Method for in-vitro evaluation on safety of cigarettes and tobaccos
A safety and tobacco technology, applied in the measurement of color/spectral characteristics, material inspection products, biological testing, etc., can solve the problems of long safety cycle, low practicability, and complex smoke composition of cigarettes, and solve the complex evaluation mechanism. , The operation procedure is simple, the evaluation result is accurate and reliable
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Embodiment 1
[0050] The present embodiment evaluates the safety of 18 kinds of cigarettes (named by letters from A to R respectively) by the following steps:
[0051] Step 1: Smoke the 18 kinds of cigarettes to be evaluated separately, and then prepare the smoke water extract CSE respectively, and detect the absorbance value OD of each CSE at 340nm 340nm , to OD 340nm As a quantitative basis for evaluating the safety of cigarettes or tobacco;
[0052] Such as figure 1 As shown, CSE is prepared as follows: a cigarette to be evaluated is sucked under standard conditions (vacuum pressure is 0.08Mpa) by vacuum pump DP-01, and the mainstream smoke produced is sequentially passed through two series-connected Collect it in a gas absorption bottle with 5mL replenishment liquid. After smoking a cigarette or tobacco to be evaluated, suck it for 10 seconds so that the replenishment liquid can fully collect the mainstream smoke. The solution is filtered through a 0.22 μm water membrane to obtain CS...
Embodiment 2
[0060] This embodiment is used to prove that the incubation time in step 3 of the method of the present invention is selected for the reason of 20min:
[0061] Add 20 μL of CSE made from cigarette A to the rat liver homogenate obtained in Example 1, and detect the inhibitory rate of CSE to TrxR activity in the rat liver homogenate at different time points, and draw the time-effect curve of the inhibitory rate, Such as Figure 5 As shown in a, it can be seen from the figure that the inhibition rate has stabilized at 20 minutes, so the selected time is 20 minutes.
[0062] In addition, the biological reagents were changed to mouse lung homogenate and TrxR pure enzyme preparations (product number: T9698, produced by sigma company), and the same test was carried out, and the results were as follows: Figure 5 As shown in b and 5c, it can be seen from the figure that when CSE inhibits different biological agents, the inhibition rate can be stabilized for 20 minutes.
Embodiment 3
[0064] This example is used to prove that the results of the method of the present invention are reliable and repeatable.
[0065] The biological reagents in Example 1 were respectively replaced with mouse lung homogenate and TrxR pure enzyme preparations, and the same test was carried out. The dose-effect curves of the inhibition rate obtained in step 3 were as follows: Figure 6 a and 6b.
[0066] Use the ratio of the S value of cigarette A and cigarette R to compare the results obtained by using three biological agents to evaluate the safety of cigarettes, such as Figure 7 As shown, it can be seen that although the ratios obtained when different biological agents are selected for evaluation are different, they have the same rule, and the S value of cigarette A is greater than the S value of cigarette R. The feasibility of the method of the present invention is proved.
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