Summary of Study ST002495

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 PR001610. The data can be accessed directly via it's Project DOI: 10.21228/M8KT5B 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.

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Study ID	ST002495
Study Title	Disrupted intestinal microbiota contributes to the pathogenesis of anorexia nervosa (Part 2)
Study Summary	Anorexia nervosa (AN) is an eating disorder with a high mortality affecting about 0.5% of women, where no evidence-based effective treatment exists. The pathogenesis likely involves genetic and environmental alterations. We hypothesized that a disrupted gut microbiota contributes to AN pathology. In analyses comparing 70 AN with 77 healthy females, we found multiple taxa, functional modules, structural variants and growth rates of bacterial gut microbiota, and viral gut microbiota that were altered in AN with parts of these perturbations linked to estimates of eating behavior and mental health. In silico, causal inference analyses implied bacterial metabolites mediated parts of the impact of altered gut microbiota on AN behavior, and in vivo, fecal microbiota transplantation from AN cases to germ-free mice induced a lower body weight and hypothalamic and adipose tissue gene expressions related to aberrant energy metabolism and eating and mental behavior.
Institute	Örebro University
Last Name	McGlinchey
First Name	Aidan
Address	Room 2217, Södra Grev Rosengatan 30, 70362 Örebro
Email	aidan.mcglinchey@oru.se
Phone	+46 0736485638
Submit Date	2022-05-18
Raw Data Available	Yes
Raw Data File Type(s)	mzdata.xml
Analysis Type Detail	LC-MS
Release Date	2023-02-27
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR001610
Project DOI:	doi: 10.21228/M8KT5B
Project Title:	Disrupted intestinal microbiota contributes to the pathogenesis of anorexia nervosa
Project Summary:	Anorexia nervosa (AN) is an eating disorder with a high mortality affecting about 0.5% of women, where no evidence-based effective treatment exists. The pathogenesis likely involves genetic and environmental alterations. We hypothesized that a disrupted gut microbiota contributes to AN pathology. In analyses comparing 70 AN with 77 healthy females, we found multiple taxa, functional modules, structural variants and growth rates of bacterial gut microbiota, and viral gut microbiota that were altered in AN with parts of these perturbations linked to estimates of eating behavior and mental health. In silico, causal inference analyses implied bacterial metabolites mediated parts of the impact of altered gut microbiota on AN behavior, and in vivo, fecal microbiota transplantation from AN cases to germ-free mice induced a lower body weight and hypothalamic and adipose tissue gene expressions related to aberrant energy metabolism and eating and mental behavior.
Institute:	Örebro University
Last Name:	McGlinchey
First Name:	Aidan
Address:	Room 2217, Södra Grev Rosengatan 30, 70362 Örebro
Email:	aidan.mcglinchey@oru.se
Phone:	+46 0736485638

Subject:

Subject ID:	SU002589
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Gender:	Male and female

Factors:

