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Sample preparation of H. pylori cholesteryl glucosides

HJ Hau-Ming Jan
YC Yi-Chi Chen
TY Tsai-Chen Yang
LO Lih-Lih Ong
CC Chia-Chen Chang
SM Sasikala Muthusamy
AA Andualem Bahiru Abera
MW Ming-Shiang Wu
JG Jacquelyn Gervay-Hague
KM Kwok-Kong Tony Mong
CL Chun-Hung Lin
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First, 50 μM azido-cholesterol was added to the culture of H. pylori. Then, Folch partitioning was performed using a chloroform/methanol (1/2) extraction for 1 h and another round of chloroform/methanol (2/1) extraction for 1 h, followed by an extraction with 0.9% KCl. The organic layer was evaporated to give a dried residue that was then subjected to a click reaction with 0.25 mM alkyne-dye (4-N-methylamino-1,8-napthalimidopropyne, abbreviated as MAN) in the presence of 1.25 mM tris(benzyltriazolylmethyl)amine (TBTA), 12.5 mM sodium ascorbate, and 0.25 mM CuSO4 at room temperature for 1 h. After the samples were dried, they were stored at −20 °C until HPLC or MS analysis.

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