V alidation of a L aboratory

I ncubator U sing Wireless

and C abled Datalogger
P art I I
By Laura Valdes-Mora
Elite MicroSource Corporation
and Pamela Hardt-English
PhF Specialists, Inc.

T
he following article describes
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
ber 31487. This is a gravity convec-
T his validation tion unit model number 6M with an
was designed operating temperature range of 25C
to 70C. The incubator has the fol-
and per for med lowing electrical characteristics:
fr om the
Volts- 120
viewpoint of a Hertz- 50/60
micr obiologist Watts- 300
with assistance The overall dimensions of the
fr om validation unit are 42 X 21 X 39 with cham-
ber dimensions 36 X 18 X 27.The
exper ts. incubator contains a dual hydraulic
thermostat, a mercury-in-glass ther-
mometer, two shelves, two heaters, double inner
doors, and double outer doors.

The incubator is a conventional unit where fresh Installation Qualification (IQ)

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-
During the IQ, personnel documented all items list-
ed in the Incubator IQ Form. Refer to Appendix A of
validation protocol, previously published in the
Journal of Validation Technol ogy.1 A second person
reviewed and approved all verifications.

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.
E quipment and Materials
Incubator
Appendix A
IQ 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-
oratory.
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 Volts 1 Phase 60 Hertz 20 Amps
Equipment Rating:
125 Volts 1 Phase 60 Hertz 5.2 Amps
Field Measurement:
122.9 Volts 1 Phase 60 Hertz 4.8 Amps
Cord and Plug NEMA Designation: 5-15 P
Wall receptacle NEMA Designation: 5-15R
Laura Valdes-Mora & Pamela Hardt-English
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. They
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.
E quipment 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 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
Laura Valdes-Mora & Pamela Hardt-English

LC 7 20713 5219 In the diagrams seen for the top and lower shelf,
LC 8 20703 4521 each number corresponds to the thermocouple and
LC 9 11542 5679 the logger placed at the location. The two devices
LC 10 11537 4553 were side-by-side for each one of the locations indi-
Windows-based PC or laptop computer to oper- cated. The box in the lower shelf with an R in the
ate data loggers center indicates the location of the chart recorder, as
External Temperature Device shown in Figure 2.
Timer
Figure 2
E quipment Picture R elative P os ition of the
See Figure 1. T hermoc ouples and
Figure 1 Wireles s S ens ors
Top S helf
E mpty C hamber S et-Up
1 2

4 5
Front

L ower S helf
6 R 7

9 10
Front
E mpty

Tests for Temperature Setting

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.
The incubator was set to 35-37Cor 36 + 1C.
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.After
closing the door and allowing TCs to stabilize, the
lowest temperature recorded was 29C, as shown in
Fi gure 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 59.2Cas
 Laura Valdes-Mora & Pamela Hardt-English

Figure 3 Figure 4
R es ults of L o w R es ults of High
Temperature S etting Temperature S etting
40.0 70.00

35.0 60.00

30.0 50.00

25.0 40.00

20.0 30.00
00.32 00:37 00:42
S etting
TC 1 Top Shelf Back Left TC 8 Bottom Shelf Center 20.00
TC 2 Top Shelf Back Right TC 9 Bottom Shelf Left Front
TC 3 Top Shelf Center TC 10 Bottom Shelf Front Right
TC 4 Top Shelf Front Left TC 13 Description
TC 5 Top Shelf Front Right TC 14 Description 10.00
TC 6 Bottom Shelf Back Left TC 15 Description
TC 7 Bottom Shelf Back Right TC 16 Description

