&EPA
United States
Environmental Protection
Agency
Wastewater Response Protocol
Toolbox:
Planning For and Responding To
Wastewater Contamination
Threats and Incidents
December 2011
Module 4:
Analytical Guide
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Table of Contents - Module 4
1 Introduction 4-1
1.1 Objectives of this Module 4-1
2 Current Laboratory Infrastructure in U.S 4-1
2.1 Environmental Chemistry Labs 4-3
2.2 Radiochemistry Labs 4-4
2.3 BiotoxinLabs 4-4
2.4 Chemical Warfare Labs 4-4
2.5 Microbiological Laboratories 4-4
3 Health and Safety 4-5
4 Analytical Approach for Unidentified Contaminants in Wastewater 4-7
5 Basic Screening for Organic and Inorganic Chemicals Using Standard Methods 4-9
6 Expanded Screening for Chemicals 4-15
6.1 Expanded Screening for Organic Compounds - Sample Preparation Techniques 4-16
6.2 Expanded Screening for Organic Compounds - Detection Methods 4-17
6.3 Expanded Screening for Inorganic Chemicals 4-19
T3
6.4 Expanded Screening for Cyanides 4-21
6.5 Expanded Screening for Biotoxins 4-21
6.6 Expanded Screening for Chemical Weapons 4-21
6.7 Basic and Expanded Screening for Radionuclides 4-22
^^>
7 Additional Recommendations for Chemical Screening of Wastewater Samples 4-24
C
8 Screening for Microbiologicals Including Unknowns 4-25
9 Forensic Implications of Sample Collection and Analysis 4-26
10 Data Analysis and Reporting 4-26
11 Summary 4-27
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Planning and Preparat
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Threat Warning
Initial Threat Evaluation
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1 Introduction
1.1 Objectives of this Module
The primary intended users of this module
include laboratory personnel and planners
who would provide analytical support
to a wastewater utility in the event of a
contamination threat. This module is intended
to be a planning tool for labs rather than
a how-to manual for use during an actual
incident. As part of planning for such an
incident, laboratories may want to prepare a
detailed 'Laboratory Guide' specific to their
needs and capabilities. Also, laboratories may
want to consider how they coordinate with
networks of other laboratories so as to provide
added capability and capacity.
The objectives of this module include:
1. Describing how laboratories can
respond to contamination events.
2. Describing special laboratory
considerations for handling and
processing emergency wastewater
samples suspected of contamination
with a harmful substance.
3. Presenting model approaches and
procedures for analysis of wastewater
samples suspected of contamination
with a known or unknown substance.
These analytical approaches are
intended to take advantage of existing
methodologies and infrastructures.
4. Encouraging planners to develop a
site-specific analytical approach and
Laboratory Guide that conforms to
the general principles of the model
approaches presented in this module.
Roles of Laboratories in Response to
Contamination Threats
While utility labs, especially at larger utilities,
may become quite involved with preliminary
screening and preliminary analysis of samples
from suspected contamination events, most
will not be able to implement all of the
analytical protocols described in Module 4.
Federal, state, and commercial labs may be
called upon to provide more sophisticated, in-
depth analyses.
2 Current Laboratory
Infrastructure in U.S.
The analytical approach described in this
module was developed under the assumption
that it would be implemented using the
existing laboratory infrastructure in this
country. EPA established the Environmental
Response Laboratory Network (ERLN) to
assist in addressing chemical, biological, and
radiological threats during nationally significant
incidents. The Water Laboratory Alliance
(WLA), which launched in October 2009, is the
water component of the ERLN and provides the
Water Sector (drinking water and wastewater
systems) with an integrated nationwide network
of laboratories. The WLA provides additional
analytical capability and capacity to an event
involving intentional and unintentional water
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contamination involving chemical, biological
and radiochemical contaminants. For more
information, visit http://www.epa.gov/erln/
water.html.
Also, the WLA has a Water Laboratory
Alliance - Response Plan (WLA-RP) (EPA
817-R-10-002, November 2010) that outlines
the processes and procedures for a coordinated
laboratory response to water contamination
incidents that may require more analytical
laboratory capability and capacity than a
typical laboratory can provide. It addresses
analytical demand during the emergency
response, remediation, and recovery phases of
a natural disaster, accident, or terrorist incident
affecting the water sector, (http://water.epa.
gov/infrastructure/watersecurity/wla/upload/
WLAResponsPlan_November2010.pdf)
EPA has constructed a Laboratory
Compendium to assist utilities and other
responders in locating appropriate labs
for analysis of contaminants during a
contamination incident. The Laboratory
Compendium is a database of laboratory
capabilities for environmental analysis in
water, air, soil, sediment, and other media.
Instructions on acquiring access to the
Laboratory Compendium are available at
the following website: http://www.epa.gov/
compendium.
The ERLN is also part of a larger federal
network of laboratories called the Integrated
Consortium of Laboratory Networks (ICLN).
The Department of Homeland Security
established the ICLN to coordinate laboratory
networks to respond to acts of terrorism and
other major incidents. ICLN is composed of
networks of Federal laboratories from U.S.
Department of Agriculture, Department of
Health and Human Services (Centers for
Disease Control and Prevention, Food and
Drug Administration), Department of Defense,
and the Environmental Protection Agency.
Analytical Goals
In responding to contamination incidents (intentional or unintentional), keep in mind the
following analytical goals or points:
Protect laboratory personnel and provide timely, accurate results.
Confirm or rule out the presence of significantly elevated levels of certain types or
classes of contaminants.
Check for the presence of additional contaminants, not just one.
Report accurate results and not misidentify an instrumental response, which could
lead to a false positive result.
Focus on harmful contaminants including radionuclides, biotoxins, pathogens, and
high concentrations of industrial chemicals.
Consider background concentrations of a contaminant in a specific location when
analyzing the data from wastewater samples.
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The networks of laboratories analyze clinical
and environmental samples for chemical,
biological, and radiological analytes associated
with terrorist as well as natural events.
It is likely that most emergency wastewater
samples will be sent for analysis on the basis
of a probable contamination threat. Samples
laboratory support for 'credible' incidents, and
specialty laboratories likely would be called
into service for 'confirmed' incidents.
Figure 4-1 and the narrative below summarize
the typical laboratory infrastructure, as
it currently exists, for the analysis of
environmental samples.
