Urine metabolomic analyses reveal metabolite disruptions in steroid hormone biosynthesis in mpox
Description
This data is the raw offline data of untargeted metabolomics in this study, including four groups of samples: MPXV group, HIV-MPXV group, HIV group, and healthy group. MPXV infection may affect the metabolic levels of patients. Through non targeted metabolomics techniques, it is known that mpox caused metabolic disorders in the body, especially downregulation of steroid hormone synthesis pathways. This study used urine samples, mpox samples were collected from hospitalized patients, HIV samples were collected from outpatient clinics, and healthy samples were recruited from volunteers. All samples were collected within 2 months. Immediately freeze the collected samples in liquid nitrogen and store them in a -80 ° C freezer for later use. After all samples are collected, they will be inactivated in a biosafety level 3 laboratory and then packaged and stored for later use. By analyzing this raw data, identified metabolites and differential metabolites can be obtained.
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1、Chemicals and reagents HPLC-grade acetonitrile (ACN) 、Isopropanol(IPA) and methanol (MeOH) were purchased from Merck (Darmstadt, Germany). MilliQ water (Millipore, Bradford, USA) was used in all experiments. Acetic acid was purchased from Sigma-Aldrich. All of the standards were purchased from Olchemlm Ltd. (Olomouc, Czech Republic). The stock solutions of standards were prepared at the concentration of 1 mg/mL in MeOH. All stock solutions were stored at -20°C. The stock solutions were diluted with MeOH to working solutions before analysis. 2、Sample preparation and extraction After the sample was thawed, the sample was vortexd for 10 seconds. 100 μL of the sample was transferred to a centrifuge tube, and mixed with 400 μL of methanol, vortexed for 10 minutes, stand on ice for 10 min and centrifuged at 12000 r/min for 5 min at 4°C. Take 400 μL of supernatant into a new centrifuge tube and concentrate it at 20°C until it is completely dry. Then the sample was redissolved with 100 μL methanol, vortex for 5 min, centrifuged at 12000 r/min for 3 min at 4°C. After centrifugation, transfer 80 μL of the supernatant for further LC-MS analysis. 3、UPLC Conditions The sample extracts were analyzed using an LC-ESI-MS/MS system (UPLC, ExionLC AD， https://sciex.com.cn /; MS, QTRAP® 6500+ System, https://sciex.com /). The analytical conditions were as follows, HPLC: column, Phenomenex Kinetex C18（1.7 µm, 100 mm×2.1 mm i.d.）; solvent system, 30% acetonitrile/water with 0.04% Acetic acid (A), 50% acetonitrile/isopropanol with 0.04% Acetic acid (B); The gradient was started at 5% B (0 min-1.0 min), increased to 90% B (1.0-10 min),maintained at 90% B (10-12.5 min), finaly ramped back to 5% B (12.6-15 min); flow rate, 0.35 mL/min; temperature, 40°C; injection volume: 5 μL. 4、ESI-MS/MS Conditions AB 6500+ QTRAP® LC-MS/MS System, equipped with an ESI Turbo Ion-Spray interface, operating in positive ion mode and controlled by Analyst 1.6 software (AB Sciex). The ESI source operation parameters were as follows: ion source, turbo spray; source temperature 550°C; ion spray voltage (IS) 5500 V（Positive）; curtain gas (CUR) were set at 35.0 psi; DP and CE for individual MRM transitions was done with further DP and CE optimization. A specific set of MRM transitions were monitored for each period according to the neurotransmitters eluted within this period.