Summary of Study ST001656

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 PR001061. The data can be accessed directly via it's Project DOI: 10.21228/M8JQ37 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.

Show all samples | Perform analysis on untargeted data
Download mwTab file (text) | Download mwTab file(JSON) | Download data files (Contains raw data)

Study ID	ST001656
Study Title	Characterization of anaphylaxis reveals different metabolic changes depending on severity and triggers - MS (part-II)
Study Summary	Background: Despite its increasing incidence, the underlying molecular processes of anaphylaxis remain unclear and there are not known biomarkers for appropriate diagnosis. The mechanism associated to the reactions still needs to be clarified in humans. The rapid onset and potentially fatal outcome in the absence of managed treatment, prevent its study and prompt obvious technical and ethical implications. Methods: Twenty episodes of anaphylaxis were analyzed. Sera was collected at different times: during the acute phase (T1), the recovery phase (T2) and around 2-3 months after the anaphylactic reaction (T0). The analysis included untargeted metabolomics combining liquid chromatography coupled to mass spectrometry (LC-MS) and proton-nuclear magnetic resonance (1H-NMR). Reactions were classified according to the trigger (food and/or drug) and severity (moderate and severe). Results: “Food T1 vs T2” and “moderate T1 vs T2” anaphylaxis comparisons showed clear metabolic patterns during the onset of an anaphylactic reaction, which differed from those induced by drugs, food+drug or severe anaphylaxis “T1 vs T2”. Moreover, the model of food anaphylaxis was able to distinguish the well-characterized IgE (beta-lactam) from non-IgE- mediated anaphylaxis (NSAIDs), suggesting a differential metabolic pathway associated with the mechanism of action. Moreover, metabolic differences between “moderate vs severe” at T1 and T0 were studied. Among the metabolites, glucose, lipids, cortisol, betaine and oleamide were observed altered. Conclusions: The results of the study provide the first evidence that different anaphylactic triggers, induce differential metabolic changes. Besides, the basal status might identify high risk patients, thus opening new ways to understand, diagnose and treat anaphylaxis.
Institute	The Centre of Metabolomics and Bioanalysis
Department	Analytical chemistry
Last Name	Obeso Montero
First Name	David
Address	Av. de Montepríncipe, s/n
Email	david.obesomontero@beca.ceu.es
Phone	607535650
Submit Date	2021-01-15
Num Groups	2 groups
Total Subjects	20
Raw Data Available	Yes
Raw Data File Type(s)	d
Analysis Type Detail	LC-MS
Release Date	2022-01-17
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR001061
Project DOI:	doi: 10.21228/M8JQ37
Project Title:	Characterization of anaphylaxis reveals different metabolic changes depending on severity and triggers.
Project Summary:	Despite its increasing incidence, the underlying molecular processes of anaphylaxis remain unclear and there are not known biomarkers for appropriate diagnosis. The mechanism associated to the reactions still needs to be clarified in humans. The rapid onset and potentially fatal outcome in the absence of managed treatment, prevent its study and prompt obvious technical and ethical implications. Twenty episodes of anaphylaxis were analyzed. Sera was collected at different times: during the acute phase (T1), the recovery phase (T2) and around 2-3 months after the anaphylactic reaction (T0). The analysis included untargeted metabolomics combining liquid chromatography coupled to mass spectrometry (LC-MS) and proton-nuclear magnetic resonance (1H-NMR). Reactions were classified according to the trigger (food and/or drug) and severity (moderate and severe). “Food T1 vs T2” and “moderate T1 vs T2” anaphylaxis comparisons showed clear metabolic patterns during the onset of an anaphylactic reaction, which differed from those induced by drugs, food+drug or severe anaphylaxis “T1 vs T2”. Moreover, the model of food anaphylaxis was able to distinguish the well-characterized IgE (beta-lactam) from non-IgE- mediated anaphylaxis (NSAIDs), suggesting a differential metabolic pathway associated with the mechanism of action. Moreover, metabolic differences between “moderate vs severe” at T1 and T0 were studied. Among the metabolites, glucose, lipids, cortisol, betaine and oleamide were observed altered. The results of the study provide the first evidence that different anaphylactic triggers, induce differential metabolic changes. Besides, the basal status might identify high risk patients, thus opening new ways to understand, diagnose and treat anaphylaxis.
Institute:	Hospital Universitari i Politècnic La Fe
Department:	Subdirección Médica, Departament de Salut València La Fe.
Last Name:	Hernández Fernández de Rojas
First Name:	Dolores
Address:	Avinguda de Fernando Abril Martorell, 106, 46026 València, Valencia, España.
Email:	hernandez_dol@gva.es
Phone:	91 372 47 00 ext. 4662

