Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA328143

Local Sample ID:	DIG008 DP1.Hour 2 plasma
Subject ID:	SU003145
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606

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Subject:

Subject ID:	SU003145
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
DIG008 DP1.Hour 2 plasma	SA328143	FL038954	DP1	Treatment Group
DIG008 DP1.Hour 2 plasma	SA328143	FL038954	Hour 2	Timepoint

Collection:

Collection ID:	CO003138
Collection Summary:	Whole blood was collected from the antecubital vein while participants were fasted, and 2-and 4-h after a mixed meal challenge in EDTA vacutainer tubes (BD Vacutainer, New Jersey, USA).
Sample Type:	Blood (plasma)

Treatment:

Treatment ID:	TR003154
Treatment Summary:	This study was approved by the Florida State University Institutional Review Board and registered on clinicaltrials.gov (NCT04877262). Healthy, adults who were normal-weight (BMI:18.5-24.9 kg/m2) or overweight (BMI: 25-29.9 kg/m2) and 23-42 y, were recruited from the greater Tallahassee, Florida area between December 2021 and May 2022 and provided informed consent prior to participating. Potential participants were excluded if they had any disease or condition known to interfere with metabolism or normal gastrointestinal function (diabetes, cardiovascular disease, kidney disease, prior bariatric surgery, suspected or known fistulas, gastrointestinal obstruction, gastrointestinal disease, colonoscopy within 3 months of the study, alcoholism, substance abuse disorders); irregular menstrual cycle in the past 6 months; weight fluctuations greater that 5% during the previous 6 months; antibiotic use within 3 months of participation; allergies or intolerances to foods included in the controlled diet; habitual use of laxatives, stool softeners, or anti-diarrheal medications (≥ 1x/week); whole-gut transit time >72 h; or were pregnant or lactating. Participants followed an early time-restricted eating (eTRE) schedule (i.e., all meals consumed in a 6-h window between 0800 and 1400) and a control eating schedule (i.e., all meals consumed in a 12-h window between 0800 and 2000) for 9-d each in randomized order. On day 6 of each study period, participants arrived in the morning to the laboratory for a mixed-meal tolerance test (MMTT). Participants spent 6 hours in the laboratory to measure fasting (0h) and postprandial (2h and 4h) plasma metabolite concentrations in response to the MMTT. Whole blood was collected in EDTA vacutainer tubes (BD Vacutainer, Franklin Lakes, New Jersey, USA) and centrifuged at 3000 RPM for 15 minutes at 4°C before aliquots were frozen at -80°C until analysis.

Treatment ID:

TR003154

Treatment Summary:

This study was approved by the Florida State University Institutional Review Board and registered on clinicaltrials.gov (NCT04877262). Healthy, adults who were normal-weight (BMI:18.5-24.9 kg/m2) or overweight (BMI: 25-29.9 kg/m2) and 23-42 y, were recruited from the greater Tallahassee, Florida area between December 2021 and May 2022 and provided informed consent prior to participating. Potential participants were excluded if they had any disease or condition known to interfere with metabolism or normal gastrointestinal function (diabetes, cardiovascular disease, kidney disease, prior bariatric surgery, suspected or known fistulas, gastrointestinal obstruction, gastrointestinal disease, colonoscopy within 3 months of the study, alcoholism, substance abuse disorders); irregular menstrual cycle in the past 6 months; weight fluctuations greater that 5% during the previous 6 months; antibiotic use within 3 months of participation; allergies or intolerances to foods included in the controlled diet; habitual use of laxatives, stool softeners, or anti-diarrheal medications (≥ 1x/week); whole-gut transit time >72 h; or were pregnant or lactating. Participants followed an early time-restricted eating (eTRE) schedule (i.e., all meals consumed in a 6-h window between 0800 and 1400) and a control eating schedule (i.e., all meals consumed in a 12-h window between 0800 and 2000) for 9-d each in randomized order. On day 6 of each study period, participants arrived in the morning to the laboratory for a mixed-meal tolerance test (MMTT). Participants spent 6 hours in the laboratory to measure fasting (0h) and postprandial (2h and 4h) plasma metabolite concentrations in response to the MMTT. Whole blood was collected in EDTA vacutainer tubes (BD Vacutainer, Franklin Lakes, New Jersey, USA) and centrifuged at 3000 RPM for 15 minutes at 4°C before aliquots were frozen at -80°C until analysis.

Sample Preparation:

Sampleprep ID:	SP003151
Sampleprep Summary:	Plasma (25 μL) was added to 1.5 mL Eppendorf tubes prior to the addition of 10 μL of 1 μM internal standard solution. Next, 750 μL chilled methanol was added and samples were vortexed 30 s before centrifugation at 15,000 × G for 10 min. Subsequently, the supernatant was collected and added to 1.5 mL high performance liquid chromatography (HPLC) amber glass vials, dried, and reconstituted in 100 μL 3:1 acetonitrile: methanol containing 1-cyclohexyl-ureido, 3-dodecanoic acid (CUDA; Sigma-Aldrich, St. Louis, MO, USA) solution. The solution was vortexed 30 s, transferred to microfilter tubes, and centrifuged at 10,000 × G for 3 min prior to transfer to a HPLC vial.
Sampleprep Protocol Filename:	La_Frano_Lab_Methods_eTRE_v2.pdf

Combined analysis:

Analysis ID	AN004968
Analysis type	MS
Chromatography type	HILIC
Chromatography system	Waters Acquity I-Class
Column	Phenomenex Luna NH2 (150 x 2mm,3um)
MS Type	ESI
MS instrument type	QTRAP
MS instrument name	ABI Sciex 4000 QTrap
Ion Mode	NEGATIVE
Units	Peak area

Chromatography:

Chromatography ID:	CH003750
Methods Filename:	La_Frano_Lab_Methods_eTRE_v2.pdf
Instrument Name:	Waters Acquity I-Class
Column Name:	Phenomenex Luna NH2 (150 x 2mm,3um)
Column Temperature:	30
Flow Gradient:	0 min 90%B, 10 min 5%B, 11 min 5%B, 13 min 90%B, 15 min 90%B
Flow Rate:	0.3 ml/min
Solvent A:	100% water with 20 mM ammonium acetate, 20 mM ammonium hydroxide
Solvent B:	10 mM ammonium hydroxide in 75:25 % v/v (volume %) acetonitrile/methanol
Chromatography Type:	HILIC

MS:

MS ID:	MS004708
Analysis ID:	AN004968
Instrument Name:	ABI Sciex 4000 QTrap
Instrument Type:	QTRAP
MS Type:	ESI
MS Comments:	Acquisition software AB Sciex Analyst and quantitation software Sciex MultiQuant. Metabolites detected using multiple reaction monitoring (MRM).
Ion Mode:	NEGATIVE
Analysis Protocol File:	La_Frano_Lab_Methods_eTRE_v2.pdf