Lower and Upper RLU Limits for Rev03182013

ATP Monitoring Programs

What are Lower and Upper RLU limits? The data below is extracted from Performance Evaluation of Various ATP Detecting
Units, a report prepared by the world’s largest third party reference
Lower and Upper RLU limits are set values used to easily evaluate laboratory1. The report provides the mean result for two products tested at
and categorize ATP test results in real time. various dilutions dried on stainless steel surfaces as well as serial dilution
detection levels of other food stuff. The process and results for both methods
Lower Limit: The maximum allowable RLU for a result to pass are described below.
the ATP test for cleanliness. A surface that reads at or below
the lower limit is categorized as a Pass result and considered Detection of ATP from Food Soiled Stainless Steel Surfaces
acceptable. Passing results are indicated with a check mark
(√) on the screen Process:
Upper Limit: A surface that reads above the upper limit is
categorized as a Fail result and considered not adequately To replicate the scenario of detecting residue on an unclean surface in a
cleaned. Fail results are indicated with an (X) on the screen. food and beverage processing facility, food suspensions were created
Caution Area: A surface that reads above the lower limit and and 10 replicates of 500 µl were added to a stainless steel 4 x 4 inch
at or below the upper limit is categorized as a Caution result. square and allowed to dry. An UltraSnap test device was used to collect
Caution results tell the user there could be something on the a sample and the device was activated. The RLU output was measured
surface, but not enough to cause a Fail result. Caution areas with SystemSURE Plus. The RLU limit in the right column is Hygiena’s
can also show a surface is trending towards unclean and may recommendations based on the data from the report.
require additional attention during the next cleaning. Caution
results are indicated with an exclamation point (!) on the Results:
screen. Some facilities elect to eliminate the Caution range. To
do this, the lower and upper limits are set to the same RLU SystemSURE Plus with UltraSnap Tests
Dry soiled stainless steel surfaces
value. With this setting, a surface that reads at or below the Product Dilution Results Mean Range Limit (RLU)
limit is categorized as a Pass result (√) and a surface that reads (RLU)
above the limit is categorized as a Fail result (X). Raw Ground 1:10 2945 2540-3644 Fail: 2000
Beef 1:1000 147 116-205 Pass: 100
2% Milk 1:1 173 145-195 Fail: 100
Hygiena luminometers come pre-programmed with a lower limit of
1:1000 8 2-16 Pass: 2
10 RLUs and an upper limit of 30 RLUs. The Pass, Caution, and Fail
ranges are as follows:

RLUs 0-10 11-30 31+ Detection of ATP from Food: Serial Dilutions Pipetted onto Swabs
Pass (√) Caution (!) Fail (X)
Process:
To eliminate the Caution area, set 10 RLU as the lower and upper
limit. The Pass and Fail ranges are as follows: Liquid food samples (orange juice and milk) were diluted using sterile
water in various dilutions. Solid food samples (ground beef and salad
RLUs 0-10 11+ greens) were first stomached using 10 g of sample in 90 ml sterile
Pass (√) Fail (X) water and then diluted using sterile water in various dilutions. All test
samples were shaken by hand for homogenization. Ten replicates of
each food suspension were then analyzed using an UltraSnap ATP
detection device by pipetting 10 μL of food suspension dilution
Why are the default limits 10 and 30? directly onto the swab bud, activating the device, and measuring with
the SystemSURE Plus luminometer to read RLU result. The RLU limit in
Hygiena recommends that Pass and Fail limits be determined by the the right column is Hygiena’s recommendations based on the data
facility and documented as to how they were determined. The default from the report.
limits of 10 (Pass) and 30 (Fail) RLUs on the luminometer are based on
Results:
years of food & beverage processing experience and third party
studies. (See data below). Hygiena recommends users validate these SystemSURE Plus with UltraSnap Tests
recommendations and adjust them to meet the needs of each facility’s Serial dilutions pipetted onto swabs
unique needs. Product Dilution Results Range (RLU) Limit (RLU)
Mean (RLU)
Bagged Mixed 1:10 164 129-195 Fail: 100
The following guidelines are typical of those found in the food and Salad Greens 1:100 36 31-43 Fail: 30
beverage industry and are based on stainless steel surfaces. Other 1:1000 13 10-15 Pass: 10
surfaces may be more difficult to clean and consequently may require 1:10000 2 1-2 Pass: 2
higher Pass/Fail limits to be set if complete bio-burden removal is not
necessary. (Refer to the following page for details on establishing 1 Kupski, Brian, et al. “Performance Evaluation of Various ATP Detecting Units.” Silliker, Inc.,
custom RLU limits.) Food Science Center Report RPN: 13922, (2010).
 Lower and Upper RLU Limits for Rev03182013

