1. Hold Time Study Protocol of Sterilized Garments for their Sterility
1.0OBJECTIVE
Objective of this protocol is to provide documented evidence to establish and verify the microbial presence on
sterilized garments upon holding up to 5 days after sterilization by using the microbial detection method.
2.0 SCOPE
The scope of this protocolis to evaluate the hold time after sterilization of garments. This shall be done by using
established microbiological method to determine the microbiological growth.
3.0 REFERENCE DOCUMENTS
Following documents are referred during preparation of the protocol
3.1 SOP for Sterility Testing
3.2 SOP for Garment Cabinet
4.0 PURPOSE OF STUDY
The purpose of this protocol is to establish the rate of microbial proliferation in the Sterilized garments for the
specified holding period after its sterilization.
5.0 RESPONSIBILITY
Responsibilities of Quality Assurance and Quality control Microbiology personnel involved in activities related to
the General validation protocol are defined below:
5.1Quality Assurance: Review and approval of protocol.

2. 5.2Quality Control: Preparation, review and execution of protocol.
6.0 PROCEDURE
6.1 Pre-requisite
6.1.1 Environmental conditions should be maintained within the established limit.
6.2 Procedure for Hold Time Study
6.2.1 Prepare a Dacron bag contains 1 pair of garments used in sterility testing area, and place 10 cut pieces (6x5
cm) of old dress in-between the dress.
6.2.2 Perform the sterilization of the bag as per the SOP for sterilization of dresses.
6.2.3 After sterilization place the Dacron bag in Garment cubicle for Hold time study.
6.2.4 Collect one cut piece from the dress bag aseptically and direct immerse into the sterilized SCD medium and
check for the sterility, this sample shall be treated as initial sample. (0 Hour)
6.2.5 Similarly collect the dress pieces from the hold bag at regular intervals of 24 hr, 48 hr, 72 hr, 96 hr, and 120
hr and perform the sterility test.
6.2.5 Incubate the samples at the specified temperature for 14 days (20-25°C for 7 days and 30-35°C for 7 days).
Note: 0 Hrs starts when the sterilized garments placed in Garment cubicle after sterilization.
7.0 ACCEPTANCE CRITERIA
This study is carried out to establish the hold time of sterilized garments after sterilization.
Microbial determination : No growth should be observed in sterility test.
8.0 CONCLUSION

3. After complete evaluation of the hold time study for sterilized garments used for sterility testing a final hold time
study summary report shall be prepared which should essentially contain discussion and conclusion which clearly
determine the hold time period for sterilized garments.
9.0 ABBREVIATIONS
9.1 SOP - Standard operating procedure
9.2 cm - Centimetre
9.3 Hrs - hours
9.4 SCD medium - Soybean Casein Digest Medium
9.5 °C - Degrees centigrade
9.6 Hr - Hour