Standard Practice for Personal Sampling and Analysis of Endotoxin in Metalworking Fluid Aerosols in Workplace Atmospheres

SIGNIFICANCE AND USE
5.1 Endotoxins in metalworking fluid aerosols present potential respiratory health hazards to workers who inhale them. Therefore, a consensus standard is needed to provide reliable data on workplace airborne endotoxin concentrations where metalworking fluids are used.  
5.2 This practice for measuring airborne endotoxin concentrations in metalworking fluid atmospheres will help to foster a better understanding of endotoxin exposure-response relationships.  
5.3 This practice facilitates comparisons of inter laboratory data from methods and field investigative studies.
SCOPE
1.1 This practice covers quantitative methods for the personal sampling and determination of bacterial endotoxin concentrations in poly-disperse metalworking fluid aerosols in workplace atmospheres. Users should have fundamental knowledge of microbiological techniques and endotoxin testing.  
1.2 Users of this practice may obtain personal or area exposure data of endotoxin in metalworking fluid aerosols, either on a short-term or full-shift basis in workplace atmospheres.  
1.3 This practice gives an estimate of the endotoxin concentration of the sampled atmosphere.  
1.4 This practice seeks to minimize inter laboratory variation but does not ensure uniformity of results.  
1.5 It is anticipated that this practice will facilitate inter laboratory comparisons of airborne endotoxin data from metalworking fluid atmospheres, particularly metal removal fluid atmospheres, by providing a basis for endotoxin sampling, extraction, and analytical methods.  
1.6 In 1997, the Occupational Safety and Health Administration (OSHA) empanelled a Standards Advisory Committee to make recommendations to the Administration regarding measures that the Administration could take to improve the health of workers exposed to metalworking fluids. A report to the Assistant Secretary of Labor for OSHA was submitted in July, 1999. Subcommittee E34.50 believes that the user community would benefit significantly if a standard method was developed to give the community guidance on a methodology for the sampling and analysis of personal airborne endotoxin exposure assessments in facilities using water-miscible metal removal fluids, based on the LAL assay or other endotoxin detection technologies as they become available.  
1.7 This practice does not attempt to set or imply limits for personal exposure to endotoxin in metalworking fluid aerosols in workplace environments.  
1.8 Units—The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.9 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

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Publication Date
30-Sep-2016
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NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation: E2144 − 11 (Reapproved 2016) An American National Standard
Standard Practice for
Personal Sampling and Analysis of Endotoxin in
Metalworking Fluid Aerosols in Workplace Atmospheres
This standard is issued under the fixed designation E2144; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope removal fluids, based on the LAL assay or other endotoxin
detection technologies as they become available.
1.1 This practice covers quantitative methods for the per-
sonal sampling and determination of bacterial endotoxin con- 1.7 This practice does not attempt to set or imply limits for
centrations in poly-disperse metalworking fluid aerosols in personal exposure to endotoxin in metalworking fluid aerosols
workplace atmospheres. Users should have fundamental in workplace environments.
knowledge of microbiological techniques and endotoxin test-
1.8 Units—The values stated in SI units are to be regarded
ing.
as standard. No other units of measurement are included in this
1.2 Users of this practice may obtain personal or area standard.
exposure data of endotoxin in metalworking fluid aerosols,
1.9 This standard does not purport to address all of the
either on a short-term or full-shift basis in workplace atmo-
safety concerns, if any, associated with its use. It is the
spheres.
responsibility of the user of this standard to establish appro-
priate safety and health practices and determine the applica-
1.3 This practice gives an estimate of the endotoxin con-
bility of regulatory limitations prior to use.
centration of the sampled atmosphere.
1.4 This practice seeks to minimize inter laboratory varia-
2. Referenced Documents
tion but does not ensure uniformity of results.
2.1 ASTM Standards:
1.5 It is anticipated that this practice will facilitate inter
D1356 Terminology Relating to Sampling and Analysis of
laboratory comparisons of airborne endotoxin data from met-
Atmospheres
alworking fluid atmospheres, particularly metal removal fluid
D4840 Guide for Sample Chain-of-Custody Procedures
atmospheres, by providing a basis for endotoxin sampling,
D5337 Practice for Flow RateAdjustment of Personal Sam-
extraction, and analytical methods.
