This validation was designed and performed from the viewpoint of a microbiologist with assistance from validation experts. The protocol used for this validation was previously published in the Journal of validation Technology. The incubator contains a dual hydraulic thermostat, a mercury-in-glass therm o m e t e r, two shelves, two heaters, double inner doors, and double outer doors.
This validation was designed and performed from the viewpoint of a microbiologist with assistance from validation experts. The protocol used for this validation was previously published in the Journal of validation Technology. The incubator contains a dual hydraulic thermostat, a mercury-in-glass therm o m e t e r, two shelves, two heaters, double inner doors, and double outer doors.
This validation was designed and performed from the viewpoint of a microbiologist with assistance from validation experts. The protocol used for this validation was previously published in the Journal of validation Technology. The incubator contains a dual hydraulic thermostat, a mercury-in-glass therm o m e t e r, two shelves, two heaters, double inner doors, and double outer doors.
the various steps regarding the Installation Qualification (IQ), Operational Qualification (OQ) and Performance Qualification (PQ) that were executed to complete the validation of a pharmaceutical micro- biology laboratory incubator. This validation was designed and per- formed from the viewpoint of a microbiologist with assistance from validation experts. The protocol used for this validation was previously published in the Journal of Valida- tion Technology. 1 Descr iption of the I ncubator Unit The incubator is a conventional unit where fresh air enters though an air intake at the bottom. The air is heated at a plenum below the chamber and flows into the chamber in a uniform flow pattern. Exhaust air is vented through a port at the top of the incubator. The unit is a Precision Thelco Incubator catalog num- ber 31487. This is a gravity convec- tion unit model number 6M with an operating temperature range of 25C to 70C. The incubator has the fol- lowing electrical characteristics: Volts- 120 Hertz- 50/60 Watts- 300 The overall dimensions of the unit are 42 X 21 X 39 with cham- ber dimensions 36 X 18 X 27. T h e incubator contains a dual hydraulic thermostat, a mercury-in-glass ther- m o m e t e r, two shelves, two heaters, double inner doors, and double outer doors. I nstallation Qualification (I Q) During the IQ, personnel documented all items list- ed in the Incubator IQ Form. Refer to Appendix A o f validation protocol, previously published in the Journal of Validation Te ch n o l og y. 1 A second person reviewed and approved all verifications. cT hi s val i dati on was desi gned and per for med fr om the vi ewpoi nt of a mi cr obi ol ogi st wi th assi stance fr om val i dati on exper ts. d Val i dati on of a L abor ator y I ncubator Usi ng Wi r el ess and Cabl ed Datal ogger Par t I I By Laura Valdes-Mora Elite MicroSource Corporation and Pamela Hardt-English PhF Specialists, Inc. O Laura Valdes-Mora & Pamela Hardt-English Purpose IQ was conducted to demonstrate that the piece of equipment was inspected, and contains all the required parts for its proper functioning. The pres- ence/absence of external equipment aides (recorder charts, etc.) was also documented. Equipment and Materials Incubator Appendix A I Q Observations Incubator Description The unit is a Precision Thelco Incubator, catalog number 31487. The temperature range it is designed for is from room temperature to 70C. The unit uses 120 volts and 5.2 amps. This incubator had not been previously validated, and was 29 years and seven months old at the time this validation was conducted. Incubator Model Number is 6M. Incubator Serial Number 22-AG-8. Incubator was purchased on July 14, 1971 from Precision Scientific. Vendor Manual is available. Location of Vendor Manual The manual can be found in the file cabinet of the microbiology lab- o r a t o r y. Drawings for the incubator are available and located in the vendor manual. Utilities required Electricity is required for this unit. The power source was found to be present and functional. A member of the maintenance department measured the Volts and the amper- age reading of the circuit