Summary of Study ST003896

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 PR002438. The data can be accessed directly via it's Project DOI: 10.21228/M8H53D 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	ST003896
Study Title	Postprandial Plasma Metabolomic Changes in 147 Individuals Following Ingestion of a Standard Mixed Meal
Study Summary	This study applied untargeted metabolomic techniques to investigate plasma metabolic profile changes in 147 individuals following consumption of a standard mixed meal. Time-series analyses identified dynamic responses in several key metabolic pathways and examined their associations with participants' metabolic health status. The results indicated considerable inter-individual differences in the amplitude and temporal patterns of metabolite changes, highlighting the significance of metabolic individuality.We found that substances such as Phenylalanine, allo-Inositol, Taurochenodesoxycholic acid, Succinic acid semialdehyde (notably listed twice), DNP-DL-Methionine, and 1,3,5-Trihydroxybenzene changed significantly after the standard mixed meal test (MMTT), and these changes were highly correlated with hormone fluctuations such as Insulin and Glucagon.
Institute	Nanjing Medical University
Last Name	Wang
First Name	Jiachen
Address	No. 300, Guangzhou Road, Nanjing
Email	jcwang@stu.njmu.edu.cn
Phone	02565306466
Submit Date	2025-04-27
Raw Data Available	Yes
Raw Data File Type(s)	mzML, raw(Thermo)
Analysis Type Detail	Other
Release Date	2025-07-30
Release Version	1

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Project:

Project ID:	PR002438
Project DOI:	doi: 10.21228/M8H53D
Project Title:	Postprandial Plasma Multi-Omics Changes in Response to a Standard Mixed Meal or Individual Macronutrients in Humans
Project Summary:	Postprandial metabolism is a complex and dynamic physiological process involving numerous metabolic pathways and biomolecular interactions. It plays a critical role in the development of chronic diseases such as obesity, diabetes, and cardiovascular disease. This study employed an interventional design in human participants to systematically compare the dynamic postprandial changes in plasma multi-omics profiles following ingestion of a standard mixed meal versus four individual macronutrients (glucose, protein, butter, and olive oil). The aim is to elucidate the specific effects of different nutrients on human postprandial metabolic networks and their potential implications for health, thereby providing foundational data to support personalized nutrition strategies and disease prevention.
Institute:	Nanjing Medical University
Last Name:	Wang
First Name:	Jiachen
Address:	No. 300, Guangzhou Road, Nanjing
Email:	jcwang@stu.njmu.edu.cn
Phone:	+86 02565306466

Subject:

Subject ID:	SU004031
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606

Factors:

Subject type: Human; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id	local_sample_id	Time
SA428656	P103A	0
SA428657	P97A	0
SA428658	P98A	0
SA428659	P99A	0
SA428660	P100A	0
SA428661	P101A	0
SA428662	P102A	0
SA428663	P104A	0
SA428664	P95A	0
SA428665	P105A	0
SA428666	P106A	0
SA428667	P107A	0
SA428668	P108A	0
SA428669	P109A	0
SA428670	P110A	0
SA428671	P96A	0
SA428672	P94A	0
SA428673	P112A	0
SA428674	P84A	0
SA428675	P78A	0
SA428676	P79A	0
SA428677	P80A	0
SA428678	P81A	0
SA428679	P82A	0
SA428680	P83A	0
SA428681	P85A	0
SA428682	P93A	0
SA428683	P86A	0
SA428684	P87A	0
SA428685	P88A	0
SA428686	P89A	0
SA428687	P90A	0
SA428688	P91A	0
SA428689	P92A	0
SA428690	P111A	0
SA428691	P113A	0
SA428692	P76A	0
SA428693	P140A	0
SA428694	P134A	0
SA428695	P135A	0
SA428696	P136A	0
SA428697	P137A	0
SA428698	P138A	0
SA428699	P139A	0
SA428700	P141A	0
SA428701	P132A	0
SA428702	P142A	0
SA428703	P143A	0
SA428704	P144A	0
SA428705	P145A	0
SA428706	P146A	0
SA428707	P147A	0
SA428708	P2A	0
SA428709	P133A	0
SA428710	P131A	0
SA428711	P114A	0
SA428712	P121A	0
SA428713	P115A	0
SA428714	P116A	0
SA428715	P117A	0
SA428716	P118A	0
SA428717	P119A	0
SA428718	P120A	0
SA428719	P122A	0
SA428720	P130A	0
SA428721	P123A	0
SA428722	P124A	0
SA428723	P125A	0
SA428724	P126A	0
SA428725	P127A	0
SA428726	P128A	0
SA428727	P129A	0
SA428728	P77A	0
SA428729	P1A	0
SA428730	P75A	0
SA428731	P28A	0
SA428732	P74A	0
SA428733	P23A	0
SA428734	P24A	0
SA428735	P25A	0
SA428736	P26A	0
SA428737	P27A	0
SA428738	P29A	0
SA428739	P20A	0
SA428740	P30A	0
SA428741	P31A	0
SA428742	P32A	0
SA428743	P33A	0
SA428744	P34A	0
SA428745	P35A	0
SA428746	P21A	0
SA428747	P19A	0
SA428748	P37A	0
SA428749	P9A	0
SA428750	P3A	0
SA428751	P4A	0
SA428752	P5A	0
SA428753	P6A	0
SA428754	P7A	0
SA428755	P8A	0

