1.0 PURPOSE:
2.0 SCOPE:
This SOP is applicable for cleaning and sterilization / depyrogenation of glassware / accessories at Microbiology laboratory of (Company name).
3.0 DEFINITIONS:
4.0 RESPONSIBILITY:
4.1 Microbiologist/Analyst: Responsible for execution of the procedure specified in this SOP.
4.2 Head Quality Control: Responsible for ensuring the compliance of the SOP.
5.0 PROCEDURE:
5.1 Health, safety and environment:
5.1.1 Use proper safety apparel such as hand gloves and safety goggles during cleaning of the accessories, while handling used culture media etc.
5.1.2 Use appropriate personal protection equipment such as rubber hand gloves, nose-mask, face shield and PVC apron while handling chromic acid.
5.1.3 Remove all markings, sticker label from the glassware / apparatus / accessories. If labels do not peel off, keep the glassware / accessories soaked in a suitable cleaning agent. (For e.g. 2% Extran solution for 10-15 minutes).
5.1.4 Wash all glassware / apparatus / accessories thoroughly with a suitable cleaning agent, rinse with potable water, and finally with purified water.
5.1.5 Dry the glassware / accessories / apparatus at 60°C for about 1 hour or until dry.
5.2 Washing of used pipettes / pipette tips:
5.2.1 Put used pipette /pipette tips into a tray / container containing any disinfectant.
5.2.2 Decontaminate by autoclaving, after autoclaving wash thoroughly with a suitable cleaning agent.
5.2.3 Rinse thoroughly with potable water and then with purified water. Dry the pipettes at 60°C for one hour or until dry.
5.3 Washing of used glassware:
5.3.1 Proceed for disposal of media as per preparation and disposal of microbial culture media SOP.
5.3.2 Wash thoroughly with a suitable cleaning agent and rinse with potable water. Finally rinse with purified water.
5.3.3 Dry the glassware at 60°C for about 1 hour or until dry.
5.4 Washing of new glassware:
5.4.1 After routine cleaning, treat with 5 % Sodium carbonate solution, followed by soaking in 1 % Hydrochloric acid solution overnight.
5.4.2 Wash with excess quantity of potable water and rinse thoroughly. Rinse with purified water and dry at 60°C for about one hour or until dry.
5.5 Washing of greasy glassware:
5.5.1 Treat greasy glassware with 5 % Sodium carbonate or soak in dilute chromic acid solution, overnight. Chromic acid is prepared as follows:
Take 100 g of Sodium dichromate and dissolve in about 50 ml of water. Add, slowly with constant stirring about 750 ml of concentrated Sulphuric acid, in an ice water-bath. Allow to cool and use.
5.5.2 Rinse thoroughly with excess of potable water and then with purified water.
5.6 Cleaning of glassware used for cytotoxic and potent materials:
5.6.1 After completion of analysis, the cytotoxic and potent materials soak the glassware’s in the deactivating agent specified in the individuals monograph or the deactivation procedure and hold the glassware till the specified time.
5.6.2 Wash all glassware / apparatus / accessories thoroughly with a suitable cleaning agent, rinse with potable water, and finally with purified water.
5.6.3 Dry the glassware / accessories / apparatus at 60°C for about 1 hour or until dry.
5.7 Sterilization / depyrogenation of glassware / apparatus / accessories:
Note: All empty glassware and accessories used for BET sampling should preferably be sterilized / depyrogenated in the hot air oven. Apparatus made of rubber / silicon /polyethylene are to be sterilized in the autoclave.
5.7.1 The type of wrapper to be used for glassware / apparatus/ accessories should depend on the type of sterilization / depyrogenation carried.
e.g. sampling scoops, spoons should be wrapped in aluminum foil and should be sterilized / depyrogenated by dry heat. Membrane filtration assembly should be wrapped in butter paper / parchment paper / any suitable wrapper for autoclaving. Empty tubes with cotton plugs should be wrapped with butter paper / parchment paper / any suitable wrapper and autoclaved, whereas empty tubes with metal caps can be wrapped with aluminum foil / any suitable wrapper and sterilized / depyrogenated with dry heat.
5.7.2 Place the apparatus in the hot air oven / autoclave and appropriate chemical indicators designed for the specific sterilization cycle shall be placed within the load.
5.7.3 For hot air oven: Set the temperature and time of the hot air oven as per validated procedure. After sterilization is over, allow to cool to the door opening temperature of the equipment before opening the hot air oven. Refer the respective equipment standard operating procedure.
5.7.4 For autoclave: Operate the autoclave as per standard operating procedure.
5.7.5 Using appropriate safety apparels, remove the sterilized / depyrogenated glassware / apparatus / accessories from the hot air oven / autoclave.
5.7.6 Use the glassware / apparatus / accessories only after it has cooled to ambient temperature.
5.7.7 Ensure that the sterilized / depyrogenated glassware / apparatus / accessories are appropriately labeled as per with the date of sterilization, load number, use before date and sign.
5.7.8 Store sterilized / depyrogenated glassware / apparatus / accessories in a clean dry place and use them within validated period.
5.7.9 Sterilization / depyrogenation record shall be maintained. The records must include details of each load such as date, load number, and items sterilized / depyrogenated. Hot air oven record must also include the time of switched on, required temperature reached at, set temperature and the time of switched off. The autoclave sterilization record must also include the quantities dispensed, temperature and pressure record.
5.8 Preparation of garments for sterilization:
5.8.1 Check the garments for absence of damage to zips/velcro tapes, string of booties, snood etc.
5.8.2 Fold the garment neatly and put it into a bag made of non-shedding material.
5.8.3 Wrap the bag in a double layer of butter paper/ parchment paper/wrapping paper and seal with tape or with steripouch sealer. Label with date of sterilization and use before date.
5.8.4 Load the garments in the autoclave.
5.8.5 Sterilize the garment in the autoclave at 121-124 oC for 30 minutes. After completion of cycle, unload the garments and keep in designated place for usage.
5.8.6 Number of sterilization cycles of each garment shall be monitored.
5.8.7 Take the garments to the respective testing areas, and then place the garments in the assigned shelf in garment cubicle/cupboard.
5.8.8 Use the garments within validated period.
6.0 ABBREVIATIONS:
% : Percentage
°C : Degree centigrade
SOP :Standard Operating Procedure
ml : Milliliter
BET : Bacterial endotoxin.