shown in Fi gure 4. It appears that the chart recorder 0.00

responds more slowly than the thermocouples to 01:08 01:13 01:18 01:23
temperature changes. S etting
Due to the fact that the safety light came on during TC 1 Top Shelf Back Left TC 8 Bottom Shelf Center
this test, it was decided to record current tempera- TC 2 Top Shelf Back Right TC 9 Bottom Shelf Left Front
TC 3 Top Shelf Center TC 10 Bottom Shelf Front Right
tures, and set the knob back to its regular setting of TC 4 Top Shelf Front Left TC 13 Description
number five. The incubator manual was reviewed, TC 5 Top Shelf Front Right TC 14 Description
TC 6 Bottom Shelf Back Left TC 15 Description
and it was found that the safety light is related to a TC 7 Bottom Shelf Back Right TC 16 Description
safety thermostat, which operates alternatively to the
control thermostat. be 26 hours, 44 minutes, and 39 seconds.
After increasing the temperature as per step num- At 03:52:40 PM, all TCs were within temperature
ber three, it took the incubator 75 minutes to come range. There were still some hot pockets left from
down to the set temperature range of 35-37C. the previous study. It took the chamber an addition-
Once all thermocouples were within the tempera- al 42 minutes to stabilize all locations.
ture range established by the protocol (32-37C), The data obtained show that all thermocouples
with the understanding that temperature readings were within the specified temperature range for 24
can range from 31.5 up to 37.4C (per rounding hours as shown in Figure 5.
rules), the 24-hour mapping study was started.
Wi reless Datalogger, Tracksense: The study start-
24-Hour Temperature Mapping ed at 08:43:00. All wireless loggers (LCs) were at the
The empty chamber test was conducted for over target temperature range at 10:07:00 as shown in
24 hours. Temperature readings were recorded every Fi gure 6. It took the sensors 1 hour and 24 minutes to
two minutes. From the time the run was started all be stabilized. This is almost double the amount of
(02:10:40 PM) to its end, the time was determined to time it took the TCs to get into temperature range.
Laura Valdes-Mora & Pamela Hardt-English

Figure 5 Figure 6
R es ults of 24-Hour E mpty R es ults of 24-Hour E mpty
C hamber Mapping C hamber Mapping
T hermoc ouples Wireles s Trac ks ens e
40.0 40.0

30.0 30.0

20.0 20.0

10.0 10.0

0.0 0.0
02:40 07:25 12:10 16:55 21:40 08:43-1 13:28-1 18:13-1 22:58-1 03:43-2
S etting S etting
TC 1 Top Shelf Back Left TC 8 Bottom Shelf Center LC 1 Top Shelf Back Left LC 6 Bottom Shelf Back Left
TC 2 Top Shelf Back Right TC 9 Bottom Shelf Left Front LC 2 Top Shelf Back Right LC 7 Bottom Shelf Back Right
TC 3 Top Shelf Center TC 10 Bottom Shelf Front Right LC 3 Top Shelf Center LC 8 Bottom Shelf Center
TC 4 Top Shelf Front Left TC 13 Description LC 4 Top Shelf Front Left LC 9 Bottom Shelf Left Front
TC 5 Top Shelf Front Right TC 14 Description LC 5 Top Shelf Front Right LC 10 Bottom Shelf Front Right
TC 6 Bottom Shelf Back Left TC 15 Description
TC 7 Bottom Shelf Back Right TC 16 Description
temperature setting test was completed immediately
In order for this to be a 24-hour run, the readings prior to the empty chamber mapping. Future mapping
needed to last until 10:07:00 on day number two. The studies should allow for more time between tests
last reading on day number two was made at either by adding two extra hours, or by running the
08:41:00, which completes the 24-hour run, but does mapping study for 30 hours in order to compensate
not map the temperature within specified ranges for for the adjustment period. There are companies that
24 hours. This is one hour and 20 minutes short of the change temperature ranges of their incubators on a
requirement. This is due to the fact of not being able regular basis, due to the variety of tests conducted and
to see the data until the study is complete. Note that the lack of incubators. In this situation, the chamber
the wireless system is being used to determine its should be allowed several hours to equilibrate prior to
suitability for validating incubators by comparing it loading it. However, this is not the case of this partic-
to the real time system. This validation study is based ular piece of equipment. Therefore, this phenomenon
on the data obtained from traditional thermocouples. observed during the 24-hour empty chamber test
should not occur during day-to-day use.
Thermocouple Test: Started at 2:40:40 and ended
at 26:52:40. Tracksense: All wireless sensors were within
All TCs reached the target temperature range at specified temperature range at 9:43:00. During the
2:40:40. During the first hour and eight minutes, there first 24 minutes, three sensors reached 37.5C. This
were 10 temperature excursions: one on TC 5 of was not observed at any other point in time during
0.1C and the other seven on TC 8. Of the excursions the 24-hour study.
on TC 8, six were of 0.1C, and one was of 0.2C. During the review of TC temperatures, the excur-
This was not observed at any other point in time dur- sions stated above were not found. The highest tem-
ing the 24-hour study. This indicates that there were perature found during this review using the comput-
still a few hot spots in the chamber after the highest er screen was 37.1C
 Laura Valdes-Mora & Pamela Hardt-English

