#METABOLOMICS WORKBENCH Elisa_Zubeldia_20220719_042035 DATATRACK_ID:3355 STUDY_ID:ST002236 ANALYSIS_ID:AN003646 PROJECT_ID:PR001426
VERSION             	1
CREATED_ON             	July 27, 2022, 9:45 am
#PROJECT
PR:PROJECT_TITLE                 	Metabolomics approach to the study of food allergy
PR:PROJECT_SUMMARY               	This project focuses on the metabolomics study of different types of food
PR:PROJECT_SUMMARY               	allergy with various cohorts.
PR:INSTITUTE                     	Institute of Applied Molecular Medicine
PR:DEPARTMENT                    	Basic Medical Sciences. Faculty of Medicine
PR:LAST_NAME                     	Pérez-Gordo
PR:FIRST_NAME                    	Marina
PR:ADDRESS                       	Avda. Monteprincipe s/n 28668 Boadilla del Monte, Madrid, España
PR:EMAIL                         	marina.perezgordo@ceu.es
PR:PHONE                         	Tlf: 91 372 47 00 ext. 14675
#STUDY
ST:STUDY_TITLE                   	The impact of IgE in gut and serum metabolomes in a murine experimental model of
ST:STUDY_TITLE                   	allergic enteritis
ST:STUDY_TYPE                    	Case-control study
ST:STUDY_SUMMARY                 	The pathological mechanism of the gastrointestinal forms of food allergies is
ST:STUDY_SUMMARY                 	less understood in comparison to other clinical phenotypes, such as asthma, and
ST:STUDY_SUMMARY                 	anaphylaxis, partly due to difficulty in the access to intestinal tissues and
ST:STUDY_SUMMARY                 	because of a highly complex interplay between microbiota and intestinal mucosa.
ST:STUDY_SUMMARY                 	Importantly, a high level of IgE is a poor prognostic factor in gastrointestinal
ST:STUDY_SUMMARY                 	allergies. This study aimed to investigate how IgE influences the development of
ST:STUDY_SUMMARY                 	intestinal inflammation and the metabolome in allergic enteritis (AE), using IgE
ST:STUDY_SUMMARY                 	knock-in (IgEki) mice expressing high levels of IgE. Ovalbumin-sensitized and
ST:STUDY_SUMMARY                 	egg-white diet fed (OVA/EW) BALB/c WT mice developed moderate AE, whereas OVA/EW
ST:STUDY_SUMMARY                 	IgEki mice induced more aggravated intestinal inflammation with enhanced
ST:STUDY_SUMMARY                 	eosinophil accumulation.
ST:INSTITUTE                     	Institute of Applied Molecular Medicine
ST:LAST_NAME                     	Zubeldia-Varela
ST:FIRST_NAME                    	Elisa
ST:ADDRESS                       	Avda. Monteprincipe s/n 28668 Boadilla del Monte, Madrid, España
ST:EMAIL                         	elisa.zubeldiavarela@ceu.es
ST:PHONE                         	Tlf: 91 372 47 00 ext. 14675
ST:NUM_GROUPS                    	4 groups: BALB/c wild type (WT) and IgE knock-in mice (C.Ighg1tm1.1Pyu) in the
ST:NUM_GROUPS                    	BALB/c background, both sensitised and non-sensitised to ovalbumin.
