A Guide To ATP Hygiene Monitoring
A Guide To ATP Hygiene Monitoring
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TABLE OF CONTENTS
QUICK START GUIDE.............................................................................................................. 4
INTRODUCTION...................................................................................................................... 5
SECTION 1: An Overview of ATP Monitoring Systems .................................................... 7
1.1 What is ATP? ............................................................................................................................... 7
1.2 Measuring ATP with bioluminescence technology .................................................. 7
1.3 Interpreting results on the luminometer ....................................................................... 8
1.4 Other uses for ATP monitoring systems ........................................................................ 9
SECTION 2: Using Your ATP Monitoring System ............................................................ 10
2.1 Proper sampling technique and activation procedure ........................................10
2.1.1 Collecting samples with the testing device .............................................10
2.1.2 Measuring ATP with SystemSURE Plus and EnSURE .........................13
2.1.3 Ensuring correct results with calibration controls ...............................14
2.2 Where to do ATP testing .....................................................................................................15
2.2.1 Establishing your facility’s test locations (control points) ................15
2.2.2 Types of contact areas .....................................................................................15
2.2.3 Diagram of typical production floor control points ............................17
2.2.4 Monitoring individual control points ..........................................................18
2.2.5 Setting up test plans..........................................................................................19
2.3 Determining RLU limits for your facility ......................................................................20
2.3.1 Setting up RLU limits .........................................................................................20
2.3.2 General Pass/Caution/Fail limits .................................................................22
2.3.3 Storing and viewing RLU limit data ............................................................23
2.4 Testing frequency ..................................................................................................................24
2.5 Corrective action procedures ...........................................................................................25
2.6 Continuous improvement...................................................................................................27
SECTION 3: Further Help ................................................................................................... 28
3.1 Glossary ......................................................................................................................................28
3.2 Typical RLU limit guidelines..............................................................................................29
3.3 Contact Hygiena .....................................................................................................................30
Hygiena - Americas ......................................................................................................30
Hygiena International ..................................................................................................30
Hygiena China .................................................................................................................30
Hygiena Online ...............................................................................................................30
Notes ..................................................................................................................................... 31
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QUICK START GUIDE
Proper Sampling Procedure
1. Identify the location to be tested and turn on the luminometer. Select the test location from the
programmed locations. Remove the ATP testing device from the outer tube. If conducting a surface
test, press firmly down on the swab tip and collect a sample from a 10 x 10 cm (4 x 4 in) area. Use a
side-to-side and up-and-down motion while rotating the swab tip.
2. Place the swab back into the swab tube. The ATP testing device is now ready to be activated or
can be left inactive for up to 4 hours. Once activated, the test must be read within 60 seconds.
3. To activate, break the plastic valve at the top of the device by bending the bulb backward and
forward. Squeeze the bulb twice to expel the liquid in the bulb to the bottom of the tube.
4. Bathe the swab bud in the liquid by shaking gently in a side-to-side motion for 5-10 seconds.
5. Place the entire test device into the luminometer and close the lid.
6. Holding the luminometer in a vertical position, press ‘OK’ to initiate reading. The test result
will appear on the screen in 15 seconds.
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INTRODUCTION
Hygiena ATP Monitoring Systems
The following are some of the benefits experienced by companies using ATP
monitoring:
Optimize cleaning chemicals, equipment and labor so that the plant can
maintain a high cleanliness level without an excessive amount of waste
Ensure product quality and avoid recalls which will protect brand image
and reputation
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Components of ATP Monitoring Systems
Hygiena ATP monitoring systems are comprised of three core components:
(1) Luminometer
(2) Test devices
(3) Data tracking software
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SECTION 1: An Overview of ATP Monitoring Systems
ATP is an energy molecule found in all plant, animal and microbial cells. It fuels
metabolic processes such as cellular reproduction, muscle contraction, plant
photosynthesis, respiration in fungi, and fermentation in yeast. All organic
matter (living or once-living) contains ATP, including food, bacteria, mold and
other microorganisms. The detection of ATP on a surface or in water therefore
indicates the presence of biological matter that may not otherwise be visible
to the eye. In industries where plant hygiene control or cleanliness is crucial,
ATP testing is an excellent tool for detecting and measuring biological matter
that should not be present after cleaning.
Hygiena ATP testing devices contain a natural enzyme found in fireflies. This
enzyme, called luciferase, produces a simple bioluminescence (light-
producing) reaction when it comes into contact with ATP. Using
bioluminescence technology, the SystemSURE Plus and EnSURE luminometers
can measure extremely low levels of ATP collected with testing devices.
Measuring the amount of bioluminescence from an ATP reaction provides an
excellent indication of surface cleanliness or water quality because the
quantity of light generated by the
reaction is directly proportional to the
amount of ATP present in the sample.
