Summary of Study ST004201

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 PR002649. The data can be accessed directly via it's Project DOI: 10.21228/M87P0Z 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	ST004201
Study Title	SCFA quantification of blood serum in microplastic exposed mice - Targeted GC/MS
Study Type	in vivo
Study Summary	To investigate the impact of ingested MPs on the gut microbiome and the metabolome, 8-week-old male and female C57/BL6 mice were orally gavaged mixed plastic (5 µm) exposure consisting of polystyrene, polyethylene, and the biodegradable/biocompatible plastic, poly(lactic-co-glycolic acid), twice a week for 4 weeks at 0, 2, or 4 mg/week (n = 8/group). Fecal pellets were collected for bacterial DNA extraction and metagenomic shotgun sequencing, and serum was subjected to targeted and untargeted metabolomics. A total of 1162 bacterial species and 1205 metabolites were evaluated for downstream analysis. MPs exposure resulted in significant sex-specific and dose-dependent changes to the gut microbiome composition along with substantial regulation of predicted metabolic pathways. Untargeted metabolomics in serum showed that a low MPs dose displayed a more prominent effect on key metabolic pathways such as amino acid metabolism, mitochondrial function, and inflammation. Additionally, SCFA-targeted metabolomics showed significant changes in neuroprotective SCFAs levels in both sexes. Our study demonstrates that microplastics dysregulate the gut microbiome and serum metabolome, highlighting potential human disease risks.
Institute	University of Washington
Department	Environmental and Occupational Health Science
Laboratory	Cui Lab
Last Name	Kim
First Name	Kyle
Address	3150 Stephanie Loop Northeast
Email	kk1109@uw.edu
Phone	3606883268
Submit Date	2025-09-10
Num Groups	6
Total Subjects	24
Num Males	12
Num Females	12
Raw Data Available	Yes
Raw Data File Type(s)	mzML, d
Analysis Type Detail	GC-MS
Release Date	2025-10-10
Release Version	1

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Project:

Project ID:	PR002649
Project DOI:	doi: 10.21228/M87P0Z
Project Title:	In vivo exposure of mixed microplastic particles in mice and its impacts on the murine gut microbiome and metabolome
Project Type:	MS quantitative analysis
Project Summary:	Microplastics (MPs) are emerging environmental contaminants due to increasing global plastic production and waste. Microplastics, defined as plastic particles less than 5 mm in diameter, are formed through degradation of larger plastics via sunlight, weathering, and microbes. These plastic compounds are widely detected in water, soil, food, as well as human stool and blood. The gut microbiome, often referred to as our second genome, is important in human health and is the primary point of contact for orally ingested microplastics. To investigate the impact of ingested MPs on the gut microbiome and the metabolome, 8-week-old male and female C57/BL6 mice were orally gavaged mixed plastic (5 µm) exposure consisting of polystyrene, polyethylene, and the biodegradable/biocompatible plastic, poly(lactic-co-glycolic acid), twice a week for 4 weeks at 0, 2, or 4 mg/week (n = 8/group). Fecal pellets were collected for bacterial DNA extraction and metagenomic shotgun sequencing, and serum was subjected to targeted and untargeted metabolomics. A total of 1162 bacterial species and 1205 metabolites were evaluated for downstream analysis. MPs exposure resulted in significant sex-specific and dose-dependent changes to the gut microbiome composition along with substantial regulation of predicted metabolic pathways. Untargeted metabolomics in serum showed that a low MPs dose displayed a more prominent effect on key metabolic pathways such as amino acid metabolism, mitochondrial function, and inflammation. Additionally, SCFA-targeted metabolomics showed significant changes in neuroprotective SCFAs levels in both sexes. Our study demonstrates that microplastics dysregulate the gut microbiome and serum metabolome, highlighting potential human disease risks.
Institute:	University of Washington
Department:	Environmental and Occupational Health Science
Laboratory:	Cui Lab
Last Name:	Kim
First Name:	Kyle
Address:	3150 Stephanie Loop Northeast, Lacey, Washington, 98516, USA
Email:	kk1109@uw.edu
Phone:	3606883268
Funding Source:	National Institute of Health (NIH), National Institute of Environmental Health Sciences (NIEHS)