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

mb_sample_id	local_sample_id	Category
SA249292	sample_0068	1 - Healthy
SA249293	sample_0067	1 - Healthy
SA249294	sample_0119	1 - Healthy
SA249295	sample_0115	1 - Healthy
SA249296	sample_0066	1 - Healthy
SA249297	sample_0073	1 - Healthy
SA249298	sample_0076	1 - Healthy
SA249299	sample_0001	1 - Healthy
SA249300	sample_0074	1 - Healthy
SA249301	sample_0120	1 - Healthy
SA249302	sample_0072	1 - Healthy
SA249303	sample_0122	1 - Healthy
SA249304	sample_0052	1 - Healthy
SA249305	sample_0050	1 - Healthy
SA249306	sample_0049	1 - Healthy
SA249307	sample_0127	1 - Healthy
SA249308	sample_0053	1 - Healthy
SA249309	sample_0054	1 - Healthy
SA249310	sample_0124	1 - Healthy
SA249311	sample_0057	1 - Healthy
SA249312	sample_0056	1 - Healthy
SA249313	sample_0114	1 - Healthy
SA249314	sample_0113	1 - Healthy
SA249315	sample_0096	1 - Healthy
SA249316	sample_0104	1 - Healthy
SA249317	sample_0094	1 - Healthy
SA249318	sample_0105	1 - Healthy
SA249319	sample_0097	1 - Healthy
SA249320	sample_0098	1 - Healthy
SA249321	sample_0101	1 - Healthy
SA249322	sample_0100	1 - Healthy
SA249323	sample_0099	1 - Healthy
SA249324	sample_0092	1 - Healthy
SA249325	sample_0091	1 - Healthy
SA249326	sample_0108	1 - Healthy
SA249327	sample_0083	1 - Healthy
SA249328	sample_0109	1 - Healthy
SA249329	sample_0110	1 - Healthy
SA249330	sample_0107	1 - Healthy
SA249331	sample_0086	1 - Healthy
SA249332	sample_0090	1 - Healthy
SA249333	sample_0088	1 - Healthy
SA249334	sample_0087	1 - Healthy
SA249335	sample_0128	1 - Healthy
SA249336	sample_0051	1 - Healthy
SA249337	sample_0021	1 - Healthy
SA249338	sample_0020	1 - Healthy
SA249339	sample_0018	1 - Healthy
SA249340	sample_0143	1 - Healthy
SA249341	sample_0024	1 - Healthy
SA249342	sample_0026	1 - Healthy
SA249343	sample_0136	1 - Healthy
SA249344	sample_0137	1 - Healthy
SA249345	sample_0129	1 - Healthy
SA249346	sample_0016	1 - Healthy
SA249347	sample_0015	1 - Healthy
SA249348	sample_0009	1 - Healthy
SA249349	sample_0007	1 - Healthy
SA249350	sample_0006	1 - Healthy
SA249351	sample_0148	1 - Healthy
SA249352	sample_0010	1 - Healthy
SA249353	sample_0011	1 - Healthy
SA249354	sample_0144	1 - Healthy
SA249355	sample_0013	1 - Healthy
SA249356	sample_0012	1 - Healthy
SA249357	sample_0031	1 - Healthy
SA249358	sample_0027	1 - Healthy
SA249359	sample_0134	1 - Healthy
SA249360	sample_0130	1 - Healthy
SA249361	sample_0133	1 - Healthy
SA249362	sample_0042	1 - Healthy
SA249363	sample_0040	1 - Healthy
SA249364	sample_0036	1 - Healthy
SA249365	sample_0034	1 - Healthy
SA249366	sample_0032	1 - Healthy
SA249367	sample_0044	1 - Healthy
SA249368	sample_0045	1 - Healthy
SA249369	sample_0037	1 - Healthy
SA249370	sample_0126	2 - Anorexia
SA249371	sample_0132	2 - Anorexia
SA249372	sample_0106	2 - Anorexia
SA249373	sample_0145	2 - Anorexia
SA249374	sample_0147	2 - Anorexia
SA249375	sample_0146	2 - Anorexia
SA249376	sample_0103	2 - Anorexia
SA249377	sample_0131	2 - Anorexia
SA249378	sample_0123	2 - Anorexia
SA249379	sample_0118	2 - Anorexia
SA249380	sample_0139	2 - Anorexia
SA249381	sample_0138	2 - Anorexia
SA249382	sample_0135	2 - Anorexia
SA249383	sample_0116	2 - Anorexia
SA249384	sample_0140	2 - Anorexia
SA249385	sample_0112	2 - Anorexia
SA249386	sample_0142	2 - Anorexia
SA249387	sample_0117	2 - Anorexia
SA249388	sample_0121	2 - Anorexia
SA249389	sample_0141	2 - Anorexia
SA249390	sample_0111	2 - Anorexia
SA249391	sample_0125	2 - Anorexia

Showing page 1 of 2 Results: 1 2 Next Showing results 1 to 100 of 148

Collection:

Collection ID:	CO002582
Collection Summary:	The metabolites listed as gut microbiota-related metabolites is based on literature mining. Serum samples were randomized, and the sample preparation was done as described previously. Shortly, 400 μL of MeOH containing internal standards (heptadecanoic acid, deuterium-labeled DL-valine, deuterium- labeled succinic acid, and deuterium-labeled glutamic acid, c= 1 μg/ml) was added to 30 μl of the serum samples which were vortex mixed and incubated on ice for 30 min after which they were centrifuged (9400 × g, 3 min) and 350 μL of the supernatant was collected after centrifugation. The solvent was evaporated to dryness and 25 μL of MOX reagent was added and the sample was incubated for 60 min at 45 °C. 25 μL of N-Methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA) was added and after 60 min incubation at 45 °C 25 μL of the retention index standard mixture (n-alkanes, c=10 μg/ml) was added. The analyses were done by Agilent 7890B gas chromatography (GC) coupled to Agilent 7200 auadrupole time-of-flight mass spectrometry (Q-TOF MS) (Agilent Technologies, Palo Alto, CA, USA). Injection volume 1 μL with 100:1 split on PTV at 70 °C, heating to 300 °C at 120 °C /min. Column: Zebron ZB- SemiVolatiles. Length: 20m, I.D. 0.18mm, film thickness: 0.18 μm. With initial Helium flow 1.2 mL/min, increasing to 2.4 mL/min after 16 mins. Oven temperature program: 50 °C (5 min), then to 270°C at 20 °/min and then to 300 at 40 °/min (5 min). EI source: 250 °C, 70 eV electron energy, 35μA emission, solvent delay 3 min. Mass range 55 to 650 amu, acquisition rate 5 spectra/s, acquisition time 200 ms/spectrum. Quad at 150 °C, 1.5 mL/min N2 collision flow, aux-2 temperature: 280 °C. Calibration curves were constructed using alanine, citric acid, fumaric acid, glutamic acid, glycine, lactic acid, malic acid, 2-hydroxybutyric acid, 3-hydroxybutyric acid, linoleic acid, oleic acid, palmitic acid, stearic acid, cholesterol, fructose, glutamine, indole-3-propionic acid, isoleucine, leucine, proline, succinic acid, valine, asparagine, aspartic acid, arachidonic acid, glycerol-3-phosphate, lysine, methionine, ornithine, phenylalanine, serine and threonine purchased from Sigma-Aldrich (St. Louis, MO, USA) at concentration range of 0.1 to 80 μg/ml. An aliquot of each sample was collected and pooled and used as quality control samples, together with National Institute of Standards and Technology (NIST) CRM1950 serum sample, an in-house pooled serum sample. The relative standard deviation of the concentrations was on average 16% for the pooled QC samples, 10% for the NIST samples. Analysis of serum bile acids and serum semipolar metabolites The sample preparation procedure was performed as described previously 80 . The plate was preconditioned with 450 μL acetonitrile before the addition of 100 μL of sample and 10 μL of PFAS and BA internal standard mixture (200 ng/mL and 1000 ng/mL respectively). Thereafter, 450 μL of acetonitrile containing 1% formic acid were added to each well and the samples extracted using a 10” vacuum manifold. The eluate was evaporated to dryness under nitrogen gas flow and reconstituted to 80 μL of MeOH/2 mM aqueous NH4AC. Chromatographic separation was carried out using an Acquity UPLC BEH C18 column (100 mm × 2.1 mm i.d., 1.7 μm particle size), fitted with a C18 precolumn (Waters Corporation, Wexford, Ireland). Mobile phase A consisted of H2O:MeOH (v/v 70:30) and mobile phase B of MeOH with both phases containing 2mM ammonium acetate as an ionization agent. The flow rate was set at 0.4 mLmin-1 with the elution gradient as follows: 0-1.5 min, mobile phase B was increased from 5% to 30%; 1.5-4.5 min, mobile phase B increased to 70%; 4.5-7.5 min, mobile phase B increased to 100% and held for 5.5 min. A post-time of 5 min was used to regain the initial conditions for the next analysis. The total run time per sample was 18 min. The dual ESI ionization source was settings were as follows: capillary voltage was 4.5 kV, nozzle voltage 1500 V, N2 pressure in the nebulized was 21 psi and the N2 flow rate and temperature as sheath gas was 11 L min-1 and 379 °C, respectively. In order to obtain accurate mass spectra in MS scan, the m/z range was set to 100-1700 in negative ion mode. MassHunter B.06.01 software (Agilent Technologies, Santa Clara, CA, USA) was used for all data acquisition. Identification of compounds was done by in-house spectral library using MS (and retention time), MS/MS information. Quantitation was based on a matrix matched calibration curve spiked with native compounds. The calibration curve consisted of concentrations ranging from 0 – 1600 ng mL -1 for BAs. The RSD for the BAs was on average 17.8 % for the QC samples and 19.4 % for the NIST samples.
Sample Type:	Blood (serum)
Storage Conditions:	-80℃