Chemical Analysis
Biological Analysis
Radiochemical
Labs
Environmental
Chemistry Labs
Environmental
Microbiology Labs
Chemical
Weapons
Biotoxins
Figure 4-1. Types of Laboratories for Analysis of Environmental Samples.
sent to a laboratory as a result of a probable
contamination threat should be treated as if
they contain a potentially harmful substance.
However, the site characterization process,
along with the threat evaluation process,
should result in most highly hazardous
samples being screened before they reach the
laboratory. Some organizations have an "All
Hazards Receipt Facility" (AHRF) which is
activated to screen unknown samples before
those samples are sent to a laboratory. From
a safety standpoint, it is important for a
laboratory to realize that it will not be expected
to determine every potential contaminant. For
instance, utility laboratories typically may
expect to receive samples from 'possible'
incidents. The utility labs may need additional
2.1 Environmental Chemistry Labs
This group includes many EPA, state, utility,
and commercial water analysis labs. Most
environmental chemistry labs are set up to
perform analysis of wastewater samples for
compliance with the Clean Water Act and/or
the Resource Conservation and Recovery Act,
as well as some state and local regulations.
Because these laboratories are typically
certified to utilize regulatory compliance
methods, unless the lab tests for a particular
analyte on a routine basis, they may not
necessarily be able to utilize a method for a
specific contaminant without advance notice.
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There are also a number of research
laboratories within the government and
academic sectors that may be available on a
limited basis. These labs may be equipped with
advanced instrumentation and highly trained
analysts who can implement exploratory
techniques.
2.2 Radiochemistry Labs
If a radioactive contaminant is suspected,
analysis should be performed by a laboratory
specifically equipped to handle such material
and analyze for a range of radionuclides.
EPA, Department of Energy (DOE), states,
and some commercial firms have labs
specifically dedicated to the analysis of
radioactive material. Information concerning
EPA's radiological emergency response and
laboratory services is available at http://
www.epa.gov/radiation/emergency-response-
overview.html. Another source of support
is the Federal Radiological Monitoring and
Assessment Center (FRMAC) operated by the
Department of Energy:
http: //www. nv. doe. gov/national security/
homelandsecurity/frmac/.
2.3 Biotoxin Labs
Currently, few laboratories are set up
specifically for the analysis of biotoxins. There
are a number of laboratories in government
and academia that perform biotoxin analysis,
usually for matrices other than wastewater
(e.g., seafood and agricultural products). It is
possible that some biotoxin analyses could
be performed in qualified environmental
chemistry labs using techniques such as gas
chromatography-mass spectrometry (GC/
MS), high performance liquid chromatography
(HPLC), immunoassay, and possibly liquid
chromatography-mass spectrometry (LC/
MS). However, this capability is not currently
widespread.
2.4 Chemical Warfare Labs
Chemical Weapons are those weapons that the
Chemical Weapons Convention (CWC) has
placed on a list known as Schedule 1. These
are toxic chemicals with few or no legitimate
uses other than for military purposes. There are
only a handful of laboratories in the U.S. that
are qualified and permitted to perform analysis
for Schedule 1 chemical weapons material.
Among other qualifications, these labs possess
appropriate analytical instrumentation, are
supplied with analytical standards of Schedule
1 chemical weapons material, and have
implemented necessary safety measures. Some
of these labs can only be accessed via certain
federal agencies such as the FBI and include
the U.S. Army Edgewood Laboratory and the
Lawrence Livermore National Laboratories.
EPA is developing capability and capacity to
analyze environmental samples potentially
contaminated with chemical warfare agents
and degradents at seven fixed laboratories and
two mobile laboratories.
2.5 Microbiological Laboratories
The analysis of waterborne pathogens will
likely be performed by an environmental
microbiology lab. Environmental microbiology
laboratories (including those of EPA, state
environmental agencies, utilities, and the
commercial sector) routinely analyze water
samples for indicators of fecal contamination
(e.g., fecal coliform bacteria, total coliform
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bacteria, and E. coif). An analytical limitation
is that specific culture analyses for waterborne
pathogens such as Salmonella spp. and
Shigella spp. are not routinely performed in
most environmental microbiology laboratories.
In the event that a contamination threat or
event involves select agents such as Bacillus
anthracis, Brucella spp., Yersiniapestis,
Francisella tularensis, and C. botulinum
toxins, among others, samples would probably
be transported by federal authorities to a lab
within the Centers for Disease Control and
Prevention Laboratory Response Network.
As discussed later in this module, the presence
of microbiological pathogens in wastewater
typically does not constitute the same health
risk as when these pathogens are found in
drinking water. Therefore, there may not be the
same need to analyze potentially contaminated
wastewaters for harmful microbes as there is
for chemical contaminants.
3 Health and Safety
It is important to realize that details important
for laboratory safety are integrated into
the Threat Evaluation (Module 2) and Site
Characterization (Module 3) processes even
though they occur outside of the laboratory
setting. The threat evaluation and site
characterization processes help to define
the hazard conditions at the site of sample
collection, identify who should collect the
samples and determine which laboratories
should analyze them.
The following are some important
considerations for the safety of personnel who
will be processing laboratory samples that
may contain unknown, possibly dangerous
substances.
Currently, laboratories should have a plan
in place to ensure worker safety. Some
laboratories may wish to treat certain
emergency wastewater samples as hazardous
material, whether they be chemical, biological,
or radiochemical in nature. They may also
decide to develop a specific health and safety
plan (HASP) to address this potential risk,
although there is currently no requirement to
do so in most cases.
Laboratory personnel involved in the handling
and analysis of wastewater samples should
have appropriate current safety training
that will allow them to adhere to applicable
regulations. Laboratories may wish to explore
some of the measures contained in regulations
for the handling of hazardous materials, such
as OSHA 1910.120 (http://www.osha.gov/
pi s/oshaweb/owadi sp. show_document?p_
table=standards&p_id=9707).
Additionally, there is health and safety
suggestions contained in various government
publications including Biosafety in
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Figure 4-2. Lab Personnel Using a Protective
Lab Hood.
Microbiological and Biomedical Laboratories,
5th Edition. National Center for Infectious
Diseases, Centers for Disease Control and
Prevention, Office of Health and Safety, 2009.
http://www.cdc.gov/biosafety/publications/
bmblS.
Analysis of potentially hazardous samples
during an emergency situation may require
additional personal protective equipment (PPE)
above that normally used in the laboratory.
