Summary of Study ST003895

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	ST003895
Study Title	Postprandial Plasma Metabolomic Changes in 24 Metabolically Healthy Individuals Following Ingestion of Four Different Isocaloric Macronutrients
Study Summary	This study included 24 metabolically healthy adults who received isocaloric oral loads of glucose, protein, butter, or olive oil. Plasma samples were collected at various time points, and untargeted metabolomic analyses were conducted to monitor dynamic changes in metabolites. The findings demonstrated significant differences in postprandial metabolite profiles induced by different macronutrients, offering key clues to the regulatory mechanisms of dietary components on metabolism.Under glucose load stimulation, 2-((2R)-2-Hydroxycyclohexyl)acetic acid, 3-(2-Ethylhexyl)phthalic acid, and Testosterone changed significantly. Under protein load stimulation, Oleamide, Monoolein, and Methyl gallate showed significant changes. Under olive oil load stimulation, Oxandrolone, 4-Nitrophenol, and 4,6-Dinitro-O-cresol changed significantly. Under butter load stimulation, 4-Nitrophenol, Oxandrolone, and 12alpha-12-Hydroxy-7,13-abietadien-18-oic acid showed significant changes.
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:	SU004030
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
SA428176	3P04A	0
SA428177	2P22A	0
SA428178	2P23A	0
SA428179	2P25A	0
SA428180	1P02A	0
SA428181	3P02A	0
SA428182	3P03A	0
SA428183	3P05A	0
SA428184	2P20A	0
SA428185	3P06A	0
SA428186	3P07A	0
SA428187	3P08A	0
SA428188	3P09A	0
SA428189	3P10A	0
SA428190	3P11A	0
SA428191	2P21A	0
SA428192	2P19A	0
SA428193	3P13A	0
SA428194	2P09A	0
SA428195	2P03A	0
SA428196	2P04A	0
SA428197	2P05A	0
SA428198	2P06A	0
SA428199	2P07A	0
SA428200	2P08A	0
SA428201	2P10A	0
SA428202	2P18A	0
SA428203	2P11A	0
SA428204	2P12A	0
SA428205	2P13A	0
SA428206	2P14A	0
SA428207	2P15A	0
SA428208	2P16A	0
SA428209	2P17A	0
SA428210	3P12A	0
SA428211	3P14A	0
SA428212	2P01A	0
SA428213	4P17A	0
SA428214	4P11A	0
SA428215	4P12A	0
SA428216	4P13A	0
SA428217	4P14A	0
SA428218	4P15A	0
SA428219	4P16A	0
SA428220	4P18A	0
SA428221	4P09A	0
SA428222	4P19A	0
SA428223	4P20A	0
SA428224	4P21A	0
SA428225	4P22A	0
SA428226	4P23A	0
SA428227	4P25A	0
SA428228	4P10A	0
SA428229	4P08A	0
SA428230	3P15A	0
SA428231	3P22A	0
SA428232	3P16A	0
SA428233	3P17A	0
SA428234	3P18A	0
SA428235	3P19A	0
SA428236	3P20A	0
SA428237	3P21A	0
SA428238	3P23A	0
SA428239	4P07A	0
SA428240	3P25A	0
SA428241	4P01A	0
SA428242	4P02A	0
SA428243	4P03A	0
SA428244	4P04A	0
SA428245	4P05A	0
SA428246	4P06A	0
SA428247	2P02A	0
SA428248	1P01A	0
SA428249	3P01A	0
SA428250	1P14A	0
SA428251	1P05A	0
SA428252	1P07A	0
SA428253	1P08A	0
SA428254	1P09A	0
SA428255	1P10A	0
SA428256	1P11A	0
SA428257	1P12A	0
SA428258	1P13A	0
SA428259	1P15A	0
SA428260	1P03A	0
SA428261	1P16A	0
SA428262	1P17A	0
SA428263	1P18A	0
SA428264	1P19A	0
SA428265	1P20A	0
SA428266	1P21A	0
SA428267	1P22A	0
SA428268	1P23A	0
SA428269	1P25A	0
SA428270	1P04A	0
SA428271	1P06A	0
SA428272	2P25D	120
SA428273	3P03D	120
SA428274	2P07D	120
SA428275	2P06D	120

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Collection:

Collection ID:	CO004023
Collection Summary:	Blood samples were collected at fasting baseline (0 min) and at 30, 60, 120, and 180 min post-consumption (a total of 480 samples) and stored at -80 C until analysis.
Sample Type:	Blood (plasma)

Treatment:

Treatment ID:	TR004039
Treatment Summary:	A cohort of healthy volunteers was recruited for the Single Macronutrient Tolerance Test (SMNTT). The inclusion criteria were: age ≥ 18 years old, no history of abnormal glucose levels (fasting plasma glucose < 6 mmol/L, 2-hour postprandial plasma glucose <7.8 mmol/L); normal body mass index (BMI, 18–24 kg/m²); no family history of diabetes; no history of malignancy, severe organ diseases, or significant acute or chronic infections; and no recent use of glucocorticoids, immunosuppressants, or medications known to significantly affect metabolism. A total of 12 healthy men and 12 healthy women, aged 18-40 years, completed the SMNTT. Participants underwent four successive SMNTT (glucose, whey protein, butter, and olive oil), each separated by a one-week interval. We used 75g of anhydrous glucose powder, 78.5g of protein powder, 40.1g butter or 33.9g olive oil for each load. Each test provided 300 kcal of energy. After a 12-hour overnight fast, participants consumed one of the four nutrients: glucose or whey protein dissolved in 250 mL of water, olive oil consumed directly, or butter accompanied by three lettuce leaves.

Treatment ID:

TR004039

Treatment Summary:

A cohort of healthy volunteers was recruited for the Single Macronutrient Tolerance Test (SMNTT). The inclusion criteria were: age ≥ 18 years old, no history of abnormal glucose levels (fasting plasma glucose < 6 mmol/L, 2-hour postprandial plasma glucose <7.8 mmol/L); normal body mass index (BMI, 18–24 kg/m²); no family history of diabetes; no history of malignancy, severe organ diseases, or significant acute or chronic infections; and no recent use of glucocorticoids, immunosuppressants, or medications known to significantly affect metabolism. A total of 12 healthy men and 12 healthy women, aged 18-40 years, completed the SMNTT. Participants underwent four successive SMNTT (glucose, whey protein, butter, and olive oil), each separated by a one-week interval. We used 75g of anhydrous glucose powder, 78.5g of protein powder, 40.1g butter or 33.9g olive oil for each load. Each test provided 300 kcal of energy. After a 12-hour overnight fast, participants consumed one of the four nutrients: glucose or whey protein dissolved in 250 mL of water, olive oil consumed directly, or butter accompanied by three lettuce leaves.

Sample Preparation:

Sampleprep ID:	SP004036
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:

SP004036

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:	CH004850
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
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:	AN006398
Analysis Type:	MS
Chromatography ID:	CH004850
Num Factors:	5
Num Metabolites:	423
Units:	Normalized counts