|
Material and Method
1.1 Sample£ºHealthin
tea offered by Chongqing Healthin Health Food
Research Institute. Treatment of sample: Pour
boiling water into 200g sample tea (water should be
20 times of sample); filter after 45 minutes and
pick up the liquid; pour the deposit again and then
combine the two liquid. Condense the liquid to
100ml. This is the formulated concentrate of the
test. £¨1ml¡Ö2g Healthin tea£©
1.2 Tested animal:
the Grade I Kunming Mouse and the SD Rat are all
provided by Sichuan Antibacterial Industry Research
Institute of the State Medical Administration.
1.3 The acute per os
test for both rats and mice: According to the method
of Horn, male and female are tested respectively.
Kunming mouse weighs 20-24g and SD rat weighs
180g-220g. Each dosage team consists five mice and
rats respectively. The tested dosage is 4.0,8.6,18.6
and 40g/kg respectively. Observe for two weeks and
record the animal's toxic reaction and death.
1.4 Mouse erythrocyte
micronucleus test: Kunming Mouse (Weighs 25-32g) is
classified into five teams random. Ten mice each
team, half are male and half are female. Three
Healthin test teams with the dosage of 5.0¡¢10.0¡¢20.0g/kg.bw
respectively. One female control group (distilled
water£© and one male control group (cyclophosphamide:
40mg/kg.bw). The tested animals were fed twice
within 0-24 hours and by the 30th hours, the mice
were killed. Take some femoral medulla, dye with
10%Giemsa liquid and then observe under oil
immersion lens. Count and record cells with
microkernel. Figure the rate of microkernel and
analyze the difference of the rate of microkernel
between the test team and the control group.
1. 530d Feeding
Experiment£ºRandom classified the weaned SD rats
(male rats weigh 125¡À20g ) into four groups, 20
rats each group with half of the female rats and
half of the male rats. Each test team fed the rats
at the dosage of 5.0¡¢10.0¡¢20.0g/kg respectively.
In the other control group, £¨The feeding dosage is
0£©. After the experiment, carry out the
serological check and histopathological check.
2 Conclusion
2.1 Acute toxicity
test£ºmouse's per os acute toxicity(see chart I)
Chart 1 mouse's per
os acute toxicity of Healthin Prevervation Tea
| Animal |
Sex |
Amount
of animal |
Method |
Dose
£¨g/kg.bw) |
Amount
of the Dead |
LD50(g/kg.bw£© |
Conclusion |
| Mouse
of Kunming Origin |
Male |
5 |
per
os |
4.0 |
0 |
>40 |
Innocuity |
| 5 |
per
os |
8.6 |
0 |
| 5 |
per
os |
18.6 |
0 |
| 5 |
per
os |
40.0 |
0 |
| Mouse
of Kunming Origin |
Female |
5 |
per
os |
4.0 |
0 |
>40 |
Innocuity |
| 5 |
per
os |
8.6 |
0 |
| 5 |
per
os |
18.6 |
0 |
| 5 |
per
os |
40.0 |
0 |
As the chart shown,
mouse and rate has the same testing result in per os
acute toxicity test. After fed with Healthin
Preservation Tea, none of the testing team is toxic.
There was no animal dead within two weeks. That is
to say, acute per os LD50 of Healthin Preservation
Tea to both mouse and the rat are more than 40g/kg.bw.
So, the sample tea is innocuous.
2.2 Mouse's cellulae
medullares micronucleus test£ºSee Chart 2.
Chart 2 Healthin
Preservation Tea's Effect on Mouse's cellulae
medullares micronucleus originating rate
| Team |
Dose
£¨g/kg.bw) |
Animal
amout
|
Observed
cells |
Microkernel
celss |
Rate
of microkernel£¨¦Î¡ÀS£¬¡ë£© |
| Neu
Control Group |
0 |
10 |
1000¡Á10 |
53 |
5.3¡À1.16 |
| Healthin
Preservation Tea |
5.0 |
10 |
1000¡Á10 |
41 |
4.1¡À1.64 |
| 10.0 |
10 |
1000¡Á10 |
30 |
3.0¡À1.05 |
| 20.0 |
10 |
1000¡Á10 |
49 |
4.9¡À1.79 |
| Cyclophosphamide |
0.04 |
10 |
1000¡Á10 |
360 |
36.0¡À10.9* |
*P<0.01
From Chart 3, we can
draw a conclusion that the rate of microkernel of
positive control group is obviously higher than the
negative control group £¨P<0.01£©.While, the
groups with the Healthin Preservation Tea is lower
than the negative ones. That is, the result of the
test is negative.
