The Procedure for Anticancer Evaluation
The target cancer cell lines were seeded in 96-well plates
(5000cells/well) with 100μL DMEM supplemented with 10% fetal bovine
serum, and cultured at 37 oC in a humidified
CO2 incubator (95% air, 5% CO2) for
24h. While the cell lines grew to 90% in logarithmic growth, the
culture medium was removed from each well, and 100 μL fresh DEME was
added to each well. Then, 10μL
different concentrations of P. nagi kernel oil solutions were
added into each well (every concentration was repeated for 5 times) and
the plates were incubated for another 48h at 37 oC.
Subsequently, 10μL CCK8 was added to each well, and the plates were
cultured at 37 oC for another 4 hours. The optical
density was measured at a wave-length of 450 nm on an ELISA microplate
reader. DMEM and DMSO solution (V/V: 10/1) was used as a negative
control. The results were expressed as the inhibition calculated at the
ratio [(1-(OD450 treated/ OD450negative control)) ×100].
3. Results and discussion
3.1 The influence of steaming time
on the yield and quality of P. nagi kernel oil
The P. nagi kernel oil was obtained through two steps: (1)
steaming the powder of P. nagi kernel; (2) squeezing the steamed
powder of P. nagi kernel. During the experiment, we found that
the time of steaming of the powder of P. nagi kernel is extremely
important for the yield and the quality of the oil, the short time
steaming does not lead to the higher yield, while long time steaming
will lead to the higher yield, but the oil also contains higher water
content, which affects the quality of the oil, making the oil very
turbid. Thus, we optimized the experimental conditions about the
steaming time, the yield and the quality of oil. The results (see totable 3 ) exhibited that the yield of oil increased with the
increasing of the steaming time, and the steaming time is 20min with the
90% yield of oil, and the oil quality is normal; even if the steaming
time is 25min with the 93% yield, the oil quality is turbid and the
yield increased only 3% compared with the 20min steaming time. Thus, we
selected steaming time 20min as the optimized condition.
Table 3 . Steaming time of P. nagi kernel, yield and oil
quality