Treatment:

Treatment ID:	TR002601
Treatment Summary:	No treatment was applied.

Sample Preparation:

Sampleprep ID:	SP002595
Sampleprep Summary:	Serum samples were randomized, and the sample preparation was done as described previously 43,79 . Shortly, 400 μL of MeOH containing internal standards (heptadecanoic acid, deuterium-labeled DL-valine, deuterium- labeled succinic acid, and deuterium-labeled glutamic acid, c= 1 μg/ml) was added to 30 μl of the serum samples which were vortex mixed and incubated on ice for 30 min after which they were centrifuged (9400 × g, 3 min) and 350 μL of the supernatant was collected after centrifugation. The solvent was evaporated to dryness and 25 μL of MOX reagent was added and the sample was incubated for 60 min at 45 °C. 25 μL of N-Methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA) was added and after 60 min incubation at 45 °C 25 μL of the retention index standard mixture (n-alkanes, c=10 μg/ml) was added. The analyses were done by Agilent 7890B gas chromatography (GC) coupled to Agilent 7200 auadrupole time-of-flight mass spectrometry (Q-TOF MS) (Agilent Technologies, Palo Alto, CA, USA). Injection volume 1 μL with 100:1 split on PTV at 70 °C, heating to 300 °C at 120 °C /min. Column: Zebron ZB- SemiVolatiles. Length: 20m, I.D. 0.18mm, film thickness: 0.18 μm. With initial Helium flow 1.2 mL/min, increasing to 2.4 mL/min after 16 mins. Oven temperature program: 50 °C (5 min), then to 270°C at 20 °/min and then to 300 at 40 °/min (5 min). EI source: 250 °C, 70 eV electron energy, 35μA emission, solvent delay 3 min. Mass range 55 to 650 amu, acquisition rate 5 spectra/s, acquisition time 200 ms/spectrum. Quad at 150 °C, 1.5 mL/min N2 collision flow, aux-2 temperature: 280 °C. Calibration curves were constructed using alanine, citric acid, fumaric acid, glutamic acid, glycine, lactic acid, malic acid, 2-hydroxybutyric acid, 3-hydroxybutyric acid, linoleic acid, oleic acid, palmitic acid, stearic acid, cholesterol, fructose, glutamine, indole-3-propionic acid, isoleucine, leucine, proline, succinic acid, valine, asparagine, aspartic acid, arachidonic acid, glycerol-3-phosphate, lysine, methionine, ornithine, phenylalanine, serine and threonine purchased from Sigma-Aldrich (St. Louis, MO, USA) at concentration range of 0.1 to 80 μg/ml. An aliquot of each sample was collected and pooled and used as quality control samples, together with National Institute of Standards and Technology (NIST) CRM1950 serum sample, an in-house pooled serum sample. The relative standard deviation of the concentrations was on average 16% for the pooled QC samples, 10% for the NIST samples. Analysis of serum bile acids and serum semipolar metabolites The sample preparation procedure was performed as described previously 80 . The plate was preconditioned with 450 μL acetonitrile before the addition of 100 μL of sample and 10 μL of PFAS and BA internal standard mixture (200 ng/mL and 1000 ng/mL respectively). Thereafter, 450 μL of acetonitrile containing 1% formic acid were added to each well and the samples extracted using a 10” vacuum manifold. The eluate was evaporated to dryness under nitrogen gas flow and reconstituted to 80 μL of MeOH/2 mM aqueous NH4AC. Chromatographic separation was carried out using an Acquity UPLC BEH C18 column (100 mm × 2.1 mm i.d., 1.7 μm particle size), fitted with a C18 precolumn (Waters Corporation, Wexford, Ireland). Mobile phase A consisted of H2O:MeOH (v/v 70:30) and mobile phase B of MeOH with both phases containing 2mM ammonium acetate as an ionization agent. The flow rate was set at 0.4 mLmin-1 with the elution gradient as follows: 0-1.5 min, mobile phase B was increased from 5% to 30%; 1.5-4.5 min, mobile phase B increased to 70%; 4.5-7.5 min, mobile phase B increased to 100% and held for 5.5 min. A post-time of 5 min was used to regain the initial conditions for the next analysis. The total run time per sample was 18 min. The dual ESI ionization source was settings were as follows: capillary voltage was 4.5 kV, nozzle voltage 1500 V, N2 pressure in the nebulized was 21 psi and the N2 flow rate and temperature as sheath gas was 11 L min-1 and 379 °C, respectively. In order to obtain accurate mass spectra in MS scan, the m/z range was set to 100-1700 in negative ion mode. MassHunter B.06.01 software (Agilent Technologies, Santa Clara, CA, USA) was used for all data acquisition. Identification of compounds was done by in-house spectral library using MS (and retention time), MS/MS information. Quantitation was based on a matrix matched calibration curve spiked with native compounds. The calibration curve consisted of concentrations ranging from 0 – 1600 ng mL -1 for BAs. The RSD for the BAs was on average 17.8 % for the QC samples and 19.4 % for the NIST samples.

