Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA174766

Local Sample ID:	ST4
Subject ID:	SU001949
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	C57BL/6 mice (Wild-type)
Age Or Age Range:	six weeks old
Gender:	Female

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Subject:

Subject ID:	SU001949
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	C57BL/6 mice (Wild-type)
Age Or Age Range:	six weeks old
Gender:	Female

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
ST4	SA174766	FL021460	TCDD	Experimental factor

Collection:

Collection ID:	CO001942
Collection Summary:	Six-week-old female wildtype (WT) C57BL/6 mice acquired from Jackson Laboratories (Bar Harbor, ME) were housed in an AAALAC-accredited specific-pathogen-free animal facility located at the grounds of the University of South Carolina School of Medicine for the entirety of all experiments. Mice within the facility were housed within polycarbonate cages containing cellulose fiber chips as bedding in a temperature and humidity-controlled environment. After a 2-week acclimatization period, the mice were divided randomly into two groups that would be administered a single 100µl intraperitoneal injection containing sterile corn oil (VEH group) or an intraperitoneal injection of 10µg/kg TCDD suspended within sterile corn oil (TCDD group). At the 72h time point following TCDD or VEH exposure, the mice were humanely euthanized by an overdose of inhaled isoflurane. Cecal content and blood sera were collected at the time of necropsy.
Sample Type:	Blood (serum)

Treatment:

Treatment ID:	TR001961
Treatment Summary:	Six-week-old female wildtype (WT) C57BL/6 mice acquired from Jackson Laboratories (Bar Harbor, ME) were housed in an AAALAC-accredited specific-pathogen-free animal facility located at the grounds of the University of South Carolina School of Medicine for the entirety of all experiments. Mice within the facility were housed within polycarbonate cages containing cellulose fiber chips as bedding in a temperature and humidity-controlled environment. After a 2-week acclimatization period, the mice were divided randomly into two groups that would be administered a single 100µl intraperitoneal injection containing sterile corn oil (VEH group) or an intraperitoneal injection of 10µg/kg TCDD suspended within sterile corn oil (TCDD group). At the 72h time point following TCDD or VEH exposure, the mice were humanely euthanized by an overdose of inhaled isoflurane. Cecal content and blood sera were collected at the time of necropsy.

Treatment ID:

TR001961

Treatment Summary:

Six-week-old female wildtype (WT) C57BL/6 mice acquired from Jackson Laboratories (Bar Harbor, ME) were housed in an AAALAC-accredited specific-pathogen-free animal facility located at the grounds of the University of South Carolina School of Medicine for the entirety of all experiments. Mice within the facility were housed within polycarbonate cages containing cellulose fiber chips as bedding in a temperature and humidity-controlled environment. After a 2-week acclimatization period, the mice were divided randomly into two groups that would be administered a single 100µl intraperitoneal injection containing sterile corn oil (VEH group) or an intraperitoneal injection of 10µg/kg TCDD suspended within sterile corn oil (TCDD group). At the 72h time point following TCDD or VEH exposure, the mice were humanely euthanized by an overdose of inhaled isoflurane. Cecal content and blood sera were collected at the time of necropsy.

Sample Preparation:

Sampleprep ID:	SP001955
Sampleprep Summary:	Cecal content and serum samples were processed according to previously described methods for the metabolomic profiling using liquid chromatography-mass spectrometry (LC-MS) with slight modifications. For cecal content, ~25 mg was aliquoted into 1.5-mL Eppendorf tubes (Hamburg, Germany) containing ~20 mg acid washed glass beads (Sigma-Aldrich, St. Louis, MO), extracted into ice-cold MeOH:water (1:1, v/v) on a Qiagen TissueLyzer at 50 Hz for 10 min (Hilden, Germany), and centrifuged at 12,000 rpm for 10 min. For blood sera, 20 µL aliquots were extracted by adding 180 µL cold MeOH, briefly vortexed, and incubated at -20 ºC for 30 min for protein precipitation. The supernatants of both matrices were dried in a CentriVap vacuum concentrator (Labconco, MO) and resuspended in ACN:water (2:98, v/v) upon analysis.

Sampleprep ID:

SP001955

Sampleprep Summary:

Cecal content and serum samples were processed according to previously described methods for the metabolomic profiling using liquid chromatography-mass spectrometry (LC-MS) with slight modifications. For cecal content, ~25 mg was aliquoted into 1.5-mL Eppendorf tubes (Hamburg, Germany) containing ~20 mg acid washed glass beads (Sigma-Aldrich, St. Louis, MO), extracted into ice-cold MeOH:water (1:1, v/v) on a Qiagen TissueLyzer at 50 Hz for 10 min (Hilden, Germany), and centrifuged at 12,000 rpm for 10 min. For blood sera, 20 µL aliquots were extracted by adding 180 µL cold MeOH, briefly vortexed, and incubated at -20 ºC for 30 min for protein precipitation. The supernatants of both matrices were dried in a CentriVap vacuum concentrator (Labconco, MO) and resuspended in ACN:water (2:98, v/v) upon analysis.

Combined analysis:

Analysis ID	AN003034
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	Thermo Vanquish
Column	Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um)
MS Type	ESI
MS instrument type	Orbitrap
MS instrument name	Thermo Q Exactive Orbitrap
Ion Mode	POSITIVE
Units	m/z

Chromatography:

Chromatography ID:	CH002247
Instrument Name:	Thermo Vanquish
Column Name:	Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um)
Chromatography Type:	Reversed phase

MS:

MS ID:	MS002821
Analysis ID:	AN003034
Instrument Name:	Thermo Q Exactive Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	fullscan *.raw data acquired by Thermo XCalibur software
Ion Mode:	POSITIVE