Viable
Particle Count By Using Settle Plate Method
1. Purpose:-
To determine airborne
microbiological contamination in clean rooms by Settle Plate Method
2. Scope:-
It
is applicable to following areas:
o Liquid Injectable area (Production Department)
o IV Infusion filling area (Production Department)
o Sterility testing Suit (Microbiology Department)
3. HSE
Statement:-
Not Applicable.
4. Responsibilities:-
i)
Manager Quality Control is responsible
to ensure that procedure & formats are followed entirely as approved.
ii)
Microbiologist is responsible to perform the
test.
iii)
Lab attendant is responsible
to assist the officer microbiologist for the preparation of test materials and
to take the plates to respective areas.
5. Materials:-
5.1 Petri plates (90mm)
5.2 Soya bean casein digest agar (SCDA)
5.3 Screw cap flask
5.4 SS-Tray
5.5 Hot plate and stirrer
6.
Definitions:-
6.1 Viable particle
count: (Also referred as Total Air borne Aerobic Microbial
(Count):-
The
recovered number of colony forming units (cfu) per unit volume of air.
6.2 Settle Plate Method:
-
This method is widely used
to qualitatively asses the environments over
prolonged
exposure times. Settle plates when exposed for four hours period can provide a
limit of detection for suitable evaluation of aseptic environment.
Flow Chart:-
8.0 Description:-
8.1 Procedure:-
8.1.1 Preparation of Settle plates:-
8.1.1.1 Weigh desired amount of agar medium
(e.g.; Nutrient agar or SCDA) in a screw cap flask.
8.1.1.2 Dissolve agar medium in calculated
volume of distilled water according to manufacturer’s
instructions. Tight the cap
of the flask and autoclave the medium at 1210C for 15min
8.1.1.3 Take clean and washed petriplates
(90mm) and sterilize them in hot air oven at 2000C for 2 hours.
8.1.1.4 Allow petriplates to cool at room
temperature.
8.1.1.5 Pour approximately 30ml molten and
autoclaved agar medium cooled at 400C sterile petriplates.
8.1.1.6 Allow agar medium to solidify.
8.1.1.7 After solidification, incubate each
petriplate in inverted position for 48-72 hours at
32.50C
8.1.1.8 Examine each petriplate for
contamination after incubation. Contaminated petriplates should be
disposed off
8.1.1.9 Use only contamination free
petriplates for the test.
8.1.1.10 Mop each petriplate from outside
with a clean duster dipped in 70% IPA and label according to sampling plan.
8.1.11 Do not label the plate from the lid
side. In case of mixing of lids, this may cause
in misinterpretation of results.
8.1.1.12
Place all the petriplates in a SS-tray properly mopped with 70% IPA and send to
the desired area for exposure.
8.1.1.13 During
transportation, do not open the lid of SS-tray.
8.1.1.14 Place the SS-tray
in Pass Through of the area under UV light.
8.1.2 Plate Exposure
8.1.2.1 Open the SS-tray in
controlled area and bring the petriplates out of it.
8.1.2.2 Place the plates in
appropriate defined positions.
8.1.2.3 Lift the lid of
petriplate and place it in downside up position on the stand.
8.1.2.4 Leave the petriplate
open for 4 hours.
8.1.2.5 Close the petriplate
after the time of exposure.
8.1.2.6 Clean the traces of
agar or condensation traces from the place where plates have been exposed with
suitable disinfectant.
8.1.2.7 Place the
petriplates back in SS-tray and Place it in Pass Through.
8.1.2.8 Transfer the SS-Tray
to Microbiological Lab.
8.1.2.9 During
transportation, do not open the lid of SS-tray.
8.1.2.10 Incubate each
petriplate in inverted position at 32.5 + 2.50C for 48-72 hours
along with an unexposed Petri plate mentioning as Negative Control.
8.1.2.11 After incubation,
record the results by using colony counter
8.2 Specifications:-
For each grade of controlled
areas or clean rooms limits are given below.
Table: Limits* for Microbial contamination
(CFU/ Plate for 4 hours) ‡
|
Sr. No.
|
Grade / Class
|
Standard Limits(cfu/plate)
|
Alert Limits(cfu/plate)
|
Action Limits(cfu/plate)
|
|||
|
At Rest
|
In Operation
|
At Rest
|
In
Operation
|
At Rest
|
In
Operation
|
||
|
1
|
A
|
<1
|
<3
|
>1
|
>1
|
>1
|
>1
|
|
2
|
B
|
<1
|
<5
|
>1
|
>1
|
>2
|
>3
|
|
3
|
C
|
<5
|
<50
|
>2
|
>3
|
>20
|
>30
|
|
4
|
D
|
<50
|
<100
|
>20
|
>30
|
>40
|
>50
|
·
Limits are bacterial count. Mold or fungal count should be considered
as zero.
8.3. Frequency:-
8.3.1 Each operating shift
8.3.2 After shut down for a weak or more
9. Records
9.1 Aseptic Area Monitoring (SPM & SAS)-
Ampoule and IV Infusion Filling Area
9.2 Aseptic Area Monitoring (SPM & SAS) Vial Filling Area
9.3 Area Monitoring
of Sterility Testing Room
10 References:-
10.1 Guidelines on the Test Methods for Environmental Monitoring for
Aseptic Dispensing Facilities, Produced by: A Working group of the Scottish
Quality Assurance Specialist Interest Group, February 2004. (For
All Classes at rest)
10.2 Sterile Production, World Health Organization). (For All Classes
at Operational Condition)
10.3 EU Requirements,FDA Requirements, WHO
Requirements on Viable Particle count
10.4 United States
11. Distribution:-
This SOP has to be distributed in below mentioned
Departments:-
|
Sr. NO
|
Distributed to
|
Received
(Current)
|
Returned
(Obsolete)
|
|
1
|
Quality Control
Department
|
|
|
|
2
|
Quality Management
Department
|
|
|
12
Revision History:-
Date Changes
N/A
|