Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA113589

Local Sample ID:	KO_DW_2
Subject ID:	SU001473
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	Samtako (Gyeonggi-do, Korea)
Gender:	Male

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Subject:

Subject ID:	SU001473
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	Samtako (Gyeonggi-do, Korea)
Gender:	Male

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
KO_DW_2	SA113589	FL014339	Olfr544 KO mouse	Genotype
KO_DW_2	SA113589	FL014339	HFD	Treatment

Collection:

Collection ID:	CO001468
Collection Summary:	Fecal samples were thawed on ice, and 50 mg of each sample was transferred to a fresh 1.5-mL Eppendorf tube. Then, 1 mL of pure methanol (Merck Millipore, Burlington, MA, USA) was added to each sample. The mixture was homogenised using a needle, sonicated for 30 min, vortexed for 10 min, and centrifuged at 16,100 × g for 10 min at 4°C. The supernatant was filtered by using a syringe filter (0.20-µm pore size; Thermo Fisher Scientific, Waltham, MA, USA). Aliquots of 500 μL of the supernatant were dried under vacuum at room temperature.
Sample Type:	Feces

Treatment:

Treatment ID:	TR001488
Treatment Summary:	The mice were fed on 60% high fat diet (HFD) for 4 weeks and subsequently randomly assigned into the test or vehicle group. HFD used in this study is rodent diet containing 60% of calories from fat (D12492, Research Diets, Inc). The mice in the test group were fed with a HFD and 0.05% (w/w) AzA for 6 weeks or a HFD and orally administered AzA (50 mg/kg body weight) for 8 weeks. The former feeding was to confirm antiobesogenic effects of AzA in diet and the latter was to investigate microbiome and metabolomic analyses. The mice in the vehicle group were administered ddH2O. The body weight and food intake of the mice were measured twice a week. Mice were fasted overnight before being sacrificed.

Treatment ID:

TR001488

Treatment Summary:

The mice were fed on 60% high fat diet (HFD) for 4 weeks and subsequently randomly assigned into the test or vehicle group. HFD used in this study is rodent diet containing 60% of calories from fat (D12492, Research Diets, Inc). The mice in the test group were fed with a HFD and 0.05% (w/w) AzA for 6 weeks or a HFD and orally administered AzA (50 mg/kg body weight) for 8 weeks. The former feeding was to confirm antiobesogenic effects of AzA in diet and the latter was to investigate microbiome and metabolomic analyses. The mice in the vehicle group were administered ddH2O. The body weight and food intake of the mice were measured twice a week. Mice were fasted overnight before being sacrificed.

Sample Preparation:

Sampleprep ID:	SP001481
Sampleprep Summary:	Methoximation and silylation were performed for qualification and relative quantification of metabolites using GC/TOF–MS, For methoximation, 10 µL of 40 mg/mL methoxyamine hydrochloride in pyridine (Sigma-Aldrich) was added to the dried sample, and the mixture was incubated at 30°C for 90 minutes. For silylation, 45 µL of N-methyl-N-trimethylsilyl-trifluoroacetamide (Fluka, Buchs, Switzerland) was added to the mixture, and the mixture was incubated at 37°C for 30 minutes.

Sampleprep ID:

SP001481

Sampleprep Summary:

Methoximation and silylation were performed for qualification and relative quantification of metabolites using GC/TOF–MS, For methoximation, 10 µL of 40 mg/mL methoxyamine hydrochloride in pyridine (Sigma-Aldrich) was added to the dried sample, and the mixture was incubated at 30°C for 90 minutes. For silylation, 45 µL of N-methyl-N-trimethylsilyl-trifluoroacetamide (Fluka, Buchs, Switzerland) was added to the mixture, and the mixture was incubated at 37°C for 30 minutes.

Combined analysis:

Analysis ID	AN002339
Analysis type	MS
Chromatography type	GC
Chromatography system	Agilent 7890B
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	Arbtrary unit

Chromatography:

Chromatography ID:	CH001714
Instrument Name:	Agilent 7890B
Column Name:	Restek Rtx-5Sil (30m x 0.25mm,0.25um)
Chromatography Type:	GC

MS:

MS ID:	MS002181
Analysis ID:	AN002339
Instrument Name:	Leco Pegasus HT TOF
Instrument Type:	GC-TOF
MS Type:	EI
MS Comments:	Mass spectra were recorded over a mass range of 85–500 m/z at an acquisition rate of 16 spectra/s. The temperatures of the ion source and transfer line of the TOF–MS were 250°C and 280°C, respectively. The electron ionisation energy was 70 eV.Chroma TOF Software C version (LECO) was used for the detection and deconvolution of peaks and mass spectra. BinBase was used for the identification and relative quantification of metabolites from preprocessed data by matching mass spectra and retention indices of peaks with the customised reference mass spectral and retention index libraries, acquired using authentic standards with identical data acquisition parameters for the Fiehn and NIST libraries. Peaks with mass spectral similarities above 700, in comparison to their authentic standards in the libraries, were regarded as authentic metabolites. Intensities of metabolites were reported as peak heights of their unique ion intensities. For management of missing values, the lowest background intensity was subtracted from the intensity of the quantified ion in its retention time region ± 5s, using MZmine.
Ion Mode:	POSITIVE