Answer Posted / praveen mewar
Dilute Sautan's Medium (Twin Pack) HiMedia code No. MM1276
Dilute Sauton's Fluid Medium is used as a diluent Mycobacterium species.
Composition**
Ingredients Gms / Litre
Part A -
Ferric ammonium citrate ( brown) 0.017
L-Asparagine 1.330
Magnesium sulphate, 7H2O 0.167
Dipotassium hydrogen phosphate 0.178
Sodium dihydrogen phosphate 0.057
Sodium chloride 0.036
Polysorbate 80 0.833
Part B -
Citric acid 0.667
Final pH ( at 25°C) 7.2±0.2
**Formula adjusted, standardized to suit performance parameters
Directions
Suspend 2.52 grams(the equivalent weight of dehydrated medium of Part A ) in 500 ml distilled water containing 20 ml
glycerol. Heat if necessary to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 20
minutes. Suspend 0.667 grams of Part B in 500 ml distilled water. Sterilize by autoclaving at 121°C for 20 minutes. After
autoclaving aseptically mix Part A and Part B solution .Cool to 45-50°C.Dispense into tubes or flasks as desired.
Principle And Interpretation
Dilute Sauton's Medium is used for determining the number of colony forming units (CFU) in vaccines of known potency
as per I.P. (1, 2).
As per the I.P. method, contents of the freeze-dried vaccine for human use are reconstituted with the diluent stated on the vial.
Contents of 5 such containers are pooled and three dilutions of the pooled vaccine are prepared using Dilute Sauton's Medium
so as to obtain an optimum of 100, 40 and 20 colonies from an inoculum of 0.2 ml. An inoculum of 0.2 ml from each dilution
is further inoculated onto L.J. Medium (MM162) slant surface and incubated at 37°C for 28 days. The vaccine passes the test
if 0.1 ml of the reconstituted vaccine contains between 1 x105 and 33 x 105 colony forming units.
Salts like ferric ammonium citrate and magnesium sulphate provide inorganic ions and nitrogen sources essential for the growth
of Mycobacteria. Asparagine is added to promote the initiation of growth and increase the growth rate. Glycerol in the medium
serves as a carbon source.
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