Air-Cleaner Performance / CASE STUDY

First-Minute Bioaerosol Testing for Active-in-Air Devices

ARE Labs used a room-pretreatment bioaerosol method to measure viable airborne microorganism reduction one minute after a biological aerosol entered a pretreated chamber.

BioaerosolAir cleanerDevice evaluation
Technical case study

What a First-Minute Endpoint Shows for Active-in-Air Devices

Abstract

This technical case study describes how ARE Labs measured near-immediate viable bioaerosol reduction in a room that was pretreated before the aerosol was introduced.

Study question

The method addressed a product-mechanism question that standard chamber decay testing does not isolate: what viable bioaerosol concentration remains one minute after a new challenge enters room air that has already been treated?

Selected findings

  • The method compared viable bioaerosol concentration after a one-minute post-aerosolization mixing period in matched control and pretreated-room conditions.
  • MS2 bacteriophage showed 90.66% one-minute net reduction, while the bacterial challenges showed lower organism-dependent reductions.
  • The case shows why active-in-air technologies may need both standard chamber testing and timing-specific room-pretreatment endpoints.
Endpoint
1 minuteViable bioaerosol concentration was measured shortly after aerosolization into a pretreated chamber.
Chamber
16 m3The study used a sealed stainless steel bioaerosol chamber under controlled conditions.
Organisms
3 challengesMS2 bacteriophage, Klebsiella aerogenes, and Staphylococcus epidermidis were used.

The method problem

Most air-treatment tests measure how quickly a device reduces particles or viable bioaerosols in a chamber over time. That remains a useful question for filtration devices, in-device UV systems, ducted treatment systems, and recirculating room purifiers because contaminated air generally must pass through the device or its treatment zone.2,3

Active-in-air technologies act at a different point in the room. Rather than waiting for contaminated air to pass through a filter, fan, duct, lamp housing, or treatment cell, these products place an active constituent into the room air before a bioaerosol event. The active may therefore already be present when a person breathes, coughs, sneezes, or talks.

That mechanism changes the test question. A standard multi-time-point decay method can measure longer-duration reduction, but it may not isolate the earliest exposure window. For active-in-air devices, the first minute after emission may matter because it is close to the point when newly airborne particles become available for inhalation.

Room pretreatment design

ARE Labs designed the room-pretreatment method around a simple but difficult measurement question: when an active-in-air product is already operating, what viable bioaerosol concentration can be measured immediately after a new challenge? The study was designed to answer a different question, not replace standard long-duration chamber testing.

The study compared two room conditions. In the control condition, the active product was not operating before the biological aerosol event. In the treatment condition, the product was operating before the bioaerosol was introduced. The objective was to measure viable bacterial and viral particles one minute after the simulated room event.

TABLE 1Room-pretreatment method parameters used in the article.
ParameterValue or description
Test conceptRoom pretreatment before bioaerosol introduction
Pretreatment period30 minutes
Bioaerosol generationCollison 24-jet nebulizer
Post-aerosolization mixing before sampling1 minute
ChamberSealed 16 m3 stainless steel bioaerosol chamber
Sampling approachAGI-30 impingers at opposite chamber locations
EnumerationSerial dilution, plating, incubation, and viable count enumeration
EndpointImmediate viable bioaerosol concentration comparison against matched controls

Holding bioaerosol generation constant was central to the design. ARE Labs used the same stock preparation, nebulization conditions, and physical generation setup in the control and treatment trials. This helped isolate the pretreatment condition as the basis for comparison rather than a change in the challenge process.

What the method measured

The room-pretreatment method measured viable bioaerosol concentration after a short, defined post-emission mixing period. Sampling followed aerosolization and the one-minute mixing period, so this was not a CADR decay endpoint. It was a direct comparison of viable concentrations between matched control and pretreated conditions.

FIGURE 1One-minute net reduction by challenge organismRoom pretreatment produced organism-dependent viable bioaerosol reduction after one minute.Bar chart showing one-minute net bioaerosol reduction of 90.66 percent for MS2, 41.75 percent for Klebsiella aerogenes, and 30.80 percent for Staphylococcus epidermidis.
View figure data as a table
organismOne-minute net reduction
MS290.66%
K. aerogenes41.75%
S. epidermidis30.8%

Source: approved ARE Labs room-pretreatment bioaerosol method summary.

  • The endpoint is a one-minute viable concentration comparison, not a CADR decay result.
  • Organism names are shortened in the chart labels for readability.

The reported one-minute net reductions varied by organism. MS2 bacteriophage showed 90.66% reduction, Klebsiella aerogenes showed 41.75% reduction, and Staphylococcus epidermidis showed 30.80% reduction. These differences keep the interpretation tied to each organism's observed response rather than reducing the findings to a single headline number.

