Summary of Study ST000121

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 PR000108. The data can be accessed directly via it's Project DOI: 10.21228/M8GW2Z This work is supported by NIH grant, U2C- DK119886.

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

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Study IDST000121
Study TitleImpact of anesthesia and euthanasia on metabolomics of mammalian tissues: studies in a C57BL/6J mouse model
Study TypeAnesthesia effect study
Study SummaryWe examined the effect of several commonly-used methods of anesthesia and euthanasia for collection of skeletal muscle, liver, heart, adipose and serum of C57BL/6J mice. The data revealed tissue-specific impacts of different anesthesia and euthanasia strategies. Based on these findings, we present a more optimal collection strategy mammalian tissues and recommend that rodent tissues intended for metabolomics studies be collected under anesthesia rather than post-euthanasia.
Institute
University of Michigan
DepartmentInternal Medicine
LaboratoryEvans Lab / Burant Lab
Last NameEvans
First NameCharles
Address6321 Brehm Tower, 1000 Wall Street, Ann Arbor MI 48105
Emailchevans@umich.edu
Phone734-232-8177
Submit Date2014-11-21
Num Groups6
Total Subjects49
Raw Data AvailableNo
Raw Data File Type(s)d
Analysis Type DetailLC-MS
Release Date2015-02-01
Release Version1
Charles Evans Charles Evans
https://dx.doi.org/10.21228/M8GW2Z
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000108
Project DOI:doi: 10.21228/M8GW2Z
Project Title:Impact of anesthesia and euthanasia on metabolomics of mammalian tissues: studies in a C57BL/6J mouse model
Project Type:Anesthesia effect study
Project Summary:We examined, for the first time using untargeted and targeted metabolomics, the effect of several commonly-used methods of anesthesia and euthanasia for collection of skeletal muscle, liver, heart, adipose and serum of C57BL/6J mice. The data revealed tissue-specific impacts of different anesthesia and euthanasia strategies. Based on these findings, we present a more optimal collection strategy mammalian tissues and recommend that rodent tissues intended for metabolomics studies be collected under anesthesia rather than post-euthanasia.
Institute:University of Michigan
Department:Internal Medicine
Laboratory:Evans lab / Burant lab
Last Name:Evans
First Name:Charles
Address:6321 Brehm Tower, 1000 Wall St.
Email:chevans@umich.edu
Phone:734-232-8177
Funding Source:National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of Health

Subject:

Subject ID:SU000140
Subject Type:Animal
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:C57BL/6J
Age Or Age Range:20 weeks
Weight Or Weight Range:26.8 +/- 2.2 g
Gender:Male
Animal Animal Supplier:Jackson Labs (Bar Harbor, ME)
Animal Housing:8 / cage
Animal Light Cycle:12 h / 12 h light/dark
Animal Feed:Standard chow, ad lib.
Animal Water:Ad lib. Access
Species Group:Mammal

Factors:

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

mb_sample_id local_sample_id Tissue
SA006504F14Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006505F16Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006506F17Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006507F13Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006508F15Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006509F12Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006510F09Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006511F10Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006512F11Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006513F23Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Cervical Dislocation
SA006514F24Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Cervical Dislocation
SA006515F22Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Cervical Dislocation
SA006516F18Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Cervical Dislocation
SA006517F25Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Cervical Dislocation
SA006518F19Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Cervical Dislocation
SA006519F20Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Cervical Dislocation
SA006520F21Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Cervical Dislocation
SA006521F40Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Continuous isoflurane
SA006522F35Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Continuous isoflurane
SA006523F37Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Continuous isoflurane
SA006524F38Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Continuous isoflurane
SA006525F41Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Continuous isoflurane
SA006526F39Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Continuous isoflurane
SA006527F34Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Continuous isoflurane
SA006528F36Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Continuous isoflurane
SA006529F05Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Isoflurane overdose
SA006530F06Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Isoflurane overdose
SA006531F07Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Isoflurane overdose
SA006532F04Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Isoflurane overdose
SA006533F08Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Isoflurane overdose
SA006534F03Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Isoflurane overdose
SA006535F01Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Isoflurane overdose
SA006536F02Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Isoflurane overdose
SA006537F28Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Ketamine
SA006538F26Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Ketamine
SA006539F29Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Ketamine
SA006540F27Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Ketamine
SA006541F30Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Ketamine
SA006542F33Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Ketamine
SA006543F32Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Ketamine
SA006544F31Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Ketamine
SA006545F46Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Pentobarbital
SA006546F47Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Pentobarbital
SA006547F49Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Pentobarbital
SA006548F45Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Pentobarbital
SA006549F43Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Pentobarbital
SA006550F42Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Pentobarbital
SA006551F48Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Pentobarbital
SA006552F44Epididymal white adipose (fat) | Anesthesia / Euthanasia method:Pentobarbital
SA006553G17Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006554G13Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006555G12Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006556G11Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006557G09Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006558G16Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006559G15Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006560G14Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006561G10Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Carbon Dioxide
SA006562G18Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Cervical Dislocation
SA006563G23Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Cervical Dislocation
SA006564G24Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Cervical Dislocation
SA006565G22Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Cervical Dislocation
SA006566G21Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Cervical Dislocation
SA006567G19Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Cervical Dislocation
SA006568G20Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Cervical Dislocation
SA006569G25Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Cervical Dislocation
SA006570G34Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Continuous isoflurane
SA006571G39Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Continuous isoflurane
SA006572G40Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Continuous isoflurane
SA006573G38Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Continuous isoflurane
SA006574G37Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Continuous isoflurane
SA006575G35Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Continuous isoflurane
SA006576G36Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Continuous isoflurane
SA006577G41Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Continuous isoflurane
SA006578G08Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Isoflurane overdose
SA006579G02Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Isoflurane overdose
SA006580G01Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Isoflurane overdose
SA006581G03Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Isoflurane overdose
SA006582G04Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Isoflurane overdose
SA006583G06Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Isoflurane overdose
SA006584G07Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Isoflurane overdose
SA006585G05Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Isoflurane overdose
SA006586G30Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Ketamine
SA006587G31Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Ketamine
SA006588G29Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Ketamine
SA006589G28Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Ketamine
SA006590G26Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Ketamine
SA006591G27Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Ketamine
SA006592G32Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Ketamine
SA006593G33Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Ketamine
SA006594G47Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Pentobarbital
SA006595G48Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Pentobarbital
SA006596G49Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Pentobarbital
SA006597G42Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Pentobarbital
SA006598G46Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Pentobarbital
SA006599G45Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Pentobarbital
SA006600G43Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Pentobarbital
SA006601G44Gastrocnemius (skeletal muscle) | Anesthesia / Euthanasia method:Pentobarbital
SA006602H16Heart | Anesthesia / Euthanasia method:Carbon Dioxide
SA006603H17Heart | Anesthesia / Euthanasia method:Carbon Dioxide
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Collection:

