A Guide to Aseptic

Manufacturing:
What to Consider
Innovate | Aseptic Manufacturing Guide

TABLE OF CONTENTS

3 | Overview

4 | Advantages, Disadvantages
and Challenges

7 | Best Packaging

9 | Plant
Layout
& Equipment

10 | Processing Options

12 | Finished Product

13 | Beyond Food Packaging:

The Innovate System

17 | Beyond Food Packaging:

Microbial Considerations

Beyond Food Packaging:

18 | Environmental Monitoring
Solutions

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Innovate | Aseptic Manufacturing Guide

Overview

Aseptic manufacturing is a complex process; understanding everything involved in the pro-

cess is essential to providing the best finished product possible. Not only does it involve
packaging decisions, plant and equipment selection, and layout and processing equipment,
it requires a clear understanding of what type of processing is best suited for the final prod-
uct. Options such as UHT vs ESL play a major role but final product testing is of critical
importance to ensure final product is safe for consumption. Implementing rapid tests for
final product release can assist in this process. This guide discusses aseptic manufacturing
options, challenges, and testing solutions to overcome these challenges.

What is aseptic manufacturing?

Aseptic manufacturing is the method used to commercially sterilize a product and fill and
seal it into a sterile container under aseptic conditions. The final filled package is hermetically
sealed to produce a final product that is impermeable to external contamination or spoilage
organisms. Therefore, it can be stored at ambient conditions for extended periods of time. In
addition, thermal processing of the food product takes place outside the packaging, allowing
it to be rapid and controlled. This is followed by cooling, before packaging. This method al-
lows the product to maintain its nutritional and sensory qualities (i.e., smell and taste).

Raw material
Aseptic Zone

Continuous heating Hold tube Continuous cooling

Package material Aseptic filler Hold tank

Finished product

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Innovate | Aseptic Manufacturing Guide

Advantages of Aseptic Manufacturing

Precision Processes
• Automated controlled filling provides reliable weight consistency
• Automatic tamper-proof sterilization assures quality confidence
• Integrated workflows offer fill-to-seal sterility

Environmental Optimization
• Steam sterilization eliminates chemical exposure and improves safety profile
• Lower transportation weight and storage needs reduces greenhouse
emissions and logistics costs
• Elimination of refrigeration reduces energy use and expense

Food Safety & Nutrition

• Maximum nutrient content provides for more healthful foods
• Enhanced palatability creates greater appeal and higher consumption
• Extended shelf stability allows for longer dating and maximum safety

Consumer Satisfaction
• Natural mouth feel offers an enhanced consumer experience
• Bacterial and contaminant elimination delivers optimal consumer safety
• Dramatically reduced waste allows for good environmental stewardship

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Innovate | Aseptic Manufacturing Guide

Disadvantages of Aseptic Manufacturing

Complexity
• Extensive process validations impact workflow and add cost
• Skilled operators require expensive, time-consuming technical training
• Aseptic workflow requirements may impact existing product/brand
quality and characteristics

Cleaning
• Clean room operations increase total costs
• Complex clean-in-place procedures introduce additional cost and
contamination risk
• Specialized training increases cost and impacts productivity

Cost
• Clean-in-place materials and solutions increase expenses
• Premium aseptic processing and packaging decrease profit
• A more skilled workforce commands higher wages

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Innovate | Aseptic Manufacturing Guide

Considerations Summary

1. Complex operations
• Difficult to validate, clean and operate processing equipment.
• Aseptic system validations takes the longest amount of time among shelf-life stable
sterilization technologies.
• Changeover to a new product involves time-consuming clean-in-place (CIP) processes.
2. Requires highly technical operational training
• Operators and maintenance workers in a clean room environment must understand
intricate control systems by undergoing detailed operator training.
3. Challenges in cleaning processing systems
• CIP systems pose challenges with piping, proper cleaner selection/concentration, and
the thermal and mechanical forces used within the piping system to ensure maximum
cleaning without damage, along with optimizing temperature and contact time.
• Each design/application will have a unique CIP procedure, adding complexity.
4. Cost of cleaning in place procedures
• CIP circuit design can be single use with the cleaning solution discarded after CIP cycle
completion or cleaning solutions can be reused/recovered for additional use.
• Single use systems are the easiest to design and control – all cleaning, sanitizing, and
rinse solutions are used only once, but prove to be costly.
• CIP processes require cleaning verification and validation.
5. Expensive aseptic processing and packaging equipment
• Equipment for aseptic processing and packaging is typically more expensive than
conventional technologies.
• Processors must build a “clean room” to house the packaging operation.
• Containers used for packaging must be hermetically sealed under a sterile environment.