These PPE requirements should be determined
during the creation of the site-specific HASP.
These may include, among others, the use of
butyl gloves and full face shields especially
during pouring and splitting of non-volatile
samples.
Appropriate hoods (Figure 4-2) and other
physical control measures should always be
utilized when handling samples containing
potentially hazardous unknown contaminants.
The laboratory should also be outfitted with
safety equipment such as eyewashes, safety
showers, spill containment devices, and
first aid kits. The laboratory should be fully
informed about the sample collection and site
investigation procedures, including any field
safety screening and rapid field testing results.
However, to reduce risks associated with
potential, undetected hazards, laboratories may
wish to screen the sample for various hazards
upon receipt at the laboratory, regardless of the
reported field safety screening results.
The water solubility of potential contaminants
sometimes contributes to their safe handling.
Steps should be taken to avoid volatilizing or
aerosolizing wastewater samples, which would
then increase the inhalation risk. Accordingly,
separatory funnel liquid-liquid extractions,
which may release aerosols when vented, are
not recommended unless laboratories utilize
appropriate hoods or other precautions.
Dilution of a hazardous wastewater sample
with laboratory-grade water helps reduce risks
associated with handling of the sample and its
analysis for chemical contaminants. Dilution,
however, may interfere with the ability to
detect and quantify contaminants. If dilution
is desired, 'log dilutions' may be utilized. For
instance, a 1/1000 dilution may be analyzed
first, followed by a 1/100 dilution if nothing
is detected in the highest dilution. These can
be followed by a 1/10 dilution, and finally the
undiluted sample.
Like dilution, reducing the volumes of sample
handled may help minimize exposure for both
chemical and biological contaminants. Certain
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analytical techniques involve using smaller
sample volumes. For example, micro-liquid
extraction utilizes only about 40 ml compared
with large volume extractions which utilize
1L or more. Selecting analytical approaches
requiring smaller volumes of sample may
help to limit risk to lab personnel dealing with
suspect samples.
Approaches to limiting the potential exposure
to unknown pathogens prior to chemical
analysis may be to irradiate (UV or gamma), or
pasteurize, the samples. Currently there is no
general consensus on proper use of irradiation
to reduce risk associated with sample handling
and analysis while maintaining the integrity
of the sample and analysis. Therefore, these
techniques for reducing pathogen exposure are
not validated methods and are experimental
at best. However, they could be utilized by
the laboratory, on portions of the sample, as
an exploratory technique. It should be noted
that UV sterilization or heat sterilization may
also alter the identity or quantity of some
chemicals.
4 Analytical Approach for
Unidentified Contaminants in
Wastewater
In the case of a complete unknown, the
problem of identifying and quantifying a
specific contaminant presents a significant
challenge. The difficulty arises from the large
number of potential contaminants of concern,
and the impracticality of screening for all of
them. To address this issue, EPA recommends
using an analytical approach for unknowns that
is based on contaminant classes derived from
a prioritization of chemicals and pathogens of
concern if present in a wastewater system.
The recommended analytical approach
for unknown contaminants in wastewater
presented in this module is comprehensive
for selected priority contaminants and
provides coverage for hundreds of additional
contaminants. The following assumptions and
principles were used in the development of this
approach:
Selection of target analytes was based
on an assessment of contaminants likely
to pose a threat to public health, public
safety, utility employee health and safety,
property, utility operations/infrastructure,
and the environment.
Existing laboratory infrastructure and
analytical methods were utilized.
Analytical procedures are tiered, with a
progression from field safety screening
and rapid field testing, through laboratory
screening, to confirmatory analysis.
Samples that cannot receive confirmatory
analysis in the lab performing the
initial testing are subsequently referred
to laboratories that can perform a
confirmatory analysis.
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The entire approach relies on the
systematic elimination of potential
contaminants, both to ensure the safety
of sampling and laboratory personnel and
to aid in identification of the unknown
contaminant.
It is also important to realize that identification
of unknown contaminants in wastewater
samples is not an exact science. This is
especially true given the difficult analytical
matrix presented by wastewater. There is no
guarantee that any combination of technology
will always yield successful identification of
unknown contaminants.
It should be emphasized that Module 4 is
not intended to represent a prescriptive how-
to laboratory manual. Rather, this model
screening procedure is intended to be a
recommended planning tool for laboratories to
formulate a Laboratory Guide specific to their
own needs and capabilities. The Laboratory
Guide for the lab dealing with emergency
samples is similar to the Emergency Response
Plan prepared by the utility in that both can be
based extensively on information presented
in the EPA Wastewater Response Protocol
Toolbox, but both should still be customized to
local needs and resources.
Also, the Water Laboratory Alliance -
Response Plan (WLA-RP) provides a
structure to coordinate laboratory capability
and capacity to prevent duplication of effort,
maximize efficiencies and effectiveness,
improve communication, and increase
analytical support. Laboratories are encouraged
to increase awareness of the WLA-RP through
notification and discussion with the state
drinking water programs and emergency
management agencies.
Additionally, EPA has recently published
additional guidance on sample collection
entitled Sampling Guidance for Unknown
Contaminants in Drinking Water (EPA 817-
R-08-003, November 2008) (see www.
epa.gov/watersecurity; search under Water
Laboratory Alliance). The guidance integrates
recommendations for pathogen, toxin,
chemical, and radiochemical sample collection,
preservation, and transport procedures to
support multiple analytical approaches for
the detection and identification of potential
contaminants in drinking water.
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5 Basic Screening for Organic
and Inorganic Chemicals Using
Standard Methods
The recommended chemical screen integrates
a number of analytical techniques to cover
abroad range of chemical classes. These
techniques include not only wet chemistry and
instrumental analysis, with which laboratories
are typically familiar, but also hand-held
equipment and commercially available test
kits, such as those based on immunoassays.
The overall screening approach for unknown
chemicals is broken into two parts, the
basic screen (Section 5) and the expanded
screen (Section 6). The basic screen utilizes
established (standardized) analytical methods
for the analysis of contaminants in wastewater.
The WLA-RP also has a section on Basic
Field/Safety Screening to assist laboratories
in procedures for dealing with unidentified
contaminants. Typically, these methods are
produced as a standard by a recognized method
development organization and contain steps
to defensibly confirm the presence and/or
quantity of specific contaminants. Table 4-1
lists several sources of standard methods.