Sampleprep ID:

SP002595

Sampleprep Summary:

Serum samples were randomized, and the sample preparation was done as described previously 43,79 . Shortly, 400 μL of MeOH containing internal standards (heptadecanoic acid, deuterium-labeled DL-valine, deuterium- labeled succinic acid, and deuterium-labeled glutamic acid, c= 1 μg/ml) was added to 30 μl of the serum samples which were vortex mixed and incubated on ice for 30 min after which they were centrifuged (9400 × g, 3 min) and 350 μL of the supernatant was collected after centrifugation. The solvent was evaporated to dryness and 25 μL of MOX reagent was added and the sample was incubated for 60 min at 45 °C. 25 μL of N-Methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA) was added and after 60 min incubation at 45 °C 25 μL of the retention index standard mixture (n-alkanes, c=10 μg/ml) was added. The analyses were done by Agilent 7890B gas chromatography (GC) coupled to Agilent 7200 auadrupole time-of-flight mass spectrometry (Q-TOF MS) (Agilent Technologies, Palo Alto, CA, USA). Injection volume 1 μL with 100:1 split on PTV at 70 °C, heating to 300 °C at 120 °C /min. Column: Zebron ZB- SemiVolatiles. Length: 20m, I.D. 0.18mm, film thickness: 0.18 μm. With initial Helium flow 1.2 mL/min, increasing to 2.4 mL/min after 16 mins. Oven temperature program: 50 °C (5 min), then to 270°C at 20 °/min and then to 300 at 40 °/min (5 min). EI source: 250 °C, 70 eV electron energy, 35μA emission, solvent delay 3 min. Mass range 55 to 650 amu, acquisition rate 5 spectra/s, acquisition time 200 ms/spectrum. Quad at 150 °C, 1.5 mL/min N2 collision flow, aux-2 temperature: 280 °C. Calibration curves were constructed using alanine, citric acid, fumaric acid, glutamic acid, glycine, lactic acid, malic acid, 2-hydroxybutyric acid, 3-hydroxybutyric acid, linoleic acid, oleic acid, palmitic acid, stearic acid, cholesterol, fructose, glutamine, indole-3-propionic acid, isoleucine, leucine, proline, succinic acid, valine, asparagine, aspartic acid, arachidonic acid, glycerol-3-phosphate, lysine, methionine, ornithine, phenylalanine, serine and threonine purchased from Sigma-Aldrich (St. Louis, MO, USA) at concentration range of 0.1 to 80 μg/ml. An aliquot of each sample was collected and pooled and used as quality control samples, together with National Institute of Standards and Technology (NIST) CRM1950 serum sample, an in-house pooled serum sample. The relative standard deviation of the concentrations was on average 16% for the pooled QC samples, 10% for the NIST samples. Analysis of serum bile acids and serum semipolar metabolites The sample preparation procedure was performed as described previously 80 . The plate was preconditioned with 450 μL acetonitrile before the addition of 100 μL of sample and 10 μL of PFAS and BA internal standard mixture (200 ng/mL and 1000 ng/mL respectively). Thereafter, 450 μL of acetonitrile containing 1% formic acid were added to each well and the samples extracted using a 10” vacuum manifold. The eluate was evaporated to dryness under nitrogen gas flow and reconstituted to 80 μL of MeOH/2 mM aqueous NH4AC. Chromatographic separation