Simplified diagram of an aseptic processing and packaging system

Raw product Bulk heating Bulk holding (T,t)

preparation

Packing material
decomtamination

Final product Aseptic filling Bulk cooling

and packaging

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Innovate | Aseptic Manufacturing Guide

Choosing the Best Packaging Technique

The biggest challenges for aseptic processing are the limitations and disadvantages encoun-
tered during actual application of this technology.
To minimize costs, cold aseptic filling has become a common technique with beverage man-
ufacturers for products including UHT milk, fruit juices and sports/energy drinks. Its flexibility
means that it is suitable for a wide range of products from fresh fruit dices and purees to mar-
inades and dairy products. This diversity has also lead to the development of a wide range
of fillers and packaging types.
Nevertheless, aseptic processing and filling has specific requirements as it combines several
elements into a single, integrated production line. This ensures microbiological safety throughout
the process from initial product treatment to sealing of the final product. Considerations include:
1. Thermal treatment
2. Cooling (where necessary)
3. Sterilization of the packaging
4. Filling and sealing under aseptic conditions

Packaging type
Typically, based on the final product, you must first determine the type of aseptic packaging
you want. Options include can systems, bottle systems, sachet and pouch systems, cup sys-
tems, carton systems, and bulk packaging systems. The packaging must not only protect the
product, but it must maintain the quality of the products. Factors to consider when selecting
the packaging material include seal strength and integrity, package shape, stiffness and du-
rability, as well as barrier properties. Product containers should be sufficiently durable to with-
stand the mechanical, chemical, and thermal stresses encountered during normal distribution.
For all these reasons, it is common to use aseptic packaging which incorporates multiple ma-
terials that are assembled either by lamination or co-extrusion processes to maximize integrity.
Common packaging generally includes barriers to light, moisture, and oxygen. In addition, the
sealant material used must be compatible with the product and the closure systems.

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Innovate | Aseptic Manufacturing Guide

Choosing the Best Packaging Technique

Next, determine what type of packaging line works the best for your product(s).
1. Film and seal
Preformed containers are sterilized, filled in an aseptic environment, and then sealed.
2. Form, film and seal
Sterilizing rolls of material which are then formed into the package in a sterile environ-
ment, followed by filling and sealing.
3. Erect, film and seal
Knocked down blanks are then erected, sterilized, filled and sealed.
4. Thermoform and seal
Roll stock is sterilized, thermoformed, filled, and sealed – all aseptically.
5. Blow, mold, fill and seal
A wide range of possible materials that are sterilized, filled, and sealed.

Choice of packaging must address the following considerations:

1. It should hold the product without leaking, thus maintaining the sterility of the product.
2. It should prevent physical damage to the product.
3. It should run effortlessly on the filling lines.
4. It should survive the packaging process.
5. It should be biologically safe, non-toxic and compatible with the foodstuff.
6. It should be resistant to environmental attack – rodents, insects, dirt, microorganisms, etc.
7. It should be tamper-proof – showing evidence when tampered with.
8. It should offer a barrier to oxygen and gas exchange (i.e., maintain the atmosphere it was packaged under).
9. It should be easy to open and handle.
10. It should communicate product and manufacturer information.
11. It should be cost efficient.
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Innovate | Aseptic Manufacturing Guide

Plant & Equipment Layout

The next consideration is the plant layout and all the equipment needed for manufacturing
and packaging a product aseptically. The facility must be constructed as to minimize oc-
cluded surfaces, be well ventilated using HEPA (High Efficiency Particulate Air) filters and be
easily cleaned, maintained, and decontaminated. The equipment used in this room is also a
consideration. The proper equipment must be selected to ensure the components can be
easily cleaned and maintained, while also remaining sturdy and non-shedding of particulates.

