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TEST REPORT TEST TYPE: Liquid Suspension Time Kill Study -Quantitative Test Based On ASTM 2315 TEST METHOD of Colloidal Silver Product at Contact time points: 30 sec, 1 min, 2 min, 5 min, 10 min, 15 min and 30 min. against following microorganisms: Bacterial cultures: 1. S. aureus 2. E. coli 3. Methicillin Resistant S. aureus 4. P. aeruginosa 5. K. pneumoniae 6. Enterococcus faecalis Fungal cultures: 1. Candida albicans 2. Aspergillus niger SAMPLE DETAILS No. of samples receipt date B. 2 x500 ml No: CAS -10PPM One 27-09-2016 SAMPLE DESCRIPTION: Two amber colored bottles.

TEST INFORMATION: Analysis Started on 01-11-2016 Analysis Completed on 12-11-2016 B. No: CAS Contact time 30 sec, 1 min, 2 min, 5 min, 10 min, 15 min and 30 min. Vol. of used Diluent Used 9.9 ml PBS Vol. of Inoculum used Neutralizer used 0.1 ml Dey Engley Neutralizing Broth METHODS USED: METHOD STANDARD: Time Kill Study -Quantitative Test Based On ASTM 2315 Test Method Based on ASTM 2315. MEDIA USED: Soyabean Casein Digest Broth Yeast Malt Broth DE Broth Soyabean Casein Digest Agar Potato Dextrose Agar Phosphate Buffered Saline (PBS) Phosphate Saline 0.85% with 0.2 % Tween 80 CULTURE USED: 1. Staphylococcus aureus ATCCC 6538 2. Klebsiella pneumoniae ATCC 10031 3. Escherichia coli ATCC 8739 4. Methicillin Resistant S. aureus ATCC 33591 5. P. aeruginosa ATCC 9027 6. Enterococcus faecalis ATCC 29212 Fungal cultures: 1. Candida albicans ATCC 10231 2. Aspergillus niger ATCC16404 3. INCUBATION CONDITIONS 35-37 C, Aerobic for 24-48 hrs for Bacteria. 25-27 C Aerobic for 72-96 hrs for Fungi.

I INITIAL INNOCULUM COUNT Staphylococcus aureus ATCC 6538 Klebsiella pneumoniae ATCC 10031 Escherichia coli ATCC 8739 Methicillin Resistant S. aureus ATCC 33591 P. aeruginosa ATCC 9027 Enterococcus faecalis ATCCC 29212 Candida albicans ATCC 10231 Aspergillus niger ATCC16404 CFU/ml 1.5 x 10 8 1.3 x 10 8 2.5 x 10 8 1.7 x 10 8 3.3 x 10 8 1.5 x 10 8 2.0 x 10 8 1.2 x 10 8

II Test Microorganism Concentration Contact Time CFU / ml Log reduction vs. at Percent Reduction vs. at 4.1 E+06 2.0E+05 1.32 95.1222 2.3E+05 1.26 94.390 S. aureus ATCC 6538 B. No: CAS 1.62 6.62 97.561 6.62 1 6.62 6.62 9.0 E+06 3.4E+03 3.42 99.9962% 6.0E+03 3.18 99.9333% Klebsiella pneumoniae ATCC 10031 B. No: CAS 5.0E+03 1.0E+02 3.25 4.95 99.9444% >99.9988% 4.0E+02 4.95 >99.9955% 1 6.95 >99.9993% 6.95 >99.9998%

Test Microorganism Concentration Contact Time CFU / ml Log reduction vs. at Percent Reduction vs. at 2.0 E+05 2.0E+04 1.00 90.00000 % 2.0E+04 1.00 90.0000% Escherichia coli ATCC 8739 B. No: CAS 1 4.5 E+06 3.0E+05 1.18 93.3333% 7.0E+05 84.44444% Methicillin Resistant S. aureus ATCC 33591 B. No: CAS 1.8E+05 7.0E+02 1.39 3.81 6.65 96.0000% 99.9844% 1 6.65 6.65

Test Microorganism Concentration Contact Time CFU / ml Log reduction vs. at Percent Reduction vs. at 4.0 E+05 1.0E+04 1.60 97.5000% 2.0E+03 2.30 99.5000% P. aeruginosa ATCC 9027 B. No: CAS 1 2.0 E+05 1.4E+04 1.15 93.0000% Enterococcus faecalis ATCC 29212 B. No: CAS 1.1E+04 4.0E+03 5.0E+03 1.26 1.70 1.60 94.5000% 98.0000% 97.5000% 4.0E+03 1.70 98.0000% 1 4.0E+03 1.70 98.0000% 3.0E+03 1.82 98.5000%

Test Microorganism Concentration Contact Time CFU / ml Log reduction vs. at Percent Reduction vs. at 3.1 E+05 2.0E+05 35.4838% 67.7419 % Candida albicans ATCC 10231 B. No: CAS 1.0E+04 3.0E+04 1.49 1.01 96.7741% 90.3225 % 2.0E+04 1.19 93.5484% 1 1.3E+04 1.38 95.8064 % 8.0E+03 1.59 97.4193% 4.0 E+05 Aspergillus niger ATCC 16404 B. No: CAS 2.0E+05 2.2E+05 3.0E+04 1.12 92.5000% 1 2.0E+05

RESULTS: Under the given test conditions: 1. The Test - B. No: CAS exhibits more than count of Staphylococcus aureus ATCC 6538 at of contact time. 2. The Test - B. No: CAS exhibits more than count of Klebsiella pneumoniae ATCC 10031 at. of contact time. 3. The Test - B. No: CAS exhibits more than count of Escherichia coli ATCC 8739 at of contact time. 4. The Test - B. No: CAS exhibits more than count of Methicillin Resistant S. aureus ATCC 33591 at of contact time. 5. The Test - B. No: CAS exhibits more than count of P. aeruginosa ATCC 9027 at of contact time. 6. The Test - B. No: CAS exhibits 98.50% reduction in the count of Enterococcus faecalis ATCC 29212 after of contact time. 7. The Test - B. No: CAS exhibits 97.42 % reduction in the count of Candida albicans ATCC 10231 after of contact time. 8. The Test - B. No: CAS exhibits 75.00% reduction in the count of Aspergillus niger ATCC16404 after of contact time. Report prepared by: MAA Checked by: AGA Disclaimer Shubhda Research Institute does not assume any responsibility as regards the origin of the sample, resultss obtained or further actions taken by you based on our results. EOD