Summary of Study ST002708

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	ST002708
Study Title	Levels of central carbon metabolites in choroid plexus as part of natural diurnal variation
Study Summary	This study employs targeted LC-MS analysis of choroid plexus (ChP) tissue to assess relative changes in the levels of intermediates from the central carbon metabolism, with additional attention to mitochondrial energy precursors (ATP/ADP, NAD(P) and NAD(P)H) at two time points in the diurnal cycle. For this purpose, 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). Two different ChP tissues were collected for analysis (LV - left ventricle and 4V - 4th ventricle). Also two separate extractions were performed: 80% MetOH based ("MetOH" study factor) and FB (MetOH with Na-ascorbate and Na-acetate additives, "FB" study factor). The two extractions are optimal for central carbon metabolites or NAD(P)/H respectively.
Institute	Boston Childrens Hospital
Department	Pathology
Laboratory	Kanarek Lab
Last Name	Petrova
First Name	Boryana
Address	330 Longwood Av
Email	boryana.petrova@childrens.harvard.edu
Phone	6173557433
Submit Date	2023-05-15
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:	SP002819
Sampleprep Summary:	ChP were extracted by brief sonication in either 200µl 80% LC/MS-grade methanol extraction solvent or 200µl of ”FB” extraction solvent (80% LC/MS-grade methanol, 20% 25 mM Ammonium Acetate and 2.5 mM Na-Ascorbate prepared in LC/MS water and supplemented with isotopically labeled internal standards (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 cleared supernatant was 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.

Sampleprep ID:

SP002819

Sampleprep Summary:

ChP were extracted by brief sonication in either 200µl 80% LC/MS-grade methanol extraction solvent or 200µl of ”FB” extraction solvent (80% LC/MS-grade methanol, 20% 25 mM Ammonium Acetate and 2.5 mM Na-Ascorbate prepared in LC/MS water and supplemented with isotopically labeled internal standards (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 cleared supernatant was 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.