Standardized methods may be selected from
an appropriate method database, such as
the Water Contaminant Information Tool
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Table 4-1: Sources of Standardized Methods
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Name
Water Contaminant
Information Tool
(worn
EPA SW-846 methods
40 CFR Parts 136 and
141
National
Environmental
Method Index
(NEMI)
Description
Contains methods compiled
from a number of sources.
May be consulted first.
Compendium of analytical
and sampling methods that
have been evaluated and
approved for use in
complying with RCRA
regulations.
Promulgated list of
defensible methods widely
accepted in the analytical
community for water and
wastewater.
On-line database containing
chemical, microbiological,
biological, toxicity, and
physical methods for
comparison.
Publisher
US EPA Office
of Water
US EPA Office
of Solid Waste
US EPA Office
of Resource
Conservation
and Recovery
and US EPA
Office of Water
US Geological
Survey and
US EPA
How to obtain
http //www.epa.gov/wcit
http://www.epa.gov/
epaoswer/hazwaste/test/
main.htm
http://ecfr.gpoaccess.gov
www nemi.gov
(WCIT) (http://www.epa.gov/wcit/). The
National Environmental Methods - Index
(NEMI) contains methods compiled from
many sources. These methods are reviewed
and selected by the National Methods and
Data Comparability Board (http://acwi.gov/
methods/). Some of these methods are EPA
wastewater methods, some are EPA SW-846
methods (Test Methods for Evaluating Solid
Waste, Physical/Chemical Methods), and
others were developed by USGS or DOE for
their environmental monitoring programs.
Also, EPA's National Homeland Security
Research Center's Standardized Analytical
Methods for Environmental Restoration
Following Homeland Security Events (SAM)
(EPA600-R-10-122, October 2010) (www.epa.
gov/sam/) identifies analytical methods to be
used by laboratories tasked with performing
analyses of environmental samples following a
homeland security event.
The basic screen is designed to capture many
of the chemical contaminants of concern
using a relatively small number of well-
defined, standardized analytical techniques
(Figure 4-3). The techniques chosen for basic
screening analysis are summarized in Table
4-2.
If the methods in this table are performed, then
the basic screen may cover a large percentage
of the priority chemical contaminants.
Furthermore, many other contaminants of
concern, but of lower priority, may be screened
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Table 4-2: Suggested Analytical Techniques for Performing the Basic Screen, Arranged by
Chemical Class
Chemical . T EPA Method ^lef" ^ te* Analyte
(general class) Analytical Technique (SW846) Act Method Ust
4U wrK ran 1 Jb
Volatiles (organic)
Semivolatiles (organic,
includes many
pesticides)
Trace metals (inorganic)
Total mercury
(inorganic, includes
organomercury
compounds)
Cyanides
Radionuclides
Purge-and-trap PID/ELCD
Purge-and-trap GC/MS
Solid-phase extraction GC/
MS
ICP-AES, ICP-MS,
graphite furnace AA
Cold vapor AA
Wet chemistry
Gross alpha, gross beta,
gross gamma
802 1B
8260B
8270D
3535A
6010
6020A
7010
747 1B
901 2 A
7110B
601
602
624
625
200.7
200.8
200.9
245.1
245.2
335.4
900.0
A
B
C
D
E
F
for as well. To increase confidence
in the results, only validated
methods should be used for the
basic screen (e.g., SW-846 or
comparable methods). Table 4-3
below lists contaminants that may
be detected by the basic screen
standardized methods listed in
Table 4-2.
Figure 4-3. Lab Personnel Using an Analytical Approach
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Table 4-3: Analyte Lists Corresponding to Table 4-2
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1 1,1,2-TetrachlorQethane
1.1.1-Tnchloroethane
1 . 1 .2,2-Tetrachloroethane
1,1 ,2-Trichloroethane
1,1-Dichloroethane
1,1-Oichloroethene
1 ,1-Dichloropropene
1 ,2,3-Trichlorobenzene
1,2,3-Trichloropropane
1 ,2,4-TrichIorobenzene
1,2,4-Trimethylben2ene
1 ,2-Dibrorno-3-chtoropropane
1 ,2"Dibromoeth3ne
1 ,2-Dichlorobenzene
1 ,2-Dichloroethane
1 ,2-Dichloropfopane
1 ,3,5-Trimethylbenzene
1.3-Oichlorobenzene
1,3-Dichloropropane
1 ,4-Oichlorobenzene
2.2-Dichloro propane
2-Chlorotoluene
2- Nitro propane
4-CWorotoluene
Acrylonilrile
Ally I chloride
Benzene
Bromo benzene
Bromochlorome thane
2.2'.3,3',4,4',6-Heptachlorob!phenyl
2,2',3,3'14,5l,616'-OctachlorobJphenyl
2,2' 3'.4,6-Pentachlorobiphenyl
2,2'.4,4',5,6!-Hexachlorobiphenyl
2,2',4,4'-Tetrachloroblphenyl
2,3-Dichlorobiphenyl
2,4,5-Trichlorobiphenyl
2,4-Dirritfotoluene
2,6-Dinitrotoluene
2-Chlorobiphenyl
a-8HC
Acenaphthylene
a-Chlordane
Alachlor
Aldrin
Anthracene
Atrazine
Azinphos methyl
b-BHC
Benz(a ) a nth ra cene
Benzo(a)pyfene
Benzo(b)fluoranthene
Benzo(g,h,i)perylene
Benzo(k)fluofanthene
bis(2-Ethylhexyl)adipate
bis(2-Ethylhexyl)phthalate
Bolster
Arsenic
Cadmium
Chromium
Cobalt
Copper
Lead
Mercury
Mercury
Free
cyanide
(see
method)
Cesium-
137
Indium-
192
Cobalt-60
Strontium-
90
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Table 4-3 (cont.): Analyte Lists Corresponding to Table 4-2
Bromodichtaromethane
Bromoform
Bromomethane
Butyl chloride
Carbon disulfide
Carbon tetrachloride
Chloroacetonitrile
Chlorobenzene
Chloroethane
Chloroform
Chloromethane
Cis- 1 ,2-Dichtoroethene
Cis-1,3-Dichloropropene
Dibrornochtoromethane
Dibromomethane
Dichlorodifluoromethane
Di ethyl elder
Ethyl methaerylate
Ethy tbenzene
Hexachlorobutadiene
Hexachloroelhane
Isopropylbenzene
Methacrylonitrile
Melhanol (solvent)
Methyt acryiate
Methyl methacrylate
Methyl tert-butyl ether
Methylene chloride
m-Xylene
Naphthalene
n-Butylbenzene
Nitrobenzene
n-Propylbenzerie
Butachlor
Butylbenzylphthalate
Chlorobenzilate
Chloroneb
Ctilorothalonil
Chlorpyrifos
Chrysene
cis-Permethrin
Coumaphos
Cyanazine
Dacthal
d-8HC
Demeton (mixed isomers)
Diazinon
Dibenz(a , h)anthracene
Dtchtorvos
Dieldrin
Diethyl phthalate
Dimethyl phthalate
