Environmental monitoring of Oral Solid Dosage area
This Document describes (Standard Operating procedure) SOP for Environmental monitoring of Oral Solid Dosage area
SOP for Environmental monitoring of Oral Solid Dosage area
I. Purpose & Scope:
- The purpose of this SOP is to lay down the procedure for Environmental monitoring of Oral Solid Dosage area
- This Standard operating procedure shall be applicable for Environmental monitoring of Oral Solid Dosage area
II. Responsibilities:
- All Quality Control and Quality Assurance personnel’s shall be responsible to follow and implement this SOP for Environmental monitoring of Oral Solid Dosage area
III. Introduction and Procedure:
Procedural part for Environmental monitoring of Oral Solid Dosage area :
Preparation of Plates:
- Prepare the media for environment monitoring, as per respective SOP.
- Add 1% Glycerin in media if required, to avoid dryness during exposure.
- Pre-incubated media should be used for environment monitoring. Aseptically check Pre-incubated media plates for any evidence of microbial contamination.
Transfer of Plates to the Area:
- Place the required number of media plates in Stainless steel (SS) Petri plate carrier and put other material in SS Dressing drum. Close the lid of SS Dressing drum.
- Transfer the SS Petri plate carrier and SS Container to the area to be monitored and transfer back to the microbiology laboratory after the completion of monitoring.
Settle plate Method :
- Use the plates of 90 mm diameter size for settle plate method.
- Use Soybean casein digest agar (SCDA) plates for bacterial count and Sabouraud chloramphenicol agar (SCA) or Sabouraud dextrose agar (SDA) for fungal count.
- Label the individual plate with the media lot number, Name of location/ Plate I.D., and date of monitoring. Transfer the plates to production area.
- Expose the media plates at the specified locations in production area. For list of location, refer the Annexure-I.
- Use the SS stands designed for settle plate method. Clean the SS stands with 70% IPA and keep them at locations specified for settle plate method.
- Place the appropriate plates at specified locations; on the SS stands. Carefully open the lid of plate and place it on the stand adjacent to the plate.
- After completion of exposure time, carefully close the plates by their lids and collect them in SS carrier.
- In Oral solid Dosage section expose the plates for two hours.
- Transfer the plates to lab and incubate them in incubator, Keep them in inverted position during incubation.
- The settle plates (SCDA plates) of General block shall be incubated at 30-35°C for 3 days for determination of total bacterial count. While SCA/SDA plates to be incubated at 20-25°C for 5 days for determination of total fungal count. Record the results in Annexure II.
Limit and Frequencies for Oral solid Dosage section :
| Sr No | Activity | Grade | Alert limit | Action limit | Maximum Limit
(cfu/plate) |
Frequency
|
| 1 | Bacterial monitoring by Settle Plate Method | A | — | — | < 1cfu/plate
|
Once in Month |
| D | 50cfu/plate |
80cfu/plate |
NMT 100cfu/plate |
Once in Month | ||
| 2 | Fungal monitoring by Settle Plate Method | A | — | — | < 1cfu/plate
|
Once in Month |
| D |
3cfu/plate |
4cfu/plate |
NMT 5cfu/plate |
Once in Month |
- Positive control: Keep one plate of this lot number as positive control by performing Growth promotion test by one of the respective culture. kept them with incubated plates.
- Negative Control: Keep one plate of this lot number as negative control by kept them With Sampling without exposed to the environment. Incubate them with incubated plate.
- Preparation of Trends : Monthly
Air sampling Method:
- Principle: Air is aspirated through a perforated lid, and impacted onto the surface of growth media in a standard 90 mm petri dish. Microorganisms impact the culture media and the colonies are counted after incubation period. The system measures the inflow of air regulates the aspirated volume to a constant value of 100 lit/min.
- Sampling procedure: Sampling volume is 1000 litres for a time period of 10 minutes. For the operation of air sampler refer the respective SOP.
- The Stainless steel sampling grid/sieve shall be autoclaved (at 121°C for 20minutes) between each sampling campaign.
- Between each sampling, sanitize the sampling grid/sieve with filtered 70%IPA and run sampler empty for approximate 30second for drying purpose.
- Incubate the SCDA agar plates at 20 -25 0C for 72 hours for TFC.
- After completion of incubation period for TFC, record the colony count and further incubate the plates at 30-35°C for 48 hours for TBC.
- Perform the Positive and Negative control of Used media.
- Count the number of colony forming unit (CFU) observed on the plate after 5 days. Total Viable Aerobic Count (TVAC) is the summation of the TBC & TFC after incubation period of 5 days. The count is expressed as CFU / M3.