All temperature devices were checked, and all were Acceptance Criterion
within the specified limit of + 2C from each other. All sites measured are within 32-37C
Note that the temperature loggers recorded different
actual clock times. The loggers were not synchronized PQ Observations
for this study, neither were they set to local time. Each A maximum load pattern was selected for the
one was set for a different time zone. Areal time elapsed PQ. In addition, the effect of opening doors for two
adjustment was made to account for the difference. different lengths of time, and the effect of a power
failure on the temperature profile were evaluated in
Performance Qualification: (PQ) this phase. Typically, these evaluations are con-
In this section it was verified that the loaded incu- ducted during the OQ, however, we chose the
bator chamber was capable of reaching and main- loaded chamber for these tests as we consider this
taining the target temperature range. scenario more realistic, therefore, more meaning-
ful.
Purpose
The PQ confirms that the loaded chamber works Loaded Chamber Description
within the specified limits of temperature through- Top Shelf:
out the chamber. The effect of opening doors for two 4* 2000 mL screw cap bottles of Lactose
different lengths of time, and the effect of a power Broth
failure on the incubators ability to get back into the 4 1000 mL screw cap bottles contain-
temperature profile were also evaluated. ing 500 mL each of Lactose Broth
1 Rack of 40 screw cap test tubes with
E quipment and Materials 10 mL each of Nutrient Broth
Precision Thelco Incubator (32-37C) 2 Baskets each containing 11 Whirlpak
Real Time Data Logger Ellab Model TM9616 bags each with 100 mL TSB**
Real Time Data Logger
Serial Number 00270110141 Lower Shelf:
Thermocouples Type T (Cu/Con) 1 Chart recorder
Wireless Data Loggers 1 Water bottle (used by laboratory for
Ellab Model TSM9803 temperature determinations)
Serial Number 2056 15 Stacks of plates- total of 72 plates- 42
Temperature Sensor Serial Number Blood Agar and 30 TSA***
LC 1 20022 6422 12 Whirlpak bags with 250 mL Lactose
LC 2 20753 5757 Broth each (in a basket)
LC 3 20770 5747 4 API strips
LC 4 20764 5770
LC 5 20804 5766 * Two (2) 2000 mL bottles were removed from
LC 6 20717 5309 the study. See observations for explanation.
LC 7 20713 5219 ** TSB= Tryptic Soy Broth
LC 8 20703 4521 *** TSA= Tryptic Soy Agar
LC 9 11542 5679
LC 10 11537 4553 Loading Pattern Diagrams: The purpose of the
diagrams in Figure 7 is to show the relative position
Windows-based PC or laptop computer to of the various items in the load. The diagrams are
operate data loggers. not to scale.
External Temperature Device (chart recorder) Numbers on each of the above rectangles denote
Timer number of plates on each stack. If the number is fol-
Agar plates and culture broths to create a maxi- lowed by mL, it denotes the volume of medium on
mum loaded chamber pattern. each container.
Laura Valdes-Mora & Pamela Hardt-English

