Summary of Study ST001141

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000762. The data can be accessed directly via it's Project DOI: 10.21228/M86103 This work is supported by NIH grant, U2C- DK119886.

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Study ID	ST001141
Study Title	Evaluation of metabolome sample preparation and extraction methodologies for oleaginous filamentous fungi Mortierella alpina
Study Type	method optimization
Study Summary	In this study, based on the method of fast filtration, we evaluated the three metabolomics analysis protocols commonly used for microbial metabolomics analysis in M. alpina and systematically optimised the metabolite extraction solvent.
Institute	Jiangnan University
Department	School of Food Science and Technology
Laboratory	State Key Laboratory of Food Science and Technology
Last Name	Hengqian
First Name	Lu
Address	1800 Lihu Ave, Wuxi, Jiangsu 214122, P.R. China.
Email	hengqianlu@163.com
Phone	+86 15006176136
Submit Date	2019-02-24
Analysis Type Detail	GC-MS
Release Date	2019-09-23
Release Version	1

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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.