THE BAKER COMPANY - BD Biosciences · THE BAKER COMPANY 161 Gatehouse Rd., Sanford, Maine 04073 USA...

THE BAKER COMPANY 161 Gatehouse Rd., Sanford, Maine 04073 USA

“Creating Immaculate Atmospheres”

Gatehouse Rd., Sanford, Maine 04073 USA (207) 324-8773 1-800-992-2537 FAX (207) 324-2632 Internet address: http// www.bakerco.com

“Creating Immaculate Atmospheres” of 11

Microbiological Testing Report 3/14/12

BioProtect IV with Influx Flow Cytometer

Model BP-504-5 BD Influx Baker Doc. #BAKT 185

Customer:

BD Biosciences

Introduction:

The safety cabinet was airflow balanced to a set point velocity of 55 feet per minute down flow and 105 feet per minute intake at an 8” sash opening prior to microbiological testing. Preliminary smoke tests were conducted to identify potential airflow turbulence which may affect cabinet performance. A smoke visualization test demonstrated turbulent air was curling around the top and when the Influx front door was in the opened position and that the equipment increased the air velocity towards the front of the cabinet. Smoke was not observed escaping the cabinet however.

BioProtect IV with BD Influx Flow Cytometer





I. NSF/ANSI Standard 49 Microbiological Testing

Purpose:

The following standardized tests were conducted to evaluate the biosafety cabinet performance with the Influx flow cytometer installed. The following tests were performed with the cytometer in an operational condition. Method:

All tests are performed to the NSF/ANSI Biosafety Standard 49- 2011. These tests determine whether aerosols will be contained within the cabinet, outside contaminants will not enter the cabinet work area, and aerosol contamination of other equipment in the cabinet will be minimized. The cabinet shall be turned on at least 30 min before the start of any microbiological test and operated continuously throughout all test procedures. The microbiological testing shall demonstrate passing results at an operating range of plus or minus 10 fpm from nominal set point as referenced in NSF/ANSI Standard 49 for Biosafety Cabinetry.

NSF Personnel Protection Test

Method: The cabinet was challenged with Bacillus Subtilis bacterial spores at 8.0 X 10^8

spores/ml for each test run. Each test was run for a total of 30 min with a 15 min nebulizer challenge. NSF requires the nebulizer to run for 5 minutes, Baker Company performs the testing at a more stringent 15 minute nebulizer challenge. No more than 10 colony forming units total shall be counted for each test on the filter from six AGI impingers for passing results, as per the NSF standard and no more than 5 colony forming units shall be counted on both slit sampler agar settling plates for passing results. Acceptance: The number of Bacillus subtilis spores (culture forming units) recovered from the 6 AGI samplers shall not exceed 10 CFU per test. Total slit-type air sampler plate counts shall not exceed 5 subtilis CFUs for a 30 min sampling period. The control plate shall be positive. A plate is “positive” when it contains greater than 300 CFU of B. subtilis.





Personnel Protection Test set up with nebulizer spore challenge at NSF-49 location

Containment results:

All Personnel Protection challenge tests were performed with the more stringent application of the sort chamber door in the open or up position and the Aerosol Management system (AMO) completely off. The assessment was to challenge the system with the worst condition in a static state.

Air flow balance set points: NSF‐49 Required Performance Zone

Test 1 (52 fpm down flow , 106 fpm Intake)

CFU COUNT CFU COUNT

Control Air slit samplers 6‐AGI samplers Results

Positive 0 0 PASS






CFU COUNT CFU COUNT


Positive 1 2 PASS


CFU COUNT CFU COUNT


Positive 0 0 PASS


CFU COUNT CFU COUNT


Positive 1 1 PASS

All required NSF/ANSI Standard-49, 2011 Personnel Protection Microbiological tests had Passing results.

