Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA217526

Local Sample ID:	Blk_MQBlk_1
Subject ID:	SU002352
Subject Type:	Water sample

Select appropriate tab below to view additional metadata details:

Subject:

Subject ID:	SU002352
Subject Type:	Water sample

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
Blk_MQBlk_1	SA217526	FL026414	NA	Sampling_Date
Blk_MQBlk_1	SA217526	FL026414	NA	Latitude
Blk_MQBlk_1	SA217526	FL026414	NA	Longitude
Blk_MQBlk_1	SA217526	FL026414	NA	Station
Blk_MQBlk_1	SA217526	FL026414	NA	Cast
Blk_MQBlk_1	SA217526	FL026414	NA	Depth
Blk_MQBlk_1	SA217526	FL026414	NA	sample_vol_filtered_L

Collection:

Collection ID:	CO002345
Collection Summary:	Samples for particulate metabolites were collected from 5 different stations surrounding the lava flowing into the ocean off the island of Hawaii, all from a depth of 5m. At each sampling location, single, duplicate, or triplicate filters were collected using either niskins attached to a conductivity, temperature, depth array (CTD) or the underway intake. Samples (10 L) were collected into polycarbonate carboys, filtered onto 147 mm 0.2 μm PTFE filters using peristaltic pumps, polycarbonate filter holders, and Masterflex PharMed BPT tubing (Cole-Parmer). Filters were flash frozen in liquid nitrogen and stored at -80°C until extraction. In addition to our samples, we filtered MilliQ water through a 0.2 μm PTFE filter and extracted this filter alongside samples as a methodological blank.
Sample Type:	Phytoplankton
Collection Method:	CTD Niskin arrays
Collection Location:	Hawai'i
Volumeoramount Collected:	10L
Storage Conditions:	-80℃

Treatment:

Treatment ID:	TR002364
Treatment Summary:	Samples (10 L) were collected into polycarbonate carboys, filtered onto 147 mm 0.2 μm PTFE filters using peristaltic pumps, polycarbonate filter holders, and Masterflex PharMed BPT tubing (Cole-Parmer). Filters were flash frozen in liquid nitrogen and stored at -80°C until extraction.

Sample Preparation:

Sampleprep ID:	SP002358
Sampleprep Summary:	Each sample was extracted using a modified Bligh-Dyer extraction. Briefly, filters were cut up and put into 15 mL teflon centrifuge tubes containing a mixture of 100 µm and 400 µm silica beads. Heavy isotope-labeled internal standards were added along with ~2 mL of cold aqueous solvent (50:50 methanol:water) and ~3 mL of cold organic solvent (dichloromethane). The samples were shaken on a FastPrep-24 Homogenizer for 30 seconds and chilled in a -20 °C freezer repeatedly for three cycles of bead-beating and a total of 30 minutes of chilling. The organic and aqueous layers were separated by spinning samples in a centrifuge at 4,300 rpm for 2 minutes at 4 °C. The aqueous layer was removed to a new glass centrifuge tube. The remaining organic fraction was rinsed three more times with additions of 1 to 2 mL of 50:50 methanol:water. All aqueous rinses were combined for each sample and ~2 mL of cold dichloromethane was added to the combined aqueous layer. Tubes were shaken and centrifuged at 4,300 rpm for 2 minutes at 4°C. The aqueous layer was removed to a new glass vial and dried under N2 gas. The remaining organic layer in the bead beating tubes was transferred into the glass centrifuge tube and the bead beating tube was rinsed two more times with cold organic solvent. The combined organic rinses were centrifuged, transferred to a new glass vial, and dried under N2 gas. Dried aqueous fractions were re-dissolved in 380 µL of water. Dried organic fractions were re-dissolved in 400 µL of 90:10 methanol:toluene. 20 µL of isotope-labeled injection standards in water were added to the aqueous fractions. Blank filters were extracted alongside samples as methodological blanks.

