Application Note

Impact of the new EN 17141 on the ISO

14698 Validation of MAS-100® Air Samplers
General Requirements and Validation

The scope of the new norm is to establish the The basis for an efficient microbial air sampler is a good
requirements, recommendations and methodology physical and biological efficacy:
for microbiological contamination control in clean
"The physical collection efficiency is the cut-off size
controlled environments. The document is limited to
(d50 value) which defines the aerodynamic equivalent
viable microbial contamination and refers to ISO 14644
particle diameter size at which the sampler collects 50
about non-viable contamination. ISO 14698 is valid
% of the particles in the air [12] [13]. The average
throughout the rest of the world while EN 17141 2020
equivalent diameters of microbe-carrying particles
will replace it in the European Union.
(MCPs) that form the cfu are generally larger than 1
This application note will evaluate if the new EN µm and a d50 value smaller than 2 µm is considered
17141 norm will affect the validation requirements of appropriate. The d50 value can be calculated for
volumetric air samplers described in ISO 14698. impaction samplers with multiple holes or those using
rectangular slits. For active air samplers based on
Comparison of EN 17141 and ISO 14698 impingement and or cyclonic operational principles,
no d50 value can be calculated. For all active
requirements on microbial air samplers
microbiological air samplers, the effects of impact
stress and the effect of the media dehydration during
ISO 14698 requirements for microbial air samplers are
the sampling period are further considerations.
more detailed, but generally match the requirements of
EN 17141. The physical collection efficiency is influenced by
both inlet or extraction efficiency and by separation
The requirements for validation are newly defined in the
efficiency. Inlet or extraction efficiency is a function
EN 17141. The supplier of the sampler shall demonstrate
of the inlet design of the sampler and its ability to
the collection efficiency of the sampler, the sampling
collect particles from the air in a representative way
techniques shall be validated, and the volumetric air flow
and transport the particles to the impaction nozzle
of the sampler shall be periodically calibrated.
or the filter. Separation efficiency is the ability of the
sampling device to separate and collect particles of
different sizes from the air stream by impaction onto
the collection medium or into the filter medium. The
physical collection efficiency is based on the physical
characteristics of the sampling device such as airflow,
orifice shape, orifice size and the number of orifices."

The life science business of Merck operates as

MilliporeSigma in the U.S. and Canada.
The following table will compare the requirements for sampling devices, especially volumetric microbial air
samplers as indicated in EN 17141 and ISO 14698-1.

Table 1. Comparison of requirements for sampling devices and microbial air samplers in EN 17141 and
ISO 14698:
Requirements EN 17141 Corresponding Requirements out of ISO 14698
Accessibility into the clean controlled environment for the sampling device • Ease of cleaning and disinfection or sterilization
• Ease of handling (weight, size) and operation (ease of
use, auxiliary equipment, dependence on vacuum pumps,
water electricity, etc.)
Effect of the sampling device on the process or environment to be monitored • Disturbance of unidirectional air flow by sampling
apparatus
• The exhaust air from the sampling apparatus should not
contaminate the environment being sampled or be re-
aspirated by the device
• Possible intrinsic addition of viable particles to the
biocontamination to be measured
Efficiency and precision of the sampling method • Collection accuracy and efficacy
.. a d50 value smaller than 2 µm is considered appropriate … / …For all active • Appropriate suction flow rate for low levels of viable
microbiological air samplers, the effects of impact stress and the effect of the airborne particles
media dehydration during the sampling period are further considerations • Appropriate impact/airflow velocity
• Sensitivity of the viable particle to the sampling procedure

Requirements and Methods for the

Validation of Microbial Air Samplers
The method for a complex validation of microbial air As an alternative, a simpler method is added to the
samplers is described in both norms with no significant complex method in order to allow on-site validation. This
differences. According to EN 17141 this complex method is a side-by-side comparison against an already
method should be preferably performed by an external qualified air sampler that has been validated to the
competent body and the supplier of the air sampler more complex biological and physical efficiency method.
shall demonstrate the collection efficiency of the The simplified test should be performed in minimum at
microbial air sampler. A re-validation of a qualified two different locations with a contamination level of in
instrument is not required. minimum 80 cfu/m3. he chosen sampling volume should
result in a count of 80 to 150 cfu per plate. A previously
The method for validation of collection efficiency is qualified instrument or membrane filtration method may
separated into the validation of the physical efficiency be used as a reference.
and the biological efficiency. The physical efficiency
demonstrates the ability of the air sampler to collect If using a qualified instrument, which is validated by
various particle sizes down small particles sizes, such the supplier according to the described method in
as 1 µm. The d50 value indicates the particles size Annex E of 17141, an on-site simplified validation as
at which 50% of the airborne particles are impacted well as a re-qualification is not required.
onto the agar surface by the microbial air sampler. The
biological efficiency is the ability of the air sampler to
safely capture organisms from the air.
A new requirement according to EN 17141 is a defined
d50 value of smaller than 2 µm, which was not
specified before in the ISO 14698-1.

2
Validation Results for MAS-100® Microbial Air Samplers
Every variance of air flow, such as flow rates or configuration of the sampling head (e.g. numbers and size of holes of
the sieve plate) might influence the physical and biological efficiency of microbial air samplers. Therefore, we validate
various instruments of the MAS-100® air sampler family at a competent external body (PHE Biosafety group in Porton
Down), England individually. The qualified instruments are indicated in figure 1.

2.
1.

4.
3.

