Summary of Study ST000222

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 PR000182. The data can be accessed directly via it's Project DOI: 10.21228/M8M888 This work is supported by NIH grant, U2C- DK119886.

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

Study ID	ST000222
Study Title	Bile acid targeted metabolomics of the small intestine in malnourished and control mice
Study Type	Targeted metabolomics
Study Summary	A total of 8 samples from 6 week old, female C57BL/6 mice, treated for 3 weeks with a malnourished diet or a control-fed isocaloric diet. Samples were taken from the small intestinal fecal content at the terminus of the ileum for targeted bile acid analysis.
Institute	University of Victoria
Department	The Uvic Proteomics and Metabolomics Innovation Centre
Last Name	Borchers
First Name	Christoph
Address	#3101-4464 Markham St Victoria, BC Canada, V8Z 7X8
Email	christoph@proteincentre.com
Submit Date	2015-06-08
Num Groups	2
Total Subjects	8*
Study Comments	*One sample failed quality control in the malnourished group, thus was not included in the study
Raw Data Available	Yes
Raw Data File Type(s)	wiff
Uploaded File Size	4.0 M
Analysis Type Detail	LC-MS
Release Date	2015-06-08
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP000243
Sampleprep Summary:	Bile-acid targeted metabolomics. Each sample was homogenized in LC-MS grade water at a ratio of 150 ?L per 10 mg raw material and with the aid of 5-mm stainless steel metal balls. Bile acids were extracted by addition of acetonitrile at a ratio of 350 ?L per 10 mg raw material followed by vortexing and sonication (1 min) in an ice-water ultrasonic bath. The samples were centrifuged. 20 ?L of the supernatants were precisely taken out and mixed with a predefined mix of 14 deuterium-labeled bile acids as the internal standards. The mixtures were subjected to phospholipid-depletion solid-phase extraction according to a validated protocol for sample cleanup and bile acid enrichment62. The flow-through fractions were collected and then dried under a gentle nitrogen flow. The dried residues were dissolved in 200 ?L of 50% methonal. 10 ?L were injected for quantitation by UPLC-MRM/MS. A Dionex UPLC system was connected to an AB Sciex 4000 QTRAP mass spectrometer which was operated in the negative ion multiple-reaction monitoring (MRM) mode and with electrospray ionization. UPLC separation was carried out on a 15 cm long C-18 UPLC column with water-acetonitrile-formic acid as the mobile phase for binary gradient elution using a developed and validated protocol for comprehensive analysis of bile acids in biological samples (Han, etc. manuscript submitted to Analytical Chemistry). The column temperature was 45 oC and the flow rate was 0.35 mL/min. 45 bile acids (including the 19 targeted bile acids) were involved in the quantitation by UPLC/scheduled MRM/MS. Concentrations of the detected bile acids were calculated with internal standard calibration from the linearly regressed standard calibration curves of individual bile acids. The lower limits of quantitation were 0.08 nmoles/mg for all the bile acids.

Sampleprep ID:

SP000243

Sampleprep Summary:

Bile-acid targeted metabolomics. Each sample was homogenized in LC-MS grade water at a ratio of 150 ?L per 10 mg raw material and with the aid of 5-mm stainless steel metal balls. Bile acids were extracted by addition of acetonitrile at a ratio of 350 ?L per 10 mg raw material followed by vortexing and sonication (1 min) in an ice-water ultrasonic bath. The samples were centrifuged. 20 ?L of the supernatants were precisely taken out and mixed with a predefined mix of 14 deuterium-labeled bile acids as the internal standards. The mixtures were subjected to phospholipid-depletion solid-phase extraction according to a validated protocol for sample cleanup and bile acid enrichment62. The flow-through fractions were collected and then dried under a gentle nitrogen flow. The dried residues were dissolved in 200 ?L of 50% methonal. 10 ?L were injected for quantitation by UPLC-MRM/MS. A Dionex UPLC system was connected to an AB Sciex 4000 QTRAP mass spectrometer which was operated in the negative ion multiple-reaction monitoring (MRM) mode and with electrospray ionization. UPLC separation was carried out on a 15 cm long C-18 UPLC column with water-acetonitrile-formic acid as the mobile phase for binary gradient elution using a developed and validated protocol for comprehensive analysis of bile acids in biological samples (Han, etc. manuscript submitted to Analytical Chemistry). The column temperature was 45 oC and the flow rate was 0.35 mL/min. 45 bile acids (including the 19 targeted bile acids) were involved in the quantitation by UPLC/scheduled MRM/MS. Concentrations of the detected bile acids were calculated with internal standard calibration from the linearly regressed standard calibration curves of individual bile acids. The lower limits of quantitation were 0.08 nmoles/mg for all the bile acids.