SOP Title: Cleaning of Used Glassware in Microbiology Section
- Objective:
To lay down the written procedure for Cleaning of Used Glassware in the Microbiology Section of
Quality Control Department.
- Scope:
This procedure is applicable for Cleaning of Glassware and other utensils used for microbiological
analysis in the Microbiology Section of Quality Control Department.
- Responsibility:
- Quality Control Department: To prepare and review the SOP. To follow the procedures laid down for Cleaning of Used Glassware in the Microbiology Section in Quality Control Department as per this SOP.
- Quality Assurance Department: To review and approve the SOP and Annexures.
- Accountability:
Head Quality Control Department, Head Quality Assurance Department.
- Procedure:
| 5.1 | Precautions: | |
| 5.1.1 | All contaminated glassware’s containing culture media and other biohazards must be sterilised in the autoclave at specified conditions, before cleaning to kill all the hazardous microorganisms. | |
| 5.1.2 | Always wear personal protective equipments such as safety goggles, gloves, nose masks during washing of glassware’s. | |
| 5.1.3 | Do not use brushes that are so worn that the spine hits the glass as a result serious scratches may result and scratched glass is more prone to break during analysis. | |
| 5.1.4 | Do not drop glassware’s from height or bang to the sink or tap. | |
| 5.1.5 | Visually check glassware’s for any cracks before washing. Broken glassware’s may cause injury. | |
| 5.1.6 | Do not shake glassware’s too much vigorously as it may get cracks by pressure. | |
| 5.1.7 | Do not collect or touch broken glass pieces with bare hands. It can cause injury. | |
| 5.1.8 | Ensure that the soap has been thoroughly removed from the glassware because soap when comes in contact with the acid can form a film of grease. | |
| 5.1.9 | Do not mix the glassware’s from the microbiology section with the chemical section since it may cause cross contamination and impact the media component as well as the growth of microorganisms. | |
| 5.1.10 | Protect cleaned glassware’s from dust by placing them in a dust free cabinets. | |
| 5.2 | General Procedure: | |
| 5.2.1 | Laboratory technician should wear safety goggles, gloves, masks before starting the washing of glassware’s. | |
| 5.2.2 | Microbiologist must decontaminate the media and other hazardous microbial waste prior to cleaning, as per the SOP, Procedure for destruction of cultures and microbial waste (used culture media). | |
| 5.2.3 | Drain off any non hazardous contents present in the flasks, beakers, test tubes etc. in the sink and the hazardous contents that have been decontaminated in the autoclave, in the waste containers labelled as ‘Decontaminated Microbiological Waste’. | |
| 5.2.4 | Rinse the glassware’s with tap water before transferring them into soap solution. | |
| 5.2.5 | It is the Microbiologist’s responsibility to drain off hazardous contents from the flask, petriplates or any other glassware before giving it for washing. | |
| 5.2.6 | Immerse the glassware’s in tub containing 2.5% solution of dettol or savlon for 3-4 hours. | |
| 5.2.7 | Remove the markings of pen marker on glassware’s with the methanol. | |
| 5.2.8 | Remove the silicon grease on glassware’s with Acetone or use fuming sulphuric acid (Oleum) for about 30 minutes and then rinse with purified water. | |
| 5.2.9 | Scrub inner and outer portions of glassware’s with scrubber or nylon brushes soaked in 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran. | |
| 5.2.10 | If thorough cleaning is not possible immediately, soak the glass wares in 1% v/v Teepol solution or any other soap solution till adhesive material is removed or dissolved. | |
| 5.2.11 | During washing all parts of glassware’s should be thoroughly scrubbed with nylon brush. | |
| 5.2.12 | Rinse the glassware’s with running tap water; allow the water to run into and over the glassware’s for short time until no foam of soap is produced on shaking. | |
| 5.2.13 | All the glassware’s should be finally rinsed with purified water twice and kept in tilted position to drain off water completely. | |
| 5.2.14 | Dry the glass wares in the glassware drying oven at 60°C ±5°C. | |