TABLE 2One-minute room-pretreatment bioaerosol reduction.
Challenge organismOrganism typeOne-minute net percent reduction
MS2 bacteriophageNon-enveloped RNA virus surrogate90.66%
Klebsiella aerogenesGram-negative bacterium41.75%
Staphylococcus epidermidisGram-positive bacterium30.80%

Values are net percent reductions from matched control and room-pretreatment trials in the approved source article.

Why this is different from CADR

CADR is generally a time-based removal or reduction metric. It is useful for devices that remove particles from room air over time, and many chamber studies calculate it from the difference between natural decay and device-assisted decay. The first-minute room-pretreatment endpoint measures something different.2,3

The room-pretreatment endpoint compares viable concentrations immediately after a bioaerosol event in control and pretreated room conditions. For active-in-air technologies, this can capture a mechanism that a conventional decay curve may dilute or miss, particularly when the intended interaction occurs in open room air.

Where standard methods still fit

Standard methods remain important because they structure testing and make results easier to compare. ANSI/AHAM AC-1 provides a uniform procedure for portable room air cleaner performance; AHAM AC-5 addresses bioaerosol reduction by portable air cleaners; and ASHRAE 241 provides a building-level framework for infectious aerosol control. Together, they give developers, building owners, regulators, and buyers a more consistent basis for comparison.1,2,3

The goal is not to replace standard testing. When a product mechanism falls outside a familiar method's assumptions, an additional method may be warranted. An active-in-air data package may therefore pair standard chamber reduction over time with a first-minute room-pretreatment study of early-event behavior.1

Regulatory and safety context

Air-treatment products that make antimicrobial or pathogen-related claims may raise regulatory questions. EPA public device guidance lists air purifiers, ozone generators, plasma generators, bipolar ionization generators, photocatalytic air-treatment devices, and certain UV lights or filters among air-treatment products that may be regulated as pesticidal devices when pesticidal claims are made.5

For active-in-air technologies, efficacy testing often needs to be considered alongside safety-related work. This may include device particle emissions testing, VOC or by-product emissions testing, ozone screening where relevant, active concentration assessment, surface-deposition review, and ventilation-sensitivity testing. A useful data package describes both the product's effect on bioaerosols and what the product releases into the room.4,5

Testing framework

  • Start with mechanism and use-case review so the method follows how the product is intended to work.
  • Add physical aerosol characterization where particle size, active distribution, persistence, or room mixing matter.
  • Use standard chamber reduction testing when long-duration control-corrected reduction or CADR-style performance is relevant.2,3
  • Use a first-minute room-pretreatment endpoint when the claim depends on an active being present before the bioaerosol event.
  • Map claims back to the exact method timing, organisms, controls, and safety evidence rather than treating all reductions as equivalent.5

Summary

This case showed why test timing must match the product mechanism. The first-minute room-pretreatment endpoint helped the client evaluate a question that standard decay testing does not isolate: what happens when a fresh bioaerosol enters air that has already been treated? ARE Labs designed the study around that mechanism, using matched controls, viable sampling, and explicit claim boundaries.

PUBLIC SOURCES

References and study evidence

  1. 1Standard 241, Control of Infectious AerosolsASHRAEPublic context for infectious aerosol control and equivalent clean-air framing.Open source
  2. 2ANSI/AHAM AC-1-2020, Method for Measuring Performance of Portable Household Electric Room Air CleanersAssociation of Home Appliance ManufacturersPublic standards context for portable household room air cleaner performance testing.Open source
  3. 3ANSI/AHAM AC-5-2023, Method for Assessing the Reduction Rate of Key Bioaerosols by Portable Air CleanersAssociation of Home Appliance ManufacturersPublic standards context for bioaerosol chamber testing of portable air cleaners.Open source
  4. 4Preventing the Spread of Respiratory Viruses in Public Indoor SpacesU.S. Environmental Protection AgencyPublic indoor air context for ventilation, filtration, and air cleaning.Open source
  5. 5Pesticide Devices: A Guide for ConsumersU.S. Environmental Protection AgencyPublic device-claim context for air-treatment products that make pesticidal or antimicrobial claims.Open source
QUESTIONS

What to know about this study

What is first-minute bioaerosol reduction testing?

It is a room-pretreatment method that measures viable bioaerosol concentration one minute after a biological aerosol enters air that has already been treated.

Does this replace CADR testing?

No. CADR-style decay testing and first-minute pretreatment testing answer different questions. Many active-in-air products may need both.2,3

Why does claim timing matter?

A one-minute endpoint, a two-hour endpoint, and a final chamber-reduction endpoint represent different test conditions. Claims should clearly identify the timing, organism, method, and controls.