The bioluminescence reaction is
immediate so results can be processed
at the testing site in seconds. Results are
expressed numerically on the
luminometer screen in Relative Light
Units (RLU).
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1.3 Interpreting results on the luminometer
The relationship between the amount of ATP collected in a sample and the
RLU result displayed on the luminometer is linear, which makes understanding
the technology very easy.
The RLU reading is directly proportional to the amount of ATP collected from
the sample. A high RLU reading indicates a large amount of ATP at the test
location. This in turn indicates improper cleaning and the presence of potential
contaminants.
Cleaning properly results in less ATP at the location. Lower ATP levels produce
smaller amounts of light output during the bioluminescence reaction and
consequently, a lower RLU reading.
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1.4 Other uses for ATP monitoring systems
In addition to routine ATP monitoring, other useful applications for ATP monitoring
systems include:
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SECTION 2: Using Your ATP Monitoring System
Before collecting a sample for testing, the surface should be visibly clean. If
any soiling or residue is apparent, re-clean the area before testing.
2. Collect a sample using the guidelines below. The test device is designed to
detect trace amounts of contamination. Collecting a sample on a visibly
dirty surface may interfere with the bioluminescence reaction and
produce an inaccurate test result.
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Incorrect swabbing technique:
Touching the swab shaft with
your finger.
X
Lightly touching the swab to the
sample area.
Collecting sample on only one
side of swab tip.
Swabbing an inadequate
surface area.
✔
No contact with the swab shaft.
Sufficient pressure to create
flex in the swab shaft. This
helps to break through any
biofilm.
Rotate the swab to collect
sample on all sides of swab tip.
Swabbing a 10x10 cm (4x4 in)
area (where possible).
a) Regular surfaces
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b) Irregular surfaces
3. Re-insert the swab into the tube. The test device is now
ready to be activated. A test with an active sample on it
can be left inactivated for up to 4 hours in this state. In
some facilities, users prefer to sample each location,
write the sample location on the swab tube, and run all
tests in a laboratory rather than at the test location. The
most common process is to activate and read the test
immediately after collecting the sample.
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5. Gently shake the device with a side-to-side motion for 5-
10 seconds, bathing the swab bud in the liquid-stable
reagent. The test is now activated and the
bioluminescence reaction is taking place. For optimal
results, the test should be run on the luminometer as
soon as possible, and within 60 seconds of activation.
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2.1.3 Ensuring correct results with calibration controls
To help ensure the accuracy of test results, the luminometer automatically self-
calibrates every time the instrument is turned on. In some facilities, a quality or risk
assessment program such as ISO or HACCP might require additional calibration
checks of the system. Hygiena offers two calibration kits that are recommended for
periodic use with your system: the Calibration Control Rod Kit for testing the
luminometer and the Positive Control Kit for testing ATP test devices.
Each kit contains a positive rod and negative rod. The positive rod
emits a very low level of constant light output that can be measured in
RLUs to verify proper calibration of the unit. The negative rod produces
zero (0) RLU and is used to check that background light is not entering
the instrument, while ensuring that the light detector is calibrating
correctly. The Calibration Control Rod Kit is good for five years of
repeated use.
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2.2 Where to do ATP testing
Test areas within your plant should be designated as “ATP control points” in
your ATP hygiene monitoring plan. By monitoring these control points you will
have reliable, real-time feedback on the cleanliness of a particular piece of
equipment or areas being tested. It’s important that ATP testing be routinely
performed on all important control points. This will ensure product quality,
identify issues immediately, and allow valuable trending data to be used to
improve plant hygiene.
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different places of to verify total cleanliness. One test
device could be used on multiple spots. If the test fails,
then re-cleaning of the entire piece of equipment is
required.
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2.2.3 Diagram of typical production floor control points
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2.2.4 Monitoring individual control points
Once control points have been identified, this information should be entered into
SureTrend software. In SureTrend software, locations may be identified by assigning
an alphanumeric name, group category, surface type, and additional notes. Group
and surface information is only displayed in the software and not displayed on the
handset. Locations may be assigned to test plans according to when they will be
tested or which locations should be tested together.
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2.2.5 Setting up test plans
Once locations and limits have been input into SureTrend software, test plans may
then be set up. See the SureTrend Users Manual for steps on setting up test plans.
Test plans are groups of locations that are tested one after each other, grouped
together, or tested on a specific day.
Here are some examples of test plans using the production floor on page 17:
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2.3 Determining RLU limits for your facility
Setting correct Pass, Caution and Fail levels is a fundamental part of running a
successful ATP monitoring program. RLU limits may vary depending on the
type of product being manufactured as well as the surface being checked.
However, the formulas used to determine Pass, Caution and Fail levels are
always the same.
