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1.0 PURPOSE
To lay down the
Procedure for Bio-burden, BET and LPC of primary packaging materials.
2.0 SCOPE
This is applicable to Microbiology lab
3.0 RESPONSIBILITY
Microbiologist
4.0 ACCOUNABILITY
Head of Department
5.0 PROCEDURE
5.1 Bio-burden:
5.1.1 Collect 10 units each of vials, rubber stopper, and seals and bottle
randomly. Collect 10 gm sample in case of aluminium foil and PVC sheet as per
SOP for sampling packing material.
5.1.2 Use clean and sterilized sampling devices to avoid external
contamination.
5.1.3 Bring sample to microbiology lab for testing.
5.1.4 Carry out test under LAF.
5.1.5 For Vials
and Bottle: Fill approximately half of the total
volume of vials and bottles aseptically with Sterile saline. Shake vials and
bottles in such a way that internal surface of vial and bottle are to be rinsed
properly.
5.1.6 Filter
aseptically whole content of each vial through sterilized filtration assembly
(0.45μ membrane filter) and rinse with 100 ml of sterilized 0.1% peptone water.
5.1.7 Lift
membrane filter with sterilized forceps and place it on pre-incubated soybean
casein digest agar plate avoiding air bubble entrapped under filter paper.
5.1.8 Incubate
plate at 20-250C for 72 hrs. followed by 30-350C for 48
hrs.
5.1.9 For Rubber
Stopper and Seals: Under aseptic condition dip sample of
seal and rubber stopper in each of 100 ml of sterile saline, shake it properly and
filter through sterilized filtration assembly (0.45μ membrane filter) and rinse
with 100 ml of sterilized 0.1% peptone water.
5.1.10 Lift
membrane filter with sterilized forceps and place it on pre-incubated soybean
casein digest agar plate avoiding air bubble entrapped under filter paper.
5.1.11 Incubate
plate at 20-250C for 72 hrs. followed by 30-350C for 48
hrs.
5.1.12 For
Aluminium Foil and PVC Foil: Prepare
sterilized swab in saline and Take aseptically swab of 5X5cm area
from the internal surface of aluminium foil and PVC sheet with the help of
sterilized 5X5cm template in such a way total 5X5 cm area is to be covered
properly.
5.1.13 Dip swab
stick in saline tube and vortex it properly and filter whole content of saline
through sterilized filtration assembly (0.45μ membrane filter) and rinse
membrane filter with 100 ml of sterilized 0.1% peptone water.
5.1.14 Lift
membrane filter with sterilized forceps and place it on pre-incubated soybean
casein digest agar plate avoiding air bubble entrapped under filter paper.
5.1.15 Incubate
plate at 20-250C for 72 hrs. followed by 30-350C for 48
hrs.
5.1.16 After
incubation take observation and record the result.
5.2 Bacterial Endotoxin Test (BET):
5.2.1 Collect
two units each of vials and fill with Lal Reagent Water (LRW) in suitable
volume and vortex it for 5-10 min. Pool sample in separate pyrogen free tube.
5.2.2 In case of
rubber stopper collect two stopper and dip in 2 ml of LRW and vortex it for
5-10 min.
5.2.3 Performed test as per SOP.
5.3 Liquid Particle Count (LPC):
5.3.1
For vials: Collect 15 vials for
20 ml capacity and two vial in case of capacity more than 20 ml under LAF as
per SOP.
5.3.2 Fill required vials with blank water.
5.3.3 Take aliquot in 100 ml particle free beaker for testing.
5.3.4 Check the particle against blank as per SOP.
5.3.5 For rubber stopper: Take wide mouth
capped bottle with particle free aliquot (100ml) for sampling.
5.3.6 Collect 20 rubber stoppers under LAF as per SOP.
5.3.7 Put rubber stopper directly into the bottle.
5.3.8 Bring rubber stoppers to the lab separate the
aliquot under LAF.
5.3.9 Check the particle against blank as per SOP.
5.4 Test frequency:
Initial three consignments of every new vendor and after every three month.6.0 ABBREVIATIONS
6.1 SOP - Standard operating procedure
6.2 LAF
- Laminar air flow bench
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