- Write the Probable number of colonies in front of actually numerated colonies.
- Record observations of Air Sampling in Annexure V.
- Preparation of Trends: Monthly
Limits and Frequency for oral solid dosage section:
| Grade | Alert Limit
(CFU/M3) |
Action Limit
(CFU/M3) |
Maximum Limit
(CFU/M3) |
Frequency |
| A | – | – | < 1cfu/m3 | Once in month |
| D | 150 cfu/m3 | 180 cfu/m3 | NMT 200cfu/m3 | Once in month |
Monitoring of Equipment Surface:
Monitoring of equipment surfaces by swab method:
- Take one swab (individually dipped in test tube containing Presterilized 10 ml 0.9% saline) for each sampling point.
- Lable the individual plate with Media lot number, Name of location/Plate I.D and date of monitoring.
- Keep the swabs in the SS Sampling kit and take them to Oral solid dosage section.
- Aseptically remove the sterile swab from its packing, moisten it with sterile saline . Aseptically carry the swabs of 5X5cm (25cm2) to the sampling sites. Swabbing shall be done using parallel horizontal strokes overlap with vertical Strokes in marked area.
Horizontal strokes Vertical Strokes
- Aseptically Transfer the swab immediately in the test tube containing Presterilized 10ml 0.9% saline..
- After sampling, spray the sampling site by 70 % IPA & carry all swabs.
- Collect the swabs and bring it to Microbiology lab.
- Vigorously shake the swab test tube for about 1 minute, discard the swab stick and filter the saline through presterilized 45 micron Membrane filter paper.
- Transfer the filter paper on the SCDA/Dey Engley neutralizing agar petriplate and incubate it at 20- 25°C for 72hrs for TFC and after that at 30-35°C for 48hrs for TBC.
- All swabs shall be treated in the same manner as per marking on them.
- Total Viable Aerobic Count (TVAC) is summation of the TFC and TBC after incubation period of 5 days. The count is expressed as CFU/25 cm². Record the result in Annexure III.
- If the count exceeds the limits inform to Department head, Quality Assurance and Production Manager and carry out an investigation.
- Preparation of Trends : Monthly
Limit and Frequencies :
| Sr. No | Activity | Grade | Alert Limit | Action Limit | Maximum Limit | Frequency |
| 1 | Equipment monitoring by surface swab method (Bacterial Count) | A | — | — | < 1cfu/25cm2 | Once in month |
| D | 30cfu/25cm2 | 40cfu/25cm2 | NMT 50cfu/25cm2 | |||
| 2 | Equipment monitoring by surface swab method (fungal Count) | A | — | — | < 1cfu/25cm2 | |
| D | — | NMT 1cfu/25cm2 | NMT 1cfu/25cm2 |
Actions to be taken in case of microbial counts exceed the trends:
- If the microbial counts exceed the trends in the critical areas, observe the count for next 3 days. Even if the counts of 3 days monitoring are within the trends investigate the cause of higher counts.
- After the necessary action is taken, the head of concern department shall fill format and forward to QA department who in turn shall forward the same to microbiology department to final review.
- If the microbial counts exceed the trends in non-critical areas, observe the count for next exposure, and confirm that the counts are within the trends.
- If the microbial counts exceed the trends in the critical areas and non-critical areas are continuously increasing, stop all the activities in the areas and initiate an investigation . Based on the investigation corrective action shall be taken.
- After the corrective action, the area shall be sanitized and monitoring will be done for 3 days and the counts shall be observed.
- After reviewing the counts of all the 3 days monitoring and ensuring that the counts are within the trends, take the area in use.
Actions to be taken in case of microbial counts exceed alert limits:
- If the environment monitoring results exceeds the alert limits, send a note to the concerned department through Q.A department . Initiate investigation in consultation with QA department to find out the cause.
- Investigate the cause with respect to;
- Area sanitization records.
- Frequency of sanitization.
- Type and concentration of disinfectants used.
- Entry and exit procedures.
- Behavior of the persons in the production area.
- Proper functioning of the AHU’s.
- All the related laboratory procedures.
- Take corrective action based on the results of above investigation. If upon investigation no cause is identified, sanitize the area thoroughly as per procedure.
- After the necessary action is taken by head of concern department and forward to QA department who in turn shall forward the same to Microbiology department to final review.
- After sanitization, monitor the area for 3 consecutive days and ensure that the counts are within the alert limits.
- If no improvements in the counts are observed, stop all the activities in the area and carry out a thorough investigation.