of the electrical outlet where the incubator is plugged in. The data were recorded as found. Utility Data Circuit Rating: 130 Vo l t s 1 Phase 60 Hertz 20 A m p s Equipment Rating: 125 Vo l t s 1 Phase 60 Hertz 5.2 A m p s Field Measurement: 122.9 Vo l t s 1 Phase 60 Hertz 4.8 A m p s Cord and Plug NEMA Designation: 5-15 P Wall receptacle NEMA Designation: 5-15R Notes: The ground wire is connected. Non-load cycle Amps = 0.5 An Uninterrupted Power Supply (UPS) is not used on this incubator. This unit is not hooked to a back-up generator. List of Standard Operating Procedures (SOPs) associated with this incubator; the SOPs were list- ed by number, title, and revision number. T h e y are not included in the article as SOPs, and are confidential documents of the company where we conducted this validation. SOPs were reviewed and were found to be appropriate. The incubator does not have a temperature display. There is an external temperature recorder in use for this unit, which is a Dickson Temptrace Re- corder Chart. The recorder chart is identified as number one and contains a replaceable seven-day chart (graph). Acceptance criteria were met. Operational Qualification: (OQ) In this section the performance of the equipment and its operational procedure was verified. The OQ of an incubator consists of an empty chamber tem- perature mapping. Purpose The OQ was performed to confirm that the empty chamber works within the specified limits of tem- perature (32 to 37C) throughout the chamber . Equipment and Materials Precision Thelco Incubator (32-37C) Real Time Datalogger - Ellab Model TM9616 Real Time Datalogger Serial Number 00270110141 Thermocouples Type T (Cu/Con) Wireless Data Loggers Ellab Model T S M 9 8 0 3 Serial Number 2056 Temperature Sensor Serial Number LC 1 20022 6422 LC 2 20753 5757 LC 3 20770 5747 LC 4 20764 5770 LC 5 20804 5766 LC 6 20717 5309 Laura Valdes-Mora & Pamela Hardt-English LC 7 20713 5219 LC 8 20703 4521 LC 9 11542 5679 LC 10 11537 4553 Windows-based PC or laptop computer to oper- ate data loggers External Temperature Device Timer Equipment Picture See Figure 1. Acceptance Criterion All sites measured are within 32-37C. OQ Observations Calibration The calibration of thermocouples was performed using a Kaye LTR-140 serial number 5C339H, owned by the pharmaceutical company where we conducted this validation. A total of 12 Thermocouples (TCs) were tested, and 10 of those were used for the tem- perature profiles. Calibrations were satisfactorily con- ducted according to the SOPs. The two extra calibrat- ed thermocouples were kept in reserve in case of a failure of one of the others. Location of Thermocouples and Loggers- Dia- grams: The purpose of the diagram is to show the relative position of the thermocouples and wireless sensors. The figure is not to scale. In the diagrams seen for the top and lower shelf, each number corresponds to the thermocouple and the logger placed at the location. The two devices were side-by-side for each one of the locations indi- cated. The box in the lower shelf with an R in the center indicates the location of the chart recorder, as shown in Figure 2. Tests for Temperature Setting The incubator was set to 35-37C or 36 + 1 C . While TCs were being placed, the incubator was opened for 20 minutes. It took 30 minutes after clos- ing the doors of the incubator for the unit to reach 35C. Note that the chart recorder was reading 32.4C. An additional reading of the recorder indicated the temperature to be 35.5C. Test of temperature setting number one (lowest temperature setting on this incubator). In order to expedite the decrease in temperature, we opened the door of the incubator for a few minutes. The temper- ature decreased to a range between 25-29C. A f t e r closing the door and allowing T C s to stabilize, the lowest temperature recorded was 29C, as shown in Fi g u re 3. Test of Temperature Setting #9 (highest safe temperature setting on this incubator). Safety light came on at 66C. Chart recorder read 49.9C, while the TC next to the recorder chart read 59.2C as Figure 2 R elative Pos ition of the Thermocouples and Wireles s S ens ors Top S helf Lower S helf Front Front 1 2 3 4 5 6 7 8 9 10 R Figure 1 E mpty Chamber S et-Up E mpty Laura Valdes-Mora & Pamela Hardt-English shown in Fi g u