Figure 1. Instruments validated according ISO 14698 and consequently to EN 17141 by an external competent body

1.1. MAS-100 NT® (instrument configuration identical to 1.3. MAS-100 Iso NT®
MAS-100 NT® with filter and MAS-100 NT Ex® with or 1.4. MAS-100 Iso MH® (2 separate validations for either 1 m or 9
without filter) m tube length between sampling head and control unit)
1.2. MAS-100 VF®

The validation of the physical efficiency takes place in better than the calculated values. The calculated d50
test room of 20 m3 with turbulent air. The test sampler of the tested microbial air samplers is 1.1 µm at a flow
and reference membrane filtration device are positioned rate of 100 liter/min and using a sieve plate with 300 x
in the same distance and height from the spinning 0.6 mm holes.
top aerosol generator. Washed spores of Bacillus
The biological efficiency is validated against a Casella
atrophaeus NCTC 10073 are mixed with different
Slit Sampler and the results are indicated in figure 3.
concentrations of potassium iodide in order to achieve
The biological efficiency is calculated by the recovery
the various particles sizes. The spores are distributed
of the ratio of cfu of a mixture of Bacillus atrophaeus
homogeneously using spinning top aerosol generator
NCTC 10073 spores and Staphylococcus epidermidis
to the sampling devices. As culture medium TSA w.
NCTC 11047.
LTHTh - ICRplus (reference number 1466830020) plates
are used. After sampling, the filter is transferred on Biological efficiency = Ratio of SE/BA sampled by test
to culture medium plates and all plates are incubated. device/Ratio of SE/BA sampled by Casella Slit sampler x100
Afterwards the CFU are counted and the recovery rate
for each particles size is calculated. The biological efficiciency of the MAS-100® air samplers is
indicated in figure 3. For all tested MAS-100® microbial air
The test results of the validation of physical efficiency samplers, the biological efficiency is above 70%.
of the instruments show a d50 value of less than 1 µm
for all test samplers (see figure 2). The test results The full test reports can be reviewed during an audit.
are comparable for all tested instruments and are

3
Figure 2. Test results of physical efficiency for various MAS-100® Figure 3. Test results of biological efficiency of various MAS-100®
microbial air samplers microbial air samplers

Conclusion References
• EN 17141 (2020): Cleanrooms and associated controlled
The requirements for volumetric microbial air environments - Biocontamination control
samplers do not differ significantly between the • ISO 14698-1 (2003): Cleanrooms and associated controlled
environments - Biocontamination control - Part 1: General principles
EN 17141 and ISO 14698-1. They should provide and methods
accurate and reliable results. • Physical and Biological Efficiency Testing of MAS-100 VF® - Techniques
described in ISO 14698-1: Report No. 14/013 A (2015) - (PHE Biosafety
The MAS-100 NT, MAS-100 VF, MAS-100 Iso NT ®
group in Porton Down)
and the MAS-100 Iso MH® have been validated • Physical and Biological Efficiency Testing of MAS-100 Iso MH® - 1m -
according to the requirements of ISO 14698-1 Techniques described in ISO 14698-1: Report No. 14/013 B (2015)
- (PHE Biosafety group in Porton Down)
and consequently to EN 17141 by an external
• Physical and Biological Efficiency Testing of MAS-100 Iso MH® - 9m -
competent body. Techniques described in ISO 14698-1: Report No. 14/013 C (2015)
- (PHE Biosafety group in Porton Down)
The physical efficiency is comparable for all tested
• Physical and Biological Efficiency Testing of MAS-100 Iso NT® -
instruments and provide a d50 value of less than Techniques described in ISO 14698-1: Report No. 14/013 D (2015)
1 µm. The biological efficiency is above 70% - (PHE Biosafety group in Porton Down)
compared to a Casella Slit Sampler. • Physical and Biological Efficiency Testing of MAS-100 NT® - Techniques
described in ISO 14698-1: Report No. 14/013 E (2015) - (PHE Biosafety
According to the validation results a re- group in Porton Down)
qualification of these instruments is not required.

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 Application Note

VHP Decontamination of MAS-100 NT®

Microbial Air Sampler
Hydrogen peroxide decontamination

Active microbial air sampling is frequently required for

cleanrooms and associated controlled environments.
Microbial contamination by the air sampler must be
avoided. Thus, they must be decontaminated. The
MAS-100 NT/NT Ex viable air sampler is specified for
use in ISO 5/GMP Grade A environments. Externally
accessible parts are designed for easy cleanability and
spray/wiping disinfection. Independent data published
by Sandle and Satyada (Sandle und Satyada 2015)
have proven effectiveness of external decontamination
of the MAS-100 NT using 70% isopropanol.
Vaporized hydrogen peroxide (VHP = vaporized
H2O2) is routinely used in critical environments to
bio-decontaminate surfaces and equipment. Here
we demonstrate that MAS-100 NT® (Figure 1) can
be decontaminated using VHP without effect on
instrument functionality.

Effects of exposure to VHP

MBV AG together with the isolator manufacturer SKAN
have performed a detailed evaluation of the portable Figure 1: MAS-100 NT® Microbial Air Sampler
microbial air sampler MAS-100 NT® regarding the
decontamination with VHP. The main functional quality
criterium of an impaction-based viable air sampler
such as the MAS-100 family instruments is its ability to the differential pressure-based mass-flow sensor
to aspirate a target air volume at a given air flow and integrated into the MAS-100 NT®.
impact the air with a constant speed onto an agar The goal was to simulate 10 years regular
filled Petri dish. The physical and biological recovery decontamination of a MAS-100 NT® microbial air
rate of an air sampler depends on this capability. Key sampler using VHP and test for air flow stability
to correct performance is a functioning airflow sensor over time. If such airflow calibrations remain within
which controls the air blower. Consequently, the best the instrument specification, then performance
indicator of performance under stress is testing airflow remains unaffected.
stability using an alternative measurement method

The life science business of Merck

operates as MilliporeSigma in the
U.S. and Canada.
Procedure for VHP resistance test