Di-n-butyl phthalate
Disulfoton
Endosulfan I
Endosulfao II
Endosulfan sulfate
Endrin
Endrin aldehyde
Elhoprop
Etridiazole
Fensulfothion
Perth ion
Fluorene
g-BHC
g-Chlordane
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Table 4-3 (cont.): Analyte Lists Corresponding to Table 4-2
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o-Xylene
Pentachloroethane
p-lsopropy (toluene
Propionitrile
p-Xylene
sec-Butylbenzene
Styrene
tert-Butylbenzene
Tetrachloroethene
Tetrahydrofuran
Toluene
trans-1 ,2-Dichloroethene
trans-1 ,3-Dichloropropene
trans-1 ,4-Dichloro-2-butene
Trichloroethene
Trichlorofluoromethane
Vinyl chloride
Heptachlor
Heptachlor epoxide
(Isomer B)
Hexachlorobenzene
Hexachlorocycbpentadiene
lndeno{1 ,2,3-cd)pyrene
Lindane
Merphos
Methoxychlor
Methyl parathion
Metolachlor
Metribuzin
Mevinphos
Mated
p,p'-DDD
p,p'-DDE
p,p'-DDT
Pentachlorophenol
Phenanthrene
Phorate
Propachlor
Pyrene
Ronnel
Simazine
Stirophos
Tokuthion
trans-Nonachlor
Trichloronate
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6 Expanded Screening for
Chemicals
The purpose of the expanded screen is to
capture chemical contaminants not picked
up by the basic screen. The expanded screen
may also more rapidly detect some analytes
covered by the basic screen. The expanded
screen should be sufficiently broad to permit
the analyst to screen for many possible
contaminants.
In practice, the expanded screen can be used in
addition to the basic screen, because the results
of the basic screen may provide a springboard
to guide the selection of techniques for the
expanded screen. For example, many of
the techniques in the basic screen rely on
chromatography and/or mass spectrometry, so
the data should be capable of being evaluated
for the presence of not only target analytes,
but also other compounds. Combining
observations from multiple basic screening
techniques may also be helpful.
Alternatively, some laboratories may choose
to utilize only the expanded screen, comprised
of potentially sensitive techniques, including
those summarized in Table 4-4. In the latter
case, preliminary results can be cautiously
used to make response decisions, but should
be followed up with confirmatory analysis
because screening techniques, including
some listed in Table 4-4, are not necessarily
definitive. Some details regarding utilization
of the expanded screening techniques are
included below to help guide the reader in the
selection of appropriate techniques relative to
wastewater analysis.
Table 4-4: Expanded Screening for Contaminants (Arranged by Class of Contaminant)
Contaminant Type Expanded Screening Technique
Organic
Inorganic
Cyanides
Biotoxin
Radiological
Chemical Warfare Agents
GC, GC/MS, HPLC, LC/MS, Immunoassay test kits
1C, AA, ICP, ICP-MS
Wet chemistry
Immunoassay test kits, GC/MS, HPLC, and LC/MS
Handheld equipment
GC/MS with direct injection, purge & trap, and SPE/SPME,
test kits, handheld equipment
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6.1 Expanded Screening for Organic
Compounds - Sample Preparation
Techniques
Organic analyses utilized in this approach
are comprised of some combination of the
following three steps: 1) extraction or recovery
of the contaminant from the wastewater
matrix; 2) separation of the compounds
through gas chromatography or liquid
chromatography; and/or 3) detection and
identification of the analyte. Preparatory and
extraction techniques for organic constituents
should be broad enough to recover a variety of
compound classes (e.g., a range of hydrophilic
properties and molecular weights). A variety
of techniques are used for detection of organic
constituents.
Regardless of the detector system employed,
there are a number of widely used sample
preparation techniques. These include the
following:
Large Volume Liquid/Liquid Extraction
(LLE)
This technique (SW846-Method 35IOC) is not
advisable for aerosolizable samples because it
requires the use of separately funnels that may
release aerosols when vented. The generation
of these aerosols may represent a larger health
hazard than other techniques, unless labs take
precautions such as appropriate hoods.
Direct Aqueous Injection
Although a powerful analytical technique, the
use of direct aqueous injection of wastewater
samples into a GC may present technical
difficulties in chromatographic separation and
could reduce the lifetime of the GC column
and the detector (Figure 4-4). While the high
concentrations of contaminants that might be
present during an emergency incident may
cause the use of direct injection of wastewater
samples to prove valuable, particularly
for initial and rapid screening of analytes,
the analytical system should be carefully
monitored for loss of performance. For all but
a few analytes, confirmatory analyses may be
required.
Figure 4-4. Lab Personnel Using Syringe to
Inject GC.
Micro Liquid-Liquid Extraction
(micro-LLE)
Liquid micro extraction involves the use
of small volumes of solvent (e.g., 2 ml) to
extract analytes from a small volume (e.g., 40
ml) of water. For the high concentrations of
contaminants that may be present during an
emergency incident, the use of micro-LLE of
aqueous samples with a suitable solvent, such
as methylene chloride, could prove particularly
valuable for initial and rapid screening of
analytes. The extraction could be immediately
followed by GC/MS analysis which can
provide qualitative identification. However,
micro-LLE may not provide adequate detection
limits for lower concentrations which may
occur at the tailing edge of a contaminant slug.
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Continuous Liquid-Liquid Extraction
(Cont LLE)
This technique, as described in SW846-
Method 3520C, may be used for the isolation
and concentration of water insoluble and
slightly soluble organics. Its use can result in
excellent detection limits, although analysis
times can be long.
