A suspension of triolein (80 mg), phosphatidylcholine (10 mg),
and sodium taurocholate (5 mg) in 9 mL of 0.1 M tris–HCl buffer
(pH 7.0) containing 0.1 M NaCl was sonicated for 10 min. This
sonicated substrate suspension (0.1 mL) in a test tube was preincubated
with 5 lL of test sample in DMSO and 95 lL of the
tris–HCl buffer for 3 min at 37 C. An aliquot of porcine pancreatic
lipase (250 lg/mL, type II, Sigma Chemical Co.) (50 lL) or the
tris–HCl buffer (50 lL) as a blank test was then added to start
the reaction. After 30 min of incubation, the test tube was immediately
immersed in boiling water for 2 min to stop the reaction, then
cooled with water. Free fatty acid concentration was determined
by a commercial kit (NEFA C-test Wako, Wako Pure Chemical
Industries Ltd.). Theasaponin E1 was isolated from C. sinensis and
used as a reference compound. IC50 was determined graphically
(N = 4).
A suspension of triolein (80 mg), phosphatidylcholine (10 mg),
and sodium taurocholate (5 mg) in 9 mL of 0.1 M tris–HCl buffer
(pH 7.0) containing 0.1 M NaCl was sonicated for 10 min. This
sonicated substrate suspension (0.1 mL) in a test tube was preincubated
with 5 lL of test sample in DMSO and 95 lL of the
tris–HCl buffer for 3 min at 37 C. An aliquot of porcine pancreatic
lipase (250 lg/mL, type II, Sigma Chemical Co.) (50 lL) or the
tris–HCl buffer (50 lL) as a blank test was then added to start
the reaction. After 30 min of incubation, the test tube was immediately
immersed in boiling water for 2 min to stop the reaction, then
cooled with water. Free fatty acid concentration was determined
by a commercial kit (NEFA C-test Wako, Wako Pure Chemical
Industries Ltd.). Theasaponin E1 was isolated from C. sinensis and
used as a reference compound. IC50 was determined graphically
(N = 4).
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