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MB Sample ID: SA096028
Local Sample ID: | QC06 |
Subject ID: | SU001398 |
Subject Type: | Invertebrate |
Subject Species: | Haliotis discus hannai |
Taxonomy ID: | 42344 |
Weight Or Weight Range: | 2.17 ± 0.56 g |
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Sample Preparation:
Sampleprep ID: | SP001406 |
Sampleprep Summary: | 50 mg of sample was taken and placed in an Eppendorf tube, followed by the addition of 1000 µl extraction solvent containing an internal target (V methanol : V acetonitrile : V water = 2 : 2 : 1, containing internal standard 2 µg/ml). The mixture was homogenized in ball mill for 4 min at 45 Hz, then ultrasound treated for 5min (incubated in ice water). After homogenization for 3 times, the mixture was incubated for 1 h at -20 °C to precipitate proteins. Centrifugation was performed at 12000 rpm for 15 min at 4 °C. A total of 825 µl supernatant was carefully transferred into a fresh Eppendorf tube, and dried in a vacuum concentrator without heating. 200 µl extraction solvent (V acetonitrile: V water = 1:1) was then added for reconstitution, followed by vortex 30 s and sonicated for 10 min (4 °C water bath), centrifuge for 15 min at 12000 rpm (4 °C). 75 µl supernatant was transferred into a fresh 2 ml LC/MS glass vial for UHPLC-QTOF-MS analysis, while another 20 µl was taken from each sample and pooled as QC samples. Six technical replicates were conducted to QC sample with 75 µL each for UHPLC-QTOF-MS analysis. |
Sampleprep Protocol Filename: | LC-MS-Q211.pdf |