re 4. It appears that the chart recorder responds more slowly than the thermocouples to temperature changes. Due to the fact that the safety light came on during this test, it was decided to record current tempera- tures, and set the knob back to its regular setting of number five. The incubator manual was reviewed, and it was found that the safety light is related to a safety thermostat, which operates alternatively to the control thermostat. After increasing the temperature as per step num- ber three, it took the incubator 75 minutes to come down to the set temperature range of 35-37C. Once all thermocouples were within the tempera- ture range established by the protocol (32-37C), with the understanding that temperature readings can range from 31.5 up to 37.4C (per rounding rules), the 24-hour mapping study was started. 24-Hour Temperature Mapping The empty chamber test was conducted for over 24 hours. Temperature readings were recorded every two minutes. From the time the run was started (02:10:40 PM) to its end, the time was determined to be 26 hours, 44 minutes, and 39 seconds. At 03:52:40 PM, all TCs were within temperature range. There were still some hot pockets left from the previous study. It took the chamber an addition- al 42 minutes to stabilize all locations. The data obtained show that all thermocouples were within the specified temperature range for 24 hours as shown in Figure 5. Wi reless Datalogge r, Tra ck s e n s e : The study start- ed at 08:43:00. All wireless loggers (LCs) were at the t a rget temperature range at 10:07:00 as shown in Fi g u re 6. It took the sensors 1 hour and 24 minutes to all be stabilized. This is almost double the amount of time it took the T C s to get into temperature range. Figure 3 R es ults of Low Temperature S etting 4 0 . 0 3 5 . 0 3 0 . 0 2 5 . 0 2 0 . 0 00.32 00:37 00:42 S etting TC 1 Top Shelf Back Left TC 2 Top Shelf Back Right TC 3 Top Shelf Center TC 4 Top Shelf Front Left TC 5 Top Shelf Front Right TC 6 Bottom Shelf Back Left TC 7 Bottom Shelf Back Right TC 8 Bottom Shelf Center TC 9 Bottom Shelf Left Fr o n t TC 10 Bottom Shelf Front Right TC 13 Description TC 14 Description TC 15 Description TC 16 Description Figure 4 R es ults of High Temperature S etting 7 0 . 0 0 6 0 . 0 0 5 0 . 0 0 4 0 . 0 0 3 0 . 0 0 2 0 . 0 0 1 0 . 0 0 0 . 0 0 01:08 01:13 01:18 01:23 S etting TC 1 Top Shelf Back Left TC 2 Top Shelf Back Right TC 3 Top Shelf Center TC 4 Top Shelf Front Left TC 5 Top Shelf Front Right TC 6 Bottom Shelf Back Left TC 7 Bottom Shelf Back Right TC 8 Bottom Shelf Center TC 9 Bottom Shelf Left Fr o n t TC 10 Bottom Shelf Front Right TC 13 Description TC 14 Description TC 15 Description TC 16 Description Laura Valdes-Mora & Pamela Hardt-English In order for this to be a 24-hour run, the readings needed to last until 10:07:00 on day number two. T h e last reading on day number two was made at 08:41:00, which completes the 24-hour run, but does not map the temperature within specified ranges for 24 hours. This is one hour and 20 minutes short of the requirement. This is due to the fact of not being able to see the data until the study is complete. Note that the wireless system is being used to determine its suitability for validating incubators by comparing it to the real time system. This validation study is based on the data obtained from traditional thermocouples. Thermocouple Test: Started at 2:40:40 and ended at 26:52:40. All TCs reached the target temperature range at 2:40:40. During the first hour and eight minutes, there were 10 temperature excursions: one on TC 5 of 0.1C and the other seven on TC 8. Of the excursions on TC 8, six were of 0.1C, and one was of 0.2C. This was not observed at any other point in time dur- ing the 24-hour study. This indicates that there were still a few hot spots in the chamber after the highest temperature setting test was completed immediately prior to the empty chamber mapping. Future mapping studies should allow for more time between tests either by adding two extra hours, or by running the mapping study for 30 hours in order to compensate for the adjustment period. There are companies that change temperature ranges of their incubators on a regular basis, due to the variety of tests conducted and the lack of incubators. In this situation, the