IN- VITRO TOXICITY EVALUATION

March 13th, 2016

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Order Description
DESSERTATION PROJECT PROTOCOL
A. INTRODUCTION
A1. Hydrogen peroxide which is used as food additives (ROS) and toxicological consideration of H2O2. 7-8 pages good reviews. How much hydrogen peroxide reaches to colon?
A2. How hydrogen peroxide absorb in the body and dose?
A3. What concentration has been used in the food?
A4. Does hydrogen peroxide found in food cytotoxic?
A5. About Caco2 cells
A6 Hypothesis
B.METHOD
B1 Tissue culture
C.AIM
D.RESULT
It should be calculate dose response in result.
E.DISCOSSION
Should be 3000 word
F.CONCLUSION
G.ABSTRACT

NOTES
I did approximately 3678 word but I need 6500 word more.
H2O2 treated with Caco2 cells
Commercial hydrogen peroxide was used as a 9.79 Moles and intermediate dilation was carried out during the preparing concentration of hydrogen peroxide.
Hydrogen peroxide concentrations
0.5 ml H2O2 + 4.5 ml PBS has been arrange for 1 Moles (hydrogen peroxide concentration).
3.3 ml has been taken from previous concentration + 1.7 ml PBS for 0.66 Moles (hydrogen peroxide concentration).
3.33 ml previous concentration +1.67 ml PBS for 0.44 Moles (hydrogen peroxide concentration)
3.29 ml previous +1.71ml PBS for 0.29 Moles (hydrogen peroxide concentration)
3.44 ml previous + 1.55 ml PBS 0.20 Moles (hydrogen peroxide concentration)
3.25 ml previous + 1.75 ml PBS 0.13 Moles (hydrogen peroxide concentration)
3.07 ml previous + 1.93 PBS 0.08 Moles (hydrogen peroxide concentration)
This MTT assay repeated 3 times. Because of there are 3 results.

Control
Just cell 0.08 moles 013 moles 0.20 moles 0.29 moles 0.44 moles 0.66 moles 1 moles Blank (no cell)
0.913 0.253 0.228 0.197 0.221 0.188 0.193 0.114 0.151
0.957 0.351 0.232 0.227 0.201 0.238 0.223 0.189 0.164
1.099 0.227 0.214 0.201 0.176 0.142 0.151 0.108 0.168
1.193 0.214 0.35 0.252 0.243 0.212 0.216 0.198 0.173
0.915 0.236 0.216 0.239 0.229 0.202 0.22 0.199 0.173
0.96 0.203 0.221 0.214 0.219 0.203 0.213 0.198 0.197
0.984 0.225 0.212 0.207 0.224 0.202 0.207 0.199 0.171
1.11 0.238 0.217 0.228 0.234 0.208 0.21 0.25 0.172
Table 1: Plate 1. MTT Absorbance Caco2 cells treated with hydrogen peroxide at 540 nm

Concentration (Moles) Average Standard Deviation CV(COEFFIENT OF VARI
Control (Just cell) 1.016375 0.130857 0.102184
0.08 Moles 0.243375 0.046071 0.189301
0.13 Moles 0.23625 0.046478 0.196733
0.20 Moles 0.2200625 0.019235 0.087184
0.29 Moles 0.218375 0.021044 0.096365
0.44 Moles 0.199375 0.027187 0.13636
0.66 Moles 0.204125 0.023351 0.114395
1 Moles 0.181875 0.0447604 0.261739
Blank (No cell) 0.171125 0.012778 0.074668
Table 2: Average, SD, and coefficient of variation of plate 1
Control Just cell 0.08
moles 0.13
moles 0.20
moles 0.29
moles 0.44
moles 0.66
moles 1
moles Blank
(no cell)
0.405 0.225 0.214 0.217 0.22 0.198 0.203 0.184 0.225
0.735 0.243 0.219 0.26 0.31 0.219 0.194 0.192 0.225
0.801 0.204 0.214 0.225 0.195 0.197 0.209 0.193 0.229
0.864 0.231 0.252 0.237 0.184 0.195 0.186 0.199 0.261
0.91 0.215 0.245 0.228 0.235 0.222 0.225 0.203 0.235
0.72 0.231 0.241 0.265 0.223 0.232 0.242 0.206 0.227
0.769 0.218 0.236 0.22 0.224 0.195 0.2 0.205 0.217
0.68 0.198 0.228 0.213 0.203 0.206 0.18 0.185 0.232
Table 3: plate 2 MTT absorbance Caco2 cells treated with hydrogen peroxide at 540 nm
Concentration (moles) Average Standard deviation Coefficient of variation (CV)
Control (just cell) 0.7355 0.153495 0.208695
0.08 moles 0.220625 0.014937 0.067705
0.13 moles 0.231125 0.014604 0.063185
0.20 moles 0.233125 0.019577 0.083977
0.29 moles 0.22425 0.038574 0.172012
0.44 moles 0.208 0.014422 0.069338
0.66 moles 0.204875 0.020427 0.099705
1 moles 0.195875 0.008692 0.044376
Blank 0.231375 0.013114 0.056679

Table 4: Averages, and coefficient of variation of plate2

Control
just cell 0.08
moles 0.13
moles 0.20
moles 0.29
moles 0.44
moles 0.66
moles 1
moles Blank
0.757 0.215 0.208 0.2 0.203 0.196 0.2 0.194 0.198
0.92 0.229 0.193 0.214 0.204 0.181 0.179 0.174 0.196
0.861 0.196 0.2 0.205 0.185 0.186 0.196 0.173 0.205
0.868 0.2 0.211 0.209 0.2 0.195 0.189 0.181 0.2
0.938 0.201 0.212 0.228 0.228 0.19 0.169 0.192 0.204
0.86 0.194 0.208 0.234 0.219 0.189 0.192 0.179 0.217
0.786 0.175 0.214 0.192 0.192 0.171 0.153 0.165 0.19
0.787 0.231 0.234 0.209 0.221 0.19 0.179 0.193 0.205
Table 5:Plate3 MTT absorbance Caco2 cells treated with hydrogen peroxide at 540 nm
Concentration Averages Standard Deviation Coefficient of variation
Control 0.847125 0.065287 0.077069
0.08 moles 0.205125 0.018879 0.092036
0.13 moles 0.21 0.011916 0.056745
0.20 moles 0.211375 0.013897 0.065745
0.29 moles 0.2065 0.014938 0.079776
0.44 moles 0.18725 0.008102 0.043269
0.66 moles 0.182125 0.01557 0.085488
1 moles 0.181375 0.01073 0.059157
Blank 0.201875 0.00799 0.039579
Table 6: Averages, and coefficient of variation of plate3

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