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MB Sample ID: SA099053
Local Sample ID: | 2004-F-UD BPA |
Subject ID: | SU001436 |
Subject Type: | Mammal |
Subject Species: | Peromyscus Californicus |
Taxonomy ID: | 42520 |
Age Or Age Range: | PND90 |
Gender: | Male and female |
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Sample Preparation:
Sampleprep ID: | SP001444 |
Sampleprep Summary: | Fecal samples were snap frozen in liqud nitrogen and no further preparation |
Processing Method: | Fecal samples were lyophilized for 24 h before weighing |
Processing Storage Conditions: | -20℃ |
Extraction Method: | For LCMS analysis lyophilized fecal sample was extracted with 80% Methanol containg 18 µg/ml of umbelliferon, sonicated and vortexed 20 seconds each, shaken at 140 rpmfor 2 h. For GCMS-polar analysis samples were extracted with water, containing 25 μg/ml ribitol, vortex for 20 seconds, sonicated for 20 seconds, incubated at 50C for 1h. For GCMS-nonpolar samples were extracted with chloroform, containing 10 μl/ml docosanol, vortex for 20 seconds, sonicated for 20 second, incubated at 50C for 1h. |
Extract Enrichment: | Solution were centrifuged at 3000g for 40 min then extracted 0.5 mL of supernatant for LCMS, 1 mL each of supernatant was collected for GCMS-polar and GCMS-nonpolar analysis. |
Extract Storage: | -20℃ |
Sample Resuspension: | For GCMS analysis extracts dried under nitrogen flow and storred at -20C then resuspended in 25 μl of pyridine containing 15 mg/ml methoxyamine hydrochloride |
Sample Derivatization: | GCMS samples were derivatized with 25 μl MSTFA (N-methyl-N-(trimethyl-silyl)trifluoroacetamide) + 1% TMCS (chlorotrimethylsilane) |
Sample Spiking: | Umbelliferon, ribitol and docosanol were used as internal standard for LCMS, GCMS-polar and GCMS-nonpolar respectively. |