Air quality control in reproductive laboratories: How important it is and how to implement, measure...

Air quality control in reproductive laboratories

How important it is and how to implement, measure and control

4th International Congress - Academy of Clinical Embryologists 18-20 September 2015, Kochi INDIA

Sandro C. Esteves, MD., PhD. Medical & Scientific Director, ANDROFERT

Andrology & Human Reproduction Clinic Campinas, BRAZIL

Outline

ANDROFERTandrofert.com.br

ANDROLOGY AND HUMAN REPRODUCTION CLINIC REFERRAL CENTER FOR MALE REPRODUCTION PROF. DR. SANDRO ESTEVES 2013 OCTOBER





1. Importance of air quality in IVF

2. How to implement

3. How to measure and control

ANDROLOGY AND HUMAN REPRODUCTION CLINIC - REFERRAL CENTER FOR MALE REPRODUCTION S ESTEVES, 2 2015

ANDROFERT

Morbeck, JARG 2015; 32(7):1019-24.

IMPORTANCE OF AIR QUALITY IN IVF


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25.0%

32.7%

14.1%

43.1%

Miscarriage rate

CPR

General IVF Population (N=468) Cleanroon IVF lab (Central HVAC with particle + VOC filtration)

Standard IVF lab equipped with portable filtration units

P=0.01

P=0.03

Esteves et al. Fertil Steril 2004


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CPR Miscarriage rate

36.9%

23.0%

47.1%

15.0%

Severe Male Factor Infertility (N=399) Standard IVF lab equipped with portable filtration units Cleanroon IVF lab (Central HVAC with particle + VOC filtration)

Esteves et al. Fertil Steril 2006



ANDROFERT

Van Voorhis et al. (F&S 2010) At least 7 of 10 consistently high-performing IVF centers in the USA used filtered laboratory air with HEPA and chemical, solid-phase filtration for VOCs

Munch et al. (JARG 2015): Inadvertent removal of carbon filters resulted in significant drop of KPI

Heitmann et al. (RBM Online 2015): Central HVAC (Particles + VOC) vs. Portable Units Higher IR (32.4% vs. 24.3%; p<0.01) Higher LBR (39.3% vs. 31.8%; p<0.05)

RECENT EVIDENCE


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Importance of Air Quality in IVF Summary

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ü High levels of VOC in unfiltered IVF lab air ü Mouse and human embryo development inversely

correlated with VOC levels ü Efficient particle and VOC removal by dedicated air

filtration systems ü Better IVF outcome (PR, IR, miscarriage) in

laboratories with air particle and VOC filtration ü Central HVAC seems to work better than portable

systems


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What regulatory agencies ask for

ANDROFERTANDROLOGY AND HUMAN REPRODUCTION CLINIC - REFERRAL CENTER FOR MALE REPRODUCTION S ESTEVES, 8 2015

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Social responsibility to protect public health

from exposure to harm, when scientific

investigation has found a plausible risk even though a scientific

consensus had not been reached

PRECAUTIONARY PRINCIPLE

*Convention on Biological Diversity, UN 1992; Ratified by 192 countries and the EU, except

USA, Andorra and South Sudan


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Region (directive)

European Union (EU directive 2004/23/EC; 2006/86/EC)

Brazil (Anvisa RDC33/2006; RDC23/2011)

Particle filtration

Equivalent to GMP Grade A air quality in the critical areas with a background environment at least equivalent to Grade D (exceptions apply)

At least equivalent to ISO class 5 (NBR/ISO 14644–1) in the critical areas

Microbial contamination

Microbial colony counts equivalent to those of Grade A as defined in the current GMC guide with a background environment at least equivalent to Grade D

Microbiological monitoring required; specifications not defined

Volatile organic compounds filtration

Not required

Ventilation systems should be equipped with filters imbedded with activated charcoal


ANDROFERTReviewed by Esteves & Bento, RBM Online 2013.

CONTAMINATION Any undesirable material or

substance that may affect the quality of final product

Contaminants are generated by

laboratory personnel,

processes, furniture and equipment


ANDROFERT

Indoor air

Personnel

Material and Equipment

Cell, tissue and fluids

1. Airborne particles Residues of cleaning agents and clothing, skin shedding; temporary suspension; 1< µm <100

2. Microorganisms Viruses, spores, bacteria; constant suspension in air; size <1 up to 8µm


ANDROFERT

Contamination

CONTAMINATION

3. Indoor Volatile Organic Compounds Chemical compounds that evaporate and react

with indoor ozone Result in submicronic particles and harmful by-products (eg. toluene, benzene, alcohols, acetone,

refrigerating gases, aldehydes) Cigarette smoke, automobile exhaust, detergents, lubricants,

dyes, furniture, rubber, plastics, resins, synthetic fibers, nylon, particle-board, plywood

Some are genotoxic/mutagenic


ANDROFERT

VOC in REPRODUCTIVE LABORATORIES

0

1000

2000

3000

Outside Air Indoor IVF

Lab Air

533

2769 5-fold increase

in VOC

Anderson et al. 1997


ANDROFERT

HOW TO PROTECT GAMETES AND EMBRYOS FROM CONTAMINATION

Bacteria and other contaminants can attach themselves to particles, a decrease in particles equates to an increase in air quality Pre-Filters: >10µ