Action to be taken in case the microbial counts exceed the action limits:
- If the environment monitoring results exceed the action limits, stop all the activities in the area immediately.
- Carry out thorough investigation of the results and of the area in which the counts are exceeded the limits.
- The laboratory investigation shall include analyst training, review of the colonies on the plates, isolation of the colonies, colony characterization and identification of the colonies.
- Note down any deficiencies observed during the laboratory investigation.
- Extend the investigation to the relevant areas and perform thorough investigation of the following.
- Review the sanitization procedure, frequency, type and concentration of disinfectant used.
- Review the daily records of the area monitoring with respect to temperature, humidity, differential pressure as well as non-viable particle count.
- Check the training records of the persons working in the area to confirm that they are trained for entry – exit procedure and also for precautions to be taken while work in the clean rooms.
- Check the proper working of the AHU’s and checks whether the validation was carried out properly as per the frequencies. Also inspect the area for any visible cracks or crevices.
- Note down any deficiencies observed during the investigation performed. Combine all the observations of laboratory as well as the area and make an investigation report to conclude the investigation. Based on the findings of the investigation, take the corrective action.
- Assess the impact of increase in the count beyond the action limit on the quality of the product. If the high counts are found in the critical areas, quarantine the products manufactured during the period and carryout additional tests, if necessary.
- Based on the review of investigation report and the results of additional tests, take the decision to either accept the product or to reject it.
- Any abnormal colonies observed shall be isolated and identified to know the source of contamination and subsequently corrective action will be taken.
- After the necessary action is taken by head of concern department and forward to QA department who in turn shall forward the same to microbiology department to final review.
- After the necessary action is taken, fumigate the area and monitor the area for 3 consecutive days and observe the plates for counts with the limits.
- After the results of all the three days monitoring are found to be within limits, the production activity can be started.
IV. Annexures:
Annexure I: List of Location for Environment Monitoring of Oral solid dosage section by Settle Plate Method.
Annexure II: Environment Monitoring Report of Oral solid dosage section by Settle Plate Method.
Annexure III: Environment Monitoring Report of Oral solid dosage section by Surface swab Method.
Annexure IV: List of Location For Environment Monitoring of Oral solid dosage section by Air Sampling Method.
Annexure V: Environment monitoring report of Oral solid dosage section by Air Sampling Method.
Annexure I
List of Location for Environment Monitoring of Oral solid dosage section by Settle Plate Method.
Page No : 1 of 1
| Plate I.D. | Location
(Plate exposed for 2 Hrs at following Location)
|
Grade | Maximum Limit
(cfu/plate) |
|
| Bacteria | Fungal | |||
| D | NMT
100cfu/plate |
NMT
5cfu/plate |
||
| D | NMT
100cfu/plate |
NMT
5cfu/plate |
||
| D | NMT
100cfu/plate |
NMT
5cfu/plate |
||
| D | NMT
100cfu/plate |
NMT
5cfu/plate |
||
| D | NMT
100cfu/plate |
NMT
5cfu/plate |
||
| D | NMT
100cfu/plate |
NMT
5cfu/plate |
||
| D | NMT
100cfu/plate |
NMT
5cfu/plate |
||
| D | NMT
100cfu/plate |
NMT
5cfu/plate |
||
| D | NMT
100cfu/plate |
NMT
5cfu/plate |
||
| D | NMT
100cfu/plate |
NMT
5cfu/plate |
||
| D | NMT
100cfu/plate |
NMT
5cfu/plate |
||
| D | NMT
100cfu/plate |
NMT
5cfu/plate |
||
| D | NMT
100cfu/plate |
NMT
5cfu/plate |
||
Annexure II
Environment Monitoring Report of Oral solid dosage section by Settle Plate Method.
Page No : 1 of 2
| Date of Sampling | Date of completion | ||
| Exposed by | Time of Exposure | ||
| Media Lot No. | / | Observed by | |
| Incubation Condition for Total Bacterial Count | 30-35°C For 72 hours | Incubator ID No | |
| Incubation Condition for Total fungal Count | 20-25°C For 5 days | Incubator ID No |
| Plate I.D. | Location
(Plate exposed for 2 Hrs at following Location)
|
Grade | Observation
TVAC (cfu/plate) |
|
| Total Bacterial Count | Total Fungal Count | |||
| D | ||||
| D | ||||
| D | ||||
| D | ||||
| D | ||||
| D | ||||
| D | ||||
| D | ||||
| D | ||||
| D | ||||
| D | ||||
| D | ||||
| D | ||||
Annexure II (Contd…)
Environment Monitoring Report of Oral solid dosage section by Settle Plate Method.