Equipment Procedure
Testing environment: Pharmaceutical Safety Isolator The MAS-100 NT instruments were positioned close to
from SKAN AG. 840 ppm H2O2, controlled by Dräger each other inside the PSI isolator. The perforated lids
sensor for high concentration (HC) hydrogen peroxide. were removed and placed next to the instrument.
Instruments tested: 2x MAS-100 NT The instruments were left turned off while exposed
to H2O2.
Airflow: 100 Standard liters per min (SLPM) ± 2.5%
The test chamber was filled with 840 ppm hydrogen
Instrument 1: SNR: 108400, usage time: 1971 h
peroxide. The chamber was vented daily, and the air
Instrument 2: SNR 108302, usage time: 1924 h samplers were checked with the DA-100 NT® digital
anemometer. The calibration (but no adjustment) was
Calibration instrument: DA-100 NT® digital vane performed automatically, and the resulting calibration
wheel anemometer (±1% reproducibility, temperature protocol was generated and saved.
and pressure compensated). MBV PC – based software
used to perform automated airflow calibration. This procedure was repeated for 12 days to simulate
approx. 10 years of weekly H2O2 decontamination
Reagents: 50% hydrogen peroxide (Solvay) (see Figure 2):
12 d * 24 h/d = approximately 288 h of VHP exposure
10 y * 50 weeks/y * 1 decontamination cycle/week =
500 decontamination cycles
288 h/500 decontamination cycles = 0.58 h/average
dwell time

Dwell time

840

Gassing stage
H2O2 conc. [ppm]

Venting stage

1 2 3 11 12
Time [days]

Initial Final
Calibration 2 Calibration 3
calibration calibration

Figure 2: Schematic of VHP exposure, calibration and decontamination cycle. The time required for the gassing and
venting stages are exaggerated in comparison to the dwell time.

2
 Proven resistance to H2O2 exposure The electronic components were visibly affected but
not damaged. To protect flow sensor and electronics
The results of the daily airflow calibration of the two H2O2 decontamination must be performed with the
MAS-100 NT® are presented in Figure 3. Throughout blower inactive.
the 12 days exposure of the air samplers to H2O2 the
airflow remained within the specification of ±2.5% Decolorization of the blue anodized aluminum parts
of the target air flow 100 SLPM. As the pneumatic (perforated lid 300 x 0.6 and rings) was apparent
performance of the air sampler during the simulated (Figure 4). The decolorization is only a cosmetic
time frame of 10 years remained unaffected, we degradation and has no effect on the functioning of the
concluded that the instrument can withstand at least air sampler.
the simulated 500 calibration cycles.

105. 0
Airflow [% of initial

Airflow specification
adjustment airlow]

102. 5

of MAS-100 NT®
100. 0

97.5

95.0
1 2 3 4 5 6 7 8 9 10 11 12

Time [days]
Instrument 1 Instrument 2 Target (100 SLPM)

Figure 3: Airflow calibration of two MAS-100 NT exposed to H2O2 over

12 days Figure 4: Blue ring before and after long-term exposure to vaporized
hydrogen peroxide. The decolorization has no influence on function and
Apart from testing the airflow calibration we may also occur when an instrument is frequently cleaned or disinfected
separately tested individual electronic and mechanical with solvents or detergents

components. These were placed inside the same test Note: The MAS-100 NT Ex® is the explosion proofed version of the
MAS-100 NT® and the results described above can be extended to
chamber and assessed optically after 12 days exposure. that product

Restrictions and recommendations

The results and conclusions of this technical note Alternatively, the perforated lid and the dust cover can
are only applicable to decontamination with VHP. No be dry-decontaminated at 180 °C for 60 minutes.
conclusions for the use of other decontamination agents
In case of dirt / blocked holes of the perforated lid:
can be made.
Clean them with a needle, or by using an ultrasonic
It is mandatory, that throughout the whole VHP bath. Do not wash aluminum parts in dishwashers.
decontamination process the instrument must be
switched off and the blower must remain inactive. References
1. Sandle, Tim, and Ravikrishna Satyada. "Assessment of the
It is not permitted to decontaminate the instrument or disinfection of impaction air sampler heads using 70% IPA, as
perforated lid with a liquid medium and then dry the part of cleanroom environmental monitoring." European Journal of
Parenteral & Pharmaceutical Sciences. Vol. 20. no. 3. 2015.
perforated lid by running the blower.
2. SKAN AG. Modular Working Isolator PSI-M. 04 12, 2019. https://
The blue anodized parts may experience decolorization skan.ch/en/products-view/44-modular-working-isolator-psi-m
due to VHP exposure. (accessed 4 12, 2019).

The perforated lid shall be unmounted during the VHP

decontamination. The perforated lid and the dust cover
can be autoclaved for 20 minutes at 121 °C.

3
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Acknowledgement
Many thanks to our partner MBV providing content and graphics
MBV AG
Industriestrasse 9
8712 Stäfa
Switzerland
www.mbv.ch

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64293 Darmstadt, Germany

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 Application Note

How to use MAS-100® Air Samplers with

BioTrends EM® System
Introduction
Environmental monitoring plays an important role in the contamination control strategy of pharmaceutical
manufacturing lines. The Environmental monitoring program consists of several main elements including viable
air, surface, and personnel monitoring, but also monitoring of total particles, temperature, relative humidity, and
Aseptic Process Simulation (APS). The data are used for routine trending. These trends should include:
• Increasing numbers of excursions
• Consecutive excursions from alert levels
• Regular but isolated excursion from action limits that may have a common cause as well as changes in
microbial flora type
• Numbers and predominance of specific organisms
Based on the huge number of samples tested, digitalization of tracking and managing data is helpful.
The MAS-100® air sampler family, irradiated ICR settle and contact plates, and ICR swabs offer a complete
portfolio to perform all viable environmental monitoring. All of which are suitable for cleanrooms as well
as isolators.
Archilex offers the BioTrends EM® software system developed for managing microbiological data of environmental
monitoring programs. The system is compatible with mobile devices having barcode readers that support
operators and/or analysts, speeding up and facilitating all the previously mentioned operations. The software is
compliant with FDA 21 CFR part 11. It is a management system for the entire microbiological monitoring process,
from planning and execution of sampling to incubation, plate reading, CFUs identification, and finally data trending
and reports.
During environmental monitoring, it is essential for the equipment and consumables used to “communicate” with
the computerized systems to complete the digital process.
The BioTrends EM® system is developed to collect necessary information from the following air samplers
(Figure 1) and culture media (Figure 2).