chamber should be allowed several hours to equilibrate prior to loading it. However, this is not the case of this partic- ular piece of equipment. Therefore, this phenomenon observed during the 24-hour empty chamber test should not occur during day-to-day use. Tracksense: All wireless sensors were within specified temperature range at 9:43:00. During the first 24 minutes, three sensors reached 37.5C. This was not observed at any other point in time during the 24-hour study. During the review of TC temperatures, the excur- sions stated above were not found. The highest tem- perature found during this review using the comput- er screen was 37.1C Figure 5 R es ults of 24-Hour E mpty Chamber Mapping Thermocouples 4 0 . 0 3 0 . 0 2 0 . 0 1 0 . 0 0 . 0 02:40 07:25 12:10 16:55 21:40 S etting TC 1 Top Shelf Back Left TC 2 Top Shelf Back Right TC 3 Top Shelf Center TC 4 Top Shelf Front Left TC 5 Top Shelf Front Right TC 6 Bottom Shelf Back Left TC 7 Bottom Shelf Back Right TC 8 Bottom Shelf Center TC 9 Bottom Shelf Left Fr o n t TC 10 Bottom Shelf Front Right TC 13 Description TC 14 Description TC 15 Description TC 16 Description Figure 6 R es ults of 24-Hour E mpty Chamber Mapping Wireles s Tracks ens e 4 0 . 0 3 0 . 0 2 0 . 0 1 0 . 0 0 . 0 08:43-1 13:28-1 18:13-1 22:58-1 03:43-2 S etting LC 1 Top Shelf Back Left LC 2 Top Shelf Back Right LC 3 Top Shelf Center LC 4 Top Shelf Front Left LC 5 Top Shelf Front Right LC 6 Bottom Shelf Back Left LC 7 Bottom Shelf Back Right LC 8 Bottom Shelf Center LC 9 Bottom Shelf Left Fr o n t LC 10 Bottom Shelf Front Right Laura Valdes-Mora & Pamela Hardt-English All temperature devices were checked, and all were within the specified limit of + 2C from each other. Note that the temperature loggers recorded diff e r e n t actual clock times. The loggers were not synchronized for this study, neither were they set to local time. Each one was set for a different time zone. Areal time elapsed adjustment was made to account for the diff e r e n c e . Performance Qualification: (PQ) In this section it was verified that the loaded incu- bator chamber was capable of reaching and main- taining the target temperature range. Purpose The PQ confirms that the loaded chamber works within the specified limits of temperature through- out the chamber. The effect of opening doors for two different lengths of time, and the effect of a power failure on the incubators ability to get back into the temperature profile were also evaluated. Equipment and Materials Precision Thelco Incubator (32-37C) Real Time Data Logger Ellab Model TM9616 Real Time Data Logger Serial Number 00270110141 Thermocouples Type T (Cu/Con) Wireless Data Loggers Ellab Model TSM9803 Serial Number 2056 Temperature Sensor Serial Number LC 1 20022 6422 LC 2 20753 5757 LC 3 20770 5747 LC 4 20764 5770 LC 5 20804 5766 LC 6 20717 5309 LC 7 20713 5219 LC 8 20703 4521 LC 9 11542 5679 LC 10 11537 4553 Windows-based PC or laptop computer to operate data loggers. External Temperature Device (chart recorder) Timer Agar plates and culture broths to create a maxi- mum loaded chamber pattern. Acceptance Criterion All sites measured are within 32-37C PQ Observations A maximum load pattern was selected for the PQ. In addition, the effect of opening doors for two d i fferent lengths of time, and the effect of a power failure on the temperature profile were evaluated in this phase. Ty p i c a l l y, these evaluations are con- ducted during the OQ, however, we chose the loaded chamber for these tests as we consider this scenario more realistic, therefore, more meaning- f u l . Loaded Chamber Description Top Shelf: 4* 2000 mLscrew cap bottles of Lactose B r o t h 4 1000 mL screw cap bottles contain- ing 500 mL each of Lactose Broth 1 Rack of 40 screw cap test tubes with 10 mL each of Nutrient Broth 2 Baskets each containing 11 Whirlpak bags each with 100 mL TSB** Lower Shelf: 1 Chart recorder 1 Water bottle (used by laboratory for temperature determinations) 15 Stacks of plates- total of 72 plates- 42 Blood Agar and 30 TSA*** 12 Whirlpak bags with 250 mL Lactose Broth each (in a basket) 4 API strips * Two (2) 2000 mLbottles were removed from the study. See observations for explanation. ** TSB= Tryptic Soy Broth *** TSA= Tryptic Soy Agar Loading Pattern Diagrams: The purpose of the