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SOP for Operation, Cleaning and Monitoring of Bio-Safety Cabinet


Standard operating procedure of cleaning and monitoring of Bio-Safety Cabinet by active air sampling and non viable particle count.

1.0  PURPOSE

       To lay down the procedure to perform the Operation of Bio-Safety cabinet. 

2.0  SCOPE

       It is applicable to Quality control department.

3.0  RESPONSIBILITY

       Microbiologist

4.0  ACCOUNTABILITY

       Head of Department

5.0  PROCEDURE

5.1  Operation of Bio-Safety cabinet

5.1.1  Switch on the main supply.
5.1.2  Ensure equipment is in a clean state.
5.1.3  Mop the bench with 70 % isopropyl alcohol / disinfectant.
5.1.4  Start blowers 30 minutes prior to the operation.
5.1.5  Switch “ON” the Ultra Violet (U.V) light for 1 hour prior to use or when required.
5.1.6  Ensure that when Ultra Violet (U.V) light is in “ON” condition, normal light should be switched “OFF”.
5.1.7  Record the differential pressure across the High efficiency Particulate Air (HEPA) filter.
5.1.8  Set the apparatus in the Bio safety cabinet.
5.1.9  Ensure Operation is done in the safe working zone.
5.1.10  Ensure the adequate protective gear is worn during the operation such as gloves, cap, and mask.

5.2  Cleaning of Bio-safety Cabinet

5.2.1  Keep the Bio-safety cabinet interior and exterior free from dust.
5.2.2  Before starting work, clean with 70% Isopropyl alcohol (IPA) / Disinfectant using a non-fibre shedding cloth.
5.2.3  Inside cleaning of the Bio-safety cabinet is recommended in the following cases
5.2.4  Before starting any work in the cabinet.
5.2.5  After working in the cabinet.
5.2.6  Whenever there is change of work program.
5.2.7  In the event of liquid spilling on the worktable.
5.2.8  Switch “ON” the Ultra Violet (UV) light for at least 15 minutes after handling the culture in Bio-Safety cabinet.
5.2.9  Before and after handling of culture, sanitize the outer surface of culture tube with disinfectant.

5.3  Shut - down procedure

5.3.1  Disconnect all utilities.
5.3.2  Turn off the burner and close the gas tap.
5.3.3  Switch off the main motor.
5.3.4  Clean down the entire area with 70 % Isopropyl alcohol / Disinfectant
5.4  Record the Details like burning of UV light, Velocity, Differential pressure, Cleaning, Instrument switched On/Off.

5.5  Monitoring of Bio-Safety cabinet

Monitoring of bio-safety cabinet can done by following ways,

5.5.1  Passive air sampling

5.5.1.1  Transfer the petriplate into pass box (LAL test room to Incubator).
5.5.1.2  Enter the respective area as per the SOP for entry and exit.
5.5.1.3  Decontaminate the external surface of the petriplates with the help of a sterile mop soaked in a filtered sporocidal agent.
5.5.1.4  Mark the petriplates at the base with the following details with marker
Media lot no
Name of the location
Date of exposure
Exposed by
5.5.1.5  Place the petriplates on the bench of bio-safety cabinet and remove the upper lid of the petriplates and keep it in inverted position.
5.5.1.6  Expose the media plate for 4 hours, After 4 hours of exposure, close the petriplate with lid. Collect the petriplate and transfer the plates into the incubator.
5.5.1.7  Incubate the exposed petriplate in inverted position in the incubator at 32.5 ± 2.5°C for 5 day.
5.5.1.8  After incubation observe and count the number of colonies on the colony counter or in the light source with the help of marker.
5.5.1.9  Note down the observation
5.5.1.10  If the counts obtained are above the limits specified below investigate the results and take necessary actions as per SOP for 
5.5.1.11  Frequency of Monitoring
Class A: Once in a day whenever there is activity.
5.5.1.12  Acceptance criteria
Action limit:  Class A: 1 CFU / plate

5.5.2  Active air sampling

5.5.2.1  Transfer the required number of media cassette, plate, and accessories into pass box (LAL test room to Incubator).
5.5.2.2  Enter the respective area as per the SOP for entry and exit.
5.5.2.3  Decontaminate the external surface of the plates with the help of a sterile mop soaked in a filtered sporocidal agent.
5.5.2.4  Mark the petriplates at the base with the following details with marker,
Media lot no
Name of the location
Date of exposure
Exposed by
5.5.2.5  Place the media cassette/petriplate on the Air sampler and operate the instrument as per SOP for Air Sampler.
5.5.2.6  After completion of sampling collect all the plates and incubate in inverted position in the incubator at 32.5 ± 2.5°C for 5 day.
5.5.2.7  After incubation observe and count the number of colonies on the colony counter or in the light source with the help of marker.
5.5.2.8  Note down the observation
5.5.2.9  If the counts obtained are above the limits specified below investigate the results and take necessary actions as per SOP
5.5.2.10  Frequency of Monitoring  Class A: Once in a day
5.5.2.11  Acceptance criteria
Action limit  Class A: 1 CFU / plate

5.5.3  Non viable particle count

5.5.3.1  Keep the non-viable particles counter in the pass box (LAL test room to Incubator).
5.5.3.2  Enter the respective area as per the SOP
5.5.3.3  Operate the sampler according to SOP for Air Sampler.
5.5.3.4  Perform the non-viable particle count at two different locations after completion of the activities.
5.5.3.5  Record the details of non viable particles count.
5.5.3.6  Frequency  Once in fifteen days.

6.0  ABBREVIATIONS

6.1  SOP - Standard operating procedure
6.2  CFU - Colony forming units
6.3  UV - Ultra violet

Also see: SOP for Cleaning and Operation of Bio-safety Cabinet


Ankur Choudhary is India's first professional pharmaceutical blogger, author and founder of Pharmaceutical Guidelines, a widely-read pharmaceutical blog since 2008. Sign-up for the free email updates for your daily dose of pharmaceutical tips.
Email: .moc.enilediugamrahp@ofni Need Help: Ask Question


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