Summary of Study ST001207

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000811. The data can be accessed directly via it's Project DOI: 10.21228/M8VD62 This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

Study ID	ST001207
Study Title	Lipidomics in the serum of cold exposed mice treated with 12-LOX inhibitor LOXBlock-1
Study Summary	We aimed to investigate whether the cold-induced release of 12-LOX products into the circulation were dependent on 12-LOX activation. We pre-treated C57BL6/J mice with the pharmacological inhibitor LOXBlock-1 or its vehicle (DMSO), and after 15 minutes we placed them under cold temperature (5C) for 4 hours. A control group was injected with DMSO and kept at room temperature for the same 4 hours. After this period of time we, collected the blood, and obtained the serum fraction that was immediately frozen and submitted for untargeted lipidomics.
Institute	Joslin Diabetes Center
Last Name	Leiria
First Name	Luiz
Address	One Joslin Place, Boston-MA, 02215
Email	luiz.leiria@joslin.harvard.edu
Phone	617-309-1967
Submit Date	2019-06-26
Num Groups	3
Study Comments	Joslin Diabetes Center affiliate of Harvard Medical School
Raw Data Available	Yes
Raw Data File Type(s)	wiff
Analysis Type Detail	LC-MS
Release Date	2019-07-17
Release Version	1

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Project:

Project ID:	PR000811
Project DOI:	doi: 10.21228/M8VD62
Project Title:	12-LOX metabolites in cold adaptation
Project Summary:	The goal of this project is to understand the role of the enzyme 12-lipoxygenase in the adaptive thermogenesis. We found this enzyme is activated by cold stimulation, then producing lipid metabolites in adipose tissue and releasing them into the circulation to regulate fuel utilisation and thermogenic pathways required for the cold adaptation.
Institute:	Joslin Diabetes Center
Department:	Integrative Physiology and Metabolism
Laboratory:	Yu-Hua Tseng lab
Last Name:	Leiria
First Name:	Luiz
Address:	One Joslin Place, Boston, MA-USA, 02215
Email:	luiz.leiri@joslin.harvard.edu
Phone:	1 6173091967

Subject:

Subject ID:	SU001274
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	C57BL6/J
Age Or Age Range:	12 weeks old
Weight Or Weight Range:	25-30 g
Gender:	Male
Animal Animal Supplier:	Jax Laboratories
Animal Housing:	Conventional
Animal Light Cycle:	regular 12h light /dark cycles
Animal Feed:	ad libitum normal chow diet
Animal Water:	ad libitum

Factors:

Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id	local_sample_id	Temperature	Treatment
SA084803	T5	22C	DMSO
SA084804	T6	22C	DMSO
SA084805	T1	22C	DMSO
SA084806	T2	22C	DMSO
SA084807	T3	22C	DMSO
SA084808	T7	22C	DMSO
SA084809	T4	22C	DMSO
SA084810	C4	5C	DMSO
SA084811	C3	5C	DMSO
SA084812	C5	5C	DMSO
SA084813	C6	5C	DMSO
SA084814	C1	5C	DMSO
SA084815	C7	5C	DMSO
SA084816	C2	5C	DMSO
SA084817	LB3	5C	LoxBlock-1
SA084818	LB1	5C	LoxBlock-1
SA084819	LB2	5C	LoxBlock-1
SA084820	LB4	5C	LoxBlock-1
SA084821	LB5	5C	LoxBlock-1

Showing results 1 to 19 of 19

Showing results 1 to 19 of 19

Collection:

Collection ID:	CO001268
Collection Summary:	Mice were anaesthetised with Avertine and blood was collected through cardiac puncture. Samples were let in room temperature for 30 minutes, and then centrifuged for 3 minutes at 14,000RPM. Serum fas collected and frozen at -20C for further lipidomics analysis.
Sample Type:	Blood (serum)
Storage Conditions:	-80℃

Treatment:

Treatment ID:	TR001289
Treatment Summary:	Mice were injected i.p. 15 minutes before the cold exposure, with 50ul of DMSO (vehicle) or LOXBlock-1. Then, the animas were placed in 5C or 22C temperature for 4 hours, until the samples were collected.

Sample Preparation:

Sampleprep ID:	SP001282
Sampleprep Summary:	Aliquots of 100 µL serum were taken and a mixture of deuterium-labeled internal standards was added to each aliquot, followed by 3x volume of sample of cold methanol (MeOH). Samples were vortexed for 5 minutes and stored at −20 °C overnight. Cold samples were centrifuged at 14,000g for 10 minutes, and the supernatant was then transferred to a new tube and 3 mL of acidified H2O (pH 3.5) was added to each sample prior to C18 SPE. The methyl formate fractions were collected, dried under nitrogen, and reconstituted in 50 µL MeOH:H2O (1:1, by vol). Samples were transferred to 0.5 mL tubes and centrifuged at 20,000g at 4 °C for 10 minutes. (35ul) of supernatant was transferred to LC–MS/MS vials for analysis using the BERG LC–MS/MS mediator lipidomics platform.

Sampleprep ID:

SP001282

Sampleprep Summary:

Aliquots of 100 µL serum were taken and a mixture of deuterium-labeled internal standards was added to each aliquot, followed by 3x volume of sample of cold methanol (MeOH). Samples were vortexed for 5 minutes and stored at −20 °C overnight. Cold samples were centrifuged at 14,000g for 10 minutes, and the supernatant was then transferred to a new tube and 3 mL of acidified H2O (pH 3.5) was added to each sample prior to C18 SPE. The methyl formate fractions were collected, dried under nitrogen, and reconstituted in 50 µL MeOH:H2O (1:1, by vol). Samples were transferred to 0.5 mL tubes and centrifuged at 20,000g at 4 °C for 10 minutes. (35ul) of supernatant was transferred to LC–MS/MS vials for analysis using the BERG LC–MS/MS mediator lipidomics platform.

Combined analysis:

Analysis ID	AN002009
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	Ekspert MicroLC 200 system
Column	Phenomenex Synergi Fusion-RP capillary C18 (150 × 0.5 mm,4um)
MS Type	ESI
MS instrument type	QTOF
MS instrument name	ABI Sciex 5600+ TripleTOF
Ion Mode	NEGATIVE
Units	Peak Area

Chromatography:

Chromatography ID:	CH001453
Instrument Name:	Ekspert MicroLC 200 system
Column Name:	Phenomenex Synergi Fusion-RP capillary C18 (150 × 0.5 mm,4um)
Chromatography Type:	Reversed phase

MS:

MS ID:	MS001862
Analysis ID:	AN002009
Instrument Name:	ABI Sciex 5600+ TripleTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	MS spectra were acquired in high-resolution mode (>30,000) using a 100-ms accumulation time per spectrum. Full-scan MS/MS was acquired in high sensitivity mode, with an accumulation time optimized per cycle. Collision energy was set using rolling collision energy with a spread of 15V. The identity of a component was confirmed using PeakView® software (SCIEX), and quantification was performed using MultiQuant™ software (SCIEX). C18SPE cartridges were purchased from Biotage. All solvents were of HPLC or LC-MS/MS grade and were acquired from Sigma-Aldrich, Fisher Scientific, or VWR International.
Ion Mode:	NEGATIVE