Fine dust filters: 1<10µ HEPA filters: 0,3µ = 99,97%

Removal of air particulates by forced air movement using positive air pressurization

through a series of filters with increased efficiency


ANDROFERT

Particle size/m3 of air: ISO 14.644-1 0.3 µM 0.5 µM 1 µM 5 µM

ISO 1 ISO 2 10 4 ISO 3 102 35 8 ISO 4 1,020 352 83

IVF lab ISO 5 10,200 3,520 832 29 ISO 6 102,000 35,200 8,320 293

OR ISO 7 352,000 83,200 2,930 Transfer ISO 8 3,520,000 832,000 29,300

ISO 9 35,200,000 8,320,000 293,000

CLEAN ROOM CLASSIFICATION


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VOCs are 100–1000x smaller than the effective pore size of HEPA filters

Spaces between the carbon particles contain a cloud of

delocalized electrons that act as electronic glue to chemical

contaminants

Alcohols and ketones are oxidized and thereby

detoxified by potassium permanganate

HOW TO PROTECT GAMETES AND EMBRYOS FROM CONTAMINATION

Removal of VOCs by forced air movement using positive air pressurization through a

series of filters containing activated carbon and potassium permanganate


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ANDROFERT

PORTABLE SYSTEMS

HEPA filter

VOC filter

Pre- filter

Central HVAC SYSTEMS


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BEFORE IMPLEMENTATION Risk assessment analysis

•  New or existing facility •  Age and size of laboratory •  Equipment/furniture (low off gassing?) •  Constructing materials •  Atmospheric air pollution •  Disposable materials (storage location) •  Human activity

No. per workspace, Use of protective clothing Cosmetics, etc.


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HOW WE IMPLEMENTED

ANDROFERTANDROLOGY AND HUMAN REPRODUCTION CLINIC - REFERRAL CENTER FOR MALE REPRODUCTION S ESTEVES, 21 2015

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AIR-HANDLING VENTILATION SYSTEM (with carbon + KMnO4 filtration)


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23 - S. Esteves, August 2015

2015

ISO 5 Cleanroom IVF lab + Chemical filtration


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ISO 5 Cleanroom IVF lab + Chemical filtration


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LAYOUT AND CONSTRUCTION

In situ wet construction Low off-gassing materials

Surfaces and Coatings Lighting fixtures

Flush-mounted and sealed Coved junctures Stainless steel & aluminum

Furniture, doors, windows, air vents, workstations


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0

50

100

before after

% TQE

0

20

40

before after

% miscarriage

0

50

before after

% LBR 2.3

3.2

Average No. Top Quality Embryos ET

Conventional lab (with portable filters) Cleanroom lab (Central HVAC with particle & VOC filtration)

P=0.01


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N=2,315 cycles

Cumulative live birth rates

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ANDROFERTANDROFERT

40.4% 48.0%

ANDROFERT, Referral Center for Male Reproduction

ET #3 (FET) 49

ET #2 (FET) 239

ET #1 (fresh) 822

50.5% +18.8%

+25.0% Female Age ≤38

ANDROFERT

332/822 63/239 17/49

HOW TO MEASURE AND CONTROL


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TEST EQUIPMENT

Air volume flow rate Termoanemometer

Air exchange rate Termoanemometer

Air and room pressure differential

Microanemometer and balometer

HEPA-filter integrity leak Aerosol generator

Airborne particle counts Electronic particle counter

Recovery performance Smoke generator

Lighting level Luxmeter

Noise level Decibelmeter Temperature and humidity

Thermometer and hygrometer

Third-part certification company (CCL, Brazil); semi-annually IEST 006.2 standards


ANDROFERT

TEST

Air volume flow rate

Air exchange rate

Air and room pressure differential HEPA-filter integrity leak Airborne particle counts Recovery performance Lighting level

Noise level Temperature and humidity


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Settle plates Mold and blood-agar Petri dishes

Surface sampling Swabs and contact plates

Five-finger glove printing Incubators, workstations, air, floor, wall

Limits Colony Forming Unites (CFU)

Settle plates (90mm

diameter) CFU/4 hours

Contact plates

(55mm diameter) CFU/plate

Glove print CFU/glove

<3 <3 <3

Microbiological Monitoring


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VOC Monitoring 1. Activated-carbon life-time

estimates Carbon tetrachloride activity method

2. Active air sampling on Tenax TA sorbent and 2,4 dinitrophenyl hydrazine

Summa canisters; Thermal desorption Gas chromatography (Limits <2 mcg/m3 TVOC)

3. Direct reading Snapshot reading using VOC probes Photo ionization detector


ANDROFERT

Esteves & Bento. RBM Online 2013

ü APPROVED


ANDROFERT

Key messages 1. Air quality (particulate matter and VOC)

associated to in vitro embryo development in animal and human studies

2. Regulatory agencies have issued Directives

with specific requirements for air quality in IVF labs based on precautionary principle

3. Air cleanness in IVF should integrate both particulate matter and VOC filtration


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Key messages (2) 4. Risk assessment analysis needed before

implementing air quality control to IVF units

5. Validation “in operational state” after AQC implementation


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6. Ideally, measure and control should be guided by third-part certification companies

Thank you

This presentation is available at http://www.slideshare.net/

sandroesteves

4th International Congress - Academy of Clinical Embryologists 18-20 September 2015, Kochi INDIA

Air quality control in reproductive laboratories: How important it is and how to implement, measure...

Health & Medicine

Transcript of Air quality control in reproductive laboratories: How important it is and how to implement, measure...