Whole lipids of cultured H. pylori SS1 were prepared by single-phase extraction as described by Tsugawa et al. (2020) (link). Briefly, 1.0 × 109 CFUs of dried bacterial cells were suspended in 100 μl chloroform. After 1 h incubation, 195 μl methanol and 5 μl EquiSPLASH (Avanti Polar Lipids) were mixed and incubated for another 2 h at room temperature. Thereafter, 20 μl of water was added, and the samples were incubated for 10 min. After extraction, samples were centrifuged at 2,000 ×g for 10 min, and the supernatants were collected. The extracted lipids were measured using the ACQUITY UPLC I class system (Waters) coupled with a TripleTOF 6600 (AB Sciex; Tsugawa et al., 2020 (link)). Liquid chromatography separation was performed using a reverse-phase column (ACQUITY UPLC BEH C18 column [2.1 mm i.d. × 50 mm, particle size 1.7 μm; Waters]), and data acquisition was performed by data-dependent MS/MS in the negative and positive ion modes. The detected lipid species were annotated using MS-DIAL (Tsugawa et al., 2015 (link)) and Peak View (AB Sciex).
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Lipid profiling of Helicobacter pylori
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Corresponding Organization : Chiba University
Other organizations : RIKEN Center for Integrative Medical Sciences, Gifu University, Tokyo Medical and Dental University, University of Birmingham, Victoria University of Wellington, Maurice Wilkins Centre, University of Auckland, University of Melbourne, Yokohama City University, Keio University, Amsterdam University Medical Centers
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- Whole lipids of cultured H. pylori SS1 were prepared by single-phase extraction as described by Tsugawa et al. (2020)
- The extracted lipids were measured using the ACQUITY UPLC I class system (Waters) coupled with a TripleTOF 6600 (AB Sciex; Tsugawa et al., 2020 (link)).
- Liquid chromatography separation was performed using a reverse-phase column (ACQUITY UPLC BEH C18 column [2.1 mm i.d. × 50 mm, particle size 1.7 μm; Waters]), and data acquisition was performed by data-dependent MS/MS in the negative and positive ion modes.
- The detected lipid species were annotated using MS-DIAL (Tsugawa et al., 2015 (link)) and Peak View (AB Sciex).
- 1.0 × 10^9 CFUs of dried bacterial cells were suspended in 100 μl chloroform.
- 195 μl methanol and 5 μl EquiSPLASH (Avanti Polar Lipids) were mixed and incubated for another 2 h at room temperature.
- 20 μl of water was added, and the samples were incubated for 10 min.
- After extraction, samples were centrifuged at 2,000 ×g for 10 min, and the supernatants were collected.
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