Ringer's Solution is used for
the preparation of suspension of samples in bacteriological studies and for use
as a diluent in dilution techniques for enumeration.Also used
in the microbial control of apparatus by the rinse and swab method.
Approximate Formula:
Ingredients
gm/ampoule
Sodium chloride
1.125
Potassium chloride
0.0525
Calcium chloride, anhydrous
0.03
Sodium bicarbonate
0.025
Final pH 6.8-7.2 at 25 ℃
*Adjusted and/or supplemented as required
to meet performance criteria
Directions:
To
prepare a 1/4-strength Ringer's Solution:
1.Take 1 piece of ampoule, shortly tap to
ensure the granules are at the bottom.
2. Bend the upper part of the ampoule until
it breaks open.
3. Suspend granules of 1 ampoule in 500ml of deionized
or distilled water. Shake to dissolve the granules completely.
4. Sterilize in an autoclave at 121℃(15
lbs.) for 15 minutes.
5.The appearance of the solution is clear and colorless (after autoclave).
Principle and Interpretation:
1/4-strength Ringer's Solution is isotonic
with bacteria and thus prevents them from being subjected to osmotic shock or
from being osmotically damaged when they are removed from their customary
environment. It is also more physiologically suitable for sensitive
microorganisms than physiological saline.
Appearance:
Dehydrated medium is a free-flowing white
granule. The prepared broth is a kind of colorless transparent liquid.
Precautions:
This medium is for laboratory use
only. Dried medium which is past shelf life, caking or colour variation cannot
be used.
Storage conditions and Shelf life:
Ringer ampoules for the preparation of Ringer's
Solution must
be stored tightly capped in the original container at 5-30°C. The dehydrated medium has a shelf life
of 3 years from date of manufacturing. Prepared medium may be stored, out of
direct light at 2-8°C.
Quality control:
Prepare
the culture medium as per label directions. Inoculate cultures into test
solution. At time zero (directly after inoculation) and after 45 minutes
incubation at 20-25℃, subculture to TSA and incubate at 30-35°C
for 18-24 hours. Determine colony counts at time zero and after 45 minutes.
Microorganism
Strains Number
Inoculum (CFU)
Recovery of 45 minutes
Clostridium
sporogenes
ATCC
19404
100-1000
±50%
of time zero counts
Staphylococcus
aureus
ATCC
6538
100-1000
±50% of time
zero counts
Bacillus subtilis
ATCC
6633
100-1000
±50% of time
zero counts
Pseudomonas aeruginosa
ATCC
9027
100-1000
±50% of time
zero counts
Candida albicans
ATCC
10231
100-1000
±50% of time
zero counts
Aspergillus niger
ATCC
16404
100-1000
±50% of time
zero counts
References:
1. Ministry of Health (1937). Bacteriological
tests for graded milk. Memo 139/Foods. London.
2. J.G. Davis, Laboratory Control of Dairy
Plant, Dairy lndustries Ltd., London (1956)
3.Internationaler Milchwirtschaftsverband
(1985). Zählung coliformer Bakterien in Milch und Milchprodukten
(Internationaler Standard FIL-IDF 73).
4.Internationaler Milchwirtschaftsverband
(1992). Milch und Milchprodukte – Vorbereitung der Untersuchungsproben und
Herstellung der Verdünnungen für mikrobiologische Untersuchungen
(Internationaler IMV-Standard 122).
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