Figure 1: From left to the right: portable air samplers MAS-100 NT® and MAS-100 VF® and installed air sampler MAS-100 Iso NT®

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operates as MilliporeSigma in the
U.S. and Canada.
Figure 2: From left to right: ICR 90 mm settle plate, ICR Contact Plate, and ICR Swab

Description of the Workflow

The BioTrends EM® software system with the use of mobile devices (PDAs) equipped with a barcode reader and
BioTrends EM® Mobile application, allows the recording of all activities performed in the field without using paper.
It can also retrieve all necessary data from the MAS-100 NT®, MAS-100 VF®, and MAS-100 Iso NT® air samplers of
interest for investigation, trending, and demonstrating the integrity of the acquired data.

Workflow
The sampling workflow with BioTrends EM® Mobile is very intuitive and can be summarized in the following steps
(see Figure 3):
1. Select the point to sample*
2. Select the MAS in use* (for active air samples)
3. Acquire the serial number of the plate (or swab) used, scanning the datamatrix code
4. Confirm the sampling by selecting “Save”
5. Repeat steps 1-4 for all points to sample. All activities are recorded in real time within the central system.
* The sampling point and the sampling device (steps 1-2) can be selected from the list or by scanning the
identification datamatrix code with the PDA.

Figure 3: BioTrends EM® Mobile acquires data about media and devices used from the scanner for datamatrix
codes integrated into the PDA.

2
After the sampling process, it is possible to download the data recorded by the MAS100® air samplers allowing the
BioTrends EM® system to combine the data from the two sources (MAS and PDA), giving the user more accurate
qualitative information.
For example, using the data from the device, it is possible to confirm the volume of sampled air.
The BioTrends EM® system can also compare the sampled amount versus the one expected and defined in the
sampling plan – highlighting any discrepancies or errors that occurred during instrument setup.
The workflow to download data from MAS in BioTrends EM® (see Figure 4) is very intuitive:
1. Connect the MAS-100® air sampler to a PC in which BioTrends EM® is installed.
2. In BioTrends EM® select the menu item
“Devices MAS Import Data”.
From this window, the user has a complete overview of the data present in the MAS memory.
3. Proceed to launch the “Import” function. The imported data are recorded in BioTrends EM® and cleaned from
the MAS memory.
4. By selecting the menu item
“Devices MAS View Data” it is possible to analyze the outcome of previously imported data and
highlight any anomalies.

Figure 4: Data from PDA and MAS-100® air samplers are collected in BioTrends EM® to perform deep analysis

3
Data Analysis with BioTrends EM® Software:
At the end of the monitoring process, it is possible to analyze data in a fast and reliable way, thanks to BioTrends
EM® software.
The software allows you to perform searches by applying filters for classes, environments, rooms, personnel, and
all the other relevant characteristics associated with the sampling points.
The automatic generation of statistics associated with the search carried out facilitates the review of the
alert limits.
The outcomes thus obtained can be used to generate trends, reports, and graphs in a few "clicks".

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 Application Note

How to achieve full data integrity

for the MAS-100 NT® air sampler
Introduction
The EDM 2020 draft of EU GMP Annex 1 has undergone •§§ 211.100 and 211.160 (requiring that certain
significant changes to incorporate quality management activities be “documented at the time of
through the introduction of the contamination control performance” and that laboratory controls be
strategy (CCS) concept. Environmental monitoring “scientifically sound”)
(EM) is the key process measuring the efficiency of
the sum of all measures implemented to minimize •§
 211.180 (requiring that records be retained as
the contamination risk in the aseptic manufacturing “original records,” “true copies,” or other “accurate
environment. The draft guidance on data integrity and reproductions of the original records”)
compliance with cGMP (draft guidance 2016) clarifies •§
 § 211.188, 211.194, and 212.60(g) (requiring
the role of data integrity in current good manufacturing “complete information,” “complete data derived
practice (cGMP) for drugs, as required in 21 CFR parts from all tests,” “complete record of all data,” and
210, 211, and 212. “complete records of all tests performed”)
”For the purposes of this guidance, data integrity refers Authorities, such as the FDA, expect data to be reliable
to the completeness, consistency, and accuracy of and accurate. Companies or Pharmaceutical companies
data. Complete, consistent, and accurate data should must implement meaningful and effective strategies
be attributable, legible, contemporaneously recorded, to manage their data integrity risk based on their
original or a true copy, and accurate (ALCOA).” process understanding and knowledge management of
Requirements for achieving data integrity are listed below: technologies used.

•§
 211.68 (requiring that “backup data are exact and For the microbiological path in environmental
complete,” and “secure from alteration, inadvertent monitoring, a rough overview of the required steps is
erasures, or loss”) indicated in figure 1. Depending on the available IT
infrastructure, these steps can be performed manually
•§
 212.110(b) (requiring that data be “stored to using paper-based documentation, using digital tools or
prevent deterioration or loss”) a hybrid.