Subject:

Subject ID:	SU001733
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	Trigger	Severity	Time
SA151870	P15_t0	Drug	Mild	Time 0
SA151871	P15_t1	Drug	Mild	Time 1
SA151872	P17_t0	Drug	Moderate	Time 0
SA151873	P1_t0	Drug	Moderate	Time 0
SA151874	P3_t0	Drug	Moderate	Time 0
SA151875	P1_t1	Drug	Moderate	Time 1
SA151876	P17_t1	Drug	Moderate	Time 1
SA151877	P3_t1	Drug	Moderate	Time 1
SA151878	P1_t2	Drug	Moderate	Time 2
SA151879	P17_t2	Drug	Moderate	Time 2
SA151880	P19_t0	Drug	Severe	Time 0
SA151881	P14_t0	Drug	Severe	Time 0
SA151882	P4_t0	Drug	Severe	Time 0
SA151883	P7_t0	Drug	Severe	Time 0
SA151884	P13_t0	Drug	Severe	Time 0
SA151885	P4_t1	Drug	Severe	Time 1
SA151886	P13_t1	Drug	Severe	Time 1
SA151887	P14_t1	Drug	Severe	Time 1
SA151888	P19_t1	Drug	Severe	Time 1
SA151889	P7_t1	Drug	Severe	Time 1
SA151890	P7_t2	Drug	Severe	Time 2
SA151891	P13_t2	Drug	Severe	Time 2
SA151892	P14_t2	Drug	Severe	Time 2
SA151893	P19_t2	Drug	Severe	Time 2
SA151894	P4_t2	Drug	Severe	Time 2
SA151895	P2_t0	Food	Moderate	Time 0
SA151896	P16_t0	Food	Moderate	Time 0
SA151897	P9_t0	Food	Moderate	Time 0
SA151898	P18_t0	Food	Moderate	Time 0
SA151899	P2_t1	Food	Moderate	Time 1
SA151900	P9_t1	Food	Moderate	Time 1
SA151901	P10_t1	Food	Moderate	Time 1
SA151902	P18_t1	Food	Moderate	Time 1
SA151903	P16_t1	Food	Moderate	Time 1
SA151904	P9_t2	Food	Moderate	Time 2
SA151905	P18_t2	Food	Moderate	Time 2
SA151906	P2_t2	Food	Moderate	Time 2
SA151907	P16_t2	Food	Moderate	Time 2
SA151908	P10_t2	Food	Moderate	Time 2
SA151909	P11_t0	Food	Severe	Time 0
SA151910	P11_t1	Food	Severe	Time 1
SA151911	P11_t2	Food	Severe	Time 2
SA151912	P5_t1	Idiopatic	Moderate	Time 1
SA151913	P5_t2	Idiopatic	Moderate	Time 2
SA151914	P12_t0	Idiopatic	Severe	Time 0
SA151915	P6_t0	Idiopatic	Severe	Time 0
SA151916	P6_t1	Idiopatic	Severe	Time 1
SA151917	P12_t1	Idiopatic	Severe	Time 1
SA151918	P12_t2	Idiopatic	Severe	Time 2
SA151919	P6_t2	Idiopatic	Severe	Time 2
SA151920	P20_t0	Other	Moderate	Time 0
SA151921	P8_t0	Other	Moderate	Time 0
SA151922	P20_t1	Other	Moderate	Time 1
SA151923	P8_t1	Other	Moderate	Time 1
SA151924	P8_t2	Other	Moderate	Time 2

