ROOM QUALIFICATION AND MONITORING AND GROWTH PROMOTION TEST (GPT) PRINCIPLES Carry out room qualifications based on requirements based on CPOB. Growth Promotion Test (GPT) is a way to prove the suitability of test media for its use. TOOLS AND MATERIALS - Sterile petri dishes, sterile test tubes, test tube rack, volume pipette, measuring cup sterile, round and straight Oose needle, glass slide - Hand wipes, tissue, tripod, asbestos gauze, Bunsen, Incubator 37°C - Sterile media Trypticase Soy Agar (TSA), MacConkey Agar (MCA), Cetrimide Agar (MSA), Sabourod Dextrose Agar (SDA) - Bacterial culture: Escherichia coli, Pseudomonas aeruginosa, and Candida albicans E. EXPERIMENT PROCEDURE MONITORING AND QUALIFICATION a. Air Samples (Active Sampling) • Determine the number of monitoring points • Perform testing according to the tool manual • Incubate at 20 - 25 ºC, for 4 days, then continue incubation at a temperature of 30 - 35 ºC for 48 hours. • Count and record the number of bacteria and fungi that grow on each cup • In one plate it is calculated and converted to m3 , for example: 10 cfu/500 L then converted to 20 cfu/m3 Note: Class A Air Sampler should be dedicated to the room b. Papar Cup (Settle Plate) area 10 m2, • Determine the number of monitoring points using the formula = ÿarea (m2 ), for example: then the number of monitoring points is 3.3 rounded up to 4 monitoring points. The location of monitoring points is determined based on risk analysis at the points that are most likely to have the greatest impact on production or test results. • Bring the sterile Petri dish and sterile TSA into laminar air flow • Cool the TSA to ± 50oC and pour it into a Petri dish 15 – 20 mL and let it solidify • Incubate the Petri dish at 35 – 37oC for 48 hours • Check for growth of microorganisms on each agar plate • Plates on which microorganisms are growing must be removed before testing • Each agar plate is labeled with the test date, agar plate number and name of the room being monitored • Place the agar plate in a plastic or stainless steel container that has been previously disinfected with a 70% alcohol solution and take it to the room to be monitored. • Personnel who will expose the culture media must be in clean laboratory attire, wear a mask and gloves • Expose the Petri dish (90 mm) for 4 hours in the specified place and location. Place the lid of the cup next to the media containing the culture medium. • Incubate at 35-37oC for 48 hours, continue incubation at 20 – 25oC for 4 days. • Count the number of bacteria and fungi that grow on each cup and record them on the form provided. • Perform trend analysis per time period c. Contact Cup (Contact Plate) • Prepare the contact plate by placing the growth medium in a mold to achieve a convex agar shape, similar to a pillow mold as shown in the following image. • Press the convex agar medium against the surface to be tested for three to five seconds, the particles on the test surface will move to the contact plate growth medium. • Incubate at 35-37oC for 48 hours, then continue incubation at 20 – 25oC for 4 days and carry out trend analysis per time period d. 5 finger gloves (finger dabs) • Prepare a cup containing sterile TSA media • Wear sterile gloves • Press the medium with each finger in the cup for three to five second • Incubate at 35-37oC for 48 hours, then continue incubation at 20 – 25oC for 4 days. Monitoring the Number of Room Particles Using the Particle Counter tool, the procedure depends on the brand and type of particle counter available. Analyze patterns of monitoring results per period Flow Rate Monitoring The flow rate is monitored using an anemometer. Perform trend analysis for each time period Room Temperature and Humidity Monitoring Measure temperature, humidity and pressure (if necessary) using a thermobarohygrometer or appropriate tool. Note: Your output : Each group carries out Trend analysis of the number of microbes, temperature and humidity for 2 months (Ask an Assistant)