Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA248908

Local Sample ID:	1_STD10
Subject ID:	SU002581
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

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Subject:

Subject ID:	SU002581
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
1_STD10	SA248908	FL031428	-	Genotype

Collection:

Collection ID:	CO002574
Collection Summary:	Serum samples from mice were collected and stored at -80°C.
Sample Type:	Blood (serum)

Treatment:

Treatment ID:	TR002593
Treatment Summary:	Thawed on ice, serum sample was mixed with 4 volume extraction solvent of ice-cold methanol containing indole-3-acetic acid-d4 and kynurenic acid-d5. Mixture was vortexed and incubated at −20 °C for 30 min. Following centrifugation at 12,000 × g for 15 min at 4℃, supernatant was collected and subsequently evaporated to dryness (Thermo Scientific, Waltham, MA) and dissolved in 10% acetonitrile containing 1 µM chlorpropamide. After centrifugation at 12,000 × g for 15 min at 4℃, supernatants were transferred to autosampler vials for LC-MS analysis.

Treatment ID:

TR002593

Treatment Summary:

Thawed on ice, serum sample was mixed with 4 volume extraction solvent of ice-cold methanol containing indole-3-acetic acid-d4 and kynurenic acid-d5. Mixture was vortexed and incubated at −20 °C for 30 min. Following centrifugation at 12,000 × g for 15 min at 4℃, supernatant was collected and subsequently evaporated to dryness (Thermo Scientific, Waltham, MA) and dissolved in 10% acetonitrile containing 1 µM chlorpropamide. After centrifugation at 12,000 × g for 15 min at 4℃, supernatants were transferred to autosampler vials for LC-MS analysis.

Sample Preparation:

Sampleprep ID:	SP002587
Sampleprep Summary:	C57BL6/J mice were obtained from Jackson Laboratory (Bar Harbor, ME), bred in-house and maintained on a chow diet. Ahr+/-, and Ahr-/- mice were kindly provided by Dr. Christopher Bradfield (University of Wisconsin-Madison) and bred using female Ahr+/- and male Ahr-/- mice. Mice were housed on corncob bedding in a temperature- and light-controlled facility and given access to food and water ad libitum. Mice were also maintained in a pathogen-free facility and treated humanely with approval from the Animal Care and Use Committee of the Pennsylvania State University and methods were carried out in accordance with approved guidelines. Mice in the 8-10-week-old range were utilized in all experiments. Serum samples obtained from mice exposed to PCB126 for 5 days was previously described. Briefly, mice were fed dough pills once every 24 h, each pill contained 24 µg/kg PCB126 for five days and mice were sacrificed on day 6.

Sampleprep ID:

SP002587

Sampleprep Summary:

C57BL6/J mice were obtained from Jackson Laboratory (Bar Harbor, ME), bred in-house and maintained on a chow diet. Ahr+/-, and Ahr-/- mice were kindly provided by Dr. Christopher Bradfield (University of Wisconsin-Madison) and bred using female Ahr+/- and male Ahr-/- mice. Mice were housed on corncob bedding in a temperature- and light-controlled facility and given access to food and water ad libitum. Mice were also maintained in a pathogen-free facility and treated humanely with approval from the Animal Care and Use Committee of the Pennsylvania State University and methods were carried out in accordance with approved guidelines. Mice in the 8-10-week-old range were utilized in all experiments. Serum samples obtained from mice exposed to PCB126 for 5 days was previously described. Briefly, mice were fed dough pills once every 24 h, each pill contained 24 µg/kg PCB126 for five days and mice were sacrificed on day 6.

Combined analysis:

Analysis ID	AN004065
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	Shimadzu 20AD
Column	Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um)
MS Type	ESI
MS instrument type	Triple TOF
MS instrument name	ABI Sciex 5600 TripleTOF
Ion Mode	POSITIVE
Units	uM

Chromatography:

Chromatography ID:	CH003011
Instrument Name:	Shimadzu 20AD
Column Name:	Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um)
Column Temperature:	55
Flow Gradient:	The initial condition were 97% A and 3 % B, increasing to 45% B at 10 min, 75% B at 12 min where it was held at 75% B until 17.5 min before returning to the initial conditions.
Flow Rate:	0.25 mL/min
Solvent A:	100% water; 0.1% formic acid
Solvent B:	100% acetonitrile; 0.1% formic acid
Chromatography Type:	Reversed phase

MS:

MS ID:	MS003812
Analysis ID:	AN004065
Instrument Name:	ABI Sciex 5600 TripleTOF
Instrument Type:	Triple TOF
MS Type:	ESI
MS Comments:	Peakview was used for data analysis.
Ion Mode:	POSITIVE