Summary of Study ST000545

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 PR000399. The data can be accessed directly via it's Project DOI: 10.21228/M8GG6B This work is supported by NIH grant, U2C- DK119886.

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

Study ID	ST000545
Study Title	CHEAR Plasma Reference Material Proficiency Test Biocrates
Study Type	Metabolomics
Study Summary	CHEAR Reference Material Plasma was provided by Emory University. The material was prepared and analyzed by way of LC-MS and Biocrates workflow employed by the Eastern Regional Metabolomics Resource Core (protocols available in metabolomics workbench). Six samples were injected of the sample reference material prepared in replicate.
Institute	RTI International
Department	Discovery-Sciences-Technology (DST)
Laboratory	RTI CHEAR Analytical Hub - Untargeted Analysis Resource Core
Last Name	Fennell
First Name	Timothy
Address	3040 E Cornwallis Road, Research Triangle Park, NC 27709
Email	fennell@rti.org
Phone	919-485-2781
Submit Date	2016-12-29
Raw Data Available	Yes
Raw Data File Type(s)	wiff
Chear Study	Yes
Analysis Type Detail	LC-MS
Release Date	2024-12-31
Release Version	1

Select appropriate tab below to view additional metadata details:

Sample Preparation:

Sampleprep ID:	SP000574
Sampleprep Summary:	Parent Aliquots were quickly prepared in the 4°C room using the Drummond pipet aid and serological pipet. 40 mL plasma were transferred to 50 mL tubes in 4°C room and capped immediately and placed on ice. If needed, the bulk plasma was mixed in between aliquots. The parent aliquots were labeled appropriately. Sub-Aliquots were prepared on ice at the bench. The parent aliquots were mixed by inverting the tube thoroughly before and in between aliquots as needed. 1 mL plasma was transferred to cryovials and capped immediately and stored on ice until sample splitting was competed. The sub aliquots were labeled appropriately. Sub-aliquots were stored at -80°C. Plasma Aliquots for LCMS platforms: Used sub-aliquot “PP_A_09” to prepare aliquots for various LCMS platforms. Allowed sub-aliquot to thaw on ice for 30 – 60 min. Vortexed aliquot briefly on vortexer, centrifuged at 4 °C for 2 minutes at 16,000 rcf. Aliquoted out 6 tubes for Biocrates from “PP_A_09”. Aliquots were stored at -80 °C until analysis. Sample preparation for Biocrates Plate: CHEAR plasma samples were thawed on ice for 30–60 min. All samples were vortexed on a multi-tube vortexer for 4 mins at 5,000 rpm and centrifuged at 4 °C for 5 minutes at 16,000 rcf before loading on the p180 plate. The samples were then placed on ice in the analysis order for sample loading on the p180 Biocrates plate. Biocrates Plate Preparation: A Biocrates p180 kit was prepared following the AbsoluteIDQ™ p180 Kit metabolomics procedure. Briefly, an internal standard mix was added to 95 of the 96 wells. Next, zero samples, QC standards and calibration standards were added to their corresponding wells. The samples (20 µL) were then added to the appropriate wells and dried for 30 minutes under nitrogen flow. The plate was derivatized using a 5% phenylisothiocyanate (PITC) solution in (1:1:1) ethanol:pyridine:water and, then, incubated for 20 minutes followed by a drying step under nitrogen flow. An extraction solvent (5 mM ammonium acetate in methanol) was added to all wells. The plate was then shaken and centrifuged. After centrifugation, 150 µL was removed and transferred to a second 96-well plate (LCMS plate). This second plate was diluted with 150 µL of HPLC grade water for a subsequent LCMS (MRM analysis) for measuring amino acids and biogenic amines. All wells in the original plate were diluted with 400 µL of flow injection analysis (FIA) Running Solvent for a FIA-MS (MRM analysis) for measuring lipids, acylcarnitines, and hexose.

Sampleprep ID:

SP000574

Sampleprep Summary:

Parent Aliquots were quickly prepared in the 4°C room using the Drummond pipet aid and serological pipet. 40 mL plasma were transferred to 50 mL tubes in 4°C room and capped immediately and placed on ice. If needed, the bulk plasma was mixed in between aliquots. The parent aliquots were labeled appropriately. Sub-Aliquots were prepared on ice at the bench. The parent aliquots were mixed by inverting the tube thoroughly before and in between aliquots as needed. 1 mL plasma was transferred to cryovials and capped immediately and stored on ice until sample splitting was competed. The sub aliquots were labeled appropriately. Sub-aliquots were stored at -80°C. Plasma Aliquots for LCMS platforms: Used sub-aliquot “PP_A_09” to prepare aliquots for various LCMS platforms. Allowed sub-aliquot to thaw on ice for 30 – 60 min. Vortexed aliquot briefly on vortexer, centrifuged at 4 °C for 2 minutes at 16,000 rcf. Aliquoted out 6 tubes for Biocrates from “PP_A_09”. Aliquots were stored at -80 °C until analysis. Sample preparation for Biocrates Plate: CHEAR plasma samples were thawed on ice for 30–60 min. All samples were vortexed on a multi-tube vortexer for 4 mins at 5,000 rpm and centrifuged at 4 °C for 5 minutes at 16,000 rcf before loading on the p180 plate. The samples were then placed on ice in the analysis order for sample loading on the p180 Biocrates plate. Biocrates Plate Preparation: A Biocrates p180 kit was prepared following the AbsoluteIDQ™ p180 Kit metabolomics procedure. Briefly, an internal standard mix was added to 95 of the 96 wells. Next, zero samples, QC standards and calibration standards were added to their corresponding wells. The samples (20 µL) were then added to the appropriate wells and dried for 30 minutes under nitrogen flow. The plate was derivatized using a 5% phenylisothiocyanate (PITC) solution in (1:1:1) ethanol:pyridine:water and, then, incubated for 20 minutes followed by a drying step under nitrogen flow. An extraction solvent (5 mM ammonium acetate in methanol) was added to all wells. The plate was then shaken and centrifuged. After centrifugation, 150 µL was removed and transferred to a second 96-well plate (LCMS plate). This second plate was diluted with 150 µL of HPLC grade water for a subsequent LCMS (MRM analysis) for measuring amino acids and biogenic amines. All wells in the original plate were diluted with 400 µL of flow injection analysis (FIA) Running Solvent for a FIA-MS (MRM analysis) for measuring lipids, acylcarnitines, and hexose.