Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA273876

Local Sample ID:	16
Subject ID:	SU002829
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

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Subject:

Subject ID:	SU002829
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
16	SA273876	FL035505	1	Donor
16	SA273876	FL035505	Control	Treatment
16	SA273876	FL035505	0	Time

Collection:

Collection ID:	CO002822
Collection Summary:	Two human stool donors were selected based on their inflammation profile which occurred prior to gut microbial community profiling. Female germ- free (GF) C57BL/6J mice, originally purchased from the Jackson Laboratory (Bar Harbor, ME) were housed and bred at the American Association for the Accreditation of Laboratory Animal Care-accredited Animal Resource Center at Montana State University. Mice were held in individually ventilated cages with sterile bedding before and after fecal transplantation from selected human stool donors. Two female mice received an inoculation with fecal material from a human donor categorized as having low or high systemic inflammation based on serum levels of six proinflammatory cytokines. Human donor stool slurry aliquots were administered to GF mice through oral gavage. Sexually mature male GF C57BL/6J mice were added to each cage approximately one week after transplantation. Male mice removed prior to birth of pups. Pups from the inoculated dams were co-housed by sex (3 – 5 mice/cage) with different microbial inoculations placed in separate isolators. Mice from each microbial inoculation were assigned to one of two juice groups: Aronia (ARO LO , = 3, ARO HI , n=5), or a sugar-matched juice (CON LO ,n=3, CON HI , n = 3). Weekly measurements of body weight and food and fluid intake were recorded. Blood samples were collected at the same interval into serum separating tubes. Whole blood was allowed to clot for 15 minutes before centrifugation at 1200 RPM for 15 minutes with resulting serum aliquoted and stored at -80ºC until analysis. After T8 sample collection, mice were euthanized via rapid CO 2 asphyxiation.
Sample Type:	Blood (serum)

Treatment:

Treatment ID:	TR002838
Treatment Summary:	At baseline (T0), regular drinking water was replaced with ARO or CON juice to begin a two-week familiarization period with the juice. ARO mice received an unpasteurized blend of Aronia juice, and CON mice received a sugar-matched beverage containing water, sorbitol, glucose, and fructose. The mice were housed in cages with free access to their respective juice. During the familiarization period, all mice received standard chow (LabDiet 5013). After the 2-week familiarization period (T2), mice began a 6-week high-fat diet, delivered ad libitum concomitant with juice consumption. The HFD (Teklad TD.96132) was chosen to induce obesity and present an inflammatory stimulus (Duan et al., 2018) . The HFD mimics a Western style diet and consisted of 40.6% fat, 40.7% carbohydrate, and 18.7% protein and was particularly rich in sugars and trans-fatty acids. All chow provided was sterilized via autoclaving or irradiation. A total of 150 mL of juice was provided per cage each week. Juice was refilled three times each week.

Treatment ID:

TR002838

Treatment Summary:

At baseline (T0), regular drinking water was replaced with ARO or CON juice to begin a two-week familiarization period with the juice. ARO mice received an unpasteurized blend of Aronia juice, and CON mice received a sugar-matched beverage containing water, sorbitol, glucose, and fructose. The mice were housed in cages with free access to their respective juice. During the familiarization period, all mice received standard chow (LabDiet 5013). After the 2-week familiarization period (T2), mice began a 6-week high-fat diet, delivered ad libitum concomitant with juice consumption. The HFD (Teklad TD.96132) was chosen to induce obesity and present an inflammatory stimulus (Duan et al., 2018) . The HFD mimics a Western style diet and consisted of 40.6% fat, 40.7% carbohydrate, and 18.7% protein and was particularly rich in sugars and trans-fatty acids. All chow provided was sterilized via autoclaving or irradiation. A total of 150 mL of juice was provided per cage each week. Juice was refilled three times each week.

Sample Preparation:

Sampleprep ID:	SP002835
Sampleprep Summary:	Samples were stored at -80 deg. C until ready for analysis. A liquid extraction was completed with a protein precipitation. Samples were then concentrated and stored dry until LCMS analysis.

Combined analysis:

Analysis ID	AN004414
Analysis type	MS
Chromatography type	HILIC
Chromatography system	Agilent 1290 Infinity II
Column	Waters ACQUITY UPLC BEH HILIC (150 x 2.1mm,1.7um)
MS Type	ESI
MS instrument type	QTOF
MS instrument name	Agilent 6530 QTOF
Ion Mode	POSITIVE
Units	area

Chromatography:

Chromatography ID:	CH003313
Instrument Name:	Agilent 1290 Infinity II
Column Name:	Waters ACQUITY UPLC BEH HILIC (150 x 2.1mm,1.7um)
Column Temperature:	40
Flow Gradient:	Linear 45-70%A
Flow Rate:	0.4mL/min
Solvent A:	Water with 0.1% formic acid
Solvent B:	Acetonitrile with 0.1% formic acid
Chromatography Type:	HILIC

MS:

MS ID:	MS004161
Analysis ID:	AN004414
Instrument Name:	Agilent 6530 QTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	Mass Hunter to acquire data. Processed with msconvert and mzMine. Annotation with SIRIUS software.
Ion Mode:	POSITIVE