi: 10.3389/fpls.2015.01127. eCollection 2015.
Comment

PubMed

Sample Information

ID	S1
Title	Catharanthus roseus “Equator White Eye”
Organism - Scientific Name	Catharanthus roseus
Organism - ID	NCBI taxonomy:4058
Compound - ID
Compound - Source
Preparation	Seeds of C. roseus “Equator White Eye” (Sakata Seeds, Co., Ltd., Kanagawa, Japan) were sown in the soil of pods [Pro-Mix BX (Premier Tech Horticulture, Inc., Rivière-du-Loup, QC, Canada): vermiculite = 2:1, supplemented with fertilizer]. The pods were set in a growth chamber at 24°C and a relative humidity of approximately 60% under a 16-h light (approximately 190 μmol s-1 m-2)/8-h dark cycle. The root parts of 6-weeks-old plants were harvested and immediately frozen in liquid nitrogen. The root samples were finally freeze-dried and stored at room temperature in a box containing silica gel. The hook parts of U. rhynchophylla were harvested in the medicinal plant garden of Chiba University (Chiba, Japan). The samples were dried and stored at room temperature in a box containing silica gel.
Sample Preparation Details ID
Comment

Analytical Method Information

ID	M1
Title	LC-FTICR-MS/MS
Method Details ID	MS1
Sample Amount	5 μL
Comment

Analytical Method Details Information

ID	MS1
Title	LC–FTICR–MS/MS
Instrument	LC, Agilent 1260 Infinity; MS, Bruker Daltonics SolariX 7.0 T
Instrument Type	LC-FTICR-MS
Ionization	ESI
Ion Mode	Positive
Description	Extraction of Metabolites The samples were extracted with 50 μL of 80% MeOH per mg dry weight using a mixer mill (MM300, Retsch) with zirconia beads for 10 min at 20 Hz and 4°C. After centrifugation for 10 min, the supernatant was filtered using an HLB μElution plate (Waters). LC–FTICR–MS/MS Analysis The solutions (5 μL) were analyzed using an LC–FTICR–MS instrument (LC, Agilent 1260 Infinity; MS, Bruker Daltonics SolariX 7.0 T). Analytical conditions were as follows. LC: column, Xselect CSH Phenyl-Hexyl (3.5 μm, 2.1 mm × 150 mm, Waters, Milford, MA, USA); solvent system, solvent A (water with 0.1% formic acid) and solvent B (acetonitrile with 0.1% formic acid); LC gradient program, 99.5% A/0.5% B at 0 min, 0.5% A/99.5% B at 30.0 min, 0.5% A/99.5% B at 45.0 min, 99.5% A/0.5% B at 45.1 min, and 99.5% A/0.5% B at 60.0 min; flow rate, 0.3 mL/min; column temperature, 35°C; wavelength, 200–600 nm; MS/MS detection; Acquisition Mass Control (mass range, m/z 100–600; estimated resolution power, 66,000 at m/z 400; transient length, 0.4893); Data Storage, save full profile spectrum, on; save FID file, on; Accumulation (average scan, 1; source accumulation, 0 s; ion accumulation time, 0.1 s; ion cooling time, 0 s; time of flight, 0.4 s); API Source [source type, electrospray ionization (ESI); capillary, 4500 V; end plate offset, -500 V]; Source Gas Tune (nebulizer 1.6 bar; dry gas, 7.0 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, 150 Vpp); Octopole (frequency, 5 MHz; RF amplitude, 350 Vpp); Collision Cell (collision voltage, -2 V; DC extract bias, 0.4 V polarity, RF frequency, 2 MHz; collision RF amplitude, 1200 Vpp); Transfer Optics (time of flight, 0.4 ms; frequency, 6 MHz, RF amplitude, 350 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, 0.6 V; back trap plate, 0.6 V; sweep excitation power, 18%); Multiple Cell Accumulations (number of ICR cell fills, 1); Gated Trapping (gated trapping mode, off); Precursor Selection (prefer list, m/z of target compound [M + H]+, mass accuracy, ±20 ppm; exclude unknown charged ions, on); Quadrupole Parameters (MS/MS isolation, on; cut off in MS, m/z 100; isolation window, m/z 5); Fragmentation Mode, quadrupole collision-induced dissociation (QCID); MS/MS Boost, on (in use); MS accumulation in MS/MS Boost, 0.1 s (in use); MS/MS accumulation in MS/MS Boost, 2 s (in use); fixed collision voltage, 30 V; polarity, positive; API high voltage, on; source quench, on.
Comment_of_details