Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA080630

Local Sample ID:	M13A7
Subject ID:	SU001231
Subject Type:	Invertebrate
Subject Species:	Metacarcinus magister
Taxonomy ID:	29965
Species Group:	Invertebrates

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Subject:

Subject ID:	SU001231
Subject Type:	Invertebrate
Subject Species:	Metacarcinus magister
Taxonomy ID:	29965
Species Group:	Invertebrates

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
M13A7	SA080630	FL012337	High	pH Treatment
M13A7	SA080630	FL012337	High	DO Treatment

Collection:

Collection ID:	CO001225
Collection Summary:	After completing the required treatment time of 30-33 days, each crab was frozen at -80C and shipped to the West Coast Metabolomics Center.
Sample Type:	Muscle Tissue

Treatment:

Treatment ID:	TR001246
Treatment Summary:	Four treatments were used:high pH, high DO (dissolved oxygen); high pH, low DO; low pH, high DO; low pH, low DO.

Sample Preparation:

Sampleprep ID:	SP001239
Sampleprep Summary:	Crabs were removed from -80°C. 15 mg of muscle tissue was removed from each crab and placed in a 2 mL eppendorph tube. 2 x 3mm grinding beads were added to each sample. 225 µL of methanol with quality control standards were added to each sample. Samples were ground for 30 seconds at 1500 rpm using a GenoGrinder. 750 µL of methyl-tertbutyl ether (MTBE) was added to each sample. Samples were vortexed for 10 seconds and then shaked at 4°C for 6 minutes. 188 µL of LC-MS grade water was added to each sample. Samples were vortexed for 10 seconds and then centrifuged for 2 minutes at 14,000 rcf. The top, organic layer was split into two 350 µL aliquots, one submitted to lipidomics analysis and one saved for backup. The bottom layer was split into two 125 µL aliquots, one submitted to GC analysis and one saved as a backup.

Sampleprep ID:

SP001239

Sampleprep Summary:

Crabs were removed from -80°C. 15 mg of muscle tissue was removed from each crab and placed in a 2 mL eppendorph tube. 2 x 3mm grinding beads were added to each sample. 225 µL of methanol with quality control standards were added to each sample. Samples were ground for 30 seconds at 1500 rpm using a GenoGrinder. 750 µL of methyl-tertbutyl ether (MTBE) was added to each sample. Samples were vortexed for 10 seconds and then shaked at 4°C for 6 minutes. 188 µL of LC-MS grade water was added to each sample. Samples were vortexed for 10 seconds and then centrifuged for 2 minutes at 14,000 rcf. The top, organic layer was split into two 350 µL aliquots, one submitted to lipidomics analysis and one saved for backup. The bottom layer was split into two 125 µL aliquots, one submitted to GC analysis and one saved as a backup.

Combined analysis:

Analysis ID	AN001928
Analysis type	MS
Chromatography type	GC
Chromatography system	Agilent 7890A
Column	Restek Rtx-5Sil (30m x 0.25mm,0.25um)
MS Type	EI
MS instrument type	GC-TOF
MS instrument name	Leco Pegasus HT TOF
Ion Mode	POSITIVE
Units	Peak Height

Chromatography:

Chromatography ID:	CH001399
Instrument Name:	Agilent 7890A
Column Name:	Restek Rtx-5Sil (30m x 0.25mm,0.25um)
Column Temperature:	50-330
Flow Rate:	1 mL/min
Injection Temperature:	50°C ramped to 250°C by 12°C s-1
Sample Injection:	0.5 uL
Oven Temperature:	50°C for 1 min, then ramped at 20°C min-1 to 330°C, held constant for 5 min
Chromatography Type:	GC

MS:

MS ID:	MS001784
Analysis ID:	AN001928
Instrument Name:	Leco Pegasus HT TOF
Instrument Type:	GC-TOF
MS Type:	EI
MS Comments:	N/A
Ion Mode:	POSITIVE
Ion Source Temperature:	250°C
Ionization Energy:	-70 eV
Dataformat:	.peg, .txt, .cdf
Resolution Setting:	17 spectra/sec
Scanning Range:	80-500 Da