Figure 7 Figure 8
R elative P os ition of the Various P ic ture of F ully L oaded C hamber
Items in the L oad

Top S helf
T1 Test Tube T2
1 1
2000 Rack
0 0
mL Basket
0 0
#1
0 0
mL mL

T3
1 1 Basket
0 0 #2
2000 0 0
mL F ull
0 0
T4 mL mL T5
the loaded chamber by two 2-liter flasks, based on
B ottom S helf previously performed work.
Following this action, it took the chamber two
T6 T7
R C 3 5 hours and 45 minutes to reach the set temperature.
A A
10 10 8 Basket Five-Minute Opened Door Test
3B 2B
A B A This incubator consists of two double doors.
4 2B During this test all doors were opened for five-min-
B T8 4 API utes. The purpose of the test is to simulate a worse case
4 3B B scenario when laboratory analysts are loading inocu-
B
6 4 lated samples and their controls into the incubator.
4
A B B During this test time, the temperature of the incu-
T9 T10 bator decreased. Temperature ranged from 22.9 to
R= Chart recorder T= Thermocouple number (a 30.2C across the loaded chamber as shown inFig-
A= TSA plates wireless sensor is next to ure 9. The chart recorder reading was 30.5C. It took
C= Water Control each thermocouple)
B= Blood Agar Plates 50.5 minutes for the loaded chamber to return to the
set temperature.
Loaded Chamber Picture
An example of a loaded chamber can be seen in Three-Hour Power Outage Test
Figure 8. The purpose of the test was to simulate the effect
of a power outage on the temperature profile of a
Maximum Load Pattern fully loaded incubator chamber. The TCs were
After loading the chamber and waiting one hour checked to ensure all sites were at the set temperature
and 45 minutes, it was observed that only 2 TCs were and proceeded to disconnect the incubator. After
within the target temperature range. We analyzed the three hours, the temperature across the chamber was
content of the load, and laboratory personnel re- reviewed and the incubator was reconnected to its
viewed the loading pattern. It was concluded that the power supply.
incubator was loaded above the worst-case scenario. There was a decrease in temperature that took the
Four 2-liter flasks are never placed into this incubator loaded chamber outside the set temperature and out-
by laboratory personnel. This was done only once side the target temperature. Temperatures ranged
during the validation of a microbial limit test, but as it from 27.7 to 29.4C as shown inFi gure 10. The chart
is not a regular procedure, it was chosen to decrease recorder was reading 28.7C.
 Laura Valdes-Mora & Pamela Hardt-English

Figure 9 Figure 10
R es ults of F ive-Minute R es ults of T hree-Hour
Opened Door Tes t Power Outage Tes t
70.00 70.00

60.00 60.00

50.00 50.00

40.00 40.00

30.00 30.00

20.00 20.00

10.00 10.00

0.00 0.00
02:34 02:44 02:54 03:04 03:14 03:24 03:34 03:36 04:31 05:26 06:21 07:16
S etting S etting
TC 1 Top Back Left TC 6 Bottom Back Left
TA 1 Top Back Left TA 5 Top Front Right TC 2 Top Back Right TC 7 Bottom Back Right
TA 2 Top Back Right TA 6 Bottom Back Left TC 3 Top Center TC 8 Bottom Center
TA 3 Top Center TA 7 Bottom Back Right TC 4 Top Front Left TC 9 Bottom Front Left
TA 4 Top Front Left TA 8 Bottom Center TC 5 Top Front Right TC 10 Bottom Front Right

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 inFi gure 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
Laura Valdes-Mora & Pamela Hardt-English

Figure 11 Figure 12
R es ults of One-Minute R es ults of 24-Hour L oaded
Open Door Tes t C hamber Temperature Mapping
T hermoc ouples
40.0