Solid-Phase Extraction (SPE)
Solid-phase extraction, sometimes referred
to as liquid-solid extraction (SW846-Method
3 53 5 A), is one of the techniques for basic
screening analysis. Like micro-LLE, SPE
extracts many contaminants, but can achieve
larger concentration factors compared with
the former technique. CIS adsorbents are
commonly used. Many other adsorbents can
also be employed to extract contaminants not
amenable to CIS adsorbents. Different elution
solvents can be used. A safety advantage
associated with SPE is that it produces few
aerosols.
Solid-Phase Microextraction (SPME)
SPME involves the use of a fiber coated with
sorbent material. The sorbent coated fiber is
exposed to either the aqueous sample or the
headspace from the sample, and the analytes
then adsorb to the coating on the fiber. After
exposure to the sample, the fiber is introduced
into the detection system (i.e., GC or HPLC).
For example, after exposure to the sample,
the SPME fiber is inserted into the injector of
a GC, and contaminants are released to the
column by thermal desorption. As with micro-
LLE, another quick screen, the detection limits
achievable via the use of SPME may only be
useful in the case of elevated contaminant
concentrations. Like SPE, SPME should
produce few aerosols.
Headspace Collection
The headspace above an aqueous sample may
be injected into a GC (SW846-Method 3810).
Commercially available equipment, interfaced
with the GC, is designed to facilitate this
analysis.
Flow Injection
In flow injection, an aqueous sample or sample
extract is injected directly into an LC/MS in
such a manner that it bypasses the LC column.
Thus the analytes are not chromatographically
separated, but the technique can prove useful
if high concentrations of a single analyte are
present, or if sample preparation is employed
that is selective for particular analytes.
6.2 Expanded Screening for Organic
Compounds - Detection Methods
In addition to the sample preparation
techniques described above, there are a number
of detection methods available for organic
chemical contaminants:
Gas Chromatography with Electron Impact
lonization Mass Spectrometry
The subsequent analysis of contaminants
extracted from wastewater may be conducted
by the use of GC/MS. When the mass
spectrometry is performed using electron
impact ionization, eluting peaks show
distinctive fragmentation patterns, which may
be used in identification, particularly through
the use of a variety of computerized tools
for library matching to ionization patterns
of known compounds. Usually, the program
performs a spectral search using a user-defined
library (such as National Institute of Standards
and Technology - NIST, EPA, Wiley, etc.) and
will report the compound with the best spectral
match as the tentatively identified compound
with an estimated concentration.
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It is desirable to examine the peaks for more
than just the analytes for which the instrument
is calibrated. The analyst may utilize a
threshold for examining unidentified peaks that
exceed 10% (height threshold) of the internal
standard.
Multidetector GC in Screening Mode
A multidetector GC is utilized for specific
analytes as an alternative, and sometimes
complement, to a mass spectrometer. The
intent of using multidetector GC in the analysis
of unknowns is primarily as a screening
tool. There are more than a dozen detectors
available including electron capture, infrared,
flame ionization, nitrogen-phosphorous
specific, thermal conductivity, etc. Various GC
detectors respond to contaminants in different
ways, and the evaluation of all the data from
the various detectors increases the selectivity,
and sometimes the sensitivity, of the analysis.
For example, flame ionization detectors
respond to a wide variety of contaminants,
but typically with low sensitivity. On the
other hand, electron capture detectors are
more sensitive and react more specifically to
halogenated compounds. The detectors may
be used in series with one GC, or in parallel
through the use of multiple GCs.
High Performance Liquid Chromatography-
Ultraviolet (UV) Detector
Analogous to multidetector GC, HPLC with
UV detection can be used to determine if
organic compounds not amenable to GC
procedures (e.g., non-volatiles or thermally
unstable compounds) are present in amounts
greater than background. Calibration and
quality control samples should be included
to provide accurate analysis. Analytical
confirmation may be necessary using
established techniques such as GC/MS,
although derivatization of the compounds may
be necessary to make them amenable to GC/
MS analysis.
High Performance Liquid Chromatography-
Mass Spectrometry (LC/MS)
Many polar hydrophilic compounds cannot
be easily extracted from an aqueous sample.
Additionally, there are contaminants of large
molecular weight (e.g., biotoxins) or thermally
unstable compounds that are not amenable to
GC analysis but can sometimes be analyzed
by LC/MS. Direct aqueous injection HPLC
allows analysis of a sample without extraction
or concentration. SPME and SPE (and other
extraction procedures) may be utilized for
compounds that can be extracted. Identification
of unknowns can be performed but there are no
standardized mass spectral libraries, as in GC/
MS. Analyst interpretation can help identify
possible compound fragments and structure.
More than a decade after its
commercialization, LC/MS is not commonly
used for water analysis, although it has proved
extremely useful for analysis of target analytes
in other industries. Nonetheless, LC/MS can
be an added tool in an expanded screen for
unknown chemicals in specific cases, and may
be useful for certain classes of pesticides, such
as carbamates.
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Tandem Mass Spectrometry (MS/MS)
Both GC and HPLC may be used in
conjunction with tandem mass spectrometry,
also known as MS/MS. Different MS/MS
instruments operate under different principles
to achieve similar results, but essentially
can be considered to be like two mass
spectrometers connected by a collision cell.
The first mass spectrometer separates ionized
molecules, which are broken apart in the
collision cell, and the resulting fragments are
separated in the second mass spectrometer.
This produces a great deal of information that
can be used to identify the original molecules,
but does not necessarily produce searchable
libraries. MS/MS is not as widely available as
MS and requires a high degree of skill.
High Resolution Mass Spectrometry
(HRMS)
GC or HPLC, combined with a high resolution
mass spectrometer, may provide exact mass
data of an eluting compound, allowing for
calculation of elemental composition of
both molecular and fragmentation ions. This
information is useful in the identification of
unknown organic compounds, especially when
the result of mass spectral library research
is not conclusive or when the standard of
a tentatively identified compound is not
available. Careful quality control procedures
are required, and the technique is not always
definitive, especially for unknown compounds,
because many compounds produce fragments
with the same exact masses.