ST:TOTAL_SUBJECTS                	29
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data
SUBJECT_SAMPLE_FACTORS           	IgEki (Only EW) 1	IgEki (Only EW) 1F	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=IgEki (Only EW) 1F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (Only EW) 1	IgEki (Only EW) 1S	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=IgEki (Only EW) 1S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (Only EW) 2	IgEki (Only EW) 2F	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=IgEki (Only EW) 2F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (Only EW) 2	IgEki (Only EW) 2S	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=IgEki (Only EW) 2S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (Only EW) 3	IgEki (Only EW) 3F	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=IgEki (Only EW) 3F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (Only EW) 3	IgEki (Only EW) 3S	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=IgEki (Only EW) 3S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (Only EW) 4	IgEki (Only EW) 4F	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=IgEki (Only EW) 4F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (Only EW) 4	IgEki (Only EW) 4S	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=IgEki (Only EW) 4S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (Only EW) 5	IgEki (Only EW) 5F	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=IgEki (Only EW) 5F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (Only EW) 5	IgEki (Only EW) 5S	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=IgEki (Only EW) 5S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (Only EW) 6	IgEki (Only EW) 6F	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=IgEki (Only EW) 6F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (Only EW) 6	IgEki (Only EW) 6S	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=IgEki (Only EW) 6S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (Only EW) 7	IgEki (Only EW) 7F	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=IgEki (Only EW) 7F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (Only EW) 7	IgEki (Only EW) 7S	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=IgEki (Only EW) 7S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (OVA_EW) 1	IgEki (OVA_EW) 1F	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=IgEki (OVA_EW) 1F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (OVA_EW) 1	IgEki (OVA_EW) 1S	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=IgEki (OVA_EW) 1S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (OVA_EW) 2	IgEki (OVA_EW) 2F	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=IgEki (OVA_EW) 2F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (OVA_EW) 2	IgEki (OVA_EW) 2S	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=IgEki (OVA_EW) 2S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (OVA_EW) 3	IgEki (OVA_EW) 3F	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=IgEki (OVA_EW) 3F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (OVA_EW) 3	IgEki (OVA_EW) 3S	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=IgEki (OVA_EW) 3S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (OVA_EW) 4	IgEki (OVA_EW) 4F	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=IgEki (OVA_EW) 4F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (OVA_EW) 4	IgEki (OVA_EW) 4S	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=IgEki (OVA_EW) 4S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (OVA_EW) 5	IgEki (OVA_EW) 5F	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=IgEki (OVA_EW) 5F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (OVA_EW) 5	IgEki (OVA_EW) 5S	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=IgEki (OVA_EW) 5S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (OVA_EW) 6	IgEki (OVA_EW) 6F	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=IgEki (OVA_EW) 6F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (OVA_EW) 6	IgEki (OVA_EW) 6S	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=IgEki (OVA_EW) 6S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (OVA_EW) 7	IgEki (OVA_EW) 7F	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=IgEki (OVA_EW) 7F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (OVA_EW) 7	IgEki (OVA_EW) 7S	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=IgEki (OVA_EW) 7S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (OVA_EW) 8	IgEki (OVA_EW) 8F	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=IgEki (OVA_EW) 8F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	IgEki (OVA_EW) 8	IgEki (OVA_EW) 8S	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=IgEki (OVA_EW) 8S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (Only EW) 1	WT (Only EW) 1F	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=WT (Only EW) 1F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (Only EW) 1	WT (Only EW) 1S	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=WT (Only EW) 1S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (Only EW) 2	WT (Only EW) 2F	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=WT (Only EW) 2F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (Only EW) 2	WT (Only EW) 2S	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=WT (Only EW) 2S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (Only EW) 3	WT (Only EW) 3F	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=WT (Only EW) 3F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (Only EW) 3	WT (Only EW) 3S	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=WT (Only EW) 3S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (Only EW) 4	WT (Only EW) 4F	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=WT (Only EW) 4F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (Only EW) 4	WT (Only EW) 4S	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=WT (Only EW) 4S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (Only EW) 5	WT (Only EW) 5F	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=WT (Only EW) 5F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (Only EW) 5	WT (Only EW) 5S	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=WT (Only EW) 5S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (Only EW) 6	WT (Only EW) 6F	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=WT (Only EW) 6F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (Only EW) 6	WT (Only EW) 6S	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=WT (Only EW) 6S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (Only EW) 7	WT (Only EW) 7F	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=WT (Only EW) 7F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (Only EW) 7	WT (Only EW) 7S	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	RAW_FILE_NAME=WT (Only EW) 7S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (OVA_EW) 1	WT (OVA_EW) 1F	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=WT (OVA_EW) 1F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (OVA_EW) 1	WT (OVA_EW) 1S	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=WT (OVA_EW) 1S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (OVA_EW) 2	WT (OVA_EW) 2F	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=WT (OVA_EW) 2F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (OVA_EW) 2	WT (OVA_EW) 2S	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=WT (OVA_EW) 2S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (OVA_EW) 3	WT (OVA_EW) 3F	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=WT (OVA_EW) 3F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (OVA_EW) 3	WT (OVA_EW) 3S	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=WT (OVA_EW) 3S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (OVA_EW) 4	WT (OVA_EW) 4F	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=WT (OVA_EW) 4F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (OVA_EW) 4	WT (OVA_EW) 4S	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=WT (OVA_EW) 4S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (OVA_EW) 5	WT (OVA_EW) 5F	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=WT (OVA_EW) 5F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (OVA_EW) 5	WT (OVA_EW) 5S	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=WT (OVA_EW) 5S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (OVA_EW) 6	WT (OVA_EW) 6F	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=WT (OVA_EW) 6F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (OVA_EW) 6	WT (OVA_EW) 6S	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=WT (OVA_EW) 6S CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (OVA_EW) 7	WT (OVA_EW) 7F	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=WT (OVA_EW) 7F CE/GC/LC Pos/LC Neg
SUBJECT_SAMPLE_FACTORS           	WT (OVA_EW) 7	WT (OVA_EW) 7S	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	RAW_FILE_NAME=WT (OVA_EW) 7S CE/GC/LC Pos/LC Neg
#COLLECTION
CO:COLLECTION_SUMMARY            	Faecal and serum samples were obtained and stored at -80°C until metabolite
CO:COLLECTION_SUMMARY            	extraction.