ATP Monitoring Programs

SystemSURE Plus with UltraSnap Tests Determining Custom RLU limits

Serial dilutions pipetted onto swabs (continued)
Product Dilution Results Range (RLU) Limit (RLU)
Mean (RLU) The method for determining and validating custom lower and upper
2% Milk 1:1 301 217-366 Fail: 200 RLU limits is outlined below. In some cases, test results from different
1:10 72 59-84 Fail: 50 control points within a facility consistently fall in a similar range. In this
1:100 13 11-15 Fail: 10
case, RLU limits may be set to the same value for all control points in
1:1000 3 3-4 Pass: 5
1:10000 2 2-2 Pass: 2
the facility. Likewise, test results from similar surfaces may
Pasteurized 1:1 5617 5331-6220 Fail: 5000 consistently fall in a similar range. In this case, the RLU limits may be
Orange Juice, 1:10 3958 3563-4448 Fail: 3000 set to the same values for control points with the same surface type.
No Pulp 1:100 437 294-541 Fail 250 Hygiena’s SureTrend data tracking software allows users to take note
1:1000 56 39-70 Pass: 30 of the surface type of each test location.
1:10000 5 2-8 Pass: 2
Raw Ground 1:10 609 453-788 Fail: 400
Beef 1:100 70 54-86 Fail: 50 1. Identify control points in the facility. These are often outlined in the
1:1000 1 0-8 Pass: 10 SSOP/HACCP programs.
1:10000 0 0-1 Pass: 2 a. Control points can be programmed into the luminometer
before testing so that results are saved with the control point
Note: ATP tests are not organism tests, thus RLU values cannot be location name, date, and time of test.
directly correlated to CFU values, nor can ATP tests be a 2. Clean surfaces to the desired level of cleanliness.
replacement for microbiological tests. UltraSnap tests detect any a. This may include a total production line breakdown.
organic material that contains ATP, which may include b. Future cleanings will be held to this level of clean as a
microorganisms. Refer to the full copy of the study for details on standard.
methodology and limit of detection for microorganisms. 3. Perform an ATP test at each control point, taking 5-10 replicate tests.
Use one of two methods.
For a full copy of Performance Evaluation of Various ATP a. Perform tests over several days.
b. Or for control points with sufficient surface area, perform
Detecting Units By Silliker, Inc., please contact Hygiena.
multiple tests from different spots at that test location. (e.g.
Conveyor belt, tank, tabletop, etc.)
If you have any questions about the information provided,
4. Calculate the lower and upper RLU limits
please contact your Hygiena representative or Hygiena’s a. Lower RLU limit: Calculate the average RLU for each
customer service department at: location based on the 5-10 test results. The average result
will be the lower RLU limit.
Phone: 888-HYGIENA (494-4362) b. Upper RLU limit: There are two options for determining the
E-mail: [email protected] upper limit.
i. Multiply the lower limit by 3
Can I use the same RLU limits for every location? ii. Or determine the standard deviation from the test
results, multiply the standard deviation by 3, and
In some cases, the same RLU limits may be set for several locations. add this to the lower limit.
Optimal RLU limits will depend on a variety of factors such as
testing environment, product residue, and cleaning process
variations. See examples below.
CONTINUOUS IMPROVEMENT: Monitoring and assessment of trends is
Surface Variations: Surfaces which are easy to clean,
crucial to finding trouble zones, correcting improper cleaning procedures
such as stainless steel or other flat, non-porous surfaces
and decreasing risk. Continuous improvement provides brand protection,
may have stricter, lower limits. Surfaces which are hard
avoids recalls and shows due diligence to auditors. If high frequencies of
to clean, such as porous, grooved, creviced or irregular Caution and Fail results are obtained with ATP monitoring, SSOP should be
surfaces like conveyor belts, may have higher limits. reviewed for ways to improve cleanliness. If low frequencies of Caution or
Environmental and industrial variations: An environment Fail results are obtained, RLU limits could be reviewed and potentially
that has more stringent cleaning requirements, like a lowered to maintain high standards and generate useful management data.
clean room, may have stricter, lower limits compared to a
food processing environment with less stringent volatility.
Surface Volatility: Surfaces which have contacted high-
risk products, such as raw meat, dairy, or leafy greens in
food processing, may require stricter RLU limits to ensure
that cleaning interventions are effective.
 Lower and Upper RLU Limits for Rev03182013

ATP Monitoring Programs

RLU Limit Worksheet NOTE: To calculate the average of your test results, add all test results together
and divide by the number of tests. The standard deviation is a calculation of the
(Tip: photocopy or print duplicates of this page if you wish to variability of your results, or the amount that results differ from the average. The
standard deviation is best calculated using a spreadsheet application, like Microsoft
record data manually. Save these documents for your records.)
Excel, to minimize calculation errors. If you would like an excel document
formatted to allow for result entry and automatic average and standard deviation
calculations, please contact your Hygiena representative or the customer service
department at 888-HYGIENA (494-4362) or [email protected]

Location
Surface Type
Test # Date Tested
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Average
St Dev (optional)
Lower Threshold
Upper Threshold

Location
Surface Type
Test # Date Tested
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10
Average
St Dev (optional)
Lower Threshold
Upper Threshold