| 5.2.15 | New glassware’s should be soaked in 1% v/v solution of hydrochloric acid or nitric acid for about 5-6 hours before washing as new glassware’s are slightly alkaline in reaction. | |
| 5.2.16 | If glassware becomes unduly cloudy or dirty or contains coagulated organic matter then glass wares should be given chromic acid treatment as follow:- | |
| 5.2.17 | Chromic acid solution: Dissolving 84g of chromium trioxide in 700 ml water and add slowly, with stirring 400 ml of sulphuric acid. (While preparing solution use personal protective equipments such as goggles, gloves, a mask as chromic acid is classified as a carcinogen). | |
| 5.2.18 | Pour the few ml of chromic acid solution on dirty glassware’s and leave them for minimum 15 minutes. Add small amount of potable water with shaking. | |
| 5.2.19 | Rinse several times with water or allow to keep in water for about one hour as the chromic acid is highly acidic in nature and finally rinse with purified water twice. | |
| 5.2.20 | Dry the glass wares in the glassware drying oven at 60°C ±5°C. | |
| 5.2.21 | Use the washed glassware within one week of the washing. After one week re-wash the same glassware’s before use even if they are not used. | |
| 5.2.22 | Store the washed and dried glassware in a designated and clean cupboard with proper labelling. | |
| 5.3 | Cleaning of Test tubes, Durham’s tubes and Centrifuge tubes: | |
| 5.3.1 | Empty the contents of the test tubes in a container labelled as ‘Decontaminated Microbiological Waste’. | |
| 5.3.2 | Durham’s tube can be retrieved while transferring the contents of test tube in the container labelled as ‘Decontaminated Microbiological Waste’. | |
| 5.3.3 | Wash the tubes under the tap and immerse the tubes in tub labelled as ‘Glassware’s for Disinfection’ containing 2.5% solution of dettol or savlon for 3-4 hours. | |
| 5.3.4 | Remove and clean the tubes with 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran. | |
| 5.3.5 | Wash with tap water until all traces of soap solution are removed. | |
| 5.3.6 | Finally rinse with distilled water or purified water and keep in a tilted position to drain off water completely. | |
| 5.3.7 | Dry the tubes in drying oven at 60°C ± 5°C to remove the moisture. | |
| 5.3.8 | Sterilise the tubes before use, as per the SOP, Procedure for operation of sterilisation autoclave. | |
| 5.3.9 | Test tubes for culture maintenance should be cleaned and stored separately. | |
| 5.4 | Cleaning of Petri plates: | |
| 5.4.1 | Remove the melted media subsequent to sterilisation from the petriplates and empty in a container labelled as ‘Decontaminated Microbiological Waste’ and wash under the tap water. | |
| 5.4.2 | Immerse the petri plates in tub labelled as ‘Glassware’s for Disinfection’ containing 2.5% solution of dettol or savlon for 3-4 hours. | |
| 5.4.3 | Remove and clean the petri plates with 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran. Use a scrubber for cleaning. | |
| 5.4.4 | Wash with tap water until all the traces of soap solution are removed. | |
| 5.4.5 | Finally rinse with distilled water or purified water and keep in an inverted position to drain off water completely. | |
| 5.4.6 | Dry the petri plates in drying oven at 60°C ± 5°C to remove the moisture. | |
| 5.4.7 | Sterilise the petri plates before use, as per the SOP, Procedure for operation of sterilisation autoclave. | |
| 5.5 | Cleaning of Pipette: | |
| 5.5.1 | Immerse the pipette in a container labelled as ‘Glassware’s for Disinfection’ containing 2.5% solution of dettol or savlon for 3 – 4 hours. | |
| 5.5.2 | Ensure that the level of disinfectant in the container is high enough to immerse the maximum portion of the pipettes. | |
| 5.5.3 | Do not drop them into the container. This may break the tips and render the pipettes useless for accurate measurements. | |
| 5.5.4 | Blocked Pipette should be immersed in a cylinder filled with dichromate cleaning solution and left for 24 hours. | |
| 5.5.5 | Remove and clean the pipettes with 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran. | |
| 5.5.6 | Wash with tap water until all the traces of soap solution are removed. | |
| 5.5.7 | Finally rinse with distilled water or purified water. | |