diagrams in Figure 7 is to show the relative position of the various items in the load. The diagrams are not to scale. Numbers on each of the above rectangles denote number of plates on each stack. If the number is fol- lowed by mL, it denotes the volume of medium on each container. Laura Valdes-Mora & Pamela Hardt-English Loaded Chamber Picture An example of a loaded chamber can be seen in Figure 8. Maximum Load Pattern After loading the chamber and waiting one hour and 45 minutes, it was observed that only 2 TCs were within the target temperature range. We analyzed the content of the load, and laboratory personnel re- viewed the loading pattern. It was concluded that the incubator was loaded above the worst-case scenario. Four 2-liter flasks are never placed into this incubator by laboratory personnel. This was done only once during the validation of a microbial limit test, but as it is not a regular procedure, it was chosen to decrease the loaded chamber by two 2-liter flasks, based on previously performed work. Following this action, it took the chamber two hours and 45 minutes to reach the set temperature. Five-Minute Opened Door Test This incubator consists of two double doors. During this test all doors were opened for five-min- utes. The purpose of the test is to simulate a worse case scenario when laboratory analysts are loading inocu- lated samples and their controls into the incubator. During this test time, the temperature of the incu- bator decreased. Temperature ranged from 22.9 to 30.2C across the loaded chamber as shown in Fig- ure 9. The chart recorder reading was 30.5C. It took 50.5 minutes for the loaded chamber to return to the set temperature. Three-Hour Power Outage Test The purpose of the test was to simulate the eff e c t of a power outage on the temperature profile of a fully loaded incubator chamber. The TCs were checked to ensure all sites were at the set temperature and proceeded to disconnect the incubator. A f t e r three hours, the temperature across the chamber was reviewed and the incubator was reconnected to its power supply. There was a decrease in temperature that took the loaded chamber outside the set temperature and out- side the target temperature. Temperatures ranged from 27.7 to 29.4C as shown in Fi g u re 10. The chart recorder was reading 28.7C. Figure 7 R elative Pos ition of the Various Items in the Load B ottom S helf T1 T2 T3 T4 T5 T6 T7 T8 T9 T 1 0 2000 mL Top S helf 2000 mL Test Tube Rack 10 A 10 B 3 A 4 B 4 A 5 A 4 B 4 B 4 B 6 B 8 A R C 2 B 2 B 3 B 3 B Basket #1 1 0 0 0 mL 1 0 0 0 mL 1 0 0 0 mL 1 0 0 0 mL Basket #2 Basket API R= Chart recorder A= TSA plates C= Water Control B= Blood Agar Plates T= Thermocouple number (a wireless sensor is next to each thermocouple) Figure 8 Picture of Fully Loaded Chamber Full Laura Valdes-Mora & Pamela Hardt-English Following the power start-up, it took one hour and 35.5 minutes for the temperature to reach the set point of 35-37C. This unexpectedly long ramp time signals that the load also dropped in temperature, thus requiring a longer time to return to the set tem- perature. One-Minute Opened Door Test: The protocol was originally written for a five- minute and a 15-minute opened door test. After per- forming the five-minute opened door test, it was determined that most of the time the incubator doors do not remain open longer than just a few minutes, typically not even reaching the five-minute mark. Based on this, it was decided to change the 15- minute test to a one-minute test in order to represent a true scenario. In addition, for this test, only one side of the double doors was opened, not both, for the same reason of mimicking actual use. During the one-minute opened door test, the tem- perature decreased and was outside the set tempera- ture in some areas of the chamber. The temperatures ranged from 29.3 to 35.2C as shown in Fi g u re 11. The chart recorder was reading 33.6C. It took 15 min- utes for the chamber to return to the set temperature once the door was closed. 24-Hour Loaded Chamber Temperature Mapping The loaded chamber test was conducted for 24 hours. Temperature readings were recorded every two minutes. The study started at 08:32:00 and ended at 32:16:00. This is 16 seconds short of precisely 24- hours, however, due to rounding rules, this small Figure 10 R es ults of Three-Hour Power Outage Tes t 7 0 . 