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MB Sample ID: SA258307
| Local Sample ID: | B2_2-NEG |
| Subject ID: | SU002671 |
| Subject Type: | Plant |
| Subject Species: | Pitcairnia flammea |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002671 |
| Subject Type: | Plant |
| Subject Species: | Pitcairnia flammea |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| B2_2-NEG | SA258307 | FL033150 | 20 min | Time |
| B2_2-NEG | SA258307 | FL033150 | MeOH;MTBE (1;3 v/v) | Solvent |
| B2_2-NEG | SA258307 | FL033150 | Sample | Sample_type |
Collection:
| Collection ID: | CO002664 |
| Collection Summary: | Leaves of cultivated Pitcairnia flammea were harvested, and immediately frozen in liquid nitrogen, followed the -80ºC storage until sample preparation. |
| Sample Type: | Leaves |
| Collection Location: | Institute of Biology (UNICAMP, Brazil) |
Treatment:
| Treatment ID: | TR002683 |
| Treatment Summary: | Leaves were macerated to fine powder using a mortar and pestle in liquid nitrogen condition. |
Sample Preparation:
| Sampleprep ID: | SP002677 |
| Sampleprep Summary: | Lipid extraction was based on Hummel et al. Method. 10 mL of solvent mixture were prepared in 3 different proportions - 1:2, 1:3 or 1:4 (v/v) -, pre-cooled at 20 ºC methanol (MeOH) and methyl tert-butyl ether (MTBE). In 2 mL tubes, 50 mg of macerated sample were added and 1 mL of the solvent mixture. The samples were incubated for 5 min under agitation at 500 rpm at 4 ºC, followed by an ultrasonication in ice-cold bath in different times (10, 20, 30 minutes). Each condition was performed in technical triplicate, totalizing 27 samples. After adding 500 µL mixture of water type I:MeOH (3:1 v/v), the samples were vortexed and centrifuged for 5 min at 4 ºC, 10000 rpm. The three phases were separated and dried in a vacuum concentrator, at ambient temperature under vacuum - alcoholic mode, and stored at -80 ºC until the chromatographic analysis. |
Combined analysis:
| Analysis ID | AN004234 | AN004235 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Scientific UltiMate™ 3000 UHPLC RSLCnano system | Thermo Scientific UltiMate™ 3000 UHPLC RSLCnano system |
| Column | Supelco Sigma-Aldrich Titan C18 column (100 mm x 2.1 mm x 1.9 µm) | Supelco Sigma-Aldrich Titan C18 column (100 mm x 2.1 mm x 1.9 µm) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | NEGATIVE | POSITIVE |
| Units | Peak area | Peak area |
Chromatography:
| Chromatography ID: | CH003142 |
| Instrument Name: | Thermo Scientific UltiMate™ 3000 UHPLC RSLCnano system |
| Column Name: | Supelco Sigma-Aldrich Titan C18 column (100 mm x 2.1 mm x 1.9 µm) |
| Column Temperature: | 40 |
| Flow Gradient: | 250 µL/min: 0-2 min: 40% B, 2-3 min: 50% B, 3-6 min: 50% B, 6.1-8 min: 70% B, 8-9 min: 100% B, 9-11 min: 100% B, 11-12 min: 40% B, 12-14 min: 40% B. |
| Flow Rate: | 250 µL/min |
| Solvent A: | 40% acetonitrile/60% water; 10 mM ammonium acetate |
| Solvent B: | 10% acetonitrile/90% isopropanol; 10 mM ammonium acetate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003981 |
| Analysis ID: | AN004234 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | - |
| Ion Mode: | NEGATIVE |
| MS ID: | MS003982 |
| Analysis ID: | AN004235 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | - |
| Ion Mode: | POSITIVE |
