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VHP Qualification Protocol

This document summarizes a validation study for the use of hydrogen peroxide vapor as a decontaminant for biosafety cabinets. The study was conducted according to the requirements of NSF/ANSI 49 Annex K, which provides a protocol for validating alternative decontamination agents. The study tested hydrogen peroxide vapor systems in multiple biosafety cabinet models and found that an average concentration of 475 ppm achieved for 53 minutes was sufficient to pass all biological indicator testing criteria across three trials per cabinet, validating hydrogen peroxide vapor as an effective biosafety cabinet decontaminant.

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Phạm Quang Huy
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2K views25 pages

VHP Qualification Protocol

This document summarizes a validation study for the use of hydrogen peroxide vapor as a decontaminant for biosafety cabinets. The study was conducted according to the requirements of NSF/ANSI 49 Annex K, which provides a protocol for validating alternative decontamination agents. The study tested hydrogen peroxide vapor systems in multiple biosafety cabinet models and found that an average concentration of 475 ppm achieved for 53 minutes was sufficient to pass all biological indicator testing criteria across three trials per cabinet, validating hydrogen peroxide vapor as an effective biosafety cabinet decontaminant.

Uploaded by

Phạm Quang Huy
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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Validation Study for the Use of

Hydrogen Peroxide Vapor as a


Decontaminant for Biosafety Cabinets
in Accordance with the Requirements
of NSF/ANSI 49
ABSA
October 22, 2012
Nick Flynn, Peter Harris
B & V Testing, Inc.
Waltham, MA
800.851.9081
www.bandvtesting.com
Presentation Overview

Hydrogen Peroxide Validation


Studies and NSF 49 Annex K
Overview
Validation Study Criteria
Validation Study Methods
Validation Study Results
Hydrogen Peroxide Studies and Annex K
Overview
Studies (2011/2012) executed under direction of NSF
49 Joint Committee on Biosafety Cabinetry Hydrogen
Peroxide Task Group and in accordance with NSF 49
Annex K, Protocol for the Validation of Alternate
Biosafety Cabinet Decontaminating Methods and
Agents
Given widespread and historical use of hydrogen
peroxide as a sterilant in biomedical research and
manufacturing environments, significant industry
interest in incorporation of a validated procedure for
BSC decon utilizing HP vapor as per NSF 49
Hydrogen Peroxide Studies and Annex K
Overview (contd.)
Three separate studies utilizing three
separate hydrogen peroxide systems:
STERIS Vaporized Hydrogen Peroxide
(dry), Bioquell vapor phase hydrogen
peroxide (wet), and AeroClave
energized hydrogen peroxide (wet
aerosolized)
B&V Testing performed validation study
for STERIS VHP 1000 ARD
NSF 49 Annex K Overview

Purpose
To establish a protocol for validating alternative
decontamination agents and systems
Objective
To demonstrate the alternative decon agent/system is at least
as effective as formaldehyde gas
Decontamination Procedure
The methodology for the decontamination procedure during
the validation study should be clearly specified prior to the
study. The protocol should clearly state the method being
validated, i.e. measured concentration/fixed mass
NSF 49 Annex K Overview (contd.)

Cabinet Selection Requirements


The study shall include two different models of
each Class II Type BSC (A1,A2,B1 and B2) with a
minimum of three trials for each BSC model
including trials with no internal BSC blower and
incorporating material compatibility testing
NSF 49 Annex K Overview (contd.)

Establishing Efficacy, Biological


Indicator (BI) Criteria
Seven pairs of BIs (A+B sample)
consisting of > 106 log
geobacillus stearothermophilus
placed within BSC
(note: Annex K indicates six
locations; HP task group
increased to seven)
NSF 49, Annex K Efficacy, BI Placement
3 pairs placed between the
filter pleats on the downstream
(clean) side of the exhaust
HEPA filter ( center and
opposite corners)
1 pair placed in the
contaminated positive pressure
plenum
1 pair placed between the filter
pleats near the center of the
upstream (dirty) side of the
downflow HEPA filter
1 pair placed on the work
surface or sidewall
1 pair placed beneath the work
surface
NSF 49, Annex K Efficacy, BI Validation
Criteria
(BI) Site for single trial
Success if either of 2 samples is negative
Failure if both samples test positive
Single trial
Pass if all seven (BI) sites pass
Conditional pass if six (BI) sites pass
Fail if 2 or more (BI) sites fail
NSF 49, Annex K Efficacy, BI Validation
Criteria (contd.)

Cabinet type/Decontamination method


validation
Minimum of 3 trials
Pass if all 3 trials pass
Pass if 3 trials have conditional or full passes
with failed sites not coinciding
Trials may be repeated if cause for failure
identified
VHP Validation Experiments

Measured concentration
methodology validated
Methodology based on established
EPA-registered sterilant validation
data and B&V pre-testing
experiments
STERIS VHP EPA registration
validation data
250 PPM @ 90 minutes
400 PPM @ 30 minutes
VHP Validation Experiments (contd.)