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Collection:

Collection ID:	CO004024
Collection Summary:	Participants venous blood samples were collected at baseline (0 min) and at 30 min, 60 min, 120 min, and 180 min post-meal and stored at -80 C until analysis.
Sample Type:	Blood (plasma)

Treatment:

Treatment ID:	TR004040
Treatment Summary:	Volunteers were recruited in Nanjing, China, via poster advertisements to participate in a standard MMTT (mixed meal tolerance test). The inclusion criteria were as follows: age ≥ 18 years old, no prior diagnosis of diabetes; no history of malignant tumors, severe organ disease, or significant acute or chronic infections; and no recent use of glucocorticoids, immunosuppressants, or other medications known to substantially affect metabolism. A total of 147 participants (77 males and 70 females, aged 18-50 years) completed the MMTT. Among them, 54 participants were of normal weight (BMI ≤ 24 kg/m²), 38 were overweight (24 kg/m² < BMI ≤ 28 kg/m²), and 55 were categorised as obese (BMI > 28 kg/m²). We utilized Ensure powder (Abbott Manufacturing Singapore Private Limited), a standardized nutritional meal replacement, which provides 57.4g of carbohydrates, 15.9g of protein, and 14g of fat per 100g, delivering 430 kcal of energy. After fasting for 12 hours, each participant was instructed to consume 84g of Ensure (360 kcal) dissolved in 250 ml of water within 3-5 minutes. Participants venous blood samples were collected at baseline (0 min) and at 30 min, 60 min, 120 min, and 180 min post-meal.

Treatment ID:

TR004040

Treatment Summary:

Volunteers were recruited in Nanjing, China, via poster advertisements to participate in a standard MMTT (mixed meal tolerance test). The inclusion criteria were as follows: age ≥ 18 years old, no prior diagnosis of diabetes; no history of malignant tumors, severe organ disease, or significant acute or chronic infections; and no recent use of glucocorticoids, immunosuppressants, or other medications known to substantially affect metabolism. A total of 147 participants (77 males and 70 females, aged 18-50 years) completed the MMTT. Among them, 54 participants were of normal weight (BMI ≤ 24 kg/m²), 38 were overweight (24 kg/m² < BMI ≤ 28 kg/m²), and 55 were categorised as obese (BMI > 28 kg/m²). We utilized Ensure powder (Abbott Manufacturing Singapore Private Limited), a standardized nutritional meal replacement, which provides 57.4g of carbohydrates, 15.9g of protein, and 14g of fat per 100g, delivering 430 kcal of energy. After fasting for 12 hours, each participant was instructed to consume 84g of Ensure (360 kcal) dissolved in 250 ml of water within 3-5 minutes. Participants venous blood samples were collected at baseline (0 min) and at 30 min, 60 min, 120 min, and 180 min post-meal.