Even with the best equipment selection, another factor is developing the regular maintenance
and qualifying/requalifying regime which will ensure minimal issues as long as equipment is
installed and maintained correctly. Tied to this basic equipment are the additional tools used
for maintenance, extraction or manufacture of the finished product. Since these all originate
from an uncontrolled environment, there is risk of introducing contamination into the produc-
tion area. Therefore, an established, verified decontamination method must be used to clean
all tools before use and properly used to test the aseptic system following maintenance or
repair. Methods must also be routinely re-evaluated for effectiveness and robustness, com-
monly known as re-validation.

An established, verified decontamination method must be used to clean all tools before use.

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Innovate | Aseptic Manufacturing Guide

Processing Options

Thermal processor
The essential elements used for selecting a thermal processor are the heating characteristics
of the food product and the inactivation kinetics of specific target microorganisms which
could be present in the food product. The product is brought to sterilization temperature and
held at that temperature for the time necessary to achieve commercial sterility. Depending on
the process type, different flow rates may dictate the proper time and heat required to ensure
aseptic processing is occurring.

For example, in continuous product flow systems, the time for which the product must be
held at the sterilization temperature is achieved in the hold section or tube – therefore, the flow
rate for the fastest particle or shortest particle retention time must be accurately determined.
Many methods are available to determine the minimum residence time to ensure sterility.
These include dye or salt injections or mathematical models. These models incorporate the
flow rate, physical dimensions and design of the hold section and the flow properties of the
product. When flow characteristics are unknown for the product, then actual measurements
must be made to determine the proper calculations required to determine commercial steril-
ity. Delivery of the energy to the food product must be controlled, monitored and recorded.

Image courtesy of Tetra Pak®

If steam injection or steam infusion is used, the addition of water increases the product vol-
ume by approximately 1% per 10°F above initial product temperature as it enters the sterilizer.
There could also be volume increase due to thermal expansion of the food. This increase in
product volume must be compensated for in the establishment of the process.

No matter what method is used for thermal processing, all must be equipped with accurate,
calibrated, reliable temperature indicating devices and dated records of calibration testing
must be kept. In addition, all devices must be inspected daily to ensure they are working
properly. Sensors for these indicators need to be located in such a way as they do not alter
product flow and in batch systems, multiple indicators must be used to ensure the entire
batch is being heated equally.

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Innovate | Aseptic Manufacturing Guide

Processing Options

Aseptic tank
The aseptic tank is used for intermediate stor-
age of aseptically treated food products. It can
be used in different ways depending on the plant
design and the capacities of various units in the
processing and packaging lines. For instance, it
can be used to store surplus product during any
stoppage, such as when the packaging machines
stop. It can also be used for simultaneous packag-
ing of two products. Volume considerations must
also be taken into consideration. Typically, tanks
are designed to hold a volume sufficient for a full
shift of packaging. Nevertheless, the optimum
tank size and arrangement must be decided for
each individual process and must be compatible
with the aseptic processing equipment and pro-
cesses, and with the packaging machinery.
Image courtesy of Tetra Pak®

Microprocessors
The use of microprocessors and microcomputers
in aseptic packaging has grown significantly over
the years. Microprocessors allow for the control
of multiple process variables simultaneously. They
have the ability to feed film into the packaging ma-
chine, convert the film into the required shape and
dimensions, heat seal all seams, and shrink-wrap
a specific number of packs into a single pack.

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Innovate | Aseptic Manufacturing Guide

Processing Options

Sterilizer for packaging

No matter what packaging material is used, it must be pre-sterilized prior to filling. Since
aseptic packaging systems are complex, care must be taken in choosing the best method
for packaging material sterilization. Typical methods used include steam, hot air, hydrogen
peroxide, UV light, or irradiation. The method chosen cannot damage the material and must
also sterilize any surfaces or equipment that come in contact with the sterilized packaging.

Aseptic filling lines

Following thermal processing, and packaging sterilization, food products are accumulated
in an aseptic surge tank prior to packaging. Surge tanks are sterilized before start-up of
product flow by steam or water and must be vented to remove all air pockets. Sterile air
over-pressure must be maintained to ensure proper operation (flow of product to the fillers).
Sterile gases such as nitrogen or carbon dioxide are used to provide overpressure and create
a sterile barrier.

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Innovate | Aseptic Manufacturing Guide

Finished Products

Storage and transport of finished product

Consideration must also be given to how finished product will be handled after aseptic filling
and sealing. Conditions must preserve the integrity of the container and not impact the safe-
ty and quality of the product. Things to consider include handling during palletizing, shrink
wrapping, temperature before stacking, humidity levels, additional labeling and dryness of
surfaces. Storage conditions must be regulated to prevent deterioration or contamination of
the product or the packaging, including maintaining a constant temperature and minimizing
moisture which can affect packaging surfaces.

