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MB Sample ID: SA327783
| Local Sample ID: | M_Cefto3g_071_2 |
| Subject ID: | SU003138 |
| Subject Type: | Bacteria |
| Subject Species: | Pseudomonas aeruginosa |
| Taxonomy ID: | 287 |
| Genotype Strain: | CW41 |
| Age Or Age Range: | NA |
| Weight Or Weight Range: | NA |
| Height Or Height Range: | NA |
| Gender: | Not applicable |
| Cell Strain Details: | hypermutable clinical isolate |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003138 |
| Subject Type: | Bacteria |
| Subject Species: | Pseudomonas aeruginosa |
| Taxonomy ID: | 287 |
| Genotype Strain: | CW41 |
| Age Or Age Range: | NA |
| Weight Or Weight Range: | NA |
| Height Or Height Range: | NA |
| Gender: | Not applicable |
| Cell Strain Details: | hypermutable clinical isolate |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| M_Cefto3g_071_2 | SA327783 | FL038910 | M_Cefto3g_71h | treatment |
Collection:
| Collection ID: | CO003131 |
| Collection Summary: | For metabolomic footprint analysis, the supernatant from centrifuged bacterial samples was collected. These bacterial samples were obtained from the hollow-fiber cartridges for all untreated control and treated arms for each of the five biological replicates at 7, 23, 47, 71, 143, and 167 h. Additional bacterial samples were collected from the cartridges at 0 and 215 h for replicates 3 to 5. Matrix blank samples (of CAMHB not containing any bacteria) were collected from the diluent media bottles every 48 h for all untreated control and treated arms. |
| Sample Type: | Bacterial cells |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR003147 |
| Treatment Summary: | Ceftolozane-tazobactam was administered to simulate steady-state concentrations of ceftolozane predicted to occur in the epithelial lining fluid of the lung in patients with CF, following daily doses of 3 g/1.5 g and 6 g/3 g via continuous infusion (10.6 and 21.3 mg/L, respectively) (21-23). Total bacterial populations were quantified on antibiotic-free CAMHA, and resistant subpopulations on CAMHA containing ceftolozane-tazobactam (12 and 20 mg/L). |
| Treatment: | continuous infusion |
| Treatment Compound: | Ceftolozane-tazobactam |
| Treatment Dose: | 3 g and 6 g |
| Treatment Dosevolume: | 10.6 and 21.3 mg/L |
Sample Preparation:
| Sampleprep ID: | SP003144 |
| Sampleprep Summary: | Each sample (25 µL) was added to 100 µL of pre-chilled methanol containing the internal standards (CHAPS, CAPS, TRIS and PIPES) at 1 µM. This mixture was vortexed, and subsequently centrifuged at 14800 x g and 4°C for 10 min. The final supernatant samples containing the extracted extracellular metabolites were stored at -80°C until LC-MS analysis was performed. |
| Processing Storage Conditions: | -80℃ |
| Extract Storage: | -80℃ |
Combined analysis:
| Analysis ID | AN004958 | AN004959 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 |
| Column | Merck SeQuant ZIC-pHILIC (150 x 4.6mm,5um) | Merck SeQuant ZIC-pHILIC (150 x 4.6mm,5um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | peak height | peak height |
Chromatography:
| Chromatography ID: | CH003742 |
| Chromatography Summary: | ZIC-pHILIC chromatography at ph9, biological replicates 3-5 |
| Methods Filename: | Metabolomics_pHILIC_Parkville_v1.pdf |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Merck SeQuant ZIC-pHILIC (150 x 4.6mm,5um) |
| Column Pressure: | 60 bar at starting conditions. 180 bar at %A |
| Column Temperature: | 25 C |
| Flow Gradient: | 0 min - 80%B, 15 min - 50%B, 18 min - 5%B, 21 min - 5%B, 24 min - 80%B, 32 min - 80%B |
| Flow Rate: | 0.3 ml/min |
| Injection Temperature: | 4 C |
| Sample Injection: | 10 uL |
| Solvent A: | 20 mM ammonium carbonate |
| Solvent B: | acetonitrile |
| Analytical Time: | 32 min |
| Capillary Voltage: | 4 kV |
| Washing Buffer: | syringe wash 50% IPA |
| Sample Loop Size: | 25 uL |
| Sample Syringe Size: | 25 uL |
| Randomization Order: | yes |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004698 |
| Analysis ID: | AN004958 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Full scan LCMS data acquired using polarity switching. raw data processed using IDEOM workflow ( 10.1007/978-1-0716-0239-3_21). |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 300 C |
| Capillary Voltage: | 4 kV |
| Collision Energy: | NA |
| Collision Gas: | NA |
| Dry Gas Flow: | 50 |
| Dry Gas Temp: | 120 |
| Ion Source Temperature: | 120 C |
| Ionization: | ESI |
| Mass Accuracy: | 3 ppm |
| Precursor Type: | [M+H]+ |
| Source Temperature: | 300 |
| Acquisition Parameters File: | Metabolomics_pHILIC_Parkville_v1.pdf |
| Analysis Protocol File: | PQMS3-MPMF-WIN-0501_LCMS_data_acquisition_for_untargeted_metabolomics_analysis.pdf |
| MS ID: | MS004699 |
| Analysis ID: | AN004959 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Full scan LCMS data acquired using polarity switching. raw data processed using IDEOM workflow ( 10.1007/978-1-0716-0239-3_21). |
| Ion Mode: | NEGATIVE |
| Capillary Temperature: | 300 C |
| Capillary Voltage: | 3.5 kV |
| Collision Energy: | NA |
| Collision Gas: | NA |
| Dry Gas Flow: | 50 |
| Dry Gas Temp: | 120 |
| Ion Source Temperature: | 120 C |
| Ionization: | ESI |
| Mass Accuracy: | 3 ppm |
| Precursor Type: | [M-H]- |
| Source Temperature: | 300 |
| Acquisition Parameters File: | Metabolomics_pHILIC_Parkville_v1.pdf |
| Analysis Protocol File: | PQMS3-MPMF-WIN-0501_LCMS_data_acquisition_for_untargeted_metabolomics_analysis.pdf |