Subject:

Subject ID:	SU004353
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Age Or Age Range:	8-12 weeks
Gender:	Male and female
Animal Animal Supplier:	Taconic Biosciences
Animal Housing:	Temperature (20–24°C) and humidity (30%–60%)
Animal Light Cycle:	12-h light/dark cycle
Animal Feed:	ad libitum
Animal Water:	ad libitum
Species Group:	C57BL/6

Factors:

Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id	local_sample_id	Sample source	Sex	Treatment
SA484384	FCTRL2	Serum	Female	Control
SA484385	FCTRL1	Serum	Female	Control
SA484386	FCTRL4	Serum	Female	Control
SA484387	FCTRL3	Serum	Female	Control
SA484388	FHIGH3	Serum	Female	High_MPs
SA484389	FHIGH4	Serum	Female	High_MPs
SA484390	FHIGH1	Serum	Female	High_MPs
SA484391	FHIGH2	Serum	Female	High_MPs
SA484392	FLOW1	Serum	Female	Low_MPs
SA484393	FLOW2	Serum	Female	Low_MPs
SA484394	FLOW3	Serum	Female	Low_MPs
SA484395	FLOW4	Serum	Female	Low_MPs
SA484396	MCTRL1	Serum	Male	Control
SA484397	MCTRL2	Serum	Male	Control
SA484398	MCTRL3	Serum	Male	Control
SA484399	MCTRL4	Serum	Male	Control
SA484400	MHIGH3	Serum	Male	High_MPs
SA484401	MHIGH2	Serum	Male	High_MPs
SA484402	MHIGH1	Serum	Male	High_MPs
SA484403	MHIGH4	Serum	Male	High_MPs
SA484404	MLOW2	Serum	Male	Low_MPs
SA484405	MLOW4	Serum	Male	Low_MPs
SA484406	MLOW3	Serum	Male	Low_MPs
SA484407	MLOW1	Serum	Male	Low_MPs
SA484408	STD_3	Serum	NA	NA
SA484409	STD_4	Serum	NA	NA
SA484410	STD_5	Serum	NA	NA
SA484411	STD_6	Serum	NA	NA
SA484412	STD_7	Serum	NA	NA
SA484413	STD_8	Serum	NA	NA
SA484414	STD_10	Serum	NA	NA
SA484415	STD_9	Serum	NA	NA
SA484416	STD_11	Serum	NA	NA
SA484417	Serum_QC_1	Serum	NA	NA
SA484418	Serum_QC_2	Serum	NA	NA
SA484419	Serum_QC_3	Serum	NA	NA
SA484420	Serum_QC_4	Serum	NA	NA
SA484421	STD_2	Serum	NA	NA
SA484422	STD_1	Serum	NA	NA

Showing results 1 to 39 of 39

Showing results 1 to 39 of 39

Collection:

Collection ID:	CO004346
Collection Summary:	Blood was collected via cardiac puncture and samples were spun and separated for serum. Frozen serum samples were first thawed overnight under 4oC, and 50 μL of each sample was placed in a 2 mL Eppendorf vial prior to metabolomic analysis
Sample Type:	Blood (serum)

Treatment:

Treatment ID:	TR004362
Treatment Summary:	8-week-old male and female C57/BL6 mice were orally gavaged mixed plastic (5 µm) exposure consisting of polystyrene, polyethylene, and the biodegradable/biocompatible plastic, poly(lactic-co-glycolic acid), twice a week for 4 weeks at 0, 2, or 4 mg/week (n = 8/group).
Treatment:	in vivo oral ingestion
Treatment Compound:	mixed microplastics (polystyrene, polyethylene, poly lactic-co glycolic acid)
Treatment Route:	gastric oral gavage
Treatment Dose:	0mg/wk, 2mg/wk, 4mg/wk
Treatment Dosevolume:	100 ul of 10mg/ml twice a week for 2mg/wk and 200ul of 10mg/ml twice a week for 4mg/wk
Treatment Doseduration:	4 weeks
Treatment Vehicle:	water