Immunoassays
There are a number of immunoassay test
kits available for organic chemicals, such as
pesticides and biotoxins. These may be useful
for screening a sample for specific unknowns
in the field or in the laboratory. These kits may
be used for speed or if instrumental methods
are not available in the lab. However, use of
these kits requires that the goals of the analysis
be planned because some kits are slower
than the instruments, especially if analytical
confirmation time is considered. Also,
appropriate training is necessary in the use
of these tests. Laboratories should be aware
of the kits' reliability and levels of detection
before using them. It is important to note that
most of these test kits are not recognized by
any standard setting organization. Not all of
these products have been studied in detail
as to their efficacy for wastewater, which
may contain interfering and/or cross reacting
substances. These problems can lead to false
positive and false negative results. In general,
a positive or negative result from one of these
test kits should be considered tentative and be
confirmed through more rigorous laboratory
analysis.
6.3 Expanded Screening for Inorganic
Chemicals
The inorganic analyses include several
analytical techniques: classical wet chemistry;
instrumental techniques such as inductively
coupled plasma mass spectrometry (ICP-
MS), inductively coupled plasma atomic
emission spectrometry (ICP-AES), and atomic
absorption (AA) spectrometry for trace metals;
and ion chromatography for anionic and
cationic contaminants.
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Like the determination of organic chemicals,
there are a number of preparation steps that
are required for the analysis of inorganic
chemicals. These vary with the methodology
being employed. To select a sample
preparation approach, it may be useful to refer
to relevant standardized methods. For instance,
if the goal is to look for trace metals not listed
in a particular method, it may be useful to refer
to a method in which a wastewater sample
of similar composition to the one in question
is prepared for metal analysis. This is not an
exact process, and some metals have certain
characteristics that may cause them to not be
amenable to a preparation technique applicable
to another. For example, a digestion method
for nickel may not be suitable for mercury
analysis. Following preparation, the samples
can be analyzed by a number of techniques,
described below:
ICP-AES or ICP-MS in Semiquantitative
Mode
Analogous to multi-detector GC and UPLC
with UV detection, the ICP-AES and ICP-MS
methods (CWA Methods 200.7 and 200.8) can
also be expanded to provide a broad screening
approach to identifying unknown trace metals.
Under the semiquantitative mode, the ICP-MS
instrument, operated in scanning mode, may be
capable of providing semiquantitative results
for more than 60 elements including major
atomic cations, metals, semi-metals, rare earth
elements and selected radionuclides (uranium
and thorium). (Note: radioactive materials
should be handled by a specialized laboratory).
Ion Chromatography
Ion chromatography forms the basis of several
EPA methods to determine ions of regulatory
interest (e.g., CWAMethod 300.1). By the
correct choice of operating conditions and ion
chromatography columns, determination of
many different types of ions have appeared in
the literature.
Wet Chemistry
Wet chemistry forms the basis of many types
of chemical test kits. The chemistry and
detectors for test kits approved for compliance
monitoring are traceable to EPA methods.
Wet chemistry techniques, through the use
of autoanalyzers, form the basis of many
types of chemical analysis for environmental
and clinical applications. Manufacturers
of these devices often provide full detailed
methodology for defensible application of wet
chemistry to a variety of analytes. Titrimetric
methods are also available to analyze
background water quality parameters such as
alkalinity.
Ion Selective Electrodes (ISE)
Ion selective electrodes (ISE, also known
as electrochemical probes) can be utilized
to analyze for some background wastewater
quality parameters. A simple example of
an ISE is the familiar pH probe for the
hydrogen ion. Other ISEs are available for a
variety of ions and may be considered (e.g.,
ammonia, calcium, chloride, fluoride, nitrate,
potassium, silver, sodium, and sulfide). Some
parameters that can be monitored by ISEs
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may be useful in characterizing the extent of
contamination or verifying the credibility of a
contamination threat as part of the rapid field
testing of wastewater procedure during site
characterization.
6.4 Expanded Screening for Cyanides
Free cyanide concentration, measured without
distillation, is useful in detecting acutely
toxic cyanide. Therefore, distillation is not
used in the rapid field tests for cyanide or for
safety screening upon the receipt of samples
in the laboratory. Distillation is required for
determination of total cyanide concentration
and is the most conservative approach with
respect to public health concerns. Distillation
may be applicable for expanded cyanide
screening.
6.5 Expanded Screening for Biotoxins
Some biotoxins have been monitored routinely
for quite a while, particularly in conjunction
with naturally occurring outbreaks of biotoxins
in marine environments. There are hundreds
of biotoxins from dozens of different plant and
animal species. Analysis of some biotoxins
may be supported by the CDC Laboratory
Response Network (LRN) laboratories. The
LRN may utilize immunoassays for screening
for botulinum toxin, ricin, and some other
biotoxins.
Immunoassay kits are commercially available
for a number of biotoxins. It is important to
note that most of these kits are not recognized
by any standard setting organization, and
potential interferences and/or cross reacting
substances in wastewater are not well studied.
Because these tests are susceptible to false
positive and negative results, a positive or
negative result should be considered tentative
and should be confirmed through a more
rigorous laboratory analysis. Confirmatory
analyses usually involve GC/MS, LC, or LC/
MS. Because biotoxins tend to be very water
soluble, LC/MS may be particularly useful for
biotoxin analysis, although specialized sample
preparation techniques may be required. The
skill of the analyst is critical for this technique
to be used effectively.
6.6 Expanded Screening for Chemical
Weapons
The term chemical weapons refers to the
substances that appear on Schedule 1 of the
Chemical Weapons Convention. The Schedule
1 agents are extremely hazardous to handle
and most environmental chemistry laboratories
do not have the facilities or the procedures in
place to handle these agents. In addition, most
of the agents are not available commercially to
prepare analytical standards for quantification.
The chemical weapons agents will need
to be analyzed by special laboratories for
confirmatory analysis.
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In the unlikely event that an environmental
chemistry laboratory receives a sample
containing a chemical weapon, screening
techniques can be used to detect the presence
of the agents in wastewater. In addition, the
laboratory should notify appropriate ICS
personnel. The best analytical approach may
be to utilize the preparatory procedures for
organic chemical analysis described above
(direct injection, micro-LLE, SPE, SPME)
followed by GC/MS for identification. This
approach may only be able to determine the
presence, not concentration, of the agent
because an analytical standard would not
be available. The standard electron impact
mass spectral libraries frequently contain
mass spectra of these compounds and can be
used for tentative identification. As an aid to
increasing confidence in chemical warfare
agents' GC/MS library matches, the NIST
has developed the Automated Mass Spectral
Deconvolution and Identification System
(AMDIS) (http://chemdata.nist.gov/mass-spc/
amdis/).
