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MB Sample ID: SA309234

Local Sample ID:A_144
Subject ID:SU002974
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606

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Subject:

Subject ID:SU002974
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
A_144SA309234FL037197high-altitude groupTreatment

Collection:

Collection ID:CO002967
Collection Summary:Fecal samples were collected from participants using fecal sampling tubes (Shenzhen Medico Biomedical Technology Co., Ltd) and were also stored at -80°C after sampling.
Sample Type:Feces

Treatment:

Treatment ID:TR002983
Treatment Summary:The study enrolled two distinct groups: a plain group (PL) comprising 67 adult males from the Chengdu plain in Sichuan, and a high-altitude group (HA) comprising 163 adult males from Tibet residing at altitudes ranging from 3500m to 4500m. They were all graduated from Chengdu Medical College and enrolled in this study with the patients' consent and the sampling process was strictly consistent.The study received approval from the Ethics Committee of the Academy of Military Medicine, Chinese Academy of Military Sciences, with the approval number AF/SC-08/02.153.

Sample Preparation:

Sampleprep ID:SP002980
Sampleprep Summary:For fecal samples, 60mg of the sample was mixed with 600μL of 80% iced methanol, which included 0.001mg/mL 4-chloro-DL-phenylalanine as an internal standard. Zirconia crushing beads were added to the mixture, and the samples were disrupted using a vibratory crusher operating at 60 hertz. Subsequently, the samples were processed by ultrasonication in an ice bath for 10 minutes and left to stand at -20℃ for 30 minutes. The supernatants were collected by centrifugation at 20,000 g under 4 C for 10 minutes and then mixed with three-fold volumes of distilled water. After ultrasonication for 3 minutes to ensure even mixing, the samples were placed in a -20℃ environment for 2 hours. Finally, the supernatants obtained from centrifugation were transferred to sample vials for metabolomics analysis

Combined analysis:

Analysis ID AN004693 AN004694
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Agilent 1290 Infinity Agilent 1290 Infinity
Column Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um)
MS Type ESI ESI
MS instrument type Triple quadrupole Triple quadrupole
MS instrument name Agilent 6495 QQQ Agilent 6495 QQQ
Ion Mode POSITIVE NEGATIVE
Units counts counts

Chromatography:

Chromatography ID:CH003534
Chromatography Summary:In this study, a newly developed precision-targeted metabolomics method with a UPLC-TQ/MS system (Agilent 1290 Infnity, Agilent Technologies, USA; Agilent 6495 QQQ, Agilent Technologies, USA) in a DMRM scan-mode was applied to analyze the metabolome of interest from trial samples (serum, liver tissues and stool). Briefly, the method was performed with an ACQUITY UPLC HSS T3 column (2.1 mm i.d. × 100 mm, 1.8 μm; Waters); mobile phase A and B were water and acetonitrile with 0.1% formic acid (v/v) respectively. The flow rate was at 0.3 mL/min and the column temperature was maintained at 40 ℃. The samples were placed in an auto-sampler maintained at 4 °C with a 5 μL injection volume. The optimized gradient-elution program, as follows: 0-2 min, 98% A; 2-10 min, 98-65% A; 10-12 min, 65-20% A; 12-14 min, 20-2% A; 14-30 min, 2% A.
Methods Filename:Chromatography.pdf
Instrument Name:Agilent 1290 Infinity
Column Name:Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um)
Column Temperature:40 ℃
Flow Gradient:0-2 minutes, 98% A 2-10 minutes, 98%-65% A 10-12 minutes, 65%-20% A 12-14 minutes, 20%-2% A 14-30 minutes, 2% A.
Flow Rate:0.3 ml/min
Solvent A:100% water + 0.1% formic acid
Solvent B:100% acetonitrile + 0.1% formic acid
Chromatography Type:Reversed phase

MS:

MS ID:MS004440
Analysis ID:AN004693
Instrument Name:Agilent 6495 QQQ
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:Agilent MassHunter Workstation Data Acquisition Agilent MassHunter
Ion Mode:POSITIVE
Analysis Protocol File:MS.pdf
  
MS ID:MS004441
Analysis ID:AN004694
Instrument Name:Agilent 6495 QQQ
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:Agilent MassHunter Workstation Data Acquisition Agilent MassHunter
Ion Mode:NEGATIVE
Analysis Protocol File:MS.pdf
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