pling Pumps
1.6 In 1997, the Occupational Safety and Health Adminis- D6629 Guide for Selection of Methods for Estimating Soil
tration (OSHA) empanelled a Standards Advisory Committee Loss by Erosion
to make recommendations to the Administration regarding E1370 Guide for Air Sampling Strategies for Worker and
measures that the Administration could take to improve the Workplace Protection
health of workers exposed to metalworking fluids. A report to E1497 Practice for Selection and Safe Use of Water-
Miscible and Straight Oil Metal Removal Fluids
the Assistant Secretary of Labor for OSHA was submitted in
July, 1999. Subcommittee E34.50 believes that the user com- E1542 Terminology Relating to Occupational Health and
Safety
munity would benefit significantly if a standard method was
developed to give the community guidance on a methodology
2.2 OSHA Standards:
for the sampling and analysis of personal airborne endotoxin 29 CFR 1910.1000 Air Contaminants
exposure assessments in facilities using water-miscible metal
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
This practice is under the jurisdiction of ASTM Committee E34 on Occupa- contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
tional Health and Safety and is the direct responsibility of Subcommittee E34.50 on Standards volume information, refer to the standard’s Document Summary page on
Health and Safety Standards for Metal Working Fluids. the ASTM website.
Current edition approved Oct. 1, 2016. Published October 2016. Originally AvailablefromU.S.GovernmentPrintingOfficeSuperintendentofDocuments,
approved in 2001. Last previous edition approved in 2011 as E2144 - 11. DOI: 732 N. Capitol St., NW, Mail Stop: SDE, Washington, DC 20401, http://
10.1520/E2144-11R16. www.access.gpo.gov.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
E2144 − 11 (2016)
29 CFR 1910.1450 Occupational Exposure to Hazardous 4.2 The sample filter is extracted into a pyrogen-free solu-
Chemicals in Laboratories tion to quantitatively release endotoxin absorbed from col-
lected metalworking fluid aerosol.
2.3 Other Documents:
Criteria Document for a Recommended Standard: Occupa-
4.3 The extract solution is subjected to quantitative endo-
tional Exposure to Metalworking Fluids
toxin analysis techniques. The measured endotoxin concentra-
NIOSH Manual of Analytical Methods (NMAM)
tion is reported in terms of endotoxin potency units per unit
volume of air sampled.
3. Terminology
5. Significance and Use
3.1 For definitions of terms in this practice relating to
sampling and analysis of atmospheres, refer to Terminology
5.1 Endotoxins in metalworking fluid aerosols present po-
D1356. For definitions of terms in this practice relating to
tential respiratory health hazards to workers who inhale them.
occupational health and safety, refer to Terminology E1542.
Therefore, a consensus standard is needed to provide reliable
data on workplace airborne endotoxin concentrations where
3.2 Definitions of Terms Specific to This Standard:
3.2.1 endotoxin, n—pyrogenichighmolarmasslipopolysac- metalworking fluids are used.
charide (LPS) complex associated with the cell wall of
5.2 This practice for measuring airborne endotoxin concen-
gram-negative bacteria.
trations in metalworking fluid atmospheres will help to foster a
3.2.1.1 Discussion—Though endotoxins are pyrogens, not
better understanding of endotoxin exposure-response relation-
all pyrogens are endotoxins. Endotoxins are specifically de-
ships.
tected through a Limulus Amoebocyte Lysate (LAL) test.
5.3 This practice facilitates comparisons of inter laboratory
3.2.2 endotoxin unit (EU), n—a biological potency unit
data from methods and field investigative studies.
equivalent to the FDA Reference Standard Endotoxin (RSE).
Currently, EC-6 is equivalent to 0.1 ng 3D 1 EU.
6. Interferences
3.2.3 field blank, n—filter/cassette unit prepared for sam-
6.1 Airborne endotoxin measurements resulting from use of
pling that is taken to the sampling site and handled in the same
LAL reagents are subject to inhibition/enhancement effects
mannerastheanalyticalfilter/cassetteunit,butthatisnotapart
from a variety of bio-molecular species and physicochemical
of the sampling process.
phenomena, such as pH, temperature, filter matrix effects,
3.2.4 Gram-negative bacteria, n—prokaryotic cells that
cationic concentrations, LAL-reactive materials (LRM), en-
have a complex cell-wall structure that stain characteristically
zyme influences, and lysate composition variability and sensi-
when subjected to the differential Gram staining procedure.
tivity (a function of different lysate processing methodologies).
3.2.5 Limulus amebocyte lysate (LAL) assay, n—a biologi-
cal assay that detects endotoxin. 7. Apparatus
3.2.6 metal removal fluids, n—the subset of metal working
7.1 Sampling:
fluids that are used for wet machining or grinding to produce
7.1.1 Sampling Unit, an apparatus consisting of a personal
the finished part.
sampling pump, a 37-mm glass fiber filter, a two-piece,
3.2.6.1 Discussion—The term most often refers to straight
closed-face plastic cassette, and flexible connecting tubing
oils and water-based fluids, such as soluble, semi-synthetic,
between the personal sampling pump and the attached cassette/
and synthetic fluids.
filter unit.