CO:SAMPLE_TYPE                   	Feces; Blood (serum)
CO:STORAGE_CONDITIONS            	-80?
#TREATMENT
TR:TREATMENT_SUMMARY             	Mice  (female, 6 to 8 weeks old) were sensitized by an intraperitoneal
TR:TREATMENT_SUMMARY             	injection with 10 µg of ovalbumin (OVA, Sigma-Aldrich) and 1 mg of Alum
TR:TREATMENT_SUMMARY             	(ThermoFisher Scientific) in 500 µl of PBS or treated only with PBS twice as
TR:TREATMENT_SUMMARY             	non-sensitized controls, at a two-week interval.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	"FAECES. Frozen faeces samples were lyophilized overnight at -80°C. Faecal
SP:SAMPLEPREP_SUMMARY            	pellets were doubly extracted (i) first with MeOH:MTBE (4:1, v/v) and (ii) with
SP:SAMPLEPREP_SUMMARY            	MeOH:H2O (4:1, v/v). Equal volumes (300 µL) of the supernatants from (i) and
SP:SAMPLEPREP_SUMMARY            	(ii) were then mixed and aliquoted for subsequent analysis. SERUM. Serum samples
SP:SAMPLEPREP_SUMMARY            	(100 µL) were deproteinized by an addition of 300 µL of cold acetonitrile (1:3
SP:SAMPLEPREP_SUMMARY            	v/v) and homogenized for 15 min in a vortex. After a 10 min ice bath, samples
SP:SAMPLEPREP_SUMMARY            	were centrifuged (16000 x g, 4°C, 20 min) and stored at -20°C until the start
SP:SAMPLEPREP_SUMMARY            	of sample treatment. The resulting supernatants were filtered through 0.22 µm
SP:SAMPLEPREP_SUMMARY            	for LC-MS, 80 µL were transferred into an analytical vial for their analysis.
SP:SAMPLEPREP_SUMMARY            	For CE-MS, 80 µL of the supernatants were evaporated to dryness using a
SP:SAMPLEPREP_SUMMARY            	Speedvac Concentrator and reconstituted in 80 µL of MilliQ® water containing
SP:SAMPLEPREP_SUMMARY            	an internal standard (0.2 mM L-methionine sulfone) and 0.1 M FA. Finally, for
SP:SAMPLEPREP_SUMMARY            	GC-MS, 120 µL of the corresponding aliquots were evaporated to dryness using a
SP:SAMPLEPREP_SUMMARY            	Speedvac Concentrator, followed by the addition of 10 µL of O-methoxyamine
SP:SAMPLEPREP_SUMMARY            	hydrochloride (15 mg/mL) in pyridine for methoximation. After gently vortexing,
SP:SAMPLEPREP_SUMMARY            	the vials were incubated in darkness at room temperature for 16 h. Then, 10 µL
SP:SAMPLEPREP_SUMMARY            	of BSTFA with 1 % TMCS (v/v) were added and samples were vortexed for 5 min,
SP:SAMPLEPREP_SUMMARY            	silylation was carried out for 1 h at 70 °C and finally 120 µL of C18:0 methyl
SP:SAMPLEPREP_SUMMARY            	ester (10 mg/L in heptane) were added as an internal standard and samples were
SP:SAMPLEPREP_SUMMARY            	mixed again by gentle vortexing. Eight blank samples were prepared for GC-MS by
SP:SAMPLEPREP_SUMMARY            	the same procedure of extraction and derivatization. "
SP:SAMPLEPREP_PROTOCOL_FILENAME  	Protocolsamples.docx
SP:PROCESSING_METHOD             	Lyophilization, homogenization, protein precipitation and metabolite extraction
SP:PROCESSING_STORAGE_CONDITIONS 	On ice
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	For LC-MS, 0.5 µL of sample were injected into a Zorbax Extend C18 (50 x 2.1
CH:CHROMATOGRAPHY_SUMMARY        	mm, 1.8 µm) maintained at 60 °C. The flow rate was set at 0.6 mL/min. . The
CH:CHROMATOGRAPHY_SUMMARY        	flow rate was set at 0.6 mL/min. The elution gradient involved a mobile phase
CH:CHROMATOGRAPHY_SUMMARY        	consisting of: (A) 0.1% formic acid (FA) in water and (B) 0.1% FA in