0 0 6 0 . 0 0 5 0 . 0 0 4 0 . 0 0 3 0 . 0 0 2 0 . 0 0 1 0 . 0 0 0 . 0 0 03:36 04:31 05:26 06:21 07:16 S etting TC 1 Top Back Left TC 2 Top Back Right TC 3 Top Center TC 4 Top Front Left TC 5 Top Front Right TC 6 Bottom Back Left TC 7 Bottom Back Right TC 8 Bottom Center TC 9 Bottom Front Left TC 10 Bottom Front Right Figure 9 R es ults of Five-Minute Opened Door Tes t 7 0 . 0 0 6 0 . 0 0 5 0 . 0 0 4 0 . 0 0 3 0 . 0 0 2 0 . 0 0 1 0 . 0 0 0 . 0 0 02:34 02:44 02:54 03:04 03:14 03:24 03:34 S etting TA 1 Top Back Left TA 2 Top Back Right TA 3 Top Center TA 4 Top Front Left TA 5 Top Front Right TA 6 Bottom Back Left TA 7 Bottom Back Right TA 8 Bottom Center Laura Valdes-Mora & Pamela Hardt-English shortage is considered acceptable as this is a 24-hour run, not a 24.0-hour run. All thermocouples were within temperature range at the start of the mapping, as shown in Fi g u re 12. No excursions were found at any point in time, thus corroborating that the excursions seen during the empty chamber test were due to hot pockets left from the highest temperature setting test. The loaded chamber maintained a tight temperature range throughout the 24-hour mapping study on a per TC basis. The lowest temperature recorded was 34.5C found on TC 8, while the highest was 36.9C found on TC 9. Tra ck s e n s e : All loggers were within temperature range at the start of the mapping as shown in Fi g u re 1 3. The study started at 09:50:00, and ended at 33:52:00, for a total of 24 hours and two seconds. There were no excursions found at any point in time. The lowest temperature recorded was 34.3C found on LC 8, while the highest was 37.0C found on LC 6. All temperature devices were checked and all were within the specified limit of + 2C from each other. Note that the temperature loggers recorded diff e r- ent actual clock times. The loggers were not syn- chronized for this study, neither were they set to local time. Each one was set for a different time zone. Compar isons Between Data Collected by T her mocouples and by Wir eless L ogger s The thermocouples used are type T, copper-con- stantan thermocouples. The quality of wire and con- struction can significantly affect the accuracy of the thermocouples. The thermocouples used in these tests are Ellabs sealed thermocouples with a stain- less steel tip. The Ellab Tracksense logger sensor is a thermister made of a Silicon chip. Each logger and sensor has a separate ID number, and is factory cal- ibrated at five points in the range of -40 to 140C. The accuracy of the Ellab thermocouples and the Ellab Tracksense sensors is +/-0.2C. Figure 11 R es ults of One-Minute Open Door Tes t 4 5 . 0 4 0 . 0 3 5 . 0 3 0 . 0 2 5 . 0 2 0 . 0 07:58 08:08 08:18 08:28 S etting TC 1 Top Back Left TC 2 Top Back Right TC 3 Top Center TC 4 Top Front Left TC 5 Top Front Right TC 6 Bottom Back Left TC 7 Bottom Back Right TC 8 Bottom Center TC 9 Bottom Front Left TC 10 Bottom Front Right Figure 12 R es ults of 24-Hour Loaded Chamber Temperature Mapping Thermocouples 4 0 . 0 3 0 . 0 2 0 . 0 1 0 . 0 0 . 0 08:32 13:17 18:02 22:47 27:32 S etting TC 1 Top Back Left TC 2 Top Back Right TC 3 Top Center TC 4 Top Front Left TC 5 Top Front Right TC 6 Bottom Back Left TC 7 Bottom Back Right TC 8 Bottom Center TC 9 Bottom Left Fr o n t TC 10 Bottom Front Right Laura Valdes-Mora & Pamela Hardt-English Can these data collection systems be used inter- changeably? Fi g u re 14 shows a summary of 10 hours of data from both the empty chamber 24-hour test and the fully loaded incubator test for both the wireless and thermocouple studies. The thermocouple data were col- lected on a real time system, EVAL, and the wireless data were collected using Ellabs Tracksense system. This summary demonstrates that both systems generated comparable results. The differences in val- ues are well within the accuracy of both the thermo- couples and the thermisters, which is +/-0.2C. When complementing thermocouples with wire- less loggers or using wireless loggers in place of ther- mocouples, the issue of real time versus historical data collection must be addressed. Thermocouples are connected to real time data loggers, i.e., the data are displayed as collected. The wireless system is