7.1.1.1 Pump, a constant-flow personal sampling pump with
3.2.7 onset time, n—time required for a change of 200 mOD
an on-board battery power source and a flow rate of 2.0 L/min
(optical density) units relative to the initial OD value.
(65 %).
3.2.8 personal sampler, n—a portable sampling instrument
7.1.1.2 Filter Cassette, pyrogen-free, closed-faced, two-
that is attached to a person to ascertain the concentration of
piece polystyrene filter holder with 4 mm inlet and outlet, with
specific constituents in the air in the person’s breathing zone.
caps.
3.2.9 pyrogen-free, adj—material(s) devoid of measurable
7.1.1.3 Filter (Membrane),pyrogen-free,glassfiber,37-mm
endotoxin activity.
diameter with a cellulose support pad.
3.2.10 pyrogen-free water (PFW), n—processed water that 7.1.1.4 Connective Tubing, flexible, appropriate inside di-
is devoid of measurable endotoxin activity.
ameter.
7.1.1.5 Soap-bubble Meter, a primary standard used for
4. Summary of Practice
sampler flow rate calibration.
4.1 A known volume of workplace air in a facility utilizing
NOTE 1—An alternative primary standard is acceptable.
metalworking fluids is drawn through a sample filter cassette
7.2 Extraction:
unit.
7.2.1 Sonicator Bath, ultrasonic/water bath apparatus with a
minimum peak frequency of 40-
...


This document is not an ASTM standard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. Because
it may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current version
of the standard as published by ASTM is to be considered the official document.
Designation: E2144 − 11 E2144 − 11 (Reapproved 2016) An American National Standard
Standard Practice for
Personal Sampling and Analysis of Endotoxin in
Metalworking Fluid Aerosols in Workplace Atmospheres
This standard is issued under the fixed designation E2144; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope
1.1 This practice covers quantitative methods for the personal sampling and determination of bacterial endotoxin concentrations
in poly-disperse metalworking fluid aerosols in workplace atmospheres. Users should have fundamental knowledge of
microbiological techniques and endotoxin testing.
1.2 Users of this practice may obtain personal or area exposure data of endotoxin in metalworking fluid aerosols, either on a
short-term or full-shift basis in workplace atmospheres.
1.3 This practice gives an estimate of the endotoxin concentration of the sampled atmosphere.
1.4 This practice seeks to minimize inter laboratory variation but does not ensure uniformity of results.
1.5 It is anticipated that this practice will facilitate inter laboratory comparisons of airborne endotoxin data from metalworking
fluid atmospheres, particularly metal removal fluid atmospheres, by providing a basis for endotoxin sampling, extraction, and
analytical methods.
1.6 In 1997, the Occupational Safety and Health Administration (OSHA) empanelled a Standards Advisory Committee to make
recommendations to the Administration regarding measures that the Administration could take to improve the health of workers
exposed to metalworking fluids. A report to the Assistant Secretary of Labor for OSHA was submitted in July, 1999. Subcommittee
E34.50 believes that the user community would benefit significantly if a standard method was developed to give the community
guidance on a methodology for the sampling and analysis of personal airborne endotoxin exposure assessments in facilities using
water-miscible metal removal fluids, based on the LAL assay or other endotoxin detection technologies as they become available.
1.7 This practice does not attempt to set or imply limits for personal exposure to endotoxin in metalworking fluid aerosols in
workplace environments.
1.8 Units—The values stated in SI units are to be regarded as standard. No other units of measurement are included in this
standard.
1.9 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility
of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory
limitations prior to use.
2. Referenced Documents
2.1 ASTM Standards:
D1356 Terminology Relating to Sampling and Analysis of Atmospheres
D4840 Guide for Sample Chain-of-Custody Procedures
D5337 Practice for Flow Rate Adjustment of Personal Sampling Pumps
D6629 Guide for Selection of Methods for Estimating Soil Loss by Erosion
E1370 Guide for Air Sampling Strategies for Worker and Workplace Protection
E1497 Practice for Selection and Safe Use of Water-Miscible and Straight Oil Metal Removal Fluids
E1542 Terminology Relating to Occupational Health and Safety
This practice is under the jurisdiction of ASTM Committee E34 on Occupational Health and Safety and is the direct responsibility of Subcommittee E34.50 on Health
and Safety Standards for Metal Working Fluids.
Current edition approved Dec. 1, 2011Oct. 1, 2016. Published December 2011October 2016. Originally approved in 2001. Last previous edition approved in 20072011
as E2144 - 01E2144 - 11.(2007). DOI: 10.1520/E2144-11.10.1520/E2144-11R16.