Storage conditions must be regulated to maintain a constant temperature.

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Innovate | Aseptic Manufacturing Guide

Above and Beyond Food Packaging

Microbiological specifications
Closely tied to storage and transport are the microbiological specifications set for testing of
the finished food products. The main objectives in aseptic processing here are to control the
number of microorganisms in the food product and to prevent the recontamination of the
food after processing. Therefore, strict microbiological specifications must be set and ad-
hered to. Things to consider are sampling procedures, methodologies for testing, and limits
for acceptance.
Two major sources of microbial contamination are:
1. Heat resistant spores present in the ingredients prior to processing and which survived.
2. Post-process contamination introduced through packaging or a failure in the integrity
of the aseptic filling system.
These are typically controlled by the process design. Nevertheless, commercial sterility test-
ing verifies that the process and controls are effective. Testing would involve incubating final
food product samples in their final packaging at elevated temperatures (30°C, 55°C, the
latter only if thermophilic organisms are a concern) for a range of 7-15 days. The samples are
then examined for signs of microbial growth by plating onto non-selective agar media, incu-
bating for 3-5 days and then examined for visible colonies. A stable product should normally
contain less than 10 colonies on a agar plate.This process is quite tedious and can cause a
bottleneck in product hold and release, taking almost 3 weeks to confirm a negative result.

Rapid Results
96 samples in <30 Minutes

Hygiena Product
Innovate Incubation: Product Release
24-48 hrs*
Method

Traditional
Microbiological Product Incubation: 7-15 days* Plate & Observe: 3-5 Days Product Release
Methods

0 2 4 6 8 10 12 14 16 18 20 22 24
DAYS
*Product incubation time depends on customer’s SOP

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 Innovate | Aseptic Manufacturing Guide

Above and Beyond Food Packaging

One alternative for testing: The Innovate System

An alternative to traditional plate testing is a rapid method utilizing ATP bioluminescence. Us-
ing this technology, Hygiena’s Innovate System reduces testing and product hold time from
10-18 days to 48 hours or less, allowing faster release of product and reducing inventory
costs. This technology is designed to detect light produced from microbial ATP while mini-
mizing background levels of non-microbial ATP and is able to test a large number of samples
at once. The Innovate System provides a microplate format to analyze up to 96 individual
samples in less than 30 minutes.
The system has been validated on a wide variety of raw materials, in-process formulations,
and finished goods. In addition to testing 96 different samples on a single microtiter plate
every 30 minutes, subsequent assays can be prepared while the system is in use to keep
high-volume operations running smoothly.
The Innovate System can be adapted to determine what is “pass” or “fail” criteria, making
it even easier for a factory worker to monitor testing – green is pass and red is fail. Sample
data from multiple instruments can be saved to a single database on the company’s secure
network, helping simplify regulatory compliance. Data can be securely viewed onsite or re-
motely, and reports can be generated for analysis.

Products test negative:

Rapid release
Reduced inventory and cost
to manufacture

Products test positive:

Rapid response
Reduced contamination costs

Learn more about the Innovate System 15

Innovate | Aseptic Manufacturing Guide

Above and Beyond Food Packaging

The Innovate System

Use of the Innovate System helps food processors minimize costs for warehouse space or
holding product while also feeling reassured that any contamination events will be identified
early. In addition, the latest advancement, Innovate Autosampler III, can be paired with the
Innovate platform to prepare up to 2,000 assays per hour for analysis in the Innovate System.
This simplifies the workflow even more and frees up valuable technician time.