Experiments at B&V lab Waltham,


MA and NuAire lab Plymouth, MN
Independent microbiology
laboratory utilized for BI testing
VHP Validation Experiments (contd.)
Cabinet preparation

Exhaust Filter BI
Placement
VHP Validation Experiments (contd.)
Cabinet preparation

H2O2
Supply sensor
Filter BI
Placement
VHP Validation Experiments (contd.)
Pre-testing experiments performed on multiple BSC models
20+ runs executed to :
Establish environmental conditions (Temp/RH)
Establish target VHP concentration and identify VHP
monitoring location
Optimize vapor distribution by identifying injection and
return port locations
Establish internal blower operation cycle, as applicable
Establish VHP injection rates
Establish VHP exposure time
VHP Validation Experiments (contd.)
Methodology Conditions:
Start temperature+humidity: > 60 F/< 70% Rh
VHP injected into the downstream (clean) side
of the exhaust HEPA filter with return through
BSC access opening for A1,A2 and B1 and via
cabinet supply intake on B2.
VHP injection rate of 3g/minute, 59% H2O2
VHP injection to > 400 ppm @ center work
surface + 30 minutes continuous VHP injection,
reversing injection/return pathway halfway
through
Where operational operate the BSC blower at a
minimum of one minute for every 15 minutes
of VHP injection
VHP BSC Validation Experiments (contd.)
Study Data

Time Required to Achieve 400 ppm

30

25
Average Time to
20 reach 400 ppm:
Minutes

23 Minutes
15

10

0
A1-1

A1-2

A2-1

A2-2

B1-1

B1-2

B2-1

B2-2
BSC Evaluated
VHP BSC Validation Experiments (contd.)
Study Data
Cumulative Injection Time

60
55
50
45
Average Total
40
Minutes

35 Injection Time :
30 53 Minutes
25
20
15
10
5
0
A1-1

A1-2

A2-1

A2-2

B1-1

B1-2

B2-1

B2-2
BSC Evaluated
VHP BSC Validation Experiments (contd.)
Study Data
Average VHP Concentration

600
550
500 Average VHP
450 Concentration:
400
350 475 ppm
ppm

300
250
200
150
100
50
0
A1-1

A1-2

A2-1

A2-2

B1-1

B1-2

B2-1

B2-2
BSC Evaluated
VHP BSC Validation Study BI Sites and Trial
Results
BSC Trial # BI Site Result Trial Result

A1-1 1 All 14 BIs Negative Pass

A1-1 2 All 14 BIs Negative Pass

A1-1 3 All 14 BIs Negative Pass

A1-2 1 All 14 BIs Negative Pass

A1-2 2 All 14 BIs Negative Pass

A1-2 3 All 14 BIs Negative Pass


VHP BSC Validation Study BI Sites and Trial
Results (contd.)

BSC Trial # BI Site Result Trial Result

A2-1 1 All 14 BI Negative Pass

A2-1 2 All 14 BI Negative Pass

A2-1 3 All 14 BI Negative Pass


Work Surface BIs A+B
A2-2 1 Positive Conditional Pass

A2-2 2 All 14 BI Negative Pass

A2-2 3 All 14 BI Negative Pass


VHP BSC Validation Study BI Sites and Trial
Results (contd.)

BSC Trial # BI Site Result Trial Result

B1-1 1 All 14 BI Negative Pass

B1-1 2 All 14 BI Negative Pass


Exhaust HEPA Center BI
B1-1 3 A Positive Pass

B1-2 1 All 14 BI Negative Pass

B1-2 2 All 14 BI Negative Pass


Downflow HEPA BI A
B1-2 3 Positive Pass
VHP BSC Validation Study BI Sites and Trial
Results (contd.)

BSC Trial # BI Site Result Trial Result

B2-1 1 All 14 BI Negative Pass


Exhaust HEPA Front
B2-1 2 Right A Positive Pass

B2-1 3 All 14 BI Negative Pass

B2-2 1 All 14 BI Negative Pass

B2-2 2 All 14 BI Negative Pass

B2-2 3 All 14 BI Negative Pass


VHP BSC Validation Study, Methodology and
Cabinet Results

VHP successfully validated for decontamination of


Class II type A1, A2, B1 and B2 cabinets including
with non-operational BSC blower
Material compatibility studies showed no adverse
impact to BSC materials
NSF task force to review data for inclusion in NSF 49
Acknowledgements

NuAire, Inc.
The Baker Company
Esco Technologies, Inc.
STERIS Corporation

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