CH:CHROMATOGRAPHY_SUMMARY        	acetonitrile (ACN). The gradient was 5% B (0–1 min), 5 to 80% B (1–7 min),
CH:CHROMATOGRAPHY_SUMMARY        	80 to 100% B (7–11.5 min), and 100 to 5% B (11.5–12 min). The system was
CH:CHROMATOGRAPHY_SUMMARY        	finally held at 5% B for 3 min to re-equilibrate the system (15 min of total
CH:CHROMATOGRAPHY_SUMMARY        	analysis time). The ESI data were acquired in both modes, ESI (+) and ESI (-),
CH:CHROMATOGRAPHY_SUMMARY        	in separate runs. The capillary voltage was set at 3,000V for ESI (+) and 4,000V
CH:CHROMATOGRAPHY_SUMMARY        	for ESI (-). The drying gas flow rate was 12 L/min at 250 °C and gas nebulizer
CH:CHROMATOGRAPHY_SUMMARY        	at 52 psi; nozzle voltage was 1000V; fragmentor voltage was 175 V; skimmer and
CH:CHROMATOGRAPHY_SUMMARY        	octopole radio frequency voltage (OCT RF Vpp) were set to 65 and 750 V,
CH:CHROMATOGRAPHY_SUMMARY        	respectively. Data were collected in the centroid mode at a scan rate of 1.5
CH:CHROMATOGRAPHY_SUMMARY        	spectra per second.
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Agilent 1290
CH:COLUMN_NAME                   	Zorbax Extend C18 (50 x 2.1 mm, 1.8 µm)
CH:COLUMN_TEMPERATURE            	60 °C
CH:METHODS_FILENAME              	Protocolmethods.docx
CH:SAMPLE_INJECTION              	0.5 µL
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
AN:DATA_FORMAT                   	.d
#MS
MS:INSTRUMENT_NAME               	Agilent 6545 QTOF
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	LC-MS analysis was performed on an Agilent HPLC system (1290 infinity II
MS:MS_COMMENTS                   	series, Agilent Technologies, Waldbronn, Germany), equipped with a degasser, two
MS:MS_COMMENTS                   	binary pumps, and a thermostated autosampler coupled with quadrupole-time of
MS:MS_COMMENTS                   	flight analyzer (Q-TOF), LC-MS (6545) system (Agilent Technologies, Waldbronn,
MS:MS_COMMENTS                   	Germany)
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Peak area
MS_METABOLITE_DATA_START
Samples	IgEki (Only EW) 1F	IgEki (Only EW) 2F	IgEki (Only EW) 3F	IgEki (Only EW) 4F	IgEki (Only EW) 5F	IgEki (Only EW) 6F	IgEki (Only EW) 7F	IgEki (OVA_EW) 1F	IgEki (OVA_EW) 2F	IgEki (OVA_EW) 3F	IgEki (OVA_EW) 4F	IgEki (OVA_EW) 5F	IgEki (OVA_EW) 6F	IgEki (OVA_EW) 7F	IgEki (OVA_EW) 8F	WT (Only EW) 1F	WT (Only EW) 2F	WT (Only EW) 3F	WT (Only EW) 4F	WT (Only EW) 5F	WT (Only EW) 6F	WT (Only EW) 7F	WT (OVA_EW) 1F	WT (OVA_EW) 2F	WT (OVA_EW) 3F	WT (OVA_EW) 4F	WT (OVA_EW) 5F	WT (OVA_EW) 6F	WT (OVA_EW) 7F	IgEki (Only EW) 1S	IgEki (Only EW) 2S	IgEki (Only EW) 3S	IgEki (Only EW) 4S	IgEki (Only EW) 5S	IgEki (Only EW) 6S	IgEki (Only EW) 7S	IgEki (OVA_EW) 1S	IgEki (OVA_EW) 2S	IgEki (OVA_EW) 3S	IgEki (OVA_EW) 4S	IgEki (OVA_EW) 5S	IgEki (OVA_EW) 6S	IgEki (OVA_EW) 7S	IgEki (OVA_EW) 8S	WT (Only EW) 1S	WT (Only EW) 2S	WT (Only EW) 3S	WT (Only EW) 4S	WT (Only EW) 5S	WT (Only EW) 6S	WT (Only EW) 7S	WT (OVA_EW) 1S	WT (OVA_EW) 2S	WT (OVA_EW) 3S	WT (OVA_EW) 4S	WT (OVA_EW) 5S	WT (OVA_EW) 6S	WT (OVA_EW) 7S
Factors	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:IgE knock-in mice | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Non-sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin	Genotype:BALB/c wild type | Ovalbumin sensitisation:Sensitised to ovalbumin
