Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA339129

Local Sample ID:	QC15
Subject ID:	SU003247
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

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Subject:

Subject ID:	SU003247
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
QC15	SA339129	FL040156	MBlood	Sample source
QC15	SA339129	FL040156	-	exposure
QC15	SA339129	FL040156	-	timepoint

Collection:

Collection ID:	CO003240
Collection Summary:	At 2, 5, 12 and 24h, dams were terminally anesthetized by subcutaneous injection of 0.2 ml of Zoletil mixture (tiletamin/zolazepam, xylazin og fentanyl) and killed by exanguination by withdrawal of heart blood into Eppendorf tubes containing 36 ml K2EDTA (N=7-9 per exposure/time point). The uterus was excised and opened. Fetuses were excised from their embryonic sac, their viability confirmed, killed by decapitation, sexed by visual inspection, and their position in the uterus noted. From each litter, the first female fetus encountered in the right uterine horn, counting from the cervix, was selected and saved for analyses. The placenta was dissected into chorion (chorionic plate, labyrinth and junctional zones) and decidua by blunt/stump dissection under stereomicroscope (Wild Heerbrugg, Switzerland)76. From dams, the liver and right lung were dissected. Dissected organs were snap frozen in liquid nitrogen and kept at -80ºC
Sample Type:	Blood (plasma)

Treatment:

Treatment ID:	TR003256
Treatment Summary:	Lipopolysaccharide (LPS; E. Coli serotype 00:55 B5 LPS (Sigma Lot nr. 025M4040V)) was diluted to the final concentration (0.02 µg/µl) in double distilled pyrogen-free water (Chem-Lab, Zedelgem, Belgium). In the morning of GD 17, the pregnant mice were semi-randomized into control and LPS treatment groups (denoted Ctrl and LPS, respectively), evenly distributing weights among the groups. Out of 80 mice in total, 74 were pregnant. Animals were placed in a whole-body inhalation chamber with an attached anaesthetic vaporizer (Penlon Sigma Delta, Abingdon, UK), delivering 3-4% isoflurane in filtered air, and were intratracheally instilled with 50 µl of vehicle (Ctrl) or 1 μg LPS in 50 µl vehicle, followed by 200μl of air. Vehicle and LPS were administered through a 0.58 mm polyethylene tube (Ref: 427411, Becton Dickinson, Brøndby, Denmark) attached to a plastic syringe. The procedure has been shown not to affect gestation, offspring viability nor growth75. After instillation, animals were returned to their cage, briefly placed on heating pads and checked upon regularly until euthanization

Treatment ID:

TR003256

Treatment Summary:

Lipopolysaccharide (LPS; E. Coli serotype 00:55 B5 LPS (Sigma Lot nr. 025M4040V)) was diluted to the final concentration (0.02 µg/µl) in double distilled pyrogen-free water (Chem-Lab, Zedelgem, Belgium). In the morning of GD 17, the pregnant mice were semi-randomized into control and LPS treatment groups (denoted Ctrl and LPS, respectively), evenly distributing weights among the groups. Out of 80 mice in total, 74 were pregnant. Animals were placed in a whole-body inhalation chamber with an attached anaesthetic vaporizer (Penlon Sigma Delta, Abingdon, UK), delivering 3-4% isoflurane in filtered air, and were intratracheally instilled with 50 µl of vehicle (Ctrl) or 1 μg LPS in 50 µl vehicle, followed by 200μl of air. Vehicle and LPS were administered through a 0.58 mm polyethylene tube (Ref: 427411, Becton Dickinson, Brøndby, Denmark) attached to a plastic syringe. The procedure has been shown not to affect gestation, offspring viability nor growth75. After instillation, animals were returned to their cage, briefly placed on heating pads and checked upon regularly until euthanization

Sample Preparation:

Sampleprep ID:	SP003254
Sampleprep Summary:	plash mix (Merck) was added to the extraction solvent and tissue samples. After centrifugation and phase separation, the apolar and polar phases were transferred to separate tubes, and the apolar phase dried under N2. *Folch, J., Lees, M. & Sloane Stanley, G. H. A simple method for the isolation and purification of total lipides from animal tissues. J. Biol. Chem. 226, 497–509 (1957).

Combined analysis:

Analysis ID	AN005133	AN005134
Analysis type	MS	MS
Chromatography type	Reversed phase	Reversed phase
Chromatography system	Thermo Vanquish	Thermo Vanquish
Column	Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um)	Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um)
MS Type	ESI	ESI
MS instrument type	QTOF	QTOF
MS instrument name	Bruker Tims TOF flex	Bruker Tims TOF flex
Ion Mode	POSITIVE	NEGATIVE
Units	Peak intensity	Peak intensity

Chromatography:

Chromatography ID:	CH003884
Chromatography Summary:	Reverse phase (C18), 10 min
Instrument Name:	Thermo Vanquish
Column Name:	Waters ACQUITY UPLC CSH C18 (100 x 2.1mm,1.7um)
Column Temperature:	55
Flow Gradient:	from 0 to 0.5 min, 40–43% B; from 0.5 to 0.7 min, 43‐65% B; from 0.7 to 0.8 min, 65-70% B; from 0.8 to 2.3 min, 70-99% B; from 2.3 to 6 min, 99% B; from 6-6.8 min, 99-40% B; from 6.8-7 min before equilibration for 3 min with the initial conditions
Flow Rate:	0.4 ml/min
Solvent A:	Acetonitrile/water (60:40); 10 mM ammonium formate; 0.1% formic acid
Solvent B:	Isopropanol/acetonitrile (90:10); 10 mM ammonium formate; 0.1% formic acid
Chromatography Type:	Reversed phase

MS:

MS ID:	MS004869
Analysis ID:	AN005133
Instrument Name:	Bruker Tims TOF flex
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	Data was acquired with Trapped ion mobility spectrometry (TIMS) activated. Bruker Metaboscape software was used for data processing. Annotatation was done using the built in "lipid search" module and Lipid blast.
Ion Mode:	POSITIVE

MS ID:	MS004870
Analysis ID:	AN005134
Instrument Name:	Bruker Tims TOF flex
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	Data was acquired with Trapped ion mobility spectrometry (TIMS) activated. Bruker Metaboscape software was used for data processing. Annotatation was done using the built in "lipid search" module and Lipid blast.
Ion Mode:	NEGATIVE