Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA206368

Local Sample ID:	WT-4
Subject ID:	SU002239
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	AppNL-G-F or wild type mice
Age Or Age Range:	7 months old
Gender:	Male and female
Animal Animal Supplier:	RIKEN Center for Brain Science
Animal Light Cycle:	12 hours light and dark cycle

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Subject:

Subject ID:	SU002239
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	AppNL-G-F or wild type mice
Age Or Age Range:	7 months old
Gender:	Male and female
Animal Animal Supplier:	RIKEN Center for Brain Science
Animal Light Cycle:	12 hours light and dark cycle

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
WT-4	SA206368	FL025286	Wild type	Model
WT-4	SA206368	FL025286	B. breve MCC1274	Intervention

Collection:

Collection ID:	CO002232
Collection Summary:	Plasma samples were obtained from AD-like model and wild-type mice orally administered B. breve MCC1274 or saline for 4 months.
Sample Type:	Blood (plasma)
Storage Conditions:	-80℃

Treatment:

Treatment ID:	TR002251
Treatment Summary:	Three-month-old AppNL-G-F and wild type mice were randomly assigned to a vehicle group and a probiotic group. The vehicle group received saline, and the probiotic group received B. breve MCC1274 (1 × 10＾9 cfu/200 μl saline/mouse) orally 5 times a week for 4 months.
Treatment Compound:	Bifidobacterium breve MCC1274
Treatment Route:	oral administration
Treatment Dosevolume:	1 × 10＾9 cfu/200 μl saline
Treatment Vehicle:	saline

Sample Preparation:

Sampleprep ID:	SP002245
Sampleprep Summary:	For CE-FTMS, 40 µL of plasma was added to 160 µL of methanol containing internal standards at 0°C, 120 µL of Milli-Q water was added and mixed thoroughly, and 240 µL of this mixture was added to a Millipore 5 kDa cutoff filter (ULTRAFREE MC PLHCC, HMT) at 9100 × g for 120 min at 4°C to remove macromolecules. The filtrate was evaporated to dryness under vacuum, dissolved in 40 µL Milli-Q water, and donated to HMT's ω Scan package using capillary electrophoresis Fourier transform mass spectrometry (CE-FTMS).For LC-TOFMS, 80 µL of plasma was added to 240 µL of 1% formic acid/acetonitrile containing internal standards at 0ºC. The mixture was centrifuged at 2,300 x g, 4ºC for 5 min, then centrifuged at 2,300 x g, 4ºC for 5 min, followed by centrifugation of a hybrid SPE phospholipid cartridge (30 mg/mL of hybrid SPE-phospholipid, 30 mg/mL of (SUPELCO) and filtered to remove phospholipids. The filtrate was then evaporated to dryness under nitrogen, dissolved in 80 μL of 50% isopropanol (v/v), and metabolomic analysis was performed using liquid chromatography time-of-flight mass spectrometry (LC-TOFMS).

Sampleprep ID:

SP002245

Sampleprep Summary:

For CE-FTMS, 40 µL of plasma was added to 160 µL of methanol containing internal standards at 0°C, 120 µL of Milli-Q water was added and mixed thoroughly, and 240 µL of this mixture was added to a Millipore 5 kDa cutoff filter (ULTRAFREE MC PLHCC, HMT) at 9100 × g for 120 min at 4°C to remove macromolecules. The filtrate was evaporated to dryness under vacuum, dissolved in 40 µL Milli-Q water, and donated to HMT's ω Scan package using capillary electrophoresis Fourier transform mass spectrometry (CE-FTMS).For LC-TOFMS, 80 µL of plasma was added to 240 µL of 1% formic acid/acetonitrile containing internal standards at 0ºC. The mixture was centrifuged at 2,300 x g, 4ºC for 5 min, then centrifuged at 2,300 x g, 4ºC for 5 min, followed by centrifugation of a hybrid SPE phospholipid cartridge (30 mg/mL of hybrid SPE-phospholipid, 30 mg/mL of (SUPELCO) and filtered to remove phospholipids. The filtrate was then evaporated to dryness under nitrogen, dissolved in 80 μL of 50% isopropanol (v/v), and metabolomic analysis was performed using liquid chromatography time-of-flight mass spectrometry (LC-TOFMS).

Combined analysis:

Analysis ID	AN003526	AN003527
Analysis type	MS	MS
Chromatography type	Normal phase	Reversed phase
Chromatography system	Agilent 1260	Agilent 1200
Column	fused silica capillary (50 μm i.d. × 80 cm total length; Polymicro Technologies, Inc., Phoenix, AZ)	ODS
MS Type	ESI	ESI
MS instrument type	Orbitrap	Triple TOF
MS instrument name	Thermo Q Exactive Plus Orbitrap	Agilent 6210 TOF
Ion Mode	UNSPECIFIED	UNSPECIFIED
Units	peak area	peak area

Chromatography:

Chromatography ID:	CH002604
Instrument Name:	Agilent 1260
Column Name:	fused silica capillary (50 μm i.d. × 80 cm total length; Polymicro Technologies, Inc., Phoenix, AZ)
Internal Standard:	H3304-1002, Human Metabolome Technologies, Inc. (HMT), Tsuruoka, Yamagata, Japan
Solvent A:	commercial electrophoresis buffer (H3301-1001; I3302-1023 for cation; anion analyses, respectively, HMT)
Chromatography Type:	Normal phase

Chromatography ID:	CH002605
Instrument Name:	Agilent 1200
Column Name:	ODS
Internal Standard:	H3304-1002, Human Metabolome Technologies, Inc. (HMT), Tsuruoka, Yamagata, Japan
Chromatography Type:	Reversed phase

MS:

MS ID:	MS003284
Analysis ID:	AN003526
Instrument Name:	Thermo Q Exactive Plus Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	The spectrometer was scanned from m/z 60 to 900 in positive mode, and from m/z 70 to 1,050 in negative mode
Ion Mode:	UNSPECIFIED

MS ID:	MS003285
Analysis ID:	AN003527
Instrument Name:	Agilent 6210 TOF
Instrument Type:	Triple TOF
MS Type:	ESI
MS Comments:	The spectrometer was scanned from m/z 50 to 1,000 and peaks were extracted using MasterHands, automatic integration software (Keio University, Tsuruoka, Yamagata, Japan)
Ion Mode:	UNSPECIFIED