Showing results 1 to 55 of 55

Showing results 1 to 55 of 55

Collection:

Collection ID:	CO001726
Collection Summary:	A prospective clinical and observational study of patients with anaphylactic reactions was performed. Patients of all ages and both sexes were recruited at outpatient clinics and the departments of Emergency, and other services at Hospital La Fe. All fulfilled clinical criteria of anaphylaxis and severity was graded following the classification by Brown, et al3. Patients were classified as food, drug or idiopathic origin, as well as in mild, moderate or severe according to the number of organs affected and clinical symptoms. The allergy evaluation was conducted by the Allergology Service of Hospital La Fe. The ethical committee approved the study protocol and all subjects were informed and provided written consent. Serum samples were taken during the acute moment of the reaction at the first moment of medical attention (< 2h, hereafter referred as ‘T1’), and after clinical recovery (approximately 2-4h later, referred to as ‘T2’). Patients were treated according to the Galaxy 2016 practical guide, using all necessary drugs to rescue them. Samples were collected, following specific standard operating procedures (SOPs)31-33. Briefly, these were collected in a vacutainer tube (Ref. 368965) and processed within the first 30 to 60 min after blood extraction. Sample aliquots were stored at -80ºC until further analyses. Subsequently, between 2-3 months after the anaphylaxis, a serum sample was taken when the allergy evaluation was performed (basal state, called ‘T0’).
Sample Type:	Blood (serum)
Storage Conditions:	-80℃

Treatment:

Treatment ID:	TR001746
Treatment Summary:	Patients were treated according to the Galaxy 2016 practical guide, using all necessary drugs to rescue them.

Sample Preparation:

Sampleprep ID:	SP001739
Sampleprep Summary:	After thawing the samples, 150 µL of cold acetonitrile (0.1% formic acid, v/v) was added to 50 µL of serum, vortex and kept at -20ºC for 30 min for protein precipitation. Then, 25 µl of cleaned supernatant were transferred to a 96 well-plate for UPLC-MS analysis. Finally, 125 µL of H2O (0.1% formic acid, v/v), and 2 µL of internal standard (IS) mix solution, containing reserpine, leucine enkephaline, caffeine-d9 and phenylalanine-d5 in H20:CH3OH (1:1, 0.1% v/v formic acid) at 20 µM were added to each sample. Blank samples were prepared by replacing serum by ultrapure H20 in order to identify potential artefacts from the tube, reagents and other materials. A quality control (QC) was prepared by mixing 10 µL from each prepared sample. Samples were randomly injected in the chromatographic system (UPLC-ToF-MS) injection a QC every 6 serum samples in order to avoid intra-batch variability, as well as to enhance quality and reproducibility. Blank analysis was performed at the beginning and at end of the sequence. Sample stability and analytical drift were investigated through the internal standard intensities.

Sampleprep ID:

SP001739

Sampleprep Summary:

After thawing the samples, 150 µL of cold acetonitrile (0.1% formic acid, v/v) was added to 50 µL of serum, vortex and kept at -20ºC for 30 min for protein precipitation. Then, 25 µl of cleaned supernatant were transferred to a 96 well-plate for UPLC-MS analysis. Finally, 125 µL of H2O (0.1% formic acid, v/v), and 2 µL of internal standard (IS) mix solution, containing reserpine, leucine enkephaline, caffeine-d9 and phenylalanine-d5 in H20:CH3OH (1:1, 0.1% v/v formic acid) at 20 µM were added to each sample. Blank samples were prepared by replacing serum by ultrapure H20 in order to identify potential artefacts from the tube, reagents and other materials. A quality control (QC) was prepared by mixing 10 µL from each prepared sample. Samples were randomly injected in the chromatographic system (UPLC-ToF-MS) injection a QC every 6 serum samples in order to avoid intra-batch variability, as well as to enhance quality and reproducibility. Blank analysis was performed at the beginning and at end of the sequence. Sample stability and analytical drift were investigated through the internal standard intensities.

