Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA331613

Local Sample ID:	WT_HC_9
Subject ID:	SU003169
Subject Type:	Mammal
Subject Species:	Mus musculus
Gender:	Male and female

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Subject:

Subject ID:	SU003169
Subject Type:	Mammal
Subject Species:	Mus musculus
Gender:	Male and female

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
WT_HC_9	SA331613	FL039213	WT	Group

Collection:

Collection ID:	CO003162
Collection Summary:	Colons were harvested from euthanized mice and flash-frozen in liquid nitrogen. For all downstream analyses, the distal region of the colon (0.5-2 cm from rectum) was used, since it is where colitis is most pronounced in these models.
Sample Type:	Colon
Storage Conditions:	-80℃

Treatment:

Treatment ID:	TR003178
Treatment Summary:	Acute colitis was induced in wild type (WT) C57BL/6 mice via addition of 1.25% DSS into the drinking water for seven days. To gain insight on chronic intestinal inflammation, we used C57BL/6 IL-10-/- mice that spontaneously develop chronic colon inflammation. This inflammation is not chemically induced, but rather is considered to be a result of a dysregulated immune system, which more resembles the clinical manifestation of chronic UC and CD patients and mimics a monogenic form of IBD20. In order to synchronize and accelerate the onset of chronic colitis, mice were exposed to 200 ppm piroxicam (Sigma-Aldrich ltd.) via supplementation in normal murine chow for 14 days. With the described experimental procedures, we obtained five groups of mice (10 mice per group) for further analysis: 1) Healthy WT; 2) Day 4 of DSS (pre-symptomatic); 3) Day 10 of DSS (acute colitis); 4) Healthy IL-10-/- (pre-symptomatic); and 5) Ill IL-10-/- (chronic colitis).

Treatment ID:

TR003178

Treatment Summary:

Acute colitis was induced in wild type (WT) C57BL/6 mice via addition of 1.25% DSS into the drinking water for seven days. To gain insight on chronic intestinal inflammation, we used C57BL/6 IL-10-/- mice that spontaneously develop chronic colon inflammation. This inflammation is not chemically induced, but rather is considered to be a result of a dysregulated immune system, which more resembles the clinical manifestation of chronic UC and CD patients and mimics a monogenic form of IBD20. In order to synchronize and accelerate the onset of chronic colitis, mice were exposed to 200 ppm piroxicam (Sigma-Aldrich ltd.) via supplementation in normal murine chow for 14 days. With the described experimental procedures, we obtained five groups of mice (10 mice per group) for further analysis: 1) Healthy WT; 2) Day 4 of DSS (pre-symptomatic); 3) Day 10 of DSS (acute colitis); 4) Healthy IL-10-/- (pre-symptomatic); and 5) Ill IL-10-/- (chronic colitis).

Sample Preparation:

Sampleprep ID:	SP003175
Sampleprep Summary:	Frozen colon samples were trimmed (15–20 mg) to fit HR-MAS ZrO2 rotor insert capacity (50 μL). Each insert was filled with 2H2O containing 5.8 mM sodium trimethylsilyl [2,2,3,3-2H4]propionate (TMSP). Inserts were covered with plug and plug-restraining screw and inserted into the 4 mm rotor for HR-MAS probe.

Analysis:

MB Sample ID:	SA331613
Analysis ID:	AN005008
Analysis Type:	NMR
Num Factors:	5
Num Metabolites:	22
Units:	AU

NMR:

NMR ID:	NM000274
Analysis ID:	AN005008
Instrument Name:	Bruker 600 MHz
Instrument Type:	FT-NMR
NMR Experiment Type:	1D-1H
Spectrometer Frequency:	600 MHz
NMR Probe:	HR-MAS TXI 1H/13C/15N probe
NMR Solvent:	H2O
NMR Tube Size:	4 mm rotor for HR-MAS probe
Pulse Sequence:	CPMG
Water Suppression:	water peak suppression-presaturation
Temperature:	277 K
Number Of Scans:	64
Dummy Scans:	4
Acquisition Time:	1.36s
Spectral Width:	12 KHz
Num Data Points Acquired:	32 k
Line Broadening:	1
Baseline Correction Method:	automatic
Chemical Shift Ref Std:	doublet at 5.24 ppm
Binned Increment:	0.02 ppm
Binned Data Excluded Range:	4.5-5.5 ppm