Trending
Operator
Prepare analysis in
transfers Operator takes Culture CFU on plates
sampling plan cased of OOS:
sampling samples media are are counted &
based on risk Idenfification of
equipment into according to transferred to reported for
microbes root
analysis manufacturing sampling plan incubator trending
cause analysis
environment
and CAPA

Figure 1: Process steps for Environmental Monitoring (classical microbiological part)

The life science business of Merck

operates as MilliporeSigma in the
U.S. and Canada.
When using air samplers and culture media, it is -C
 ontemporaneous: “The evidence of actions, events or
important to fulfil the requirements for completeness, decisions should be recorded as they take place. ….”
consistency, and accuracy of data.
-O
 riginal: “The original record can be described as
The air samplers must be qualified in order to deliver the first-capture of information, whether recorded
reliable and efficient sampling results. The MAS-100 on paper (static) or electronically (usually dynamic,
NT® air sampler provides an accurate flow rate of 100 depending on the complexity of the system) ….”
SLPM ± 2.5%. This is verified on a regular basis by
a calibration process. The calibration tool is directly -A
 ccurate: “Ensuring results and records are accurate
traceable to standards, such as ISO 17025 or NIST. is achieved through many elements of a robust
Furthermore, the MAS-100 NT® air sample is validated pharmaceutical quality system. This can be comprised of:
together with ICR settle plates according to ISO 14698 •e
 quipment-related factors such as qualification,
(now replaced by EN 17141) in European countries for calibration, maintenance, and computer validation.
physical and biological efficiency. A d50 of less than
0.8 µm and a biological efficiency of more than 80% •p
 olicies and procedures to control actions and
is experimentally proven by an external, independent behaviors, including data review procedures to
laboratory. verify adherence to procedural requirements.

Reducing human error is key to maintaining data •d

 eviation management including root cause
integrity. This is achieved by designing an easy to use, analysis, impact assessments and CAPA
with an intuitive interface air sampler. • t rained and qualified personnel who understand the
importance of following established procedures and
Compliance documenting their actions and decisions.”
The MAS-100 NT® microbial air sampler is able to -C
 omplete: ”All information that would be critical
achieve full compliance following one of the two to recreating an event is important when trying to
methods listed below: understand the event. …”
1. Treat the instrument as a simple probe. -C
 onsistent: “Good Documentation Practices should
be applied throughout any process, without exception,
2. I ntegrate the instrument in a 21 CFR part 11 compliant, including deviations that may occur during the process.
fully computerized, EM management system. This includes capturing all changes made to data.“
In the EM workflow, the operation and documentation -E
 nduring: “Records … need to remain intact and
of active air sampling should follow the ALCOA+ accessible as an indelible/durable record throughout
principles, which are also described by the PIC/S the record retention period.”
Guidance on Good Practices for Data Integrity in
regulated GMP/GDP Environments (Draft 3 Nov 2018): - A
 vailable: “Records must be available for review
at any time during the required retention period,
-A
 ttributable: “It should be possible to identify the accessible in a readable format to all applicable
individual or computerized system that performed the personnel who are responsible for their review
recorded task. The need to document who performed whether for routine release decisions, investigations,
the task / function, is in part to demonstrate that trending, annual reports, audits or inspections”
the function was performed by trained and qualified
personnel. This applies to changes made to records
as well: corrections, deletions, changes, etc.” Treating the MAS-100 NT® air sampler as
-L
 egible: “All records must be legible – the information a simple probe
must be readable for it to be of any use. …” Environmental monitoring trends are generated by
the CFU counts and not the air sampler data. The
air sampler, therefore, may act as a simple probe,
comparable to a pump, which impacts a defined and
precise volume of air on the agar surface. The most
important feature for routine use of the air sampler is
high accuracy of the flow rate and the precision of the
collected volume of air.
In order to avoid any human errors by the operator
and to reduce the documentation workload, the MAS-
100 NT® air sampler offers the probability to limit the
operator action just to a one button-use, meaning
loading the plate into the instrument and pressing start
for sampling. The sample volume can be fixed in this
state. The required configuration to do so is shown
in Figure 2. Using this protected mode will ease the
process to fulfill the ALCOA+ rules indicated above
MAS-100 NT® air samplers manufactured by MBV AG, Switzerland.
www.mbv.ch while following all other rules for good documentation
practice.

2
 Protected mode: Checkbox <<Active>> Switches the
protected mode for the instrument on.
This function blocks manual intervention on the instrument.
The only accessible functions on the instrument are now
reduced to starting and stopping a sampling, and the user
has only read access.

Figure 2: Protection of the MAS-100 NT® air sampler from changing settings by the operator (see software manual of MAS-100 NT®)

Integrate the MAS-100 NT® air samplewith Novatek´s EM software

Frequently, software and hardware interfaces are MAS-100 NT® sampler integration with Novatek
complicated, require too much user interaction, and Environmental Monitoring Software provides a
in the worst case require more time and verification completely compliant solution by removing the
than manual entry. Additionally, when the equipment manipulations from the device and controlling it from
manufacturer updates drivers or models, the interface the computerized systems with full traceability, user
must as well be updated. Novatek International access controls, user rights and time stamps.
(Montreal, Canada) has market leading interfaces that
Novatek Environmental Monitoring software uploads the
are standard and use the latest technologies.
sampling protocols to the MAS-100 NT® instrument via
The partnership and co-development agreement a secure bidirectional integration. The operator follows
between Novatek and Merck KGaA Darmstadt, the instructions on the device and takes the samples
Germany helped create the first bidirectional interface, as per defined Standard Operating Procedures. Once
fully compliant to CFR-21 part 11, with the MAS-100 the sampling is completed, the operator connects the
NT® microbial air sampler. MAS-100 NT® sampler into Novatek EM Software and
the system automatically downloads the data (sampling
Currently there is a strong trend towards full
start and stop time, date, volume of air, etc.). The
automation of EM sample planning, data analysis and
operator must provide a reason for any sample that is
trending. Automation will reduce time required to
not taken.
collect, report and trend the data while complying to
data integrity requirements and eliminating the need
for paper-based double checks.