For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM Standards
volume information, refer to the standard’s Document Summary page on the ASTM website.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
E2144 − 11 (2016)
2.2 OSHA Standards:
29 CFR 1910.1000 Air Contaminants
29 CFR 1910.1450 Occupational Exposure to Hazardous Chemicals in Laboratories
2.3 Other Documents:
Criteria Document for a Recommended Standard: Occupational Exposure to Metalworking Fluids
NIOSH Manual of Analytical Methods (NMAM)
3. Terminology
3.1 For definitions of terms in this practice relating to sampling and analysis of atmospheres, refer to Terminology D1356. For
definitions of terms in this practice relating to occupational health and safety, refer to Terminology E1542.
3.2 Definitions of Terms Specific to This Standard:
3.2.1 endotoxin, n—pyrogenic high molar mass lipopolysaccharide (LPS) complex associated with the cell wall of
gram-negative bacteria.
Available from U.S. Government Printing Office Superintendent of Documents, 732 N. Capitol St., NW, Mail Stop: SDE, Washington, DC 20401, http://
www.access.gpo.gov.
Available from U.S. Department of Health and Human Services, Public Health Service, Centers for Disease Control and Prevention, National Institute for Occupational
Safety and Health, 4676 Columbia Pkwy., Cincinnati, OH 45226.
3.2.1.1 Discussion—
Though endotoxins are pyrogens, not all pyrogens are endotoxins. Endotoxins are specifically detected through a Limulus
Amoebocyte Lysate (LAL) test.
3.2.2 endotoxin unit (EU), n—a biological potency unit equivalent to the FDA Reference Standard Endotoxin (RSE). Currently,
EC-6 is equivalent to 0.1 ng 3D 1 EU.
3.2.3 field blank, n—filter/cassette unit prepared for sampling that is taken to the sampling site and handled in the same manner
as the analytical filter/cassette unit, but that is not a part of the sampling process.
3.2.4 Gram-negative bacteria, n—prokaryotic cells that have a complex cell-wall structure that stain characteristically when
subjected to the differential Gram staining procedure.
3.2.5 Limulus amebocyte lysate (LAL) assay, n—a biological assay that detects endotoxin.
3.2.6 metal removal fluids, n—the subset of metal working fluids that are used for wet machining or grinding to produce the
finished part.
3.2.6.1 Discussion—
The term most often refers to straight oils and water-based fluids, such as soluble, semi-synthetic, and synthetic fluids.
3.2.7 onset time, n—time required for a change of 200 mOD (optical density) units relative to the initial OD value.
3.2.8 personal sampler, n—a portable sampling instrument that is attached to a person to ascertain the concentration of specific
constituents in the air in the person’s breathing zone.
3.2.9 pyrogen-free, adj—material(s) devoid of measurable endotoxin activity.
3.2.10 pyrogen-free water (PFW), n—processed water that is devoid of measurable endotoxin activity.
4. Summary of Practice
4.1 A known volume of workplace air in a facility utilizing metalworking fluids is drawn through a sample filter cassette unit.
4.2 The sample filter is extracted into a pyrogen-free solution to quantitatively release endotoxin absorbed from collected
metalworking fluid aerosol.
4.3 The extract solution is subjected to quantitative endotoxin analysis techniques. The measured endotoxin concentration is
reported in terms of endotoxin potency units per unit volume of air sampled.
5. Significance and Use
5.1 Endotoxins in metalworking fluid aerosols present potential respiratory health hazards to workers who inhale them.
Therefore, a consensus standard is needed to provide reliable data on workplace airborne endotoxin concentrations where
metalworking fluids are used.
5.2 This practice for measuring airborne endotoxin concentrations in metalworking fluid atmospheres will help to foster a better
understanding of endotoxin exposure-response relationships.
E2144 − 11 (2016)
5.3 This practice facilitates comparisons of inter laboratory data from methods and field investigative studies.
6. Interferences
6.1 Airborne endotoxin measurements resulting from use of LAL reagents are subject to inhibition/enhancement effects from
a variety of bio-molecular species and physicochemical phenomena, such as pH, temperature, filter matrix effects, cationic
concentrations, LAL-reactive materials (LRM), enzyme influences, and lysate composition variability and sensitivity (a function
of different lysate processing methodologies).
7. Apparatus
7.1 Sampling:
7.1.1 Sampling Unit, an apparatus consisting of a personal sampling pump, a 37-mm glass fiber filter, a two-piece, closed-face
plastic cassette, and flexible connecting tubing between the personal sampling pump and the attached cassette/filter unit.
7.1.1.1 Pump, a constant-flow personal sampling pump with an on-board battery power source and a flow rate of 2.0 L/min
(65 %).
7.1.1.2 Filter Cassette, pyrogen-free, closed-faced, two-piece polystyrene filter holder with 4 mm inlet and outlet, with
...

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