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MB Sample ID: SA151863
| Local Sample ID: | P12_t2 |
| Subject ID: | SU001732 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Male and female |
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Subject:
| Subject ID: | SU001732 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Male and female |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| P12_t2 | SA151863 | FL017842 | Idiopatic | Trigger |
| P12_t2 | SA151863 | FL017842 | Severe | Severity |
| P12_t2 | SA151863 | FL017842 | Time 2 | Time |
Collection:
| Collection ID: | CO001725 |
| 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: | TR001745 |
| Treatment Summary: | Patients were treated according to the Galaxy 2016 practical guide, using all necessary drugs to rescue them. |
Sample Preparation:
| Sampleprep ID: | SP001738 |
| 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. |
Analysis:
| MB Sample ID: | SA151863 |
| Analysis ID: | AN002702 |
| Analysis Type: | NMR |
NMR:
| NMR ID: | NM000202 |
| Analysis ID: | AN002702 |
| Instrument Name: | Bruker 500MHz spectrometer |
| Instrument Type: | FT-NMR |
| NMR Experiment Type: | 1D-1H |
| Spectrometer Frequency: | 500MHz |