Compliance to requirements of 21 CFR Part 11 and Data integrity when using MAS-100 NT® air sampler with
Novatek EM Software are listed below:

DESCRIPTION DATA INTEGRITY & 21 CFR PART 11 NOVATEK EM AND MAS-100 NT® INTEGRATION
11.1 Once the record is deemed official and approved, it will be blocked and
Records to be trustworthy, reliable, and generally Equivalent to cannot be further modified. User cannot delete other users’ data; this is
paper records. permission controlled and is for administrative users only. Full audit trail for
the integration and data transfer.
11.1 E Novatek Software Applications source code and Client System Validation are
Computer systems (including hardware and software), controls, readily available for inspection by any regulatory authority including but not
and attendant documentation maintained under this part shall be limited to the FDA.
readily available for, and subject to, FDA Inspection.
11.10 (a) As per the regulations (21 CFR part 11, GAMP 5) and quality requirements
Validation of systems to ensure accuracy, reliability, consistent the system will be fully validated to assure the quality of the system and the
intended performance, and the ability to discern invalid or altered integrity of the records contained within the system
records.
11.10 (b) The software hardware is validated to produce complete copies in human
The ability to generate accurate and complete copies of records readable and electronic forms. All sections can be printed for review.
in both human readable and electronic form suitable for
inspection, review, and copying by the agency.
11.10 (c) Once the data is approved and deemed official it cannot be altered except
Protection of records to enable their accurate and ready retrieval by an authorized person. In the case where data is altered this change is
throughout the records retention period. tracked in the system wide audit trails. All changes are logged automatically.
Data is maintained throughout the record retention period as per the
predicate regulations.

3
DESCRIPTION DATA INTEGRITY & 21 CFR PART 11 NOVATEK EM AND MAS-100 NT® INTEGRATION
11.10 (d) Novatek Software has a fully integrated user access management feature
Limiting system access to authorized individuals. which limits access to the system and the MAS-100 NT®. No person without
a valid login can access the application.
11.10 (e) There are audit trail entries made while operators are creating or modifying
Use of secure, computer-generated, time-stamped audit trails to records using the MAS-100 NT®. Each time data is saved to the database,
independently record the date and time of operator entries and the name of the user and the date/time stamp is automatically saved to the
actions that create, modify, or delete electronic records. Record audit trail for the data in question.
changes shall not obscure previously recorded information
11.10 (f) Operation system checks include the transfer of protocol from Novatek EM
Use of operational system checks to enforce permitted to MAS-100 NT®. Sampling as per MAS-100 NT® instructions and transfer of
sequencing of steps and events, as appropriate. data from MAS-100 NT® to Novatek EM securely. Within the Novatek system
completing data entry prior to approving data is enforced. Data does not
display in certain grids until it is approved. This sequencing of events is
inherent throughout the system.
11.10 (g) Only users with active logins and valid passwords can log into the system
Use of authority checks to ensure that only authorized to access the MAS-100 NT®. The system password must consist of a
individuals can use the system, electronically sign a record, minimum six characters. After a number of unsuccessful attempts to access
access the operation or computer system input or output device, the system, the user ID becomes inactive and an email is sent to the
alter the record, or perform the operation at hand. administrator urgently and immediately advising of potential tampering. Only
the System Administrator can re-activate the user.
11.10 (h) The data is tracked to the user who captured it along with the date and time
Use of device (e.g., terminal) checks to determine, as stamp. The authenticity of the data is checked via passwords each time data
appropriate, the validity of the source of data input or is saved.
operational instruction.
11.10 (l) All personnel will be fully trained to their role in the system. This training will
Determination that persons who develop, maintain, or use be conducted and documented according to SOP on training.
electronic record/electronic signature systems have the
education, training, and experience to perform their assigned
tasks.
11.10 (k) System documentation will be stored in a document control/management
Use of appropriate controls over systems documentation
including: system. A change control procedure for computerized systems is in place to
(1) Adequate controls over the distribution of, access to and use ensure correct version of the documentation (IQ, OQ and user manual) are
of documentation for system operation and maintenance. used.
(2) Revision and change control procedures to maintain an
audit trail that documents time sequenced development and
moderation of systems documentation.
11.30 The ability to control user access via user ID and password is in the Novatek
Persons who use open systems to create, modify, maintain, EM software. Critical data such as passwords are encrypted in the database.
or transmit electronic records shall employ procedures and
controls designed to ensure the authenticity, integrity, and, as
appropriate, the confidentiality of electronic records from the
point of their creation to the point of their receipt
11.50 (a) In the Novatek EM software MAS-100 NT® integration:
Signed electronic records shall contain information associated
(1) The name of person who entered the data is shown on the screen.
with the signing that clearly indicates all of the following:
(2) The date and time of entry is automatically attached to the data.
(1) The printed name of the signer;
(3) Once reviewed the data will be blocked from further changes by the
(2) The date and time of signature execution;
users. Only the authorized person can change the data. All operations will be
(3) The meaning (such as review, approval, responsibility, or logged.
authorship) associated with the signature.
11.50 (b)
The items identified above (11.50 (a)) shall be subject to the
same controls as for electronic records and shall be included as
part of any human readable form of the electronic record.
11.70 In the Novatek EM software MAS-100 NT® integration:
Electronic signatures and hand-written signatures executed to The password along with username will ensure that signatures cannot be
electronic records shall be linked to their respective electronic excised, copied. No two people can have the same username hence there
records to ensure that the signatures cannot be excised, copied, is no possibility of transferring the authority. Once the person leaves the
or otherwise transferred to falsify an electronic record by company, his username will be deactivated by the system admin. In addition,
ordinary means. after a default time-period the password expires. All electronic signatures are
linked to their respective printed records in that the name, date and time and
a manual sign offline is included on all reports where required.
11.100 (a) In the Novatek EM software MAS-100 NT® integration
Each electronic signature shall be unique to one individual and does not accept two people with the same username.
shall not be reused by, or reassigned to, anyone else.

