Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA078327

Local Sample ID:	SM1-1
Subject ID:	SU001205
Subject Type:	Fungi
Subject Species:	Mortierella alpina
Taxonomy ID:	64518

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Subject:

Subject ID:	SU001205
Subject Type:	Fungi
Subject Species:	Mortierella alpina
Taxonomy ID:	64518

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
SM1-1	SA078327	FL011947	SM1	Treatment

Collection:

Collection ID:	CO001199
Collection Summary:	Mycelia were collected by vacuum filtration at 120 h using a paper filter (Whatman no. 1) and washed with 4°C 0.9% NaCl at room temperature, the sampling time was less than 45s. After quick-freezing in liquid nitrogen, the frozen cell pellets were further ground into powder (fresh biomass) and stored at -80°C for further use.
Sample Type:	fungi

Treatment:

Treatment ID:	TR001220
Treatment Summary:	1.Sample preparation: Two forms of biomass, including fresh weight and freeze-dried, have been used for the extraction of metabolites. Three forms of quantified biomass, including fresh weight, freeze-dried and cell debris, have been used for data normalisation. 2. We create a pooled culture and split to produce identical samples for the different methods tested. Before extraction, three ways(fresh sample,freeze-dried sample, freeze-dried sample(water added)) were adopted to prepare the metabolomics analytical sample.To test the effect of the water contained in the fresh biomass on the extraction of metabolites from M. alpina, we repeated the freeze-dried biomass with additional water added to the account for the water content in fresh biomass. 2. Five solvent mixtures (SM) were used for the extraction of the intracellular metabolites of M. alpina. The five SM were: SM1, MTBE:methanol:water (20:6:5, vol:vol:vol) at -20°C; SM2, methanol:acetonitrile:water (2:2:1, vol:vol:vol) at -20°C; SM3, methanol:water (8:2, vol:vol) at -20°C; SM4, methanol:water (1:1, vol:vol); and SM5, Ethanol:water (3:1, vol:vol) at -20°C. 3. The mycelia were harvested at 24h and 120h for metabolomics analysis.

Treatment ID:

TR001220

Treatment Summary:

1.Sample preparation: Two forms of biomass, including fresh weight and freeze-dried, have been used for the extraction of metabolites. Three forms of quantified biomass, including fresh weight, freeze-dried and cell debris, have been used for data normalisation. 2. We create a pooled culture and split to produce identical samples for the different methods tested. Before extraction, three ways(fresh sample,freeze-dried sample, freeze-dried sample(water added)) were adopted to prepare the metabolomics analytical sample.To test the effect of the water contained in the fresh biomass on the extraction of metabolites from M. alpina, we repeated the freeze-dried biomass with additional water added to the account for the water content in fresh biomass. 2. Five solvent mixtures (SM) were used for the extraction of the intracellular metabolites of M. alpina. The five SM were: SM1, MTBE:methanol:water (20:6:5, vol:vol:vol) at -20°C; SM2, methanol:acetonitrile:water (2:2:1, vol:vol:vol) at -20°C; SM3, methanol:water (8:2, vol:vol) at -20°C; SM4, methanol:water (1:1, vol:vol); and SM5, Ethanol:water (3:1, vol:vol) at -20°C. 3. The mycelia were harvested at 24h and 120h for metabolomics analysis.

Sample Preparation:

Sampleprep ID:	SP001213
Sampleprep Summary:	The obtained supernatants were vacuum dried at 30°C prior to derivatisation. The extraction solution served as a negative control to subtract from the background spectrum. The dried extract was derivatised. Briefly, the vacuum dried samples were resuspended in 100 μL of MeOX (10 mg/mL) in pyridine and incubated at 37°C for 90 min in a block heater. Then, 40 μL of MSTFA+1%TMCS was pipetted into each sample and incubated at 37°C for another 30 min. The resulting solution from each sample was collected and transferred into gas chromatography (GC) vials for analysis.

Sampleprep ID:

SP001213

Sampleprep Summary:

The obtained supernatants were vacuum dried at 30°C prior to derivatisation. The extraction solution served as a negative control to subtract from the background spectrum. The dried extract was derivatised. Briefly, the vacuum dried samples were resuspended in 100 μL of MeOX (10 mg/mL) in pyridine and incubated at 37°C for 90 min in a block heater. Then, 40 μL of MSTFA+1%TMCS was pipetted into each sample and incubated at 37°C for another 30 min. The resulting solution from each sample was collected and transferred into gas chromatography (GC) vials for analysis.

Combined analysis:

Analysis ID	AN001874
Analysis type	MS
Chromatography type	GC
Chromatography system	Thermo Trace 1310
Column	RTX-5MS
MS Type	EI
MS instrument type	Triple quadrupole
MS instrument name	Thermo TSQ8000_evo
Ion Mode	POSITIVE
Units	Peak Area

Chromatography:

Chromatography ID:	CH001355
Instrument Name:	Thermo Trace 1310
Column Name:	RTX-5MS
Chromatography Type:	GC

MS:

MS ID:	MS001730
Analysis ID:	AN001874
Instrument Name:	Thermo TSQ8000_evo
Instrument Type:	Triple quadrupole
MS Type:	EI
MS Comments:	The “raw” format files were converted to “abf ” format with the ABF converter. The MSDIAL3.40 equipped with DB_FiehnBinbase-FiehnRI database was used for peaks exaction, retention time adjustment, peak alignment, deconvolution analysis and identification.All metabolite annotations were checked again by using the the TraceFinder 3.3 software (Thermo Fisher Scientific, Waltham, MA, USA), Xcalibur 3.1 and NIST MS search.
Ion Mode:	POSITIVE