2.3 30d Feeding
Experiment
In the course of the
emperiment, no abnomity or toxicosis or death of the
tested animals happened.
2.3.1 Effect of
Healthin Preservation Tea on Rat. See Chart 3.
Chart 3 Rat's Weight
Change in 30-day Feeding Experiment
| Sex |
Dose
£¨g/kg£© |
Amount
of animal |
Average
weight£¨g,¦Î¡ÀS£© |
| Before |
After |
Weight
increase |
| Male |
0 |
10 |
124¡À11.3 |
308¡À32.5 |
184¡À32.8 |
| 5.0 |
10 |
125¡À10.7 |
300¡À58.7 |
175¡À53.2 |
| 10.0 |
10 |
122¡À7.0 |
291¡À46.3 |
169¡À42.0 |
| 20.0 |
10 |
126¡À12.3 |
299¡À40.4 |
173¡À32.5 |
| Female |
0 |
10 |
122¡À12.0 |
213¡À16.1 |
90.8¡À15.7 |
| 5.0 |
10 |
124¡À9.2 |
209¡À26.5 |
85.2¡À20.4 |
| 10.0 |
10 |
126¡À5.8 |
205¡À16.0 |
79.4¡À11.7 |
| 20.0 |
10 |
126¡À8.8 |
202¡À20.2 |
78.1¡À16.5 |
From Chart 3, we can
draw a conclusion that there is no obvious
difference of the weight increase between the groups
£¨P>0.05£©.
2.3.2 Effect of
Healthin Preservation Tea on Availability Factor of
the Rat. See Chart 4.
Chart 4 Effect of
Healthin Preservation Tea on Availability Factor of
the Rat
| Sex |
Dose
£¨g/kg£© |
Amount
of animal |
Average
weight increase
£¨g,¦Î¡ÀS£©
|
Average
feeding volume
£¨g/30d.Ö»£©
|
Availability
factor
£¨%£©
|
Statistic
t
|
P
Value |
| Male |
0 |
10 |
184¡À32.8 |
909.0 |
20.2¡À3.61 |
|
¡¡ |
| 5.0 |
10 |
175¡À53.2 |
891.0 |
19.6¡À5.97 |
0.27 |
>0.05 |
| 10.0 |
10 |
169¡À42.0 |
900.0 |
18.8¡À4.67 |
0.42 |
>0.05 |
| 20.0 |
10 |
173¡À32.5 |
874.5 |
19.7¡À3.71 |
0.30 |
>0.05 |
| Female |
0 |
10 |
90.8¡À15.7 |
594.0 |
15.3¡À2.64 |
|
¡¡ |
| 5.0 |
10 |
85.2¡À20.4 |
594.0 |
14.3¡À3.43 |
0.73 |
>0.05 |
| 10.0 |
10 |
79.4¡À11.7 |
588.0 |
13.5¡À1.99 |
1.91 |
>0.05 |
| 20.0 |
10 |
78.1¡À16.5 |
588.0 |
13.3¡À2.78 |
1.65 |
>0.05 |
From Chart 4, we can
draw a conclusion that there is no obvious
difference of the availability factor between the
groups £¨P>0.05£©.
2.3.3 Hematological
Examination. See Chart 5
Chart 5 Result of
Hematological Examination of 30-day Feeding
Experiment of Healthin Preservation Tea
Dose
£¨g/kg£© |
Amount of
Animal |
Hemoglobin
£¨g/L£© |
Erythrocyte
£¨X1012/L£© |
Leucocyte
£¨X109/L£© |
| 0 |
20 |
141¡À9.72 |
5.64¡À0.389 |
13.5¡À6.09 |
| 5.0 |
20 |
142¡À11.3 |
5.66¡À0.454 |
11.6¡À3.27 |
| 10.0 |
20 |
146¡À12.8 |
5.85¡À0.512 |
12.0¡À5.10 |
| 20.0 |
20 |
138¡À9.6 |
5.54¡À0.383 |
11.7¡À4.20 |
There is no obvious
difference of Hb¡¢RBC¡¢WBCfrom the control group£¨P>0.05£©
2.3.4 Effect of
Healthin Preservation Tea on Rat's WBC. See Chart 6.
