1-Description :
White to off white injectable suspension, free from particulate matter. filled in amber glass vial, closed
with rubber closure, sealed with tear off seal, neatly labeled and packed in a unit carton.
2-Identification :
complies as per method followed in assay preparation
3-PH:Measure the pH on calibrated PH meter, which should be in range 5.0 to 8.0
4- Average volume filled per vial:
Collect all 5 vials, individually take up the contents of each vial, measure in ml in a calibrated cylinder, It should be
in range as 100 ml to 102 ml.
5-Assay (Contents of Total Penicillin G):
Take 1.0ml of injectable suspension (eq. to 115.96 mg of total penicillin G) and dilute to 100ml in water. Take 2 ml
from the in 250 ml stoppered titration flask and add 5 ml of 1 N Sodium Hydroxide Solution. .Add 6 ml of 1N
Hydrochloric Acid and 10 ml of 0.01 N Iodine Solution. Cover, shake well and allow to stand in the dark for 25
minutes. Titrate with 0.01 N Sodium Thiosulphate Solution using Starch Solution as an indicator. End point will be
colourless. Proceed blank without sample same as above mentioned procedure. Each ml of 0.01N Iodine Solution
vs is equivalent to 0.003344 gm of Penicillin G.
x x 0.003344 x 100 = %
0.00232 of the stated amount
6-BACTERIAL ENDOTOXIN TEST:
Sensitivity 0.125EU/ml
Preparation of dilutions
AS is,1/333, 1/666 MVD is 1/666.4
+ve control std. bacterial Endotoxins 2λ
-ve control TAL water ( Endotoxin free water )
6.1 Procedure:-
(for single test vial)
Reconstitute the TAL/LAL reagent ampoules with 0.1ml of TAL water, mark/identify each ampoule with relative
dilution serial. Add 0.1ml of each of prepared dilution in to reaction ampoule. In +ve control add 2λ of std. bacterial
Endotoxin, in –ve control add 0.1ml of TAL water, mix gently and keep at 37 oC ± 1oC for 60 minutes ± 2 minutes.
Any gel formation indicates the presence of Endotoxin of 0.125EU/ml or more than 0.125EU/ml.
For multitest vial, reconstitute the TAL/LAL reagent vial as per instructions mentioned on vial
Calculation Reciprocal of dilution x sensitivity = EU/ml
Limit: NMT: 83.3 E.U/ml
7-STERILITY TEST:
7.1- Sampling
Take the number of units as 2% or 20 ampoule /vials or whichever is less of the whole one batch randomly from
different trolleys and different trays after the autoclave process is complete.
7.2- Apparatus
7.2.1-Enough media bottles containing sterilized medium solution properly Stoppered.
7.2.2-Glass Filtration Assemblies washed and sterilized (at 121 C˚ for 30 min).
7.2.3-SS filtration flask chain attached with vacuum line through collection Tank (of filtered liquid).
7.2.4-Forceps and scissors sterilized wrapped in aluminum foil (at 121 C˚ for 30 minutes).
7.2.5-Sterilized (at 121˚C for 30 minutes) parachute overhaul with cap, sterile Gloves.
7.2.6-Spirit lamp, marker for identification on media bottles.
7.2.7-Incubators set at temperature 32.5˚C±2.5˚C and 22.5˚C±2.5˚C.
7.2.8-Room temperature should not exceed 25˚C, differential pressure should not be less than 0.10 inches of water
gauge in sterility test room (i-e LAF room) to maintain the integrity of sterile room.
7.3-Preparation of Fluid Thioglycollate Medium
STAR LABORATORIES
Sample Preparation:
Take 1ml of injectable suspension( eq. to 200 mg of DihydroStreptomycin ) in 100 ml volumetric flask,and make
up the volume with Distilled water (2mg/ml)
Take 1.0 ml in 25ml volumetric flask; add 1 ml of Nitroprusside reagent ( equal volume of 10 % NaOH, 10 %
potassium ferricyanide and 10 % Nitroprusside solutions) and stay for 10 minutes at room temperature.
Measure the absorption of sample and standard preparations at 490 nm on U.V Spectrophotometer and Calculate
the contents of Dihydrostreptomycin.
X 200 = mg/ml
100