4
DESCRIPTION DATA INTEGRITY & 21 CFR PART 11 NOVATEK EM AND MAS-100 NT® INTEGRATION
11.100 (b)Before an organization establishes assigns, certifies, In the Novatek EM software MAS-100 NT® integration tracks the ownership
or otherwise sanction an individual's electronic signature, or any of each electronic signature in that all user information is recorded. This
element of such electronic signature, the organization shall verify information includes First name, Last name, Date of Birth, Employee number,
the identity of the individual. and Date of Hire.
11.100 (c)Persons using electronic signatures shall, prior to or
at the time of such use, certify to the agency that the electronic
signatures in their system.
11.200 (a)(1) In the Novatek EM software MAS-100 NT® integration:
Electronic signatures that are not based upon biometrics shall: There is a username and password associated with each person using
the software. The username and password make up the two components
Employ at least two distinct identification components such as an required to be considered an electronic signature.
identification code and password.
11.200 (a)(1)(i) In the Novatek EM software with MAS-100 NT® integration, at initial login
both the username and the password are required to access the system.
When an individual executes a series of signings during a single, (Two components)
continuous period of controlled system access, the first signing
shall be executed using all electronic signature components; At each subsequent saving of data within the login session, the software asks
subsequent signings shall be executed using at least one for the password to verify the authenticity of the person entering the data.
electronic signature component that is only executable by, and (One component)
designed to be used only by, the individual.
A user cannot enter and save data if a different user opened the program.
11.200 (a)(1)(ii) Upon saving, only the password of the user who opened the application is
accepted.
When an individual executes one or more signings not performed
during a single, continuous period of controlled system access, Each person enters the software once with a unique username and password.
each signing shall be executed using all the electronic signature Concurrent sessions are disallowed by the application.
components.
Signatures can be used only by their genuine owner.
11.200 (a)(3) In the Novatek EM software, MAS-100 NT® integration is designed such that
only the user can know both components of an electronic signature; the
Electronic signatures that are not based upon biometrics shall: username and password.
Be administered and executed to ensure that attempted use
of an individual’s electronic signature by anyone other than its
genuine owner requires collaboration of two or more individuals.
Controls for identification codes and passwords. In the Novatek EM software with MAS-100 NT® integration, when a user is
added to the system, the system enforces a password change at first logon.
(a) Maintaining the uniqueness of each combined identification
code and password, such that no two individuals have the same Each user has a unique name. The software does not accept two people with
combination of identification code and password. the same username.
There is a username and password associated with each person using the
software.
11.300 (b) In the Novatek EM software with MAS-100 NT® integration, the password
expires within a predefined time limit. The user is requested to enter a new
Ensuring that identification code and password issuances are password after the expiration of the password.
periodically checked, recalled or revised (e.g., to cover such
events as password aging). The same password cannot be used twice.
11.300 (d) In the Novatek EM software with MAS-100 NT® integration the password
consists of an adequate level of complexity.
Use of transaction safeguards to prevent unauthorized use
of passwords and/or identification codes, and to detect and After a number of unsuccessful attempts, the user ID becomes inactive. Only
report in an immediate and urgent manner any attempts at the System Administrator can reactivate the user.
their unauthorized use to the system security unit, any, as
appropriate, to organizational management. An email is triggered to immediately and urgently inform the system
administrator of the potential illegal activity.
Following loss management procedures to electronically In the Novatek EM software with MAS-100 NT® integration any user can
de-authorize lost, stolen, missing, or otherwise potentially be deactivated from the system, ensuring that there is no possibility of
compromised tokens, cards, and other devices that bear or unauthorized data entry.
generate identification code or password information, and to
issue temporary or permanent replacements using suitable,
rigorous controls.

5
 Request a demo at: SigmaAldrich.com/em-pharma-info

To place an order or receivetechnical assistance

Order/Customer Service: SigmaAldrich.com/order
Safety-related Information: SigmaAldrich.com/safetycenter
Merck KGaA
Frankfurter Strasse 250
64293 Darmstadt, Germany

For contact information for your For Technical Services, please visit,
country at SigmaAldrich.com/offices SigmaAldrich.com/techservice

SigmaAldrich.com/environmentalmonitoring

© 2021 Merck KGaA, Darmstadt Germany and/or its affiliates. All Rights Reserved. Merck, Millipore and the vibrant M are trademarks MK_AN7600EN Ver. 0.0
of Merck KGaA, Darmstadt Germany. MAS-100 NT® is a trademark of MBV AG, Switzerland. All other marks are the property of their 36585
respective owners. More information on our branded products and services on www.milliporesigma.com. 06/2021
 Application Note

Biological efficiency of the MAS-100 NT®

air sampler with TSA w. LTHThio sedi. ICR
Settle Plates
ISO 14698-1 describes standard methods to measure biological efficiency. For biological efficiency validation,
biological contamination in cleanrooms and associated the use of casein-peptone soymeal-peptone containing
controlled environments. In areas that are used for agar is recommended. As the validation depends on the
manufacturing of safe pharmaceutical products, the types of microorganisms present, the formulation and
control of biological contamination is mandatory. Part 1 quality of the culture medium also has an influence on
of ISO 14698 specifies methods which can be used to the results.
monitor risk zones in cleanrooms or give information
MAS-100® air samplers are sieve impaction systems
about sources of risks in the zones1.
based on the Anderson impaction principle. They use
Annex A describes how to determine airborne biological 90–100 mm standard Petri dishes, are easy to handle and
contamination in sterile production. For active air compact. They allow an appropriate suction flow rate,
sampling, impaction, impingement or filtration impact velocity and collection accuracy and efficacy.
samplers are recommended devices.
Annex B gives guidance on how to validate air
samplers. It is split into two parts, the physical and the