Chart 6 Effect of
Healthin Preservation Tea on Rat's WBC
| Dose£¨g/kg) |
Amount
of Animal |
Acidophilic
% |
Neuter
% |
Lymphatic
% |
Monokaryon
% |
| 0 |
20 |
0.90¡À1.21 |
19.0¡À11.2 |
79.3¡À11.6 |
0.76¡À0.83 |
| 5.0 |
20 |
1.15¡À1.90 |
18.2¡À6.66 |
79.2¡À7.53 |
1.45¡À1.05 |
| 10.0 |
20 |
1.15¡À0.88 |
18.0¡À8.15 |
78.7¡À6.91 |
1.15¡À0.93 |
| 20.0 |
20 |
0.70¡À0.82 |
18.4¡À8.09 |
79.8¡À8.04 |
1.30¡À1.03 |
2.3.5 Result of
Telophase Biochemical Examination. See Chart 7
Chart 7 Result of
Telophase Biochemical Examination
|
Dose
ml/kg
|
Amount of Animal
|
Aminotransierase
U/L
|
Urea mitrogen
mmol/L
|
Creatinine
¦Ìmol/L
|
Cholesterin
mmol/L
|
Glycerin
mmol/L
|
Blood sugar
mmol/L
|
Total protein
g/L
|
Albumin
g/L
|
| 0 |
20 |
91.4¡À17.7 |
8.51¡À1.67 |
88.2¡À9.56 |
2.03¡À0.37 |
1.18¡À0.25 |
3.52¡À0.40 |
73.9¡À5.14 |
31.1¡À2.05 |
| 5.0 |
20 |
95.9¡À13.1 |
7.21¡À1.79* |
80.0¡À10.0* |
2.24¡À0.32 |
1.19¡À0.46 |
3.57¡À0.69 |
73.3¡À5.68 |
30.7¡À3.69 |
| 10.0 |
20 |
94.2¡À16.8 |
6.40¡À1.97** |
75.9¡À3.55** |
2.20¡À0.31 |
1.27¡À0.31 |
3.79¡À0.50 |
75.4¡À4.06 |
32.1¡À2.46 |
| 20.0 |
20 |
96.8¡À16.0 |
6.52¡À0.96** |
74.9¡À5.85** |
2.15¡À0.27 |
1.31¡À0.27 |
3.88¡À0.69 |
74.1¡À3.59 |
30.2¡À1.79 |
From Chart 7, there
is no toxicological change in the examination.
2.3.6 Effect of
Healthin Preservation Tea on Rat's £¨X¡ÀS£©
| Sex |
Dose
(ml/kg) |
Amount
of animal |
HSI
(¡ë) |
KSI
(¡ë) |
SSI
(¡ë) |
TSI
(¡ë) |
| Male |
0 |
10 |
30.4¡À2.47 |
7.44¡À0.49 |
4.41¡À1.18 |
- |
| 5.0 |
10 |
31.7¡À3.04 |
7.77¡À0.81 |
3.92¡À0.74 |
- |
| 10.0 |
10 |
30.1¡À2.54 |
7.26¡À0.52 |
3.92¡À1.74 |
- |
| 20.0 |
10 |
31.9¡À41.6 |
7.82¡À0.78 |
5.12¡À1.33 |
- |
| Female |
0 |
10 |
33.6¡À4.56 |
6.92¡À0.46 |
4.46¡À0.87 |
10.0¡À1.84 |
| 5.0 |
10 |
33.0¡À3.57 |
7.22¡À0.57 |
3.48¡À1.09 |
10.7¡À2.13 |
| 10.0 |
10 |
32.0¡À2.23 |
6.72¡À0.39 |
3.97¡À0.85 |
10.0¡À1.90 |
| 20.0 |
10 |
30.8¡À1.85 |
6.82¡À0.355 |
4.27¡À1.13 |
10.2¡À1.21 |
There is no obvious
difference between the two groups.(P<0.05)
¡¡ |