Collection ID:CO000124
Collection Summary:4 tissues plus blood serum were collected and immediately following euthanasia or anesthesia induction, frozen in liquid nitrogen and stored at -80C until analysis
Sample Type:Skeletal muscle (gastrocnemius), Liver, Heart, Epididymal white adipose (fat), blood serum
Collection Method:Surgical dissection
Collection Time:As soon as possible following anesthesia/euthanasia (all tissues collected within 3 minutes)
Storage Conditions:-80 C

Treatment:

Treatment ID:TR000142
Treatment Summary:Mice were prepared or tissue collection by one of 6 modes of anesthesia or euthanasia, then tissues were rapidly surgically collected and frozen
Animal Anesthesia:One of 6 methods: 1) Cervical dislocation, 2) Carbon dioxide inhalation, 3) Isoflurane overdose, 4) continuous isoflurane, 5) ketamine, 6) pentobarbital
Animal Fasting:5 hours
Animal Endp Euthanasia:All tissue collection were terminal studies; in methods 1-3, animals were euthanized prior to tissue collection; in methods 4-6, animals were under deep anesthesia while tissues were collected.

Sample Preparation:

Sampleprep ID:SP000137
Sampleprep Summary:Frozen tissue samples were pulverized in a liquid nitrogen-chilled mortar and pestle, then sonicated in cold extraction solvent (7:2:1 methanol:chloroform:water) at an approximate ratio of 30 mg tissue (wet mass) /1 mL of solvent. After allowing samples to rest on ice 5 minutes, they were centrifuged to pellet precipitated protein and cell debris; the supernatant was placed in autosampler vials for direct LC-MS analysis. For serum, extraction solvent (1:1:1 methanol:acetonitrile:acetone) was added to serum at a 4:1 solvent:serum ratio, vortexed to mix, then centrifuged and supernatant recovered for LC-MS analysis.
Sampleprep Protocol Filename:PROTOCOLS_mouse_anesthesia.pdf
Processing Method:Cryo-pulverization + sonication in extraction solvent

Combined analysis:

Analysis ID AN000203
Analysis type MS
Chromatography type HILIC
Chromatography system Agilent 1200
Column Phenomenex Luna NH2 (150 x 2.1mm,3um)
MS Type ESI
MS instrument type TOF
MS instrument name Agilent 6220 TOF
Ion Mode NEGATIVE
Units Peak area

Chromatography:

Chromatography ID:CH000135
Chromatography Summary:Polar metabolites were analyzed by HILIC-MS using an aminopropyl stationary phase
Instrument Name:Agilent 1200
Column Name:Phenomenex Luna NH2 (150 x 2.1mm,3um)
Column Pressure:150 bar
Column Temperature:25 C
Flow Gradient:80-0% B over 20 minutes, hold 4 minutes, return to 80%B
Flow Rate:0.25 mL/min
Sample Injection:25 uL
Solvent A:100% water; 5 mM ammonium acetate, pH 9.9
Solvent B:100% acetonitrile
Analytical Time:32 min
Chromatography Type:HILIC

MS:

MS ID:MS000166
Analysis ID:AN000203
Instrument Name:Agilent 6220 TOF
Instrument Type:TOF
MS Type:ESI
Ion Mode:NEGATIVE
Capillary Voltage:3500
Collision Gas:N2
Dry Gas Flow:10 L/min
Dry Gas Temp:350 C
Gas Pressure:20 psig
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