Hygiena also provides general recommended RLU limits, which meet most
manufacturers’ hygiene standards. Examples of some typical limit guidelines
specific to the food and beverage industry can be found in Typical RLU limit
guidelines, in Section 3.2.
1. Clean product surfaces to the level that daily cleaning procedures should
achieve.
2. Conduct an ATP test at each location. Take 5-10 replicate tests at each
cleaned location, being sure not to swab the same exact surface area
more than once.
Correct Incorrect
All replicate tests can be done on the cleaned surface or equipment after
one cleaning or over the course of several days.
3. To determine the Pass limit: Calculate the average RLU for each location
based on the 5-10 test results. To do this, add all test results, and then
divide the sum by the number of tests taken. The resulting number is your
average RLU. This number is your Pass limit.
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4. To determine the Fail limit:
The resulting number is your Fail limit. If the Fail limit is zero (0), it is
acceptable to set the Pass limit to the system default of 10 RLU.
Occasionally, blank ATP test devices may emit up to 2 RLU.
5. To determine the Caution* limit: The area between the Pass and Fail
limits is the Caution range.
*NOTE: The Caution range is sometimes useful for trend analysis and eliminating an excessive
amount of re-cleans when an ATP program is implemented. Caution results can be viewed as
a warning and more attention should be given to this location the next time cleaning is done.
Often, a caution one day will be a Pass the next day. However, users may opt to forgo the
Caution range and set the Fail limit to the same RLU as Pass limit. Any result over the Pass
limit is then considered a Fail result.
The following table uses example data to illustrate average RLU and Pass/
Caution/Fail limits using the multiplication method:
Average
Test 1 2 3 4 5 6 7 8 9 10
RLU
Routine
10 15 8 19 10 13 17 14 15 11 13.2
cleaning
The data in this example would then yield the following RLU limits:
0-13 14 - 35 36+
If a user chose to eliminate the Caution zone, RLU limits for Pass and Fail
would both be set to 13 to yield the following RLU limits:
Pass Fail
0-13 14+
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NOTE: Different surfaces have different levels of risk, and therefore may
require different RLU limits. For example, porous plastic or rubber surfaces
may be more difficult to clean than stainless steel surfaces, and therefore
produce higher ATP test results. In this case, the user may choose to: 1) set
RLU limits higher for those harder-to-clean areas; or 2) intensify cleaning to
bring ATP test results on those surfaces in line with other control points.
For facilities that opt not to set their own RLU limits, Hygiena offers general
guidelines. These are common limits used for ATP hygiene monitoring, and are
based on plate count and ATP correlation studies.
SystemSURE Plus 10 11 – 29 30
EnSURE 20 21 – 59 60
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2.3.3 Storing and viewing RLU limit data
Once RLU limits are established, they can be programmed into SureTrend
software and then uploaded from the software to the luminometer. For
information on how to do this, refer to the SureTrend User’s Manual. RLU limits
can also be manually entered into the luminometer (consult the Luminometer
Operator’s Manual for details).
(Pass) - For any RLU reading that is less than or equal to the
Pass limit. A Pass result indicates the surface is clean.
(Caution) - For any RLU reading that is greater than the Pass
limit and less than the Fail limit. A Caution result
indicates the surface may not have been adequately
cleaned.
(Fail) - For any RLU reading that is greater than the Fail limit.
A Fail result indicates the surface is dirty or
contaminated.
For information on what steps should be taken after obtaining Pass, Caution
or Fail results, see Corrective action procedures in Section 2.5.
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2.4 Testing frequency
Once control points have been established and ATP RLU limits have been set,
a sampling frequency plan should be developed.
Sampling frequency should be based on the level of risk associated with the
control point being tested. Factors determining the level of risk include:
Critical (high-risk) control points should be tested on a daily basis, after each
cleaning. You may also choose to test after product line changes, after shift
changes, and or any time prior to start-up. Lower-risk control points may not
need to be tested as frequently.
Testing should be done after cleaning a surface, but prior to the application of
a sanitizer if possible. Product residue left on the surface after cleaning inhibits
sanitizers from working correctly. Sanitizer is most effective when surfaces are
free of all residues. Following this Clean-Test-Sanitize procedure prevents
wasting sanitizer and time it takes to re-apply sanitizer.
NOTE: Some sanitation procedures may require ATP testing to be done after
sanitizing, because of equipment turnover time. In such cases, follow
sanitizer’s proper dwell time and concentration levels before ATP testing.
Commonly used sanitizers at working strength should not affect Hygiena’s
ATP tests. Some acid-based sanitizers at high concentrations could slightly
reduce output signal. For a list of sanitizers that could affect the ATP
bioluminescence reaction, contact Hygiena.
