Dirty Equipment Hold Time Study Protocol

QUALIFICATION TEST METHOD

Pr-requisite:

Prior of conducting/ executing the protocol, following pre requisites must be full filled.

• Environmental conditions should be maintained within the limit.
• Personal involved in the handling, storage and testing of sample should be trained.

Qualification test program:

Following procedure should be followed for determining the hold time of the equipment’s.

• Dirty equipment holding, sampling and testing for change in microbiological properties.

Swab sampling method:

Two types of parameters will be used to evaluate the samples.

• Chemical parameter (UV absorbance)
• Microbiological parameters (Total aerobic microbial count).

Swabbing method shall be used for the collection of chemical/ microbiological samples.

Chemical:

• For chemical swab sampling cotton swab bud shall be used for swabbing.
• The swab head should be moist with purified water and then swabbed on 5cm x 5cm (25cm2). In case of circular location cover the maximum possible area since the swabbing can’t be done using the standard size of 5cm x 5cm (25cm2).
• The swabbing shall be done using approx. 5 horizontals 5 parallel vertical strokes.
• Dip the swab in 5 ml of methanol and sent to the QC for analysis for the analysis.

Microbiological Swab:

• Sterile cotton bud shall be used for swabbing site where the chances of microbial contamination.
• Pre moisten the swab in sterile 0.9% saline solution.
• Carefully swab the equipment on surface area of 5cmx5cm (25cm2).
• The swabbing shall be done using approx. 5 horizontals 5 vertical strokes to take swab by assuming that the entire area is swabbed to get the maximum recovery and immediately place in labelled test tube.
• Analyze each swab sample as per defined procedure in cleaning validation protocol.

Procedure for Microbial Analysis

• Take 4 sterile Petri dishes and aseptically, Transfer 1 ml of swab sample into each Petri dish.
• Add 20 ml sterile soybean Casein Digest Agar to two plates and 20 ml sabouraud dextrose agar to remaining two plates
• Incubate SCDA 30-35 for 5 days (bacteria) and SDA 20-25 for 5 days (fungus).
• At the end of incubation period, count the No. of colonies formed and report the results obtained.
• Limit: TMC should not be more than 50 CFU/25 cm2

Note:

Use sterile hand gloves and mask during swabbing.

Swabbing site shall not be repeated as previous location.

Microbiological swabbing should be always performed prior to chemical swabbing as per the sampling plan to negate risk of contamination.

EXPERIMENTAL PLAN:

After completion of the batch keep the equipment without cleaning for 3 days initially and collect the samples daily without cleaning as per the time mentioned in this protocol. After the final sample cleaning of the tank shall be performed and samples shall be collected and send to QC for analysis, samples shall be collected as per the sampling plan.

After cleaning samples shall be collected as per the sampling plan.

Storage: During the study the equipment shall be maintained in closed condition.

Sampling intervals and locations:

• After completion of the batch tank shall be kept on hold without cleaning for 3 days initially.
• After that microbial sampling shall be carried from designated locations daily for 3 days.
• The chemical swab samples shall be collected on final day after the cleaning activity.

Chemical swab locations: (Final Day)

S. No.	Location	Justification
1	Sample from out let	-
2	Surface of the Lid	Difficult to clean
3	Inner surface of the tank	Difficult to clean

Microbial swab locations: (initial to third day)

S. No.	Location	Justification
1	Inner surface of the tank	Difficult to clean
2	Surface of the Lid	Difficult to clean

ACCEPTANCE CRITERIA:

This study is carried out to establish the hold time of equipment’s without cleaning. Following acceptance criteria shall be followed to evaluate the efficacy of hold time before cleaning.

Microbial Determination:

Acceptance limit for total microbial count is based on in house specification for bioburden of equipment and pathogen, it is based on in house specification for purified water.

S. No.	Swab sample	Acceptance limit
1	Total microbial count (TBC & TFC)	NMT 50 cfu/25sq.cm.

 

Acceptance limit for chemical testing is based on our in house specification for purified water.

Acceptance criteria:

Residual content NMT 78.749 mcg/ml at 282 nm by UV photometry (UV absorbance NMT 2.798)

EVALUATION OF THE RESULTS:

Results shall be documented as an annexure and shall be attached with final report.  Evaluation of results shall be carried out to derive a time limit for the clean equipment hold time study.

DOCUMENTATION:

• Results and reports shall be compiled which shall contain the following:
• Summary report
• Validation protocol
• Test reports
• Summary report shall contain the following:
• Approval
• Objective
• Acceptance criteria
• Brief validation methodology
• Evaluation of results
• Final result summary
• Deviation, failure investigation reports and corrective actions (if any)