Standard operating procedures for monitoring sedimentation bacteria in clean rooms

1. Purpose: 

To accurately monitor the number of sedimentation colonies in the clean area of the clean room, ensure that the number of microorganisms in the air of the production and inspection areas meets the requirements, and ensure product quality.

2. Scope: 

Applicable to the clean area of the company.

3. Responsible person: 

The sedimentation bacteria monitoring personnel of the biological testing group shall implement this standard.

4. Reason for change: 

To further improve the operating procedures to make them more standardized and reasonable.

5. Content

5.1 Cleaning and disinfection of sampling vessels

• First wash the culture dish with clean water several times, then use a suitable brush and detergent to wash repeatedly, then rinse with drinking water until there are no bubbles, and finally wash with purified water and dry

• Put the culture dish into the stainless steel culture dish tube and then put it into the electric constant temperature blast drying oven at 250℃ for dry heat sterilization for one hour.

5.2 Instruments and equipment

• Clean bench, electric constant temperature blast drying oven, constant temperature incubator, stainless steel culture dish tube, culture dish, alcohol lamp.

• Culture medium: nutrient agar culture medium, rose bengal sodium agar culture medium.

5.3 Preparation of culture medium plates

• Heat the culture medium to melt, cool to 45°C, and operate under sterile conditions to inject the culture medium into the culture plate, about 15ml~18ml per plate.

• After the agar solidifies, invert the culture medium plate and incubate it in a 30~35°C (and 23~28.℃ incubator for 48 hours and 72 hours respectively. Ensure that the culture medium plate is sterile before use. Store the prepared culture medium plate in an environment of 4~8℃.

5.4 Sampling method: Place the prepared culture plate at the predetermined sampling point, open the culture plate cover to expose the culture medium surface for a specified time, and then cover the culture plate.

5.5 Culture

• After all sampling is completed, invert the culture plate.

• Place it in a 30~35℃, 23~28℃ constant temperature incubator for 48 hours and 72 hours respectively: 5.5.3 For each batch of culture medium, perform control culture, and the number of culture plates in each batch shall not be less than two. 5.6 Number of sampling points and minimum number of culture plates.

5.6 Number of sampling points

• Arrangement of sampling points.

• The test point in the work area is 0.8~1.5m from the ground (slightly higher than the work surface).

• Additional test points can be added at key equipment or key work activity ranges, and the sampling points should be arranged evenly to avoid excessive concentration of sampling points in a certain local area and excessive sparseness in another local area.

Number of culture dishes per ten square meters

5.7 Calculation results: The number of colonies in each culture dish is obtained by counting method. The average number of colonies of sedimentation bacteria at each measuring point is calculated by formula. Average colony count M=(M1+M2+M3+…+Mn)/n

Where, M--average colony count

M1--Colony count of culture dish No. 1

M2--Colony count of culture dish No. 2

Mn--Colony count of culture dish No. n

N--Total number of culture dishes

5.8 Production workshop inspection

• Sampling status and time.

• Static: 30 minutes; Dynamic: The production process is inspected all day, and the culture dish is changed every 4 hours.

5.9 Monitoring frequency

During the production of products, the cattle production workshop shall be monitored according to the cleanliness level of the clean area.

• For the 100-level and 10,000-level areas, each workshop shall be monitored every day, but the sampling volume and number of samples can be reduced.

• Surface contamination and human bacteria testing shall be conducted every day during the production of sterile drugs.

• Each workshop in the 100,000-level area shall be monitored once a week.

• Each workshop in the 300,000-level area shall be monitored once every two weeks.

5.10 The Quality Inspection Department shall conduct regular spot checks on each workshop: static for 30 minutes; dynamic for 4 hours.

The monitoring of the workshop before the resumption of production after the suspension of production shall be carried out in a timely manner according to the inspection request form.

Cleanliness level and standards

If all the test results meet the assessment standards, a qualified report will be issued. If individual rooms do not meet the standards, a non-qualified report will be issued, requiring immediate measures until qualified.