Sample Preparation:

Sampleprep ID:	SP004037
Sampleprep Summary:	Plasma samples were thawed on ice, and 50 µL of each sample was mixed with four volumes of methanol containing a mixed isotope internal standard solution. The mixture was shaken at 1000 rpm for 10 minutes, followed by vortex mixing. Samples were then centrifuged at 15,000 rpm for 10 minutes at 4°C. After centrifugation, the supernatant was transferred to a protein precipitation plate for filtration, and an equal volume of the filtrate was transferred to a new centrifuge tube for freeze-drying at low temperature. The freeze-dried sample was recombined with 200 μL recombinant solvent and uploaded to 10 μL mass spectrometry for quantitative analysis. Internal standards used in this assay included: D-Fructose-13C6 (CIL, USA), L-Tryptophan-d5(CIL, USA), citric acid d4(CIL, USA), L-lysine d4(CIL, USA), Benzoic acid d5(CIL, USA), [2H5]-5-Hydroxyindole-3-acetic acid (TRC, CA), N-benzoyl-D-phenylalanine (Sigma-Aldrich, GER), DNOP-d4 (Nafushengwu, CHA). The concentration of each internal standard in the final internal standard mixed solution was 0.05 mmol/L. Finally, 10 µl internal standard mixed solution was added to each sample for sample preparation. Metabolic extracts were separated on BEH C18 column (Waters, SKU: 186002352, 100 mm × 2.1 mm, 1.7 μm) using a 12-minute gradient with organic phase increased from 2% to 100% at 40℃ (solvent A: 0.1% formic acid, 2 mM ammonium formate in water; solvent B: 0.1% formic acid, 2 mM ammonium formate in methanol).

Sampleprep ID:

SP004037

Sampleprep Summary:

Plasma samples were thawed on ice, and 50 µL of each sample was mixed with four volumes of methanol containing a mixed isotope internal standard solution. The mixture was shaken at 1000 rpm for 10 minutes, followed by vortex mixing. Samples were then centrifuged at 15,000 rpm for 10 minutes at 4°C. After centrifugation, the supernatant was transferred to a protein precipitation plate for filtration, and an equal volume of the filtrate was transferred to a new centrifuge tube for freeze-drying at low temperature. The freeze-dried sample was recombined with 200 μL recombinant solvent and uploaded to 10 μL mass spectrometry for quantitative analysis. Internal standards used in this assay included: D-Fructose-13C6 (CIL, USA), L-Tryptophan-d5(CIL, USA), citric acid d4(CIL, USA), L-lysine d4(CIL, USA), Benzoic acid d5(CIL, USA), [2H5]-5-Hydroxyindole-3-acetic acid (TRC, CA), N-benzoyl-D-phenylalanine (Sigma-Aldrich, GER), DNOP-d4 (Nafushengwu, CHA). The concentration of each internal standard in the final internal standard mixed solution was 0.05 mmol/L. Finally, 10 µl internal standard mixed solution was added to each sample for sample preparation. Metabolic extracts were separated on BEH C18 column (Waters, SKU: 186002352, 100 mm × 2.1 mm, 1.7 μm) using a 12-minute gradient with organic phase increased from 2% to 100% at 40℃ (solvent A: 0.1% formic acid, 2 mM ammonium formate in water; solvent B: 0.1% formic acid, 2 mM ammonium formate in methanol).

Chromatography:

Chromatography ID:	CH004851
Chromatography Summary:	Samples were analyzed using a Thermo Scientific™ Vanquish™ UHPLC coupled with Q-Exactive™ PLUS mass spectrometer (ThermoFisher, USA) under data-dependent acquisition (DDA) mode. A full survey scan was obtained at a 70,000 resolution for the m/z range of 70 to 1050 and followed by 10 MS/MS scans in HCD mode at a 17,500 resolution. Internal standard retention times and alignment as well as coefficients of variation (CVs) of metabolite quantification values were checked daily to ensure data quality including chromatography performance, mass calibration, and extraction efficiency.
Instrument Name:	Waters Acquity
Column Name:	Waters ACQUITY UPLC CSH C18 (150 x 2.1mm,1.7um)
Column Temperature:	40 °C
Flow Gradient:	the organic phase was increased from 2% to 100% in 12 min, and the remaining 6 min was used to flush and equilibrate the column
Flow Rate:	0.3 mL/min
Solvent A:	100% water
Solvent B:	100% methanol
Chromatography Type:	Reversed phase

Analysis:

Analysis ID:	AN006399
Analysis Type:	MS
Chromatography ID:	CH004851
Num Factors:	3
Num Metabolites:	423
Units:	Normalized counts