NSF Product Protection Test:

Method:

The cabinet was challenged with Bacillus Subtilis spores at 8.0 X 10^6 spores/ml for each test run. Each test was run for a total of 30 min with a 15 min nebulizer challenge. NSF requires the nebulizer to run for 5 minutes, Baker Company performs the testing at a more stringent 15 minute nebulizer challenge. No more than 5 colony forming units total on all settling plates shall be counted for each test for passing results, as per the NSF standard. Acceptance: The number of Bacillus subtilis spores (culture forming units) on agar settling plates shall not exceed 5 CFU for each test. The control plate shall be positive. A plate is “positive” when it contains greater than 300 CFU of B.subtilis.





Product Protection results:

All Product Protection challenge tests were performed with the more stringent application of the sort chamber door in the open or up position and the Aerosol Management system (AMO) operating in the high setting mode. The assessment was to challenge the system with the worst condition in a static state.

Air flow balance set points: NSF‐49 Required Performance Zone


Control CFU COUNT Results

Positive 2 PASS



Positive 0 PASS



Positive 1 PASS



Positive 1 PASS

All required NSF/ANSI-49, 2011 Product Protection Microbiological tests had Passing results





Note: All Product Protection tests had 150mm agar settling Petri dishes placed at the bottom of the sort chamber and at the level of the waste collector as shown. No spores or culture forming units were detected in any of the testing. Plates were clean of any contamination.

Nebulizer placed in required NSF-49 location

Alternative Sort Chamber Testing

Another condition of testing was performed at the sort/stream chamber only. Agar sampling plates placed inside the chamber for product protection with AMO system on high setting and the nebulizer placed inside the chamber for personnel protection with AMO system none operational which was identified as the most extreme conditions for safety with sort chamber door in the opened position.





Personnel Protection testing with Nebulizer challenge inside sort chamber

Personnel Protection Testing: sort chamber door open

Test 1 (52 fpm down flow , 106 fpm Intake) AMO SYSTEM OFF

CFU COUNT CFU COUNT


Positive 0 8 PASS

Test 2 (52 fpm down flow , 106 fpm Intake) AMO OPERATING ON HIGH SETTING

CFU COUNT CFU COUNT


Positive 1 0 PASS





Personnel Protection testing with Nebulizer callenge in the NSF-49 position

Personnel Protection Testing:

Test 3 (52 fpm down flow , 106 fpm Intake) AMO SYSTEM OFF

CFU COUNT CFU COUNT


Positive 3 1 PASS

Test 4 (52 fpm down flow , 106 fpm Intake) AMO OPERATING ON HIGH SETTING

CFU COUNT CFU COUNT


Positive 1 1 PASS





Product Protection testing with door open and AMO system on high setting

Product Protection Testing:


AMO Operating on high setting‐sort door open


Positive 3 PASS

Test 2 (52 fpm down flow , 106fpm Intake)

AMO SYSTEM OFF ‐ SORT DOOR OPEN


Positive 3 PASS

AMO SYSTEM OFF ‐ SORT DOOR CLOSED



Positive 0 PASS

All sort chamber testing had Passing results.





Conclusion:

The above test results provided evidence that the Baker Company BioProtect IV Cabinet with the BD Influx flow cytometer passed the required NSF/ANSI Standard-49 biosafety standards. The cabinet proved safe in every condition of challenge during standard operational conditions of the cytometer. The opened door to the sort/stream chamber and challenge cylinder in place showed that safety was not compromised during daily end user functions.

Worker Comfort Testing

Noise level Test

The noise level at nominal set point was 63.0db with a background of 53 db with the cytometer operating inside the cabinet. BioProtect Cabinet operating only 62.4db

Acceptance: Overall noise level in front of the cabinet shall not exceed 67 db when measured where the ambient sound level is 57db. Measurement on the “A” weighing scale.

Microbiological Testing

Robert A Thibeault

THE BAKER COMPANY - BD Biosciences · THE BAKER COMPANY 161 Gatehouse Rd., Sanford, Maine 04073 USA...

Documents

Transcript of THE BAKER COMPANY - BD Biosciences · THE BAKER COMPANY 161 Gatehouse Rd., Sanford, Maine 04073 USA...