SE61:/S11/M103

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Sample Set Information

ID SE61
Title LC-MS analysis of parsley
Description Comprehensive analysis of metabolites in parsley (Petroselium crispum, Paramount) was performed by several combination of sample preparation and analytical conditions.

Parsley was hydroponically grown in a greenhouse (S11-S13) or a cultivation shelf (S21-S26) using control medium and stable isotope (15N or 34S)-containing medium.

Authors Sakurai N, Suda K (Kazusa DNA Research Insitute)
Reference Akimoto N, Ara T, Nakajima D, Suda K, Ikeda C, Takahashi S, Muneto R, Yamada M, Suzuki H, Shibata D and Sakurai N (2017) FlavonoidSearch: A system for comprehensive flavonoid annotation by mass spectrometry. Scientific Reports 7: 1243
Comment


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Sample Information

ID S11
Title Shoot of greenhouse grown parsley
Organism - Scientific Name Petroselinum crispum, Paramount
Organism - ID NCBI taxonomy:4043
Compound - ID
Compound - Source
Preparation Hydroponically grown in a greenhouse. Shoot was harvested.
Sample Preparation Details ID SS1
Comment

Sample Preparation Details Information

ID SS1
Title Cultivation in a greenhouse
Description The parsley seeds (Kaneko Seeds, Co. LTD. KS100-542, Gunma, Japan) were germinated on watered sand. The 20 days-old seedlings were transplanted on baked foamed clay pebbles (Hydroball, Toshi Engei Col. LTD, Saitama, Japan) in a green house, and grown 63 days with culture medium containing 3 mM KNO3, 1 mM Ca(NO3)2, 0.5 mM MgSO4, 0.5 mM NH4H2PO4, 50 µM Fe-EDTA, 49 µM H3BO3, 9 µM MnSO4, 0.8 µM ZnSO4, 0.3 µM CuSO4, and 0.1 µM Na2MoO4. Shoot was harvested, ground to powder in liquid nitrogen by mortar and pestle, and stored at -80 ºC until use.
Comment_of_details

Analytical Method Information

ID M103
Title Method 3: ESI Negative, Data dependent MS2 scan (FT, IT)
Method Details ID MS13
Sample Amount 5 mg / 20 ul injection
Comment [MassBase ID] MDLC1_48107


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The raw (binary) and near-raw (text) files of this analysis are available at MassBase.

Analytical Method Details Information

ID MS13
Title LC-FT-MS, ESI, Negative (method 3)
Instrument Agilent1100 HPLC (Agilent), LTQ-FT (Thermo Fisher Scientific)
Instrument Type LC-FTICR-MS
Ionization ESI
Ion Mode Negative
Description Frozen powder of plant material was extracted with three times volumes of methanol containing 25 uM of 7-hydroxy-5-methylflavone as an internal standard. After homogenization using a Mixer Mill MM 300 (Quiagen) at 25 Hz for 2 min twice, homogenates were centrifuged (17,400 g, 5 min, 4 degree Celsius). The supernatant was filtered through 0.2 um PTFE membrane (Millipore). Hydrophobic compounds in the filtrate were removed by absorbing to C18 silica column (MonoSpin C18, GL Science, Tokyo, Japan).

Mock sample was prepared with the same procedure without adding the plant material.

20 ul of methanol solution was applied to a TSK-gel column ODS-100V (4.6 x 250 mm, 5 um; TOSOH). Water (HPLC grade; solvent A) and acetonitrile (HPLC grade; solvent B) were used as the mobile phase with 0.1% v/v formic acid added to both solvents. The gradient program was as follows: 3% B (0 min), 97% B (90 min), 97% B (100 min), 3% B (100.1 min), and 3% B (107 min). The flow rate was set to 0.25 ml/min (0-100 min) and 0.5 ml/min (100.1-107 min). The flow rate was set to 0.5 ml/min, and the column oven temperature was set at 40 degree Celsius. Compounds are detected in ESI-negative mode in the range m/z 100-1500. Multistage MSn analyses were carried out using collision induced dissociation (CID) in a linear ion trap detector at a normalized collision energy of 35.0% and an isolation width of 4.0 (m/z), and monitored by both ion trap detector and FT-ICR detector at 25,000 (at m/z 400) mass resolution. The ESI setting was as follows: spray voltage 4.0 kV and capillary temperature 300 degree Celsius. To monitor HPLC elution, a photodiode array detector was used in the wavelength range 200-650 nm.

Mass scan events were set as follows (referred as method 3): full mass scan with FT-ICR in resolution 100,000 and MS2 scans for the most intense 5 ions of full mass scan with ion trap (IT). A dynamic exclusion setting was applied as follows: repeat count, 2; repeat duration, 30 sec; exclusion list size, 500; margin, 10 ppm, and exclusion duration, 30.

The data were acquired by Xcalibur software version 2.07 (Thermo Fisher Scientific).

Comment_of_details [column] TSK-gel ODS-100V (4.6 x 250 mm, 5 micrometer; TOSOH)

[gradient] Solvent A: water containing 0.1% v/v formic acid; Solvent B: acetonitrile containing 0.1% v/v formic acid; Gradient: 3% B (0 min), 97% B (90 min), 97% B (100 min), 3% B (100.1 min), and 3% B (107 min) [total separation time] 107 min

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