Sampleprep ID:

SP002358

Sampleprep Summary:

Each sample was extracted using a modified Bligh-Dyer extraction. Briefly, filters were cut up and put into 15 mL teflon centrifuge tubes containing a mixture of 100 µm and 400 µm silica beads. Heavy isotope-labeled internal standards were added along with ~2 mL of cold aqueous solvent (50:50 methanol:water) and ~3 mL of cold organic solvent (dichloromethane). The samples were shaken on a FastPrep-24 Homogenizer for 30 seconds and chilled in a -20 °C freezer repeatedly for three cycles of bead-beating and a total of 30 minutes of chilling. The organic and aqueous layers were separated by spinning samples in a centrifuge at 4,300 rpm for 2 minutes at 4 °C. The aqueous layer was removed to a new glass centrifuge tube. The remaining organic fraction was rinsed three more times with additions of 1 to 2 mL of 50:50 methanol:water. All aqueous rinses were combined for each sample and ~2 mL of cold dichloromethane was added to the combined aqueous layer. Tubes were shaken and centrifuged at 4,300 rpm for 2 minutes at 4°C. The aqueous layer was removed to a new glass vial and dried under N2 gas. The remaining organic layer in the bead beating tubes was transferred into the glass centrifuge tube and the bead beating tube was rinsed two more times with cold organic solvent. The combined organic rinses were centrifuged, transferred to a new glass vial, and dried under N2 gas. Dried aqueous fractions were re-dissolved in 380 µL of water. Dried organic fractions were re-dissolved in 400 µL of 90:10 methanol:toluene. 20 µL of isotope-labeled injection standards in water were added to the aqueous fractions. Blank filters were extracted alongside samples as methodological blanks.

Combined analysis:

Analysis ID	AN003701	AN003702	AN003703
Analysis type	MS	MS	MS
Chromatography type	HILIC	HILIC	GC
Chromatography system	Q Exactive™ Plus Hybrid Quadrupole-Orbitrap	Q Exactive™ Plus Hybrid Quadrupole-Orbitrap	Q Exactive™ Plus Hybrid Quadrupole-Orbitrap
Column	SeQuant ZIC-pHILIC (150 x 2.1mm,5um)	SeQuant ZIC-pHILIC (150 x 2.1mm,5um)	Waters Acquity UPLC HSS Cyano (100 x 2.1mm,1.8um)
MS Type	ESI	ESI	ESI
MS instrument type	Orbitrap	Orbitrap	Orbitrap
MS instrument name	Thermo Q Exactive HF hybrid Orbitrap	Thermo Q Exactive HF hybrid Orbitrap	Thermo Q Exactive HF hybrid Orbitrap
Ion Mode	POSITIVE	NEGATIVE	POSITIVE
Units	Normalized peak area	Normalized peak area	Normalized peak area

Chromatography:

Chromatography ID:	CH002742
Chromatography Summary:	See attached summary
Methods Filename:	Ingalls_Metabolomics_LC_HOT-LAVA.txt
Instrument Name:	Q Exactive™ Plus Hybrid Quadrupole-Orbitrap
Column Name:	SeQuant ZIC-pHILIC (150 x 2.1mm,5um)
Column Temperature:	30
Flow Rate:	0.15 ml/min
Solvent A:	85% acetonitrile/15% water; 10 mM ammonium carbonate
Solvent B:	15% acetonitrile/85% water; 10 mM ammonium carbonate
Chromatography Type:	HILIC

Chromatography ID:	CH002743
Chromatography Summary:	See attached summary
Methods Filename:	Ingalls_Metabolomics_LC_HOT-LAVA.txt
Instrument Name:	Q Exactive™ Plus Hybrid Quadrupole-Orbitrap
Column Name:	Waters Acquity UPLC HSS Cyano (100 x 2.1mm,1.8um)
Chromatography Type:	GC

MS:

MS ID:	MS003451
Analysis ID:	AN003701
Instrument Name:	Thermo Q Exactive HF hybrid Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	See protocol
Ion Mode:	POSITIVE
Analysis Protocol File:	Ingalls_Metabolomics_MS_HOT-LAVA.txt

MS ID:	MS003452
Analysis ID:	AN003702
Instrument Name:	Thermo Q Exactive HF hybrid Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	See protocol
Ion Mode:	NEGATIVE
Analysis Protocol File:	Ingalls_Metabolomics_MS_HOT-LAVA.txt

MS ID:	MS003453
Analysis ID:	AN003703
Instrument Name:	Thermo Q Exactive HF hybrid Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	See protocol
Ion Mode:	POSITIVE
Analysis Protocol File:	Ingalls_Metabolomics_MS_HOT-LAVA.txt