Sample Preparation:

Sampleprep ID:	SP004359
Sampleprep Summary:	For serum samples, 20 μL of each sample was mixed with 30 μL aqueous NaOH (0.1M in water), 20 μL IS (hexanoic acid-6,6,6-d3; 200 µM) and 430 μL MeOH in a 1.5 mL Eppendorf tube. The pH value for the mixture was 9. Samples were then vortexed for 10 s and stored at -20 ºC for 20 min. After centrifugation at 14,000 RPM for 10 min at 4 ºC, 450 μL supernatant was removed into a new Eppendorf tube. The samples were dried under vacuum at 37 ºC for 120 min using a CentriVap Concentrator (Labconco, Fort Scott, KS). Each sample was first derivatized with 40 µL of methoxyamine hydrochloride solution in pyridine (MeOX, 20 mg/mL) under 60 ℃ for 90 min. Next, 60 µL of MTBSTFA was added, and the mixture was incubated under 60 ℃ for 30 min. Then the sample was vortexed for 30 s, followed by centrifugation at 14,000 rpm for 10 min. Finally, 70 µL supernatant was collected into a new glass vial for GC-MS analysis.

Sampleprep ID:

SP004359

Sampleprep Summary:

For serum samples, 20 μL of each sample was mixed with 30 μL aqueous NaOH (0.1M in water), 20 μL IS (hexanoic acid-6,6,6-d3; 200 µM) and 430 μL MeOH in a 1.5 mL Eppendorf tube. The pH value for the mixture was 9. Samples were then vortexed for 10 s and stored at -20 ºC for 20 min. After centrifugation at 14,000 RPM for 10 min at 4 ºC, 450 μL supernatant was removed into a new Eppendorf tube. The samples were dried under vacuum at 37 ºC for 120 min using a CentriVap Concentrator (Labconco, Fort Scott, KS). Each sample was first derivatized with 40 µL of methoxyamine hydrochloride solution in pyridine (MeOX, 20 mg/mL) under 60 ℃ for 90 min. Next, 60 µL of MTBSTFA was added, and the mixture was incubated under 60 ℃ for 30 min. Then the sample was vortexed for 30 s, followed by centrifugation at 14,000 rpm for 10 min. Finally, 70 µL supernatant was collected into a new glass vial for GC-MS analysis.

Chromatography:

Chromatography ID:	CH005304
Chromatography Summary:	GC-MS experiments were performed using an Agilent 7820A gas chromatography system coupled to an Agilent 5977B mass spectrometer (Agilent Technologies, Santa Clara, CA). Chemical derivatives in the samples were separated using an HP-5 ms capillary column coated with 5% phenyl-95% methylpolysiloxane (30 m×250 µm i.d., 0.25 µm film thickness, Agilent Technologies). 1 µL of each sample was injected, and the solvent delay time was set to 5 min. The initial oven temperature was held at 60 ℃ for 1 min, ramped up to 325 ℃ at a rate of 10 ℃/min, and finally held at 325 ℃ for 10 min. Helium was used as the carrier gas at a constant flow rate of 20 mL/min through the column.
Chromatography Comments:	The temperatures of the front inlet, transfer line, and electron impact (EI) ion source were set at 250 ℃, 290 ℃, and 230 ℃, respectively
Instrument Name:	Agilent 7820A
Column Name:	Agilent HP5-MS (30m x 0.25mm, 0.25 um)
Column Temperature:	The initial oven temperature was held at 60 ℃ for 1 min, ramped up to 325 ℃ at a rate of 10 ℃/min, and finally held at 325 ℃ for 10 min
Flow Gradient:	N/A
Flow Rate:	20 mL/min
Injection Temperature:	60 ℃
Solvent A:	N/A
Solvent B:	N/A
Oven Temperature:	60 ℃ for 1 min, ramped up to 325 ℃ at a rate of 10 ℃/min, held at 325 ℃ for 10 min
Chromatography Type:	GC

Analysis:

Analysis ID:	AN006987
Analysis Type:	MS
Chromatography ID:	CH005304
Num Factors:	7
Num Metabolites:	20
Units:	Relative abundance