Sample Information
ID
|
S01
|
Title
|
Chlamydomonas reinhardtii C-9
|
Organism - Scientific Name
|
Chlamydomonas reinhardtii
|
Organism - ID
|
NCBI taxonomy:3055
|
Compound - ID
|
|
Compound - Source
|
|
Preparation
|
Chlamydomonas reinhardtii C-9 mt- are grown in the TAP culture medium in an incubator with Light at all times condition (28 degree C, 150rpm). The strain is provided from Dr Hideya Fukuzawa, Kyoto Univ.
|
Sample Preparation Details ID
|
|
Comment
|
[KomicMarket ID] KSBA_28
|
Analytical Method Details Information
ID
|
MS1
|
Title
|
LC-Orbitrap-MS ESI positive method 1
|
Instrument
|
Agilent1100 HPLC (Agilent), LTQ-Orbitrap (Thermo Fisher Scientific)
|
Instrument Type
|
LC-Orbitrap-MS
|
Ionization
|
ESI
|
Ion Mode
|
Positive
|
Description
|
Harvested sample is frozen by liquid N2 and resulting powder (100mg) are solved in 500uL 80% methanol solution. Cells are homogenized by TissueLyser (QUIAGEN) and centrifuged (15000rpm, 10min, 4 degree C). 20uL of the supernatant (300uL) is injected into HPLC after 0.2um membrane filter treatment. HPLC conditions: Agilent 1100 series (Agilent), Column: TSKgel Amide-80 (4.6 x 250 mm, 5 micrometer; TOSOH), Solvent: A; 0.1% formic acid aq. B; ACN (addition 0.1% formic acid fc.), Gradient: (B);80 to 45% (0.0 to 30.0 min), 45 to 35% (30.0 to 30.1 min), 35% (30.1 to 35.0 min), 35% to 80% (35.0 to 35.1 min), 80% (35.1 to 42.0 min), Column temp.: 40 degree C, Flow rate=0.8mL/min, PDA: 200-600 nm (2 nm step). Orbitrap-MS conditions: Filter 1: FTMS + c norm !corona !pi res=60000 o(150.0-2000.0); 2: FTMS + c norm !corona !pi res=7500 Dep MS/MS Most intense ion from (1); 3: FTMS + c norm !corona !pi res=7500 Dep MS/MS 2nd most intense ion from (1).
|
Comment_of_details
|
|
Data Analysis Details Information
ID
|
DS3
|
Title
|
PowerGet analysis for detection of all peaks (C4)
|
Description
|
Raw data files are converted to text file by MSGet software without cut off value and peaks are extracted from the text files by PowerFT with parameters (intensity cut off in peak detection=20000, margin of ion grouping in peak detection=5, peak selection filter is default, intensity cut off in peak assignment=1000).
|
Comment_of_details
|
|