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MB Sample ID: SA078257

Local Sample ID:OPC-12
Subject ID:SU001197
Subject Type:Mammal
Subject Species:Rattus norvegicus
Taxonomy ID:10116
Genotype Strain:ZDF rats
Gender:Not applicable

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Subject:

Subject ID:SU001197
Subject Type:Mammal
Subject Species:Rattus norvegicus
Taxonomy ID:10116
Genotype Strain:ZDF rats
Gender:Not applicable

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
OPC-12SA078257FL01192217-week-old ZDF ratGenotype
OPC-12SA078257FL0119226Treatment
OPC-12SA078257FL0119226Duration (weeks)

Collection:

Collection ID:CO001191
Collection Summary:After euthanasia by exsanguination under isoflurane anesthesia, a piece of liver was taken from the left lateral lobe on the treated rats, weighed at approximately 50mg, and immediately frozen in liquid nitrogen.
Sample Type:Liver
Storage Conditions:-80℃

Treatment:

Treatment ID:TR001212
Treatment Summary:The baseline control, the liver samples of ZDF rats were taken from the baseline control group of 11-week-old ZDF rats. And at the same time, the other 2 comparing groups were started on oral administration of OPC-163493 or vehicle solution for 6-weeks. After 6-week dosing, liver samples were taken from both groups, and the liver metabolites of all three groups including the baseline controls were analyzed.
Treatment Dose:0mg/kg/day, 6mg/kg/day

Sample Preparation:

Sampleprep ID:SP001205
Sampleprep Summary:The frozen liver samples were plunged into 50% acetonitrile/Milli-Q water containing internal standard. The sample was homogenized and then centrifuged. Subsequently, 800 uL of upper aqueous layer was filtered through a 5-kDa cutoff filter. The filtrate was centrifugally concentrated and re-suspended in 50 uL of Milli-Q water.
Processing Storage Conditions:4℃
Extract Storage:-80℃
Sample Resuspension:50 uL Mili-Q

Combined analysis:

Analysis ID AN001862 AN001863
Analysis type MS MS
Chromatography type CE CE
Chromatography system Agilent 7100 CE Agilent 7100 CE
Column Fused silica capillary, i.d. 50 μm × 80 cm Fused silica capillary, i.d. 50 μm × 80 cm
MS Type ESI ESI
MS instrument type Other Triple quadrupole
MS instrument name Agilent 6210 TOF Agilent 6460 QQQ
Ion Mode UNSPECIFIED UNSPECIFIED
Units Concentration (nmol/g tissue) Concentration (nmol/g tissue)

Chromatography:

Chromatography ID:CH001348
Chromatography Summary:capillary electrophoresis was connected with time-of-flight mass spectrometry (CE-TOFMS) for cation analysis and tandem mass spectrometry (CE-MS/MS) for anion.
Instrument Name:Agilent 7100 CE
Column Name:Fused silica capillary, i.d. 50 μm × 80 cm
Chromatography Type:CE

MS:

MS ID:MS001722
Analysis ID:AN001862
Instrument Name:Agilent 6210 TOF
Instrument Type:Other
MS Type:ESI
MS Comments:The spectrometer was scanned from m/z 50 to 1,000. Peaks were extracted using automatic integration software MasterHands (Keio University, Tsuruoka, Japan) in order to obtain peak information including m/z, migration time for CE-TOFMS measurement (MT) and peak area. Signal peaks corresponding to isotopomers, adduct ions, and other product ions of known metabolites were excluded, and remaining peaks were annotated with putative metabolites from the HMT metabolite database based on their MTs and m/z values determined by TOFMS. The tolerance range for the peak annotation was configured at ±0.5 min for MT and ±10 ppm for m/z. In addition, peak areas were normalized against those of the internal standards and then the resultant relative area values were further normalized by sample amount.
Ion Mode:UNSPECIFIED
  
MS ID:MS001723
Analysis ID:AN001863
Instrument Name:Agilent 6460 QQQ
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:The spectrometer was scanned from m/z 50 to 1,000. Peaks were extracted using MasterHands, automatic integration software (Keio University, Tsuruoka, Yamagata, Japan) and MassHunter Quantitative Analysis B.04.00 (Agilent Technologies) in order to obtain peak information including m/z, peak area, and migration time (MT). Signal peaks were annotated according to the HMT metabolite database based on their m/z values with the MTs. The peak area of each metabolite was normalized with respect to the area of the internal standard and metabolite concentration was evaluated by standard curves with three-point calibrations using each standard compound.
Ion Mode:UNSPECIFIED
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