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2.5 Corrective action procedures
According to the needs of the facility, the user has the option of setting up a
more extensive corrective action plan. For example, users may decide that
control points which produce Fail results should be re-tested until 3
consecutive days of Pass results are achieved. If the control point does not
successfully achieve 3 consecutive days of Pass results, cleaning procedures,
personnel, or RLU limits should be re-evaluated.
A flow chart on the next page illustrates general recommended cleaning and
corrective action procedures.
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Flow Chart: Suggested Cleaning, Test and Correction Action Procedures
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2.6 Continuous improvement
Hygiena ATP monitoring systems are designed to aid organizations striving for
continuous improvement of standards. Continuous improvement ensures
excellence in product quality, while reducing inefficiencies, avoiding recalls,
and showing due diligence and compliance to auditors and customers.
If trends show high numbers of Caution and Fail results, standard sanitation
operating procedures (SSOP’s) should be reviewed for ways to improve
protocols. If low numbers of Caution and Fail results are obtained, Pass/Fail
limits could be reviewed and potentially lowered, creating a higher cleaning
standard which would create a cleaner facility.
Pass, Caution, and Fail limits should also be re-evaluated every year and
whenever procedural or equipment changes are made. As cleaning
procedures become more efficient and effective, limits should be adjusted to
ensure your facility is operating to its maximum potential.
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SECTION 3: Further Help
3.1 Glossary
Bioluminescence
Bioluminescence is a chemical reaction that produces light when ATP comes into
contact with the enzyme luciferase. Hygiena’s ATP testing devices use
bioluminescence technology (combining ATP with luciferase) to produce a light
output that can be detected and measured by the luminometer.
Biofilm
Biofilm occurs when microorganisms work together as a population and secrete a
sticky polymer to form a solid matrix attached to a surface. Once a biofilm is
established it is very difficult to eliminate. Biofilm can cause poor product quality
and/or lost product due to contamination.
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3.2 Typical RLU limit guidelines
The following guidelines are typical of those found in food and beverage
manufacturing. For recommended RLU limits in other environments, such as
food service, hospitality, restaurants, and food manufacturing, contact
Hygiena.
Dairy
Stainless Steel <5 6–7 >8 <10 11-15 >16
Products
Juice
Stainless Steel < 10 11 – 29 > 30 <20 21-59 >60
products
Water
Stainless Steel <5 6–9 > 10 <10 11-19 >20
bottling
Brewing
Stainless Steel < 15 16 – 29 > 30 <30 31-59 >60
equipment
CIP Rinse
Stainless Steel <5 N/A >5 <10 N/A >10
Water
Raw meat
Slaughter Porous Plastic < 100 101- 199 > 200 < 200 201- 399 >400
Butchery < 50 51-99 > 100 < 100 101-199 >200
Cooked meat Stainless Steel < 25 26-49 > 50 < 50 51-99 >100
Shellfish Stainless Steel < 100 101 - 199 > 200 < 200 201 - 399 >400
Cheese
Stainless Steel <5 6-9 >10 < 10 11 - 19 >20
processing
General food
Stainless Steel < 10 11-29 > 30 < 20 21-59 >60
processors
Vegetable Porous Plastic < 100 101 – 149 > 150 < 200 201 – 299 >300
Cooked
Stainless Steel <10 11-29 >30 < 20 21-59 >60
products
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3.3 Contact Hygiena
For any questions about ATP monitoring systems, or more information about
Hygiena products, please contact Hygiena at:
Hygiena - Americas
941 Avenida Acaso
Camarillo, CA
USA 93012
Tel: +1-805-388-8007 x300
Fax: +1-805-388-5531
info@[Link]
Hygiena International
Unit 11 WENTA Business Centre
Colne Way, Watford, Hertfordshire
WD24 7 ND. UK
Tel: +44 (0)1923 818821
Fax: +44 (0)1923 818825
enquiries@[Link]
Hygiena China
Neiwailain Building Suite 21A3
No. 518 Shangcheng Road
Pudong New District, Shanghai, China
Tel: +86-21-51321081
Fax: +86-21-51321081
enquiries@[Link]
Hygiena Online
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Notes
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Hygiena - Americas Hygiena International Hygiena China
941 Avenida Acaso Unit 11 WENTA Business Centre Neiwailain Building Suite 21A3
Camarillo, CA Colne Way, Watford, Hertfordshire No. 518 Shangcheng Road
USA 93012 WD24 7 ND. UK Pudong New District, Shanghai, China
Tel: +1-805-388-8007 x300 Tel: +44 (0)1923 818821 Tel: +86-21-51321081
Fax: +1-805-388-5531 Fax: +44 (0)1923 818825 Fax: +86-21-51321081
info@[Link] enquiries@[Link] enquiries@[Link]
[Link]
REVB042013
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