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MB Sample ID: SA150510
| Local Sample ID: | DRB18invivo_8 |
| Subject ID: | SU001715 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Age Or Age Range: | 4 weeks mice |
| Gender: | Male |
| Animal Animal Supplier: | Jackson Laboratory |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001715 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Age Or Age Range: | 4 weeks mice |
| Gender: | Male |
| Animal Animal Supplier: | Jackson Laboratory |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| DRB18invivo_8 | SA150510 | FL017685 | DRB18 | Treatment |
Collection:
| Collection ID: | CO001708 |
| Collection Summary: | A549 lung cancer xenograft tumors were obtained after sacrificing the mice. The tumors were then prepared according to the protocol described and metabolomics analysis was performed. |
| Sample Type: | Tumor cells |
| Collection Method: | Tumors were homogenized in a Beadbud homogenizer (Benchmark scientific) in a mixture of LC-MS grade ice-cold methanol and water (1:1; v/v). The supernatant was collected and sonicated in a water bath incubator for 15 minutes, followed by centrifugation at 13,000 rpm for 10 minutes and collection of the supernatant. |
| Collection Location: | Homogenization vials |
| Collection Frequency: | Once |
| Collection Duration: | 4-5 minutes |
| Volumeoramount Collected: | 1ml |
| Storage Conditions: | -80℃ |
| Collection Vials: | Polypropylene 1.5 ml tubes |
| Storage Vials: | Polypropylene 1.5 ml |
Treatment:
| Treatment ID: | TR001728 |
| Treatment Summary: | Male NU/J nude mice of 3 to 4 weeks of age were purchased from The Jackson Laboratory and were fed with the Irradiated Teklad Global 19% protein rodent diet from Harlan Laboratories. The protocol for cell injection, treatment administration, weekly tumor measurement, animal euthanasia and final tumor measurements were performed as described previously (unless stated otherwise) (31). Tumor cell–injected mice were randomly divided into 2 groups: control group (n = 10) treated with PBS/DMSO (1:1, v/v) and 10 mg/kg (body weight) DRB18 treatment group (n = 10) dissolved in PBS/DMSO solution (1:1, v/v). Mice were given intraperitoneal injection with either PBS/DMSO vehicle or compound DRB18 (10 mg/kg) thrice a week for 5 weeks. |
| Treatment Protocol Filename: | prats1988_20201207_Invivo_collection_and_preparation_method.docx |
| Treatment: | Anticancer compound in xenograft tumors via i.p. |
| Treatment Compound: | Control (DMSO/PBS; 1:1 v/v) and DRB18 (10mg/kg body weight dissolved in DMSO/PBS; 1:1 v/v) |
| Treatment Route: | i.p. |
| Treatment Dose: | 10 mg/kg body weight |
| Treatment Dosevolume: | 100ul |
| Treatment Vehicle: | DMSO/PBS (1:1; v/v) |
Sample Preparation:
| Sampleprep ID: | SP001721 |
| Sampleprep Summary: | Tumors were homogenized in a Beadbud homogenizer (Benchmark scientific) in a mixture of LC-MS grade ice-cold methanol and water (1:1; v/v). The supernatant was collected and sonicated in a water bath incubator for 15 minutes, followed by centrifugation at 13,000 rpm for 10 minutes and collection of the supernatant. Supernatants collected from in vitro and in vivo extraction were then lyophilized using a speed vacuum evaporator. The samples were then dissolved into a mixture of LC-MS grade acetonitrile/water (1:1; v/v) for analysis. |
| Sampleprep Protocol Filename: | prats1988_20201207_Invivo_collection_and_preparation_method.docx |
| Processing Method: | Metabolite extraction; Quenching; speed vacuum evaporation |
| Processing Storage Conditions: | -80℃ |
| Extraction Method: | Quenching with ice cold methanol |
| Extract Enrichment: | Speed vaccum evaporator |
| Sample Resuspension: | Acetonitrile/waster (1:1) |
Combined analysis:
| Analysis ID | AN002680 |
|---|---|
| Analysis type | MS |
| Chromatography type | Unspecified |
| Chromatography system | none |
| Column | none |
| MS Type | ESI |
| MS instrument type | QTOF |
| MS instrument name | Agilent 6545 QTOF |
| Ion Mode | POSITIVE |
| Units | Normalized abundances |
Chromatography:
| Chromatography ID: | CH001974 |
| Chromatography Summary: | The entire LC/MS-MS experiment was performed in the Campus Chemical Instrumentation Center’s Mass Spectrometry and Proteomics facility at The Ohio State University. Lyophilized samples were dissolved in equal amounts of LC-MS grade water and acetonitrile and run with LC/MS-MS analysis, using an untargeted metabolomics approach by utilizing Agilent Q-TOF 6545 mass spectrometer connected to an Agilent 1290 UHPLC system with a Poroshell 120 SB-C18 (2 x 100 mm, 2.7 µm particle size) column. The LC gradient consisted of solvent A, H2O with 0.1 % Formic acid, and solvent B, 100 % acetonitrile at a 200 µL/min flow rate with an initial 2 % solvent B with a linear ramp to 95 % B at 15 min, holding at 95% B for 1 minutes, and back to 2 % B from 16 min and equilibration of 2 % B until min 32. A 5 µL volume sample was injected for each run and the top 5 ions were selected for data-dependent analysis with a 15 second exclusion window. |
| Instrument Name: | none |
| Column Name: | none |
| Column Pressure: | 800 bar |
| Column Temperature: | 40 |
| Flow Gradient: | an initial 2 % solvent B with a linear ramp to 95 % B at 15 min, holding at 95% B for 1 minutes, and back to 2 % B from 16 min and equilibration of 2 % B until min 32 |
| Flow Rate: | 200 µL/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | Unspecified |
MS:
| MS ID: | MS002479 |
| Analysis ID: | AN002680 |
| Instrument Name: | Agilent 6545 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Avin_C18_Pos.m methods was used. The data acquisition and processing was performed by masshunter software (Agilent Technologies). |
| Ion Mode: | POSITIVE |