45.0
30.0

40.0
20.0

35.0
10.0

30.0
0.0
08:32 13:17 18:02 22:47 27:32
25.0 S etting
TC 1 Top Back Left TC 6 Bottom Back Left
TC 2 Top Back Right TC 7 Bottom Back Right
20.0 TC 3 Top Center TC 8 Bottom Center
TC 4 Top Front Left TC 9 Bottom Left Front
07:58 08:08 08:18 08:28 TC 5 Top Front Right TC 10 Bottom Front Right
S etting
The lowest temperature recorded was 34.3C found on
TC 1 Top Back Left TC 6 Bottom Back Left
TC 2 Top Back Right TC 7 Bottom Back Right LC 8, while the highest was 37.0C found on LC 6.
TC 3 Top Center TC 8 Bottom Center All temperature devices were checked and all
TC 4 Top Front Left TC 9 Bottom Front Left
TC 5 Top Front Right TC 10 Bottom Front Right
were within the specified limit of + 2C from each
other.
shortage is considered acceptable as this is a 24-hour Note that the temperature loggers recorded differ-
run, not a 24.0-hour run. ent actual clock times. The loggers were not syn-
All thermocouples were within temperature range chronized for this study, neither were they set to local
at the start of the mapping, as shown in Fi gure 12. time. Each one was set for a different time zone.
No excursions were found at any point in time,
thus corroborating that the excursions seen during C ompar isons B etween Data C ollected by
the empty chamber test were due to hot pockets left T her mocouples and by W ir eless L ogger s
from the highest temperature setting test.
The loaded chamber maintained a tight temperature The thermocouples used are type T, copper-con-
range throughout the 24-hour mapping study on a per stantan thermocouples. The quality of wire and con-
TC basis. The lowest temperature recorded was struction can significantly affect the accuracy of the
34.5C found on TC 8, while the highest was 36.9C thermocouples. The thermocouples used in these
found on TC 9. tests are Ellabs sealed thermocouples with a stain-
less steel tip. The Ellab Tracksense logger sensor is
Tracksense: All loggers were within temperature a thermister made of a Silicon chip. Each logger and
range at the start of the mapping as shown in Fi gure sensor has a separate ID number, and is factory cal-
13. The study started at 09:50:00, and ended at ibrated at five points in the range of -40 to 140C.
33:52:00, for a total of 24 hours and two seconds. The accuracy of the Ellab thermocouples and the
There were no excursions found at any point in time. Ellab Tracksense sensors is +/-0.2C.
 Laura Valdes-Mora & Pamela Hardt-English

Figure 13 data were collected using Ellabs Tracksense system.

This summary demonstrates that both systems
R es ults of 24-Hour L oaded generated comparable results. The differences in val-
C hamber Temperature Mapping ues are well within the accuracy of both the thermo-
Wireles s Trac ks ens e couples and the thermisters, which is +/-0.2C.
40.0 When complementing thermocouples with wire-
less loggers or using wireless loggers in place of ther-
mocouples, the issue of real time versus historical
30.0 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-
20.0 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-
10.0
mine the start time on a test, such as the 24-hour fully
loaded incubator, because the temperature of the incu-
0.0
bator might drop during loading.
09:50 14:35 19:20 24:05 28:50 With a wireless system, there are no wires to get
S etting tangled and assembly can proceed quickly. The Ellab
LC 1 L # 20022 S # 6422 LC 6 L # 20717 S # 5309 wireless data loggers are more stable and sealed, and
LC 2 L # 20753 S # 5757 LC 7 L # 20713 S # 5219 need less frequent calibration than thermocouples.
LC 3 L # 20770 S # 5747 LC 8 L # 20703 S # 4521 Thermocouples are frequently not sealed, and there-
LC 4 L # 20764 S # 5770 LC 9 L # 11542 S # 5679
LC 5 L # 20804 S # 5766 LC 10 L # 11537 S # 4553 fore subject to corrosion and degradation that will
affect the calibration. It must be also kept in mind
Can these data collection systems be used inter- that not all thermocouples and wireless data loggers
changeably? Fi gure 14 shows a summary of 10 hours of are as accurate as the sensors used in this study.
data from both the empty chamber 24-hour test and the
fully loaded incubator test for both the wireless and Summary of Deviations:
thermocouple studies. The thermocouple data were col- The following are the deviations that occurred dur-
lected on a real time system, EVAL, and the wireless ing the execution of this validation. All deviations from