Combined analysis:

Analysis ID	AN002703	AN002704
Analysis type	MS	MS
Chromatography type	Reversed phase	Reversed phase
Chromatography system	Agilent 6550	Agilent 6550
Column	Waters Acquity BEH C18 (100 x 2mm,1.7um)	Waters Acquity BEH C18 (100 x 2mm,1.7um)
MS Type	ESI	ESI
MS instrument type	QTOF	QTOF
MS instrument name	Agilent 6550 QTOF	Agilent 6550 QTOF
Ion Mode	POSITIVE	NEGATIVE
Units	Intensity	intensity

Chromatography:

Chromatography ID:	CH001995
Chromatography Summary:	The chromatographic separation was performed by using an UPLC BEH C18 (100 x 2.1 mm, 1.7 µm, Waters, Wexford, Ireland) column from Waters (Wexford, Ireland). Autosampler and column temperatures were set to 4°C and 40°C, respectively, and the injection volume was 5 µL. Mobile phase A and B consisted of H20 and acetonitrile, respectively, both containing 0.1% formic acid. The gradient elution lasted 14 minutes at a flow rate of 400 µl min-1. The mobile phase A was maintained at 98% for 1 min, and then decreased until 75% in 2 minutes, 50% in 3 minutes and 5% in 3 more minutes. 95% of mobile phase B was held for 3 min and then, a 0.55 min gradient was used to return to the initial conditions, which were held for 2.5 min to full column recovery.
Instrument Name:	Agilent 6550
Column Name:	Waters Acquity BEH C18 (100 x 2mm,1.7um)
Column Temperature:	40
Flow Gradient:	The mobile phase A was maintained at 98% for 1 min, and then decreased until 75% in 2 minutes, 50% in 3 minutes and 5% in 3 more minutes. 95% of mobile phase B was held for 3 min and then, a 0.55 min gradient was used to return to the initial conditions, which were held for 2.5 min to full column recovery.
Flow Rate:	400 ul/min
Solvent A:	100% water; 0.1% formic acid
Solvent B:	100% acetonitrile; 0.1% formic acid
Chromatography Type:	Reversed phase

MS:

MS ID:	MS002500
Analysis ID:	AN002703
Instrument Name:	Agilent 6550 QTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	Full scan MS data from 50 to 1700 m/z with a scan frequency of 6 Hz was collected both in positive and negative electrospray ionization modes (ESI + and ESI -, respectively). The following ESI parameters were used: gas temperature, 200°C; drying gas, 14 l min-1; nebulizer, 60 psi; sheath gas temperature, 350°C; sheath gas flow, 11 l min-1. Automatic MS spectra recalibration was carried out introducing a reference standard containing m/z 149.0233, m/z 121.0508 and m/z 922.0097 into the source via a reference sprayer valve during the analysis.
Ion Mode:	POSITIVE

MS ID:	MS002501
Analysis ID:	AN002704
Instrument Name:	Agilent 6550 QTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	Full scan MS data from 50 to 1700 m/z with a scan frequency of 6 Hz was collected both in positive and negative electrospray ionization modes (ESI + and ESI -, respectively). The following ESI parameters were used: gas temperature, 200°C; drying gas, 14 l min-1; nebulizer, 60 psi; sheath gas temperature, 350°C; sheath gas flow, 11 l min-1. Automatic MS spectra recalibration was carried out introducing a reference standard containing m/z 149.0233, m/z 121.0508 and m/z 922.0097 into the source via a reference sprayer valve during the analysis.
Ion Mode:	NEGATIVE