histor- ical, i.e., the data are downloaded and displayed at the conclusion of the test. Since the data cannot be seen, preliminary tests might need to be conducted to deter- mine the start time on a test, such as the 24-hour fully loaded incubator, because the temperature of the incu- bator might drop during loading. With a wireless system, there are no wires to get tangled and assembly can proceed quickly. The Ellab wireless data loggers are more stable and sealed, and need less frequent calibration than thermocouples. Thermocouples are frequently not sealed, and there- fore subject to corrosion and degradation that will a ffect the calibration. It must be also kept in mind that not all thermocouples and wireless data loggers are as accurate as the sensors used in this study. Summary of Deviations: The following are the deviations that occurred dur- ing the execution of this validation. All deviations from Figure 13 R es ults of 24-Hour Loaded Chamber Temperature Mapping Wireles s Tracks ens e 4 0 . 0 3 0 . 0 2 0 . 0 1 0 . 0 0 . 0 09:50 14:35 19:20 24:05 28:50 S etting LC 1 L # 20022 S # 6422 LC 2 L # 20753 S # 5757 LC 3 L # 20770 S # 5747 LC 4 L # 20764 S # 5770 LC 5 L # 20804 S # 5766 LC 6 L # 20717 S # 5309 LC 7 L # 20713 S # 5219 LC 8 L # 20703 S # 4521 LC 9 L # 11542 S # 5679 LC 10 L # 11537 S # 4553 Figure 14 Data Collection Comparis on Between Thermocouples and Wireles s Dataloge rs All Values are in Degrees Centigrade Type of Data File T i m e M i n i mu m M a x i mu m Av e r age Av e r age Delta T Av e r age Te s t N a m e / S y s t e m I n t e rva l for all for all of Minimu m of Maximum ( R a n ge of all (10 M i n i mu m M a x i mu m Va l u e s Va l u e s f ro m Va l u e s H o u rs ) Va l u e s Va l u e s Av e r age ) Empty Eval1.dt 6:00:00- 36.3 37.4 36.5 37.1 0.3 36.8 Chamber 4/EVAL 16:00:00 Track1.dt 6:00:00- 36.3 37.4 36.7 37.1 0.2 36.8 4/Tracksense 16:00:00 Loaded Eval 22.dt 12:00:00- 35.5 36.9 35.9 36.3 0.3 36.1 Chamber 4 22:00:00 Track2.dt 12:00:00- 35.4 36.8 35.9 36.3 0.5 36.2 4/Tracksense Laura Valdes-Mora & Pamela Hardt-English the original protocol were justified and approved. There was no reference available for the Kaye dry bath. Equipment was sent out to the manufactur- er for routine calibration. The internal bath reference was used for all calibrations of the dry bath per- formed for this validation. The protocol indicated that the chamber should be allowed to be at temperature range for at least 30 minutes prior to conducting the five-minute opened door test. Based on OQ data, it was shown that the incubator is capable of maintaining the temperature within the range for extended periods; therefore wait- ing 30 minutes would not have made a diff e r e n c e . A 15-minute Opened Door Test was described on step 15 of the PQ protocol. This was changed to a one- minute Opened Door Test, as typically incubator doors are opened for not more than a couple of minutes. T h e new test length was considered to be reflective of a real situation. The final post calibration of the thermocouples had to be repeated because there was not enough data col- lected to confirm the high temperature calibration at 40C. The EVA L Datalogger collects and displays data every two seconds, but the program was set to save data every two minutes, and the system was shut down before the final data were stored. Therefore, the calibration was repeated at Star Services, Hayward, California, using a Kaye Liquid bath at two tempera- tures, 30 and 40C to confirm that the thermocouples were within specifications. Conclusions I Q It is concluded that the unit was installed proper- ly. All necessary components were found to be pre- sent, and all the required SOPs were found to be in place. The utilities (power) supplied to the unit, as per testing, were found to meet the requirements. OQ All test results met the protocol acceptance crite- ria. The unit is capable of maintaining the target temperature. PQ All test results met the protocol acceptance crite- ria. Based on data obtained from the opened door tests and power failure, it can be concluded that nor- mal laboratory practices are acceptable, and that extending the incubation time for the same length of time of the power failure is an acceptable solution. Note that time should be calculated from the