Table 1 Cell disruption methods used in metabolome analysis.
From: Comparison of bacteria disintegration methods and their influence on data analysis in metabolomics
Disruption method | Extraction method | Organism | Amount of biomass | Analytical method | References |
|---|---|---|---|---|---|
Freeze–thaw (× 3) | Chloroform/methanol/water (1:3:1) | P. aeruginosa | ∼108 CFU/ml (OD600 = 0.5) | HPLC/LC–MS | |
Ultrasonic bath 15 min. 70 °C | Methanol/water/chloroform (3:3:2) | P. aeruginosa | 150 mg wet biomass | GC/MS | |
60% ethanol at 78 °C for 2 min, liquid nitrogen freezeing | 3 ml ethanol (60%) | P. aeruginosa | 4·108 CFU, 1 ml OD600 1.0 | TOF–MS | |
Vortexed with methanol | Methanol/water/chloroform (5:5:8) | P. aeruginosa | 300 mg wet biomass | 1H NMR | |
Freeze–thaw (× 3) in 50% methanol | Methanol/water (1:1) | K. pneumoniae | ∼8·108 CFU/ml (OD550 = 0.7) | 1H NMR | |
Cryostat (∼ − 50 °C) | 100% methanol | K. pneumoniae | (OD600 = 0.4–0.6) | LC/MS | |
Homogenization with PBS and sonication bath 30 min | PBS buffer | K. pneumoniae | 300 ml, OD600 = 0.7–0.9 | 1H NMR | |
Freeze–thaw (× 3) in 50% methanol, liquid nitrogen freezeing 1 min | Methanol/water (1:1) | B. cereus | 50 mg | GC/TOF–MS | |
Boiling in water for 15 min | Water | C. glutamicum | 1–4 mg | GC/MS | |
Ultrasonic bath 15 min 70 °C in methanol | Methanol/water/chloroform (3:3:2) | C. glutamicum | 5·1010 CFU | GC/MS | |
Incubation with solvents in − 20 °C for 4 h | Methanol/water/chloroform (1:1:2) | C. glutamicum | 20–50 mg wet biomass | LC/MS–MS | |
Freeze–thaw (×3) in 50% methanol | Methanol/water (1:1) | E. faecalis | 50 ml of culturebroth | GC/MS | |
Sonication: sequence (6 s/4 s) for 6 min and bath for 20 min | Methanol/water/chloroform (4:1:1) | E. coli | 5 ml, OD600 = 1.0 | GC–MS | |
Freeze–thaw (×3) in methanol, liquid nitrogen freezeing | 100% methanol | E. coli | 20 ml, 108 CFU/ml | 1H NMR |
