Sterile Saline (0.9% w/v NaCl)
Dissolve 9.0gm of NaCl in 1000ml water and dispense 9.0ml of the solution into 20ml test tubes. Autoclave the tubes as per the validated cycle and cool to room temperature.
Sterile saline with polysorbate 80:
Dissolve 9.0gm of NaCl in 1000ml water and add .5gm of tween 80 to the resulting saline solution, dispense 9.0 ml of the solution into a number of 20ml test tubes. Autoclave the tubes
Preparation of Inoculums and its Standardization:
1) Take loop full of culture from working slant of respective organism and streak it on the respective agar medium. The ATCC microbial culture strains were revived and sub-cultured on the slants of Soybean casein digest agar and Sabouraud dextrose agar for the growth of bacteria …show more content…
The volume of the suspension inoculum used is between 0.5% and 1.0% of the volume of the product.
The concentration of test microorganisms that is added to the product are such that the final concentration of the test preparation after inoculation is between 1 × 105 and 1 × 106 CFU per ml of the product. Each sample tube was inoculated with one of the prepared and standardized inoculum in such a way that after inoculation the final concentration of microorganism remains between 1×105 and 1×106 CFU/ml.
The initial concentration of viable microorganisms in each test preparation is estimated based on the concentration of microorganisms in each of the standardized inoculum as determined by the plate-count method.
All the inoculated tubes were incubated at 20-25oC for 28 days and viable counts were periodically determined by pour-plate method at 0, 7, 14, and 28 days subsequent to the inoculation.
Determine by the plate-count procedure the number of CFU present in each test preparation for the applicable …show more content…
2 Topically used products made with aqueous bases or vehicles, nonsterile nasal products, and emulsions, including those applied to mucous membranes.
3 Oral products other than antacids, made with aqueous bases or vehicles.
4 Antacids made with an aqueous base. Table 2: Criteria for Tested Organisms
Category Microbes Criteria
1 Bacteria Not less than 1.0 log reduction from the initial calculated count at 7 days, not less than 3.0 log reduction from the initial count at 14 days, and no increase from the 14 days’ count at 28 days. Yeast and Molds No increase from the initial calculated count at 7, 14, and 28 days.
2 Bacteria Not less than 2.0 log reduction from the initial count at 14 days, and no increase from the 14 days’ count at 28 days. Yeast and Molds No increase from the initial calculated count at 14 and 28 days.
3 Bacteria Not less than 1.0 log reduction from the initial count at 14 days, and no increase from the 14 days’ count at 28 days. Yeast and Molds No increase from the initial calculated count at 14 and 28 days.
4 Bacteria, Yeast, and Molds No increase from the initial calculated count at 14 and 28
Dissolve 9.0gm of NaCl in 1000ml water and dispense 9.0ml of the solution into 20ml test tubes. Autoclave the tubes as per the validated cycle and cool to room temperature.
Sterile saline with polysorbate 80:
Dissolve 9.0gm of NaCl in 1000ml water and add .5gm of tween 80 to the resulting saline solution, dispense 9.0 ml of the solution into a number of 20ml test tubes. Autoclave the tubes
Preparation of Inoculums and its Standardization:
1) Take loop full of culture from working slant of respective organism and streak it on the respective agar medium. The ATCC microbial culture strains were revived and sub-cultured on the slants of Soybean casein digest agar and Sabouraud dextrose agar for the growth of bacteria …show more content…
The volume of the suspension inoculum used is between 0.5% and 1.0% of the volume of the product.
The concentration of test microorganisms that is added to the product are such that the final concentration of the test preparation after inoculation is between 1 × 105 and 1 × 106 CFU per ml of the product. Each sample tube was inoculated with one of the prepared and standardized inoculum in such a way that after inoculation the final concentration of microorganism remains between 1×105 and 1×106 CFU/ml.
The initial concentration of viable microorganisms in each test preparation is estimated based on the concentration of microorganisms in each of the standardized inoculum as determined by the plate-count method.
All the inoculated tubes were incubated at 20-25oC for 28 days and viable counts were periodically determined by pour-plate method at 0, 7, 14, and 28 days subsequent to the inoculation.
Determine by the plate-count procedure the number of CFU present in each test preparation for the applicable …show more content…
2 Topically used products made with aqueous bases or vehicles, nonsterile nasal products, and emulsions, including those applied to mucous membranes.
3 Oral products other than antacids, made with aqueous bases or vehicles.
4 Antacids made with an aqueous base. Table 2: Criteria for Tested Organisms
Category Microbes Criteria
1 Bacteria Not less than 1.0 log reduction from the initial calculated count at 7 days, not less than 3.0 log reduction from the initial count at 14 days, and no increase from the 14 days’ count at 28 days. Yeast and Molds No increase from the initial calculated count at 7, 14, and 28 days.
2 Bacteria Not less than 2.0 log reduction from the initial count at 14 days, and no increase from the 14 days’ count at 28 days. Yeast and Molds No increase from the initial calculated count at 14 and 28 days.
3 Bacteria Not less than 1.0 log reduction from the initial count at 14 days, and no increase from the 14 days’ count at 28 days. Yeast and Molds No increase from the initial calculated count at 14 and 28 days.
4 Bacteria, Yeast, and Molds No increase from the initial calculated count at 14 and 28