was carried out using an Acquity UPLC BEH C18 column (100 mm × 2.1 mm i.d., 1.7 μm particle size), fitted with a C18 precolumn (Waters Corporation, Wexford, Ireland). Mobile phase A consisted of H2O:MeOH (v/v 70:30) and mobile phase B of MeOH with both phases containing 2mM ammonium acetate as an ionization agent. The flow rate was set at 0.4 mLmin-1 with the elution gradient as follows: 0-1.5 min, mobile phase B was increased from 5% to 30%; 1.5-4.5 min, mobile phase B increased to 70%; 4.5-7.5 min, mobile phase B increased to 100% and held for 5.5 min. A post-time of 5 min was used to regain the initial conditions for the next analysis. The total run time per sample was 18 min. The dual ESI ionization source was settings were as follows: capillary voltage was 4.5 kV, nozzle voltage 1500 V, N2 pressure in the nebulized was 21 psi and the N2 flow rate and temperature as sheath gas was 11 L min-1 and 379 °C, respectively. In order to obtain accurate mass spectra in MS scan, the m/z range was set to 100-1700 in negative ion mode. MassHunter B.06.01 software (Agilent Technologies, Santa Clara, CA, USA) was used for all data acquisition. Identification of compounds was done by in-house spectral library using MS (and retention time), MS/MS information. Quantitation was based on a matrix matched calibration curve spiked with native compounds. The calibration curve consisted of concentrations ranging from 0 – 1600 ng mL -1 for BAs. The RSD for the BAs was on average 17.8 % for the QC samples and 19.4 % for the NIST samples.

Combined analysis:

Analysis ID	AN004093
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	Agilent 7890B
Column	Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um)
MS Type	ESI
MS instrument type	QTOF
MS instrument name	Agilent 7200 QTOF
Ion Mode	NEGATIVE
Units	Raw output from software

Chromatography:

Chromatography ID:	CH003031
Instrument Name:	Agilent 7890B
Column Name:	Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um)
Column Temperature:	70
Flow Gradient:	The flow rate was set at 0.4 mLmin-1 with the elution gradient as follows: 0-1.5 min, mobile phase B was increased from 5% to 30%; 1.5-4.5 min, mobile phase B increased to 70%; 4.5-7.5 min, mobile phase B increased to 100% and held for 5.5 min. A post-time of 5 min was used to regain the initial conditions for the next analysis. The total run time per sample was 18 min
Flow Rate:	0.4ml/min
Solvent A:	70% water/30% methanol
Solvent B:	100% methanol
Chromatography Type:	Reversed phase

MS:

MS ID:	MS003840
Analysis ID:	AN004093
Instrument Name:	Agilent 7200 QTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	MassHunter B.06.01 software (Agilent Technologies, Santa Clara, CA, USA) was used for all data acquisition. Identification of compounds was done by in-house spectral library using MS (and retention time), MS/MS information. Quantitation was based on a matrix matched calibration curve spiked with native compounds. The calibration curve consisted of concentrations ranging from 0 – 1600 ng mL -1 for BAs. The RSD for the BAs was on average 17.8 % for the QC samples and 19.4 % for the NIST samples.
Ion Mode:	NEGATIVE