Page No : 2 of 2
Acceptance Criteria:
| Activity | Grade | Alert Limit | Action Limit | Maximum Limit |
| Total Bacterial Count | A | — | — | <1cfu/plate |
| D | 50cfu/plate | 80cfu/plate | NMT100cfu/plate | |
| Total Fungal Count | A | — | — | <1cfu/plate |
| D | 3cfu/ plate | 4cfu/plate | NMT5cfu/plate |
Positive Control:——————————- Negative Control: ——————————-
——————————- ——————————-
Conclusion: The Microbial count of Environment monitoring by Settle plate method are found Satisfactory/Not Satisfactory.
Observed By: Checked By: Approved By :
Date: Date: Date :
Annexure III
Environment Monitoring Report of Oral solid dosage section by Surface swab Method.
Page No : 1 of 1
| Plate I.D. |
Location :
Date of Sampling :————- Performed By : ——————- Media lot No:——————- Date of Completion :——————- |
Grade | Incubation condition:
(20-250C/72 hours) Date:——————— Incubator I.D.:———— Observed On:————– |
Incubation condition:
(30 -350C/48 hours) Date:——————– Incubator I.D.:———- Observed On:———— |
||
| Bacteria
(cfu/25cm2) |
Fungal
(cfu/25cm2) |
Bacteria
(cfu/25cm2) |
Fungal
(cfu/25cm2) |
|||
| D | ||||||
| D | ||||||
| D | ||||||
| D | ||||||
| D | ||||||
| D | ||||||
| D | ||||||
| D | ||||||
| Activity | Grade | Alert Limit | Action Limit | Maximum Limit |
| Total Bacterial Count | A | — | — | <1cfu/25cm2 |
| D | 30cfu/25cm2 | 40cfu/25cm2 | NMT50cfu/25cm2 | |
| Total Fungal Count | A | — | — | <1cfu/25cm2 |
| D | — | NMT1cfu/25cm2 | NMT1cfu/25cm2 |
Positive Control:——————————- Negative Control: ——————————-
Conclusion: The Microbial count of Environment monitoring by Surface Swab method are found Satisfactory/Not Satisfactory.
Observed By: Checked By: Approved By :
Date: Date: Date :
Annexure IV
List of Location For Environment Monitoring of Oral solid dosage section by Air Sampling Method.
Page No : 1 of 1
| Plate I.D. |
Location (Sampling Volume is 1000lit for 10min sampling period)
|
Grade |
Alert Limit (CFU/M3) |
Action Limit (CFU/M3) |
Maximum Limit (CFU/M3) |
| D | 150cfu/m3 | 180cfu/m3 | 200cfu/m3 | ||
| D | 150cfu/m3 | 180cfu/m3 | 200cfu/m3 | ||
| D | 150cfu/m3 | 180cfu/m3 | 200cfu/m3 | ||
| D | 150cfu/m3 | 180cfu/m3 | 200cfu/m3 | ||
| D | 150cfu/m3 | 180cfu/m3 | 200cfu/m3 | ||
| D | 150cfu/m3 | 180cfu/m3 | 200cfu/m3 |
Annexure V
Environment monitoring report of Oral solid dosage section by Air Sampling Method.
Page No : 1 of 1
| Plate I.D. | Location
(Sampling Volume is 1000lit for 10min sampling period)
Date of Sampling :————- Performed By : ——————- Media lot No:——————- Date of Completion :————– |
Grade | Incubation condition:
(20-250C/72 hours) Date:——————— Incubator I.D.:————- Observed On:————— |
Incubation condition:
(30 -350C/48 hours) Date:——————– Incubator I.D.:————- Observed On:————- |
||
| Air Sampling time: | ||||||
| Bacteria
(cfu/m3) |
Fungal
(cfu/m3) |
Bacteria
(cfu/m3) |
Fungal
(cfu/m3) |
|||
| D | ||||||
| D | ||||||
| D | ||||||
| D | ||||||
| D | ||||||
| D | ||||||
| Grade | Alert Limit
(CFU/M3) |
Action Limit
(CFU/M3) |
Maximum Limit
(CFU/M3) |
| A | – | – | < 1cfu/m3 |
| D | 150 cfu/m3 | 180 cfu/m3 | NMT 200cfu/m3 |
Positive Control:——————————- Negative Control: —————————-
Conclusion: The Total Viable aerobic count of Environment monitoring by Air Sampling method are found Satisfactory/Not Satisfactory.
Observed By : Checked By: Approved By :
Date: Date: Date :