In the unlikely event that chemical weapons
agents are present, the expanded screen for
organic chemicals is procedurally designed to
reduce risk to personnel handling the sample,
namely through reduction of aerosols. As
with any organic chemical, an additional
way to reduce risk would be through sample
dilution. The laboratory may first start with
the most dilute sample (1/1,000) and if
nothing is detected may proceed to analyze
the next dilution (1/100), followed by the 1/10
dilution, and lastly the undiluted sample. If
the laboratory proceeds through the undiluted
sample and nothing is detected, it may be that
the sample is a non-detect for the chemical
weapon that would be captured by the screen.
If chemical weapons agents are identified
in the screen, proper notifications should
be made to the Incident Commander or
appropriate official within the ICS structure.
Also notify law enforcement who may be able
to gain access to laboratory resources that can
confirm the presence of the chemical weapons
agent. EPA is developing the capability and
capacity at seven fixed laboratories and two
mobile laboratories to analyze environmental
samples potentially contaminated with
chemical warfare agents and degradents. Other
notifications may be required by applicable
laws and regulations.
6.7 Basic and Expanded Screening for
Radionuclides
Screening for radionuclides is somewhat
different than screening for other chemical
contaminants since radionuclides can be
characterized by both the type of radiation they
emit as well as their exact chemical identity.
Accordingly, initial screening for radionuclides
may involve measurement of gross
radioactivity. However, any initial screening
that indicates the presence of a radionuclide
should be followed by analytical confirmation
of the chemical identity. A schematic for
radionuclide screening is shown in Figure 4-5.
The results of field testing for radioactivity
should be compared to background levels to
determine whether the site may have been
contaminated with radioactive material.
The analysis for gross alpha and beta radiation
may be conducted as a screening method for
alpha and beta particle activities in wastewater
and used to determine if specific radiological
analyses are needed. Preliminary analysis can
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THREAT;
Preform Field Testing
for Radioactivity
r *»
<-'" Field test positive for~
*.__ radioactivity? __,.*'
Does lab policy **»v
require screening for
-.^ radioactivity? ,-''
Preform laboratory
screening analysis for
alpha, beta and gamma
radiation
Screening results ''T
positive? j.-''
V
Additional analysis for
radionuclides not
required
Preform analysis for
specific radionuclides
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first be conducted in the field using appropriate
field portable or hand-held devices, but may
be verified in the laboratory. As part of their
safety plan, laboratories may wish to screen
samples upon arrival for gamma radiation
using appropriate technologies such as hand
held detectors.
If the presence of radioactive material is
indicated by the initial screening, specific
radioisotopes may be determined by
radiochemical specific procedures, using
techniques with which radiation labs are
already familiar. These procedures often
involve separation of the radionuclide from
the sample by precipitation techniques, and
subsequent determination by a gas flow
proportional counting system or scintillation
detector system for alpha and beta emitters
and an appropriate gamma detector for gamma
emitters. For example, strontium-89 and
strontium-90 can be precipitated as carbonates
from the sample. Additional precipitation steps
allow separation from other radionuclides and
interferences.
Due to the unique nature of radionuclide
analysis, some laboratories have developed in-
house procedures for radionuclide analysis that
make use of their special skills and capabilities
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to enhance the speed of analysis, especially
since some standardized methods are not
rapid methods. For example, one standardized
method for radioactive strontium in water
recommends a two-week in-growth period
for obtaining the yttrium isotope from the
purified strontium. Modification of the method
produces much faster results. Reduction
in analysis time could be accomplished by
measuring the total amount of an element's
radionuclide, not the isotopic distribution.
Also, for some isotopes, faster results may be
obtained by simply reducing the volume of
water processed.
It must be emphasized that radiochemical
analysis should be performed only by licensed,
specialty laboratories, and the need for such
analysis should be indicated by the field
screening equipment for alpha, beta, and
gamma emitters, or other specifics of the
incident, such as threats.
As described above, the basic screen is
rather comprehensive because it requires
identification of the specific radionuclide if
indicated by the screens for gross alpha, beta,
and gamma radiation. Therefore, the expanded
screen is designed to capture radionuclides that
do not fall into the energy range of the gross
radionuclide screen for gross alpha and beta.
Fortunately, these radionuclides have specific
standardized methods designed for their
analysis, and radionuclide labs may also have
additional reliable methods at their disposal for
their analysis.
Two other techniques that may be particularly
useful for radionuclide analysis are gamma
spectroscopy, which can directly identify the
gamma emitting radionuclide, and inductively
coupled plasma mass spectrometry (ICP-MS).
Principal considerations in the use of both of
these techniques include detection limits and
availability of instrumentation.
7 Additional Recommendations
for Chemical Screening of
Wastewater Samples
Unlike drinking water analysis, wastewater
analysis is complicated by the high solids
content of samples. This is especially true for
raw sewage as well as primary effluent and
mixed liquor from the wastewater treatment
process. Solids residue is much less of a factor
in secondary or tertiary effluent from the
treatment chain.
The following practical observations and
suggestions may help to overcome the
analytical challenges posed by the difficult
wastewater matrix:
The purge and trap extraction/
concentration method can be utilized
without modification to introduce volatile
organic compounds into a GC or GC/MS.
Because the sample itself does not come
into contact with the sensitive components
of the analytical system, there should be
no fouling potential for the GC or GC/MS
even when raw sewage, primary effluent,
or mixed liquor samples are analyzed.
Solid phase extraction can be used directly
on secondary or tertiary effluent samples.
The extract can then be analyzed by GC,
GC/MS, or other appropriate techniques.
When screening raw sewage, mixed
liquor, and primary effluent samples, the
samples can be filtered through a 0.45um
membrane filter to remove residue. The
filtrate can then be extracted by solid
phase extraction and the extract analyzed
by HPLC, GC, GC/MS, or other methods.
The filter retentate from the step above can
also be digested via Soxhlet extraction
using SW-846 methods 3540C or 3541. If
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necessary, the extract can subsequently be
purified using a gel-permeation clean-up
method such as SW-846 method 3640A.
The product of this preparatory step can
then be analyzed using GC, GC/MS, or
other techniques.
8 Screening for Microbiologicals
Including Unknowns