| 5.5.8 | Dry the pipettes in drying oven at 60°C ± 5°C to remove the moisture. | |
| 5.5.9 | Sterilise the pipettes before use, as per the SOP, Procedure for operation of sterilisation autoclave. | |
| 5.6 | Cleaning of Flask for media preparation: | |
| 5.6.1 | Empty the contents of the Flasks in a container labelled as ‘Decontaminated Microbiological Waste’. | |
| 5.6.2 | Wash under the tap and immerse the flasks in tub labelled as ‘Glassware’s for Disinfection’ containing 2.5% solution of dettol or savlon for 3-4 hours. | |
| 5.6.3 | Remove and clean the flasks with 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran. | |
| 5.6.4 | Wash with tap water until all the traces of soap solution are removed. | |
| 5.6.5 | Finally rinse with distilled water or purified water and keep in a tilted position to drain off water completely. | |
| 5.6.6 | Dry the flasks in drying oven at 60°C ± 5°C to remove the moisture. | |
| 5.6.7 | Sterilise the pipettes before use, as per the SOP, Procedure for operation of sterilisation autoclave. | |
| 5.7 | Cleaning of Dropper pipette, tips for micropipette: | |
| 5.7.1 | Wash under the tap and immerse the dropper pipettes and tips in a container labelled as ‘Glassware’s for Disinfection’ containing 2.5% solution of dettol or savlon for 3-4 hours. | |
| 5.7.2 | Remove and clean the dropper pipettes and tips with 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran. | |
| 5.7.3 | Wash with tap water to remove all traces of soap solution. | |
| 5.7.4 | Finally rinse with purified water. | |
| 5.7.5 | Dry the dropper pipettes and tips in drying oven at 60°C ± 5°C to remove the moisture. | |
| 5.7.6 | Tips for micropipette can be used for analysis, after sterilisation as per the SOP, Procedure for operation of sterilisation autoclave. | |
| 5.8 | Cleaning of Microscopes Slides and Cover slips: | |
| 1. New Slides: | ||
| 5.8.1 | Place slides overnight in a beaker of 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran. | |
| 5.8.2 | Rinse each slide under tap water until all traces of soap solution are removed. | |
| 5.8.3 | Wipe the slides, one at a time with a soft cloth. Place on blotting paper or filter paper that doesn’t shed any fibres and allow to dry. | |
| 5.8.4 | Pack the slides in stacks of 10 or 20 groups in paper. | |
| 2. Dirty slides: | ||
| 5.8.5 | Oil is removed by rubbing with blotting paper. One or two drops of alcohol can be used to remove residual oil. | |
| 5.8.6 | Cover slips are removed using a needle or forceps. | |
| 5.8.7 | Now soak in beaker of 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran. | |
| 5.8.8 | If slide have been used for infected specimen, they should be placed in disinfectant solution of 2.5% solution of dettol or savlon for 3-4 hours. | |
| 3. Cover slips: | ||
| 5.8.9 | Immerse the cover slips in a beaker of 1% v/v (1 ml in 100 ml) of commercially available liquid soap solution such as Teepol, Labolein or Extran. | |
| 5.8.10 | Soak for 2-3 hours, shaking gently from time to time. | |
| 5.8.11 | Rinse the beaker with cover slips under tap water. | |
| 5.8.12 | Finally rinse with distilled water or purified water. | |
| 5.8.13 | Drain the cover slips out on a gauze pad. | |
| 5.8.14 | Dry in hot air oven at 60°C till drying and keep in a clean petri dish. | |
- Definitions/Abbreviations:
- Definitions :
- Hazardous waste: A hazardous waste is a waste that poses substantial or potential threats to health or the environment.
- Non-hazardous waste: Non-hazardous waste means the waste that is not harmful to health and the environment.
- Carcinogens: A substance or agent that can cause cells to become cancerous by altering their genetic structure so that they multiply continuously and become malignant.
- Sterilization: Sterilizationis a term referring to a process that eliminates or kills all forms of life, including transmissible agents (such as fungi, bacteria, viruses, spore forms, etc.)
- Abbreviations :
| Abbreviation | Expansion |
| SOP | Standard Operating Procedure |
| QC | Quality Control |
| QA | Quality Assurance |
| % | Percent |
| v/v | Volume by volume |
| °C | Degree centigrade |
| g | grams |
| ml | Milliliter |
*Note – Ready to use SOP available in “DOWNLOAD” Section.