3-ketosphinganine	4060681	3738249	3873905	4084926	5228101	3373071	2482899	5104726	3237822	2448251	3804802	4948846	5634634	7124331	3500685	3774725	4028202	6802534	4855048	3496035	2890015	6408355	11487152	18293734	9148964	8275366	6282773	6055370	9797724	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Campesterol	326622	263556	296561	328889	286516	337992	296987	423334	410033	398003	430252	385130	525909	221004	320173	173366	244666	188998	294646	224241	152182	236437	229585	197786	216516	175209	159639	100581	265861	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Cholesterol	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	389260	189858	172615	48078	267741	331861	297018	312287	261003	248150	291215	159221	296913	328447	139332	134492	27346	61432	36127	182531	27197	289752	29008	116715	31639	34863	29556	222461	257019
D-erythro-Sphingosine C-20	1115243	741246	950660	1099922	1073915	1196034	712177	613215	850793	1341408	1170073	1275075	1294213	720916	1323557	781846	840579	480124	516769	417841	299259	690036	1225273	1253279	603887	769359	536390	770801	1118950	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
L-Lysine	631229	513361	506977	557421	536932	374681	465706	253938	728838	422551	442683	648521	438448	830816	790917	568740	488325	464638	684245	575127	588782	602380	880153	943668	1058216	792585	968926	973731	875837	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
L-Proline	1349578	1155915	1137215	925537	1209891	810301	965661	738379	1018577	921833	910998	1621338	853803	1218609	1176250	1277917	1083156	1190788	1888218	1175855	1122674	1917834	3904542	3716979	3419897	2422564	2791354	2560946	3037069	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
L-threo-Sphingosine C-18	1961831	1281297	2061506	1773603	2341811	2375960	1380151	1747772	3741785	4886231	5327192	7504572	5245007	2251904	6000302	1277352	1623049	1115010	1284863	988177	648225	1253871	2859928	2502951	1081878	1248938	988300	1439682	2284585	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
L-Tyrosine	399953	368837	312718	352157	342265	301377	308945	292421	402629	208813	253341	450155	272993	318100	292562	229453	234825	198315	299304	226015	240692	324814	306015	285971	310045	308232	323262	264199	277695	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Oleamide	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	789219	569326	4345482	779436	8141848	12387125	447895	15658831	15840139	10459224	13440841	4742280	11254017	10968036	1173952	3128262	306396	643972	322576	3576342	304192	12709618	372679	4612135	427662	328956	326051	10161570	4863142
Palmitic amide	0	0	265543	224748	225870	182372	0	0	206034	220554	190639	244041	239884	251026	167601	236918	309239	190551	0	224768	0	157485	0	0	0	0	0	174238	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
PC(14:0/0:0)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	280312	381662	330198	365828	595633	277528	333214	127925	112791	126431	210538	85196	133620	184880	148143	133567	177248	167162	196946	168273	186294	245806	189936	117378	159592	260807	137450	395402	330297
PC(16:0/0:0)	5746147	4167727	8348776	2179490	4400364	4490898	2740052	3575679	2509933	879443	2465646	1524296	3950572	427151	1696689	1461127	2046314	1598621	2132653	2050993	1725322	1416705	904838	770801	778195	992737	1959731	1122640	820028	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
PC(18:2/0:0)	574742	301680	418779	310071	283727	243431	258355	652244	389362	191416	417647	326260	479555	63311	334908	125970	139044	178857	188523	116602	164291	138966	156597	151805	153821	110952	129966	86015	126789	64079337	68007255	69923277	94633471	128875215	74092519	50948745	45374898	36615166	33327676	62561738	25328043	36279402	61728580	34042566	40374732	35086075	28281663	36724307	49174271	30395416	57399371	35347016	32311694	23848877	36052265	20349866	84989809	77614348
