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MB Sample ID: SA099057
| Local Sample ID: | 1800-M-UD BPA |
| Subject ID: | SU001436 |
| Subject Type: | Mammal |
| Subject Species: | Peromyscus Californicus |
| Taxonomy ID: | 42520 |
| Age Or Age Range: | PND90 |
| Gender: | Male and female |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001436 |
| Subject Type: | Mammal |
| Subject Species: | Peromyscus Californicus |
| Taxonomy ID: | 42520 |
| Age Or Age Range: | PND90 |
| Gender: | Male and female |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 1800-M-UD BPA | SA099057 | FL014039 | UD BPA | Exposure |
| 1800-M-UD BPA | SA099057 | FL014039 | M | Gender |
Collection:
| Collection ID: | CO001431 |
| Collection Summary: | Fecal boli were collected at PND 90, snap frozen in LN2, stored in -80 freezer until processing |
| Sample Type: | Feces |
| Collection Method: | Snap freezen in LN2 right after excretion |
| Collection Location: | Animal Science Research Center, University of Missouri, Columbia |
| Collection Frequency: | 1 time from each animal |
| Volumeoramount Collected: | 3-4 boli |
| Storage Conditions: | -20℃ |
| Collection Vials: | 2mL vials |
| Storage Vials: | 2mL vials |
Treatment:
| Treatment ID: | TR001451 |
| Treatment Summary: | 2 weeks before breeding and during gestation, dams were fed estrogen-free AIN 93G or AIN 93G diet containing LD BPA, UD BPA or genistein untill one male and one female pups weaned. F1 pups fed on AIN 93G diet after weaning. |
| Treatment: | diet |
| Treatment Compound: | Genistein and Bisphenol A |
| Treatment Route: | oral |
| Treatment Dose: | Genistein: 250 mg/kg feed weight; BPA: 5 mg/kg feed weight for LD BPA and 50 mg/kg feed weight for UD BPA group |
| Treatment Doseduration: | 2 weeks before breeding and untill one male and one female pups weaned. |
| Treatment Vehicle: | AIN control diet |
| Animal Endp Tissue Coll List: | Fecal |
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. |
Combined analysis:
| Analysis ID | AN002266 | AN002267 | AN002268 |
|---|---|---|---|
| Analysis type | MS | MS | MS |
| Chromatography type | Reversed phase | GC | GC |
| Chromatography system | Waters Acquity I-Class | Agilent 6890N | Agilent 6890N |
| Column | Waters Acquity BEH C18 (150 x 2mm,1.7um) | DB-5MS (J&W Scientific) 60m | DB-5MS (J&W Scientific) 60m |
| MS Type | ESI | EI | EI |
| MS instrument type | QTOF | Single quadrupole | Single quadrupole |
| MS instrument name | Bruker maXis Impact qTOF | Agilent 5973 | Agilent 5973 |
| Ion Mode | POSITIVE | POSITIVE | POSITIVE |
| Units | intensity | intensity | intensity |
Chromatography:
| Chromatography ID: | CH001665 |
| Instrument Name: | Waters Acquity I-Class |
| Column Name: | Waters Acquity BEH C18 (150 x 2mm,1.7um) |
| Column Pressure: | >11000 psi |
| Column Temperature: | 60 |
| Flow Gradient: | Solvent B (acetonitrile) was increased from 5% to 70% over 30 min, then to 95% over 3 min, held at 95% for 3 min, then returned to 5% for equilibrium |
| Flow Rate: | 0.560 ml/min |
| Internal Standard: | Umbelliferon |
| Retention Time: | 1 to 35 min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% acetonitrile |
| Analytical Time: | 40 min |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH001666 |
| Chromatography Summary: | 5:1 split for GCMS-polar injection with 1uL injection volume. |
| Instrument Name: | Agilent 6890N |
| Column Name: | DB-5MS (J&W Scientific) 60m |
| Column Pressure: | variable |
| Column Temperature: | variable between 80-315 |
| Flow Rate: | 1mL/Min of Helium gas |
| Internal Standard: | Ribitol |
| Retention Time: | 9 to 61 min |
| Sample Injection: | 5:1 split injection with 1uL injection volume |
| Analytical Time: | 61 min |
| Chromatography Type: | GC |
| Chromatography ID: | CH001667 |
| Chromatography Summary: | 1:1 split for GCMS-nonpolar injection with 1uL injection volume. |
| Instrument Name: | Agilent 6890N |
| Column Name: | DB-5MS (J&W Scientific) 60m |
| Column Pressure: | variable |
| Column Temperature: | variable between 80-315 |
| Flow Rate: | 1mL/Min of Helium gas |
| Internal Standard: | Docosanol |
| Retention Time: | 9 to 61 min |
| Sample Injection: | 1:1 split injection with 1uL injection volume |
| Analytical Time: | 61 min |
| Chromatography Type: | GC |
MS:
| MS ID: | MS002110 |
| Analysis ID: | AN002266 |
| Instrument Name: | Bruker maXis Impact qTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Bruker Hystar and otof for acquisition and Metaboscape 4.0 and MS-DIAL for data analysis |
| Ion Mode: | POSITIVE |
| Collision Energy: | Varied, depended on procursor m/z |
| Collision Gas: | Nitrogen |
| Dry Gas Flow: | 12 l/min |
| Dry Gas Temp: | 250 |
| Fragmentation Method: | CID |
| Gas Pressure: | 43.5 psi of nebulizer gas |
| Dataformat: | .d |
| Nebulizer: | Nitrogen |
| MS ID: | MS002111 |
| Analysis ID: | AN002267 |
| Instrument Name: | Agilent 5973 |
| Instrument Type: | Single quadrupole |
| MS Type: | EI |
| MS Comments: | Agilent Chemstation software for data acquisition and AMDIS and MetIdea for data analysis |
| Ion Mode: | POSITIVE |
| Helium Flow: | 1mL/min |
| Ion Source Temperature: | 230 |
| Ionization Energy: | 70ev |
| Reagent Gas: | Helium |
| Source Temperature: | 230 |
| Dataformat: | .d |
| MS ID: | MS002112 |
| Analysis ID: | AN002268 |
| Instrument Name: | Agilent 5973 |
| Instrument Type: | Single quadrupole |
| MS Type: | EI |
| MS Comments: | Agilent Chemstation software for data acquisition and AMDIS and MetIdea for data analysis |
| Ion Mode: | POSITIVE |
| Helium Flow: | 1mL/min |
| Ion Source Temperature: | 230 |
| Ionization Energy: | 70ev |
| Reagent Gas: | Helium |
| Source Temperature: | 230 |
| Dataformat: | .d |
