Summary of Study ST002707

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 PR001678. The data can be accessed directly via it's Project DOI: 10.21228/M8T146 This work is supported by NIH grant, U2C- DK119886.

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

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

Study ID	ST002707
Study Title	Levels of T3 (triiodothyronine) and T4 (thyroxine) in CSF (cerebrospinal fluid) and plasma as part of natural diurnal variation
Study Summary	This study employs targeted LC-MS analysis of CSF and plasma to assess relative changes in the levels of thyroid hormone (T3: triiodothyronine and T4: thyroxine) at two time points in the diurnal cycle. For this purpose, wild-type CD1 mice were kept in a circadian cabinet housing with 12-hour light cycle (7 a.m. on/ 7 p.m. off) and tissues were collected consistently at an a.m and a p.m time point (9 a.m. and 9 p.m). CSF was collected from adult (3 months old). Internal standards (13C-labelled T3 and T4) as well as calibration curves were used to estimate the respective T3 and T4 concentration in the examined biofluids.
Institute	Boston Children's Hospital, Harvard Medical School
Laboratory	Kanarek Lab
Last Name	Petrova
First Name	Boryana
Address	Enders 1116.2 300 Longwood Ave
Email	Boryana.Petrova@childrens.harvard.edu
Phone	6179197352
Submit Date	2023-05-15
Num Groups	2
Total Subjects	27
Raw Data Available	Yes
Raw Data File Type(s)	raw(Thermo)
Analysis Type Detail	LC-MS
Release Date	2023-05-28
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP002818
Sampleprep Summary:	CSF was collected from adult (3 months old) wild-type CD1 mice. Samples were placed on wet ice, then spun 1000xg for 10 minutes at 4C. The supernatant was collected. Per condition, 5-10µL of fresh, cleared CSF or 5µl serum was extracted in 4:6:3 chlorophorm:methanol:water mixture supplemented with isotopically labeled T3 and T4 (Cambridge Isotope Laboratories, CLM-7185-C and CLM-8931-PK ) as well as isotopically labelled 17 amino acids and isotopically labelled reduced glutathione (Cambridge Isotope Laboratories, MSK-A2-1.2 and CNLM-6245-10). After centrifugation for 10 min at maximum speed on a benchtop centrifuge (Eppendorf) the top, hydrophilic layer (top phase) was transferred to a new tube, dried using a nitrogen dryer and reconstituted in 20 µl water (supplemented with QReSS, Cambridge Isotope Laboratories, MSK-QRESS-KIT) by brief vortexing. Extracted metabolites were spun again and cleared supernatant was transferred to LC-MS micro vials. A small amount of each sample was pooled and serially diluted 3- and 10-fold to be used as quality controls throughout the run of each batch. I calibration curve of unlabeled T3 and T4 standards was run per experiment for concentration calculations.

Sampleprep ID:

SP002818

Sampleprep Summary:

CSF was collected from adult (3 months old) wild-type CD1 mice. Samples were placed on wet ice, then spun 1000xg for 10 minutes at 4C. The supernatant was collected. Per condition, 5-10µL of fresh, cleared CSF or 5µl serum was extracted in 4:6:3 chlorophorm:methanol:water mixture supplemented with isotopically labeled T3 and T4 (Cambridge Isotope Laboratories, CLM-7185-C and CLM-8931-PK ) as well as isotopically labelled 17 amino acids and isotopically labelled reduced glutathione (Cambridge Isotope Laboratories, MSK-A2-1.2 and CNLM-6245-10). After centrifugation for 10 min at maximum speed on a benchtop centrifuge (Eppendorf) the top, hydrophilic layer (top phase) was transferred to a new tube, dried using a nitrogen dryer and reconstituted in 20 µl water (supplemented with QReSS, Cambridge Isotope Laboratories, MSK-QRESS-KIT) by brief vortexing. Extracted metabolites were spun again and cleared supernatant was transferred to LC-MS micro vials. A small amount of each sample was pooled and serially diluted 3- and 10-fold to be used as quality controls throughout the run of each batch. I calibration curve of unlabeled T3 and T4 standards was run per experiment for concentration calculations.