PE (18:1/0:0)	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	418972	339366	439557	485504	618943	455146	359225	280402	308121	259007	393802	114913	265379	364782	179581	146611	175060	199786	226536	211287	143192	372797	376629	150197	283228	255122	255018	389283	316738
PE(14:0/0:0)	770517	903210	979537	1003286	546326	697410	861698	1051814	1895496	767171	1394229	1722469	1588730	994037	664046	1111962	1052647	796564	833307	1195443	397314	1217664	2488450	1559601	1672925	2310330	2288691	2199548	2113394	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
PE(15:0/0:0)	5537493	6714058	9118425	7583051	4580857	6398122	6987969	6032761	11528501	5613009	10191906	8945011	12659974	7440376	3649398	11887272	11421575	5279256	6892263	12336844	1985538	12312974	15584531	13000085	11817190	14599722	13117777	10792852	16364840	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
PE(17:0/0:0)	721347	611761	687954	2492935	515984	972992	848127	639446	609563	250829	484363	894987	402138	476703	1143803	5777320	4099189	3199733	2554317	3813282	917375	2215848	4458763	1131787	3522565	5778771	3131659	2835598	4883012	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
PE(17:1/0:0)	482623	694752	371454	616450	278241	453443	325113	275495	330913	96577	156439	369940	273796	72820	216294	811831	590888	465895	383986	556517	178263	342313	486571	391942	565964	843738	456524	420105	640907	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
PG(16:0/0:0)	1647187	2379914	1887718	618399	528357	987441	749844	2473026	1052676	776554	760995	1290372	1275009	59674	484239	472699	496273	437233	242386	451318	538978	372634	581157	40198	392880	577489	456109	515095	457706	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Sitosterol	405010	346818	347921	324710	260650	338496	378803	650251	501530	422579	490011	360289	613709	249100	304061	290162	303280	255547	302485	308417	218167	323032	265027	256616	278458	188681	207020	159028	312295	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Spermidine	405978	347333	524880	589927	595671	366504	508039	96204	251202	168617	354715	1156994	1111565	2048897	1198869	411969	365394	310010	330617	251140	295405	218656	1101227	1564987	1048042	816573	805887	757729	760189	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Sphinganine	5812685	2520239	3931269	5978080	8298733	6367377	3495311	4829989	3440257	5032009	5555426	3500263	7235604	1995623	4394173	2139015	1974040	1975880	4304989	1670760	821631	2768997	2661958	3211958	1500635	1899601	1868123	3087024	3470035	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Sphingosine	4186080	2810706	4667310	3905635	5096186	5470690	2965964	3798106	8306211	10950087	12129609	16921732	11830107	5121924	14010060	2881432	3586577	2448110	2678680	2249168	1489229	2703767	6507267	5828644	2452565	2789666	2183105	2965714	5248158	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	HMDB ID	PubChem CID	KEGG number	Metabolite_ID (CAS# number)	LipidMaps
3-ketosphinganine
Campesterol
Cholesterol
D-erythro-Sphingosine C-20
L-Lysine
L-Proline
L-threo-Sphingosine C-18
L-Tyrosine
Oleamide
Palmitic amide
PC(14:0/0:0)
PC(16:0/0:0)
PC(18:2/0:0)
PE (18:1/0:0)
PE(14:0/0:0)
PE(15:0/0:0)
PE(17:0/0:0)
PE(17:1/0:0)
PG(16:0/0:0)
Sitosterol
Spermidine
Sphinganine
Sphingosine
METABOLITES_END
#END