AMCEN LAB SDN BHD ( W) No.18, Lorong Talang 9, Perai, Penang. Tel: Fax:

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1 CUSTOMERS VENTURE SDN BHD Certificate No : AMC/nU1604/033 Lots.3A.010 & 3A.009 & 3A.011 & 3A.015, Sample Log Code : nu1604/022 4th Floor,Endah Parade, Sample Received Date : 19-Apr-2016 No.1,Jalan 1/149E,Taman Sri Endah, Issuance Date : 15-Jun-2016 Bandar Baru Sri Petaling, Kuala Lumpur. Tel: Fax : Attention : Mr.Dickson Cheng Sample Description: Cure Liquefaction Silver 10ppm Water ( ) Test Summary: It is a cell-based colorimetric assay and a quantitative method to determine the cell viability. Yellow tetrazolium MTT is reduced to purple formazan by NADH released from mitochondrial activity in viable cells. Absorbance values of samples will be measured by ELISA reader. Objective: To measure the in-vitro cytotoxic effect of samples on breast cancer cell lines MCF-7 and MDA-MB-231. Page 1 of 8

2 Test procedures: Harvest the MCF-7 cells (Human Breast Adenocarcinoma) and MDA-MB-231 (Human Caucasian Breast Adenocarcinoma) Plate the cells in 96 well plate. Incubate overnight. Treatment of cells with water sample. Incubate the treated cells for 24 hours. Add MTT Reagent to each wells. Incubate for 4 hours. Remove media from each well. Resuspend the cells with DMSO. Obtain absorbance value from ELISA Reader. Result: MCF-7 Control Before Treated Page 2 of 8

3 MCF -7 After Treated with Silver Water Page 3 of 8

4 MDA Control Before Treated Page 4 of 8

5 MDA After Treated with Silver Water Page 5 of 8

6 Viability of cells (%) AMCEN LAB SDN BHD ( W) Table : Viability of Cancer Cell (MDA-MB-231 & MCF-7) Versus Concentration of Silver Water MDA-MB-231 MCF concentration (10ppm/ml) MDA OD1 OD2 OD3 OD average STDEV 10ppm ppm ppm ppm ppm ppm ppm ppm ppm Control Page 6 of 8

7 MDA OD average STDEV 10ppm ppm ppm ppm ppm ppm ppm ppm ppm MCF OD1 OD2 OD3 OD average STDEV 10ppm ppm ppm ppm ppm ppm ppm ppm ppm Control MCF OD average STDEV ppm ppm ppm ppm ppm ppm ppm ppm ppm Page 7 of 8

8 Conclusion: The testing on silver water against two breast cancer cells i.e MDA-MB-231 and MCF7 have been successfully done and the solution inhibited both cells with IC50 values of IC 50= 0.3ppm (for MDA) and IC50=0.088 ppm (For MCF7). The results of the present study showed potent cytotoxic effects on MCF-7 cells and MDA-MB-231. The criteria of cytotoxicity for the crude extract, as established by the U.S. National Cancer Institute (NCI), is an IC 50 < 20 ppm in the preliminary assay. Yeow Liang Ming General Manager B. Sc., M.Sc., M.B.A. M1737/4047/2000 Page 8 of 8