Figure 14
Data C ollection C omparis on B etween Thermocouples and Wireles s Dataloge rs
A ll Values are in Degrees C entigrade
Type of Data F ile T ime Mi n i mu m Ma 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 te m I n te rva l for all for all of Minimu m of Maximum ( R a n ge of all
(10 Mi n i mu m Ma 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 difference.
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. The
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 EVAL 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.
C onclusions
IQ
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).
C onclusion
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-
ed).
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.
 Laura Valdes-Mora & Pamela Hardt-English

by phone at 408-275-0161, by fax at 408-280-0979,

A bout the Author s or by email at phfspec@ pacbell.net.
L a ura Va lde s -Mora serves as Technical S ervices Dir-
ector, acting as P rincipal C ons ultant and National Suggested R eading
S pe a ker on US P Microbiology Updates and related The following publications were consulted during the preparation of the
topics at E lite MicroS ource C orporation, a R es ource protocol, execution of the work, and preparation of the final report.
C enter for P harmaceutical and B iotechnology Mic ro-
Cloud, P.A., Validating a Laboratory Incubator,B i ophar m30,
biology. S he received a B. S . de gree in Biology from the pp. 39-42. November (1997).
U niversity of P uerto R ico in 1982, and an M.S . de gre e Phil Cloud, Pharmaceutical Equipment Validation: The
in Microbiology from North C arolina S tate University in Ultimate Qualification Guidebook (Interpharm Press, Buffalo
Grove, IL1998).
1 9 8 6 .M s. Va lde s -Mora was a Director at AAI P ha rma , Cloud, P., How to Develop and Manage Qualification Proto-
was forme rly in charge of production of bacteria l cols for FDACompliance. (Interpharm Press, Buffalo Grove,
IL 1999).
endospores at AMS C O, and has been the P resident
of the P harmaceutical Microbiology F orum (P MF )
s ince 1994. Ms. Valdes -Mora can be reached by R efer ences
phone at 850-763-5453, by fax at 850-913-9735, or by 1. Institute of Validation Technology. Journal of Val i dati on
Technology. Vol. 8 No. 1, November 2001. pp. 63-72.
e-mail at E MS ource@ aol.com.
Pamela Hardt-E nglish, is P res ident of PhF S pecial- A r ticle A cr onym L isting
is ts. S he has M.S . de grees in Food S cience and
IQ: Installation Qualification
A gric ultural E ngineering from the University of C alifor-
LC: Wireless Loggers
nia (Davis ) and a B. A . in C omputer S cience from the
OQ: Operational Qualification
Univers ity of C alifornia (B erkeley). For the las t 23
years at P hF S pecialists and F MC C orporation, Ms.
PQ: Performance Qualification
Hardt-E nglis h has s erved as a recognized F DA and RTD: Real Time Datalogger
U S DA process authority, instituted HAC C P progra ms, SOP: Standard Operating Procedure
conducted thermal proces s ing s tudies , and valida- TC: Thermocouple
tions of autoclave s, incubators, and other chambers. UPS: Uninterrupted Power Supply
S he has des igned and taught numerous food pro- TSA: Tryptic Soy Agar
cess ing courses. Ms. Hardt-E nglish can be reached TSB: Tryptic Soy Broth