Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA247794

Local Sample ID:	36-week db/db+atRA-1
Subject ID:	SU002564
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	db/m; db/db; db/db+retinol; db/db+retinoic acid; db/db+AAV9-RDH10; RDH10-cKO; RDH10fl/fl
Age Or Age Range:	32-week; 36-week; 21-week
Gender:	Male

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Sample Preparation:

Sampleprep ID:	SP002570
Sampleprep Summary:	Sample preparation Homogenized the heart tissue in 200μL of cold NaCl solution (0.9%) for 5 seconds, add 100μL retinol-D4 (IR-23012, IsoSciences, Ambler, PA, USA) as internal standard for Rol and atRA, and retinyl acetate (46958, Sigma-Aldrich, St. Louis, MO) as internal standard for retinyl esters, then homogenize for another 5 seconds. For Rol and at RA, 1mL formic acid n-hexane solution (1%) was added to the homogenizing to facilitate phase separation by a 5-minutes vortex and 5-min centrifugation (16,200 g). The supernatants were collected in a new tube and blown dry with nitrogen followed by re-dilution with 100μL 70% methanol. For retinyl esters, 24 μL 0.5 M NaOH solution was added to tissue homogenate, followed by 1 mL n-hexane. The mixture was votexed 5 min and centrifuged at 16,200 g for 5 min. The supernatants were collected in a new tube and blown dry with nitrogen followed by re-dilution with 100μL 100% methanol. The processes above should be protected from the light.

Sampleprep ID:

SP002570

Sampleprep Summary:

Sample preparation Homogenized the heart tissue in 200μL of cold NaCl solution (0.9%) for 5 seconds, add 100μL retinol-D4 (IR-23012, IsoSciences, Ambler, PA, USA) as internal standard for Rol and atRA, and retinyl acetate (46958, Sigma-Aldrich, St. Louis, MO) as internal standard for retinyl esters, then homogenize for another 5 seconds. For Rol and at RA, 1mL formic acid n-hexane solution (1%) was added to the homogenizing to facilitate phase separation by a 5-minutes vortex and 5-min centrifugation (16,200 g). The supernatants were collected in a new tube and blown dry with nitrogen followed by re-dilution with 100μL 70% methanol. For retinyl esters, 24 μL 0.5 M NaOH solution was added to tissue homogenate, followed by 1 mL n-hexane. The mixture was votexed 5 min and centrifuged at 16,200 g for 5 min. The supernatants were collected in a new tube and blown dry with nitrogen followed by re-dilution with 100μL 100% methanol. The processes above should be protected from the light.