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Updated: May 15, 2026
Purification and Visualization of Lipopolysaccharide from Gram-negative Bacteria by Hot Aqueous-phenol Extraction
Published on: May 28, 2012
[Removal of oral Prevotella intermedia Endotoxin by octyl phenyl polyoxyethylene ether extraction method]
Ai-wu Wang1, Yan Liu, Kong-xin Hu
1Department of Stomatology, Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, China.
Objective:
To investigate an effective purification method for removing endotoxin from Prevotella intermedia.
Methods:
The main protein ingredients of bacteria prepared from ammonium sulfate precipitation were further treated with octyl phenyl polyoxyethylene ether (Triton X-114), and then processed at 4°C, 37°C and 25°C. The obtained aqueous phase after at least two more cycle repeated operations was assayed for endotoxin by Western blotting, LAL-clotting method, in vitro cell stimulation and in vivo animal experiments.
Results:
Western blotting and LAL-clotting method demonstrated that the reduction in endotoxin level was greater than 99.99% and recovery of the proteins after endotoxin removal was greater than 90% with Triton X-114 treatment for 3 cycles. The cytokines expression level was lower in both in vitro cell stimulation and in vivo animal experiments than in untreated group (P < 0.05).
Conclusions:
The extraction method provides a new choice for endotoxin removal from large volumes of the oral Prevotella intermedia.
