Summary of Study ST001361

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 PR000931. The data can be accessed directly via it's Project DOI: 10.21228/M8BT4S This work is supported by NIH grant, U2C- DK119886.

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

Study ID	ST001361
Study Title	Serum tryptophan metabolomics in CKD
Study Type	MS quantitative analysis
Study Summary	Serum was processed using a targeted metabolomics platform for quantifying tryptophan metabolites as a number of these metabolites are well establish uremic toxins.
Institute	University of Florida
Last Name	Ryan
First Name	Terence
Address	1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA
Email	ryant@ufl.edu
Phone	352-294-1700
Submit Date	2020-04-02
Num Groups	2
Total Subjects	16
Num Males	8
Num Females	8
Raw Data Available	Yes
Analysis Type Detail	LC-MS
Release Date	2020-12-31
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP001443
Sampleprep Summary:	Twenty-five microliters of each mouse serum was spiked with 5 µL internal standards (IS) solution consisted of tryptophan ¹³C₁₁, serotonin D4, kynurenine D4, kynurenic Acid D5, xanthurenic acid D4, anthranilic acid ¹³C₆, indoxyl sulfate ¹³C₆, p-cresol sulfate D7 and 3-indole-acetate D7. Metabolite extraction was done by protein precipitation using 200 µl of 8:1:1 Acetonitrile: Methanol: Acetone with 0.1% formic acid. Further protein precipitation was allowed by incubating the samples at 4°C for 20 min. Samples were placed in an ultrasonic bath for 10 min and then centrifuged at 20 000 xg for 5min at 4°C to pellet the protein. 190 µl supernatant was transferred from each sample into clean tube and dried under a gentle stream of nitrogen at 30°C. The dried extracts were re-suspended with 25 µL water with 0.1% formic acid. Resuspension was allowed at 4°C for 10 -15 min then samples were centrifuged at 20000 xg for 5 min at 4°C. Supernatants were transferred into clean LC-vials for targeted LC-MS quantitation on a Thermo Q-Exactive Oribtrap mass spectrometer with Dionex UHPLC and autosampler.

Sampleprep ID:

SP001443

Sampleprep Summary:

Twenty-five microliters of each mouse serum was spiked with 5 µL internal standards (IS) solution consisted of tryptophan ¹³C₁₁, serotonin D4, kynurenine D4, kynurenic Acid D5, xanthurenic acid D4, anthranilic acid ¹³C₆, indoxyl sulfate ¹³C₆, p-cresol sulfate D7 and 3-indole-acetate D7. Metabolite extraction was done by protein precipitation using 200 µl of 8:1:1 Acetonitrile: Methanol: Acetone with 0.1% formic acid. Further protein precipitation was allowed by incubating the samples at 4°C for 20 min. Samples were placed in an ultrasonic bath for 10 min and then centrifuged at 20 000 xg for 5min at 4°C to pellet the protein. 190 µl supernatant was transferred from each sample into clean tube and dried under a gentle stream of nitrogen at 30°C. The dried extracts were re-suspended with 25 µL water with 0.1% formic acid. Resuspension was allowed at 4°C for 10 -15 min then samples were centrifuged at 20000 xg for 5 min at 4°C. Supernatants were transferred into clean LC-vials for targeted LC-MS quantitation on a Thermo Q-Exactive Oribtrap mass spectrometer with Dionex UHPLC and autosampler.