time the incubator reaches the temperature range (see rec- ommendations). Conclusion The incubator was tested and verified to operate properly according to the manufacturer and protocol requirements. This unit is therefore considered vali- dated. Recommendations In order to have enough data collected, one should consider running the temperature mappings with a safety factor of six hours, that is, for a 24-hour run, conduct a 30-hour mapping study, especially if wire- less sensors will be used. The validation presented here was completed in three days of continuous work. Scheduling and exe- cuting the work in this manner is taxing and compli- cated. At times, one will be working very early or very late hours. Careful pre-scheduling is highly rec- ommended. Consider adding extra time to the sched- ule for unforeseen situations (example: chamber may not get to temperature in the time you have predict- e d ) . Older incubators that do not have a digital temper- ature display can benefit from having one installed. Digital temperature displays are available; they have a Real Time Datalogger (RTD) which is placed in the incubator either through a port (if available) or through the top vent. Microbiologists should consider that after open- ing the incubator doors, the equipment loses heat, causing a decrease in temperature, which typically falls outside the teststemperature range. Based on this, incubation times should be recorded from the time the incubator is at temperature range, not from the time samples are placed in the incubator. Caution must be exercised when reading temper- ature from an external chart recorder. Data from this validation clearly showed that chart recorders are slow in capturing accurate temperature. J Laura Valdes-Mora & Pamela Hardt-English About the Author s L a u ra Va l d e s - M o ra serves as Technical S ervices Dir- ector, acting as Principal Consultant and National S p e a ker on US P Microbiology Updates and related topics at E lite MicroS ource Corporation, a R esource Center for Pharmaceutical and Biotechnology M i c r o- b i o l o g y. S he received a B. S. d e gree in Biology from the U n i versity of Puerto R ico in 1982, and an M.S. d e gr e e in Microbiology from North Carolina State University in 1 9 8 6 . M s. Va l d e s - M o ra was a Director at AAI P h a rm a , was fo rm e rly in charge of production of bacteri a l endospores at AMS CO, and has been the President of the Pharmaceutical Microbiology Forum (PMF) since 1994. Ms. Valdes-Mora can be reached by phone at 850-763-5453, by fax at 850-913-9735, or by e-mail at E MS ource@ aol.com. Pamela Hardt-E nglish, is President of PhF S pecial- i s t s. S he has M.S. d e grees in Food Science and A gri c u l t u ral E ngineering from the University of Califo r- nia (Davis) and a B. A . in Computer S cience from the University of California (Berkeley). For the last 23 years at PhF Specialists and FMC Corp o ration, Ms. Hardt-E nglish has served as a recognized FDA and U S DA process authori t y, instituted HACCP progra m s, conducted thermal processing studies, and valida- tions of autoclave s, incubators, and other chambers. S he has designed and taught numerous food pro- cessing courses. M s. Hardt-E nglish can be reached by phone at 408-275-0161, by fax at 408-280-0979, or by email at phfspec@ pacbell.net. Suggested R eading The following publications were consulted during the preparation of the protocol, execution of the work, and preparation of the final report. Cloud, P.A., Validating a Laboratory Incubator, B i o p h a r m30, pp. 39-42. November (1997). Phil Cloud, Pharmaceutical Equipment Validation: T h e Ultimate Qualification Guidebook (Interpharm Press, Buffalo Grove, IL1998). Cloud, P., How to Develop and Manage Qualification Proto- cols for FDACompliance. (Interpharm Press, Buffalo Grove, IL 1999). R efer ences 1. Institute of Validation Te c h n o l o g y. Journal of Va l i d a t i o n Technology. Vol. 8 No. 1, November 2001. pp. 63-72. IQ: Installation Qualification LC: Wireless Loggers OQ: Operational Qualification PQ: Performance Qualification RT D : Real Time Datalogger S O P : Standard Operating Procedure T C : T h e r m o c o u p l e UPS: Uninterrupted Power Supply TSA: Tryptic Soy Agar TSB: Tryptic Soy Broth Ar ticle Acr onym L isting