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MB Sample ID: SA328808
Local Sample ID: | M_Cefto_3g_167h_E |
Subject ID: | SU003150 |
Subject Type: | Bacteria |
Subject Species: | Pseudomonas aeruginosa |
Taxonomy ID: | 287 |
Genotype Strain: | CW41 |
Age Or Age Range: | NA |
Weight Or Weight Range: | NA |
Gender: | Not applicable |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU003150 |
Subject Type: | Bacteria |
Subject Species: | Pseudomonas aeruginosa |
Taxonomy ID: | 287 |
Genotype Strain: | CW41 |
Age Or Age Range: | NA |
Weight Or Weight Range: | NA |
Gender: | Not applicable |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
M_Cefto_3g_167h_E | SA328808 | FL039045 | M_Cefto_3g_167h | treatment |
Collection:
Collection ID: | CO003143 |
Collection Summary: | A hypermutable P. aeruginosa clinical isolate, CW41, was challenged with ceftolozane-tazobactam (Zerbaxa®, MSD, Australia) in the HFIM (C3008-1 cartridges; FiberCell Systems Inc., Frederick, MD, USA), in five biological replicates performed across two studies. The first study, with replicates 1 and 2, was conducted over 167 h and the second study, with replicates 3, 4 and 5, over 215 h. Briefly, the studied isolate was characterized as susceptible to ceftolozane-tazobactam (MIC 4 mg/L), and MDR (i.e. resistant to at least 1 antibiotic from each of ≥3 antibiotic classes) (17-20). The HFIM studies used cation-adjusted Mueller Hinton broth (CAMHB) and agar (CAMHA) [Becton Dickinson & Co., Sparks, MD, USA, with 25.0 mg/L Ca2+ and 12.5 mg/L Mg2+]. 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). |
Sample Type: | Bacterial cells |
Treatment:
Treatment ID: | TR003159 |
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 Compound: | Ceftolozane-tazobactam |
Treatment Dose: | 3 g and 6 g |
Treatment Dosevolume: | 10.6 and 21.3 mg/L |
Sample Preparation:
Sampleprep ID: | SP003156 |
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 (Figure S2B). |
Processing Method: | IDEOM |
Processing Storage Conditions: | -80℃ |
Extract Storage: | -80℃ |
Combined analysis:
Analysis ID | AN004977 | AN004978 |
---|---|---|
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: | CH003758 |
Chromatography Summary: | ZIC-pHILIC chromatography at pH 9 using pos neg switching |
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 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 |
Solvent A: | 20 mM ammonium carbonate |
Solvent B: | acetonitrile |
Washing Buffer: | syringe wash 50% IPA |
Chromatography Type: | HILIC |
Chromatography ID: | CH003759 |
Chromatography Summary: | ZIC-pHILIC chromatography at pH 9 using pos neg switching |
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 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 |
Solvent A: | 20 mM ammonium carbonate |
Solvent B: | acetonitrile |
Washing Buffer: | syringe wash 50% IPA |
Chromatography Type: | HILIC |
MS:
MS ID: | MS004717 |
Analysis ID: | AN004977 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | polarity switching used, resolution 35 k, full scan |
Ion Mode: | POSITIVE |
Capillary Temperature: | 300 C |
Capillary Voltage: | 3.5 kV |
Collision Energy: | NA |
Dry Gas Flow: | 50 |
Dry Gas Temp: | 120 |
Ion Source Temperature: | 120 C |
Mass Accuracy: | 3 ppm |
Precursor Type: | [M+H]+ |
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: | MS004718 |
Analysis ID: | AN004978 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | polarity switching used, resolution 35 k, full scan |
Ion Mode: | NEGATIVE |
Capillary Temperature: | 300 C |
Capillary Voltage: | 4 kV |
Collision Energy: | NA |
Dry Gas Flow: | 50 |
Dry Gas Temp: | 120 |
Ion Source Temperature: | 120 C |
Mass Accuracy: | 3 ppm |
Precursor Type: | [M-H]- |
Acquisition Parameters File: | Metabolomics_pHILIC_Parkville_v1.pdf |
Analysis Protocol File: | PQMS3-MPMF-WIN-0501_LCMS_data_acquisition_for_untargeted_metabolomics_analysis.pdf |