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MB Sample ID: SA220283
Local Sample ID: | Blank1_C |
Subject ID: | SU002378 |
Subject Type: | Water sample |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002378 |
Subject Type: | Water sample |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
Blank1_C | SA220283 | FL027457 | CX-SPE | Extraction_Approach |
Blank1_C | SA220283 | FL027457 | NA | Location |
Collection:
Collection ID: | CO002371 |
Collection Summary: | Samples for dissolved metabolites from four unique environmental locations (Lake Washington (LW), Puget Sound (PS), Station ALOHA in the North Pacific Subtropical Gyre (ALOHA), and the North Pacific (NP)) from depths of 15 m for ALOHA and NP samples, 8 m for the PS sample, and 1 m for the LW sample. Samples were processed by filtering the water through 147 mm 0.2 μm PTFE filters using peristaltic pumps (ALOHA, NP, PS samples) and or glass vacuum filtration onto a 47 mm 0.2 μm PTFE filter. The filtrated was collected and stored in 2 L acid-washed polycarbonate bottles (ALOHA, NP, PS samples) or 45 mL acid-washed polypropylene falcon tubes at -20 C for between 1 month and 4 years until analysis. |
Sample Type: | seawater |
Storage Conditions: | -20℃ |
Treatment:
Treatment ID: | TR002390 |
Treatment Summary: | Not Applicable |
Sample Preparation:
Sampleprep ID: | SP002384 |
Sampleprep Summary: | Cation-Exchange Solid Phase Extraction (CX-SPE) Sample Volume: 40 mLs Solid Phase Resin/Column: Strong cation-exchange resin (Dowex 50WX8; H+ form, 100-200 mesh, Sigma-Aldrich, Vienna, Austria) Solvents/Reagents: 1 M NH3, 3 M HNO3 Brief Procedure: 35 g of resin was added to a glass chromatography column with a fritted disk and a PTFE stopcock. The resin was equilibrated with 50 mL water, 100 mL of 1M NH3, 50 mL water, 100 mL 3M HNO3, and 50 mL H2O. The samples were acidified with HNO3 to pH 2 and heavy isotope-labeled internal standards added. The sample was loaded onto column, allowed to stand for 5 minutes, and then drained from the column. The column was then rinsed with 50 mL water. Approximately 200 mL of 1M NH3 was added to column. Ammonia eluted from column in 10 mL fractions. The pH of each fraction checked by dabbing a small drop of sample onto a pH strip with a combusted glass Pasteur pipette. The alkaline front (the 10 mL fraction where the pH increases from approximately 2-4 to 9-11), the fraction before, and the two fractions after were collected, combined, and dried down under N2 gas in a water bath of approximately 32 degrees C. Dried fractions were redissolved in 380 uL of water. 20 uL of isotope-labeled injection standards in water were added to the fractions. Columns were regenerated for reuse through the addition of 50 mL of water, 100 mL 3M HNO3, and 50 mL of water. When not in use, columns were stored completely filled with 0.01 M HNO3. Water was extracted alongside samples as methodological blanks. PPL-Solid Phase Extraction (PPL-SPE) Sample Volume: 40 mLs Solid Phase Resin/Column: Agilent Bond Elut PPL cartridge, 1 g bed mass, 6 mL volume Solvents/Reagents: methanol, 0.01 M HCl, Brief Procedure: Sample acidified with HCl to pH 2 and heavy isotope-labeled internal standards added. Column prepped by adding 1 cartridge volumes of each of methanol followed by 0.01 M HCl. Sample loaded onto columns using a peristaltic pump at a flow rate of 10 mL/min Column rinsed with 2 cartridge volumes 0.01 M HCl. Sample eluted with 1 cartridge volume methanol and dried down under N2 gas. Dried fractions were redissolved in 380 uL of water. 20 uL of isotope-labeled injection standards in water were added to the fractions. Water was extracted alongside samples as methodological blanks. |
Combined analysis:
Analysis ID | AN003744 | AN003745 | AN003746 |
---|---|---|---|
Analysis type | MS | MS | MS |
Chromatography type | HILIC | HILIC | Reversed phase |
Chromatography system | Waters Acquity I-Class | Waters Acquity I-Class | Waters Acquity I-Class |
Column | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) | Waters Acquity HSS Cyano (1.8um,2.1 mm x 100 mm) |
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 | Peak Area | Peak Area | Peak Area |
Chromatography:
Chromatography ID: | CH002774 |
Chromatography Summary: | See attached summary |
Methods Filename: | Ingalls_Lab_LC_Methods_2.txt |
Instrument Name: | Waters Acquity I-Class |
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: | CH002775 |
Chromatography Summary: | See attached summary |
Methods Filename: | Ingalls_Lab_LC_Methods_2.txt |
Instrument Name: | Waters Acquity I-Class |
Column Name: | Waters Acquity HSS Cyano (1.8um,2.1 mm x 100 mm) |
Column Temperature: | 35 |
Flow Rate: | 0.4 ml/min |
Solvent A: | 100% water; 0.1% formic acid |
Solvent B: | 100% acetonitrile; 0.1% formic acid |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS003491 |
Analysis ID: | AN003744 |
Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | See Protocol |
Ion Mode: | POSITIVE |
Analysis Protocol File: | CXSPE_Method_Paper_MS_Methods.txt |
MS ID: | MS003492 |
Analysis ID: | AN003745 |
Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | See Protocol |
Ion Mode: | NEGATIVE |
Analysis Protocol File: | CXSPE_Method_Paper_MS_Methods.txt |
MS ID: | MS003493 |
Analysis ID: | AN003746 |
Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | See Protocol |
Ion Mode: | POSITIVE |
Analysis Protocol File: | CXSPE_Method_Paper_MS_Methods.txt |