Documented Proof of Performance

Customers have put the Innovate System to the test. A large dairy manufacturer partnered
with us to test various types of dairy products from both animal and plant sources for low
levels of contamination (manually introduced for the study). A wide range of organisms were
tested depending on the type of milk product. Microorganisms used included Salmonella,
E. coli, Enterobacter, Pseudomonas, Bacillus, Staphylococcus, Saccharomyces, Aspergillus
and Clostridium.
In all cases, for all microorganisms tested (except for Clostridium sporogenes in ESL oat
milk), the Innovate System was able to detect low spike levels (~10 CFU per pack) at a 24
hour time mark. RLU values in the spiked products exceeded the uncontaminated product
RLU thresholds, set as three times the baseline values. Read the details of each study here:

Oat Milk Soy Milk ESL Milk UHT Milk

Other customers have asked us about testing more highly acidic beverages, such as fruit
juices, tomato juice and protein drinks for the presence of low levels of contamination. In
these cases, all organisms tested were detected within 24-48 hours of incubation. Read the
full report here.
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Innovate | Aseptic Manufacturing Guide

Above and Beyond Food Packaging

Other Microbial Considerations

Additional problems that pose a risk of microbiological contamination include inefficient em-
ployee hygiene practices, ineffective training, the development of biofilms, ineffective use of
cleaners and disinfectants, lack of sanitary equipment design, ineffective application of sani-
tation principles, reactive rather than routine maintenance, contamination of raw materials or
the presence of niche environmental sites. These must be addressed by identifying the po-
tential areas of concern and collecting environmental samples of the areas to test for possible
microbial contamination. This includes monitoring rollers on conveyors, spacing between
close-fitting metal or plastic parts, seams on rubber seals around piping or doors and any
other possible niche. In addition, raw materials must be sourced from reputable suppliers,
stored properly, and involve a kill-step during processing to eliminate any pathogens.
Visual monitoring of these risks can help minimize the presence of these potential sources of
contamination. However, it is not enough. Each processing facility must have an environmen-
tal and food-contact surface testing program in place. It must be designed to detect trends
that indicate a potential loss of sanitation control has occurred so timely corrective actions
can be taken. The program must include sample sites for testing, sampling frequency, and a
quick way to analyze results as soon as they become available.

Collect samples to test for possible microbial contamination.

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Innovate | Aseptic Manufacturing Guide

Above and Beyond Food Packaging

Environmental monitoring solutions

A rapid way to test the environment is to perform simple surface environmental monitoring of
equipment, work areas, floors, and walls. The fastest and simplest way to do this is through
swabbing of high-risk surfaces with swabbing devices that can be quickly read in an envi-
ronmental monitoring device. Hygiena’s EnSURE™ Touch monitoring system works perfect
for this application.
EnSURE Touch works much like a smartphone. Select the proper swabbing device, collect
a sample using the prewetted swab, activate the device and get a reading in 15 seconds.
Swabbing devices such as UltraSnap™ can detect very low levels of ATP (below 1 fmole) –
the level of ATP is correlated to the level of bioorganic load, a direct measurement of potential
contamination. Hygiena also offers SuperSnap™ devices for the detection of extremely low
levels of ATP (down to 0.1 fmole) allowing it to be used as an allergen cross-contamination
prevention tool. If specific allergens are of concern, rapid testing can be performed with our
AlerTox® sticks which provide rapid results in 10 minutes with no cross-reactivity for a num-
ber of allergens from egg to casein to peanut.
Many aseptic processing facilities also have concerns about Listeria contamination in their
food products. Listeria tends to be more prevalent under cooler conditions but still can
become a problem in an aseptic processing facility. Rapidly identifying this organism can
prevent product discards, production delays, and reassure product is safe. BAX® System
Listeria PCR Assays allow for the detection of various Listeria species, including L. mono.
All tests are AOAC-RIPTM approved and provide results within hours after enrichment.
No matter what aseptic processing system you employ, keep in mind that constant monitor-
ing of processes will ensure a final food product that is safe for consumer consumption. Be-
ing proactive is essential to streamlined processes and maximizes product output and profit.

ALLERGEN
DETECTED

ALLERGEN
FREE

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Innovate | Aseptic Manufacturing Guide

Conclusion

Aseptic manufacturing requires multiple considerations

To decide the best option for your finished product, you must first consider the advantag-
es and disadvantages of the processing method– is it the proper method for your finished
product? Next, you must understand packaging options based on how the product will be
used by consumers. Alternative processing options are available as well and pose their own
challenges. Last, you must consider how you will ensure the final product passes all microbial
testing options and how to ensure your manufacturing environment is clean. This will ensure
your product is safe for consumers once it leaves your facility.

Facility
Equipment

Validation
Aseptic Process
Finish
Processing
Product
Testing
Personnel
Control &
Validation

Learn more about product quality testing and

environmental monitoring products at hygiena.com

Learn More

www.hygiena.com INN-EB-4001-REVA 19