Table 1. Physical and biological efficiency of the MAS-100® family (results derived from
independent validation of MAS-100® family acc. ISO 14698 with TSA + LTHTh Cat. No. 146683)
Characteristics Iso-MH (1) Iso-MH (9) Iso NT VF
Physical Efficiency for particle size of 0.8 µm 60.41 62.58 60.20 n/a n/a
Physical Efficiency for particle size of 1.0 µm n/a n/a n/a 78.77 84.18
Physical Efficiency for particle size of 1.3 µm 71.68 82.60 76.91 84.10 85.81
Physical Efficiency for particle size of 2.2 µm 96.90 91.54 91.96 91.76 94.22
Physical Efficiency for particle size 5.4-6 µm 99.04 93.27 90.24 92.65 99.65
Biological Efficiency 76.74 73.74 78.08 82.62 76.78

We offer a variety of TSA – ICR formulations with and without neutralizers. Table 2 lists the ICR articles used in this application note
MAS-100 instruments manufactured by MBV AG, Switzerland, www.mbv.ch, MBV. Air. Nothing else.

Materials and Methods

Table 2. TSA ICR test plates for air sampling

Cat. No. Product Name Format
146786* TSA+LTHThio sedi- ICR Ready-to-Use**
146683* TSA+LTHTh sedi- ICRplus Lockable, Ready-to-Use
available in 20 and 120 pack sizes;
*

media with a lower concentration of sodium thiosulfate compared to 146683

**

The life science business of Merck

operates as MilliporeSigma in the
U.S. and Canada.
Tryptic Soy Agar + LTHThio sedi. – ICR and + LTHTh outlets etc. and to ensure a good distance between the
- ICRplus in 90 mm settle plates are designed for the individual samplers.
determination of the total aerobic microbial count in
Active air sampling was performed in a non-controlled
active or passive air monitoring as well as glove prints
environment with high human movement in order to
in Isolators and Clean Rooms.
detect a broad range of real airborne microorganisms
In a test room, two MAS-100 NT® instruments (Ref. and further to achieve higher microbial counts
No. 109191) were placed on two tables. Each sampler for comparison. Two independent test trials were
was placed in this specific position for the whole trial performed on consecutive two days in the same place,
(figure 1). Positions were chosen to achieve symmetry on the same table and at the same time (in total 14
in the room with respect to walls, ventilation inlets/ plates per agar type).

Figure 1. Sampling concept for active air sampling. Two MAS-100 NT® instruments were placed in different
positions on two tables in an uncontrolled environment outside of cleanroom conditions
MAS-100 instruments manufactured by MBV AG, Switzerland, www.mbv.ch, MBV. Air. Nothing else.

For both test trials, agar plates were tested in a After sampling, all plates were incubated for 2 days at
specific order (table 3). Before sampling of a 1000 32.5 ±2.5 °C. All lockable plates were incubated in the
liter volume every sampling step consisted of a three locked ("closed") position. After incubation, all visible
minutes delay. In case a reduced sampling volume of colonies were counted and corrected by the Feller table.
500 liters (run 7 on day 1) or 250 Liter (run 7 on day
2) the final CFU measurements were scaled-up to a
1000 liter sampling volume.

Table 3. Order of sampling (day 1 and 2).

A and B stands for the different agar plates that were used.
Position 1 Position 2
Run 1 A B
Run 2 B A
Run 3 A B
Run 4 B A
Run 5 A B
Run 6 B A
Run 7 A B

2
Results and Conclusion
The single colony counts within this non-controlled environment varied between 33 and 97 CFU, whereby the two
different ICR plate media formulations showed comparable recoveries, as shown in figure 2.

Figure 2. CFU/m3 recovered with MAS-100 NT® in a non-controlled environment on 2 consecutive days for the settle plates TSA w. LTHTh - ICR+
and TSA w. LTHThio sedi. - ICR
MAS-100 instruments manufactured by MBV AG, Switzerland, www.mbv.ch, MBV. Air. Nothing else.

The mean recovered CFU per 1 m³ for each medium are shown in table 4 below.

Table 4. Average cfu/m3 out of 14 runs (7 runs per day)

Cat.No. Product Average recovery (cfu/m3)
146786 TSA + LTHThio sedi. – ICR 57
146683 TSA + LTHTh - ICR plus 60

3
The results indicate that there was no significant difference between the recoveries obtained with
the 2 different TSA media. It can be concluded that the biological efficiency, which was validated
for the MAS-100 NT® instrument with the TSA w. LTHTh - ICR+ (Cat. No. 146683) settle plate,
was also maintained when using TSA w. LTHThio sedi. -ICR (Cat. No. 146786) which contains a
lower concentration of sodium thiosulfate.

Literature and further readings

1. ISO 14698-1(2003): Cleanrooms and associated controlled environments - Biocontamination
control - Part 1: General principles and methods
2. Application Note: Active air sampling with Tryptic Soy Agar – ICR (Cat. No. 146001, 146050,
146069 and 146683)

Request a demo at: SigmaAldrich.com/em-pharma-info

To place an order or receivetechnical assistance

Order/Customer Service: SigmaAldrich.com/order
Safety-related Information: SigmaAldrich.com/safetycenter
Merck KGaA
Frankfurter Strasse 250
64293 Darmstadt, Germany

For contact information for your For Technical Services, please visit,
country at SigmaAldrich.com/offices SigmaAldrich.com/techservice

SigmaAldrich.com/environmentalmonitoring

© 2020 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved. Merck, the vibrant M and Millipore are trademarks of MK_AN6446EN Ver. 1.0
Merck KGaA, Darmstadt, Germany or its affiliates. All other trademarks are the property of their respective owners. Detailed information 32380
on trademarks is available via publicly accessible resources. 09/2020