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MB Sample ID: SA289385
Local Sample ID: | QC03-NEG |
Subject ID: | SU002856 |
Subject Type: | Plant |
Subject Species: | Pitcairnia flammea |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002856 |
Subject Type: | Plant |
Subject Species: | Pitcairnia flammea |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
QC03-NEG | SA289385 | FL035677 | Pooled QC | Collection locale |
Collection:
Collection ID: | CO002849 |
Collection Summary: | Pitcairnia flammea individuals were collected in different altitudes locations and named according to the local. PAP was collected at 2140 m, MAR 2037 m, PES 1496 m, ITA 1139 m, MAC 959 m, COR 425 m, RAN 25 m and UBA 20 m. Then, the plants were cultivated at the Institute of Biology (UNICAMP, Brazil). The mid-region of the youngest fully expanded leaves of five to seven individuals per population of healthy plants with visually appealing leaves 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: | TR002865 |
Treatment Summary: | Leaves were macerated to fine powder using a mortar and pestle in liquid nitrogen condition. |
Sample Preparation:
Sampleprep ID: | SP002862 |
Sampleprep Summary: | Lipid extraction was based on Hummel et al. Method. 60 mL of solvent mixture was prepared with pre-cooled (-20ºC) methanol (MeOH, grade HPLC, LiChrosolv® Reag. Ph. Eur.) and methyl-tert-butyl-ether (MTBE, grade HPLC, purity 99,9%, Sigma-Aldrich) in proportion (1:3 v/v). In a 2 mL tube, 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 (Microtube Shaking Incubator AccuTherm, Labnet International, Inc.), followed by an ultrasonication (Branson 5800 Ultrasonic Bath, Emerson, Danbury, USA) in ice-cold bath in 10 minutes. 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 (Hettich Zentrifugen Mikro 220R, Tuttlingen, DE). The three phases were separated and dried in a vacuum concentrator (Concentrator Plus, Eppendorf AG, Hamburg, DE), at ambient temperature under vacuum - alcoholic mode, and stored at -80 ºC until the chromatographic analysis. |
Sampleprep Protocol Filename: | SamplePrep-BromeliadLipidomics.pdf |
Combined analysis:
Analysis ID | AN004459 | AN004460 |
---|---|---|
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 (100 x 2.1mm, 1.9um) | Supelco Sigma-Aldrich Titan C18 (100 x 2.1mm, 1.9um) |
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: | CH003348 |
Methods Filename: | UntargetedRP-BromeliadLipidomics.pdf |
Instrument Name: | Thermo Scientific UltiMate™ 3000 UHPLC RSLCnano system |
Column Name: | Supelco Sigma-Aldrich Titan C18 (100 x 2.1mm, 1.9um) |
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: | MS004206 |
Analysis ID: | AN004459 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | - |
Ion Mode: | NEGATIVE |
Analysis Protocol File: | UntargetedRP-BromeliadLipidomics.pdf |
MS ID: | MS004207 |
Analysis ID: | AN004460 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | - |
Ion Mode: | POSITIVE |
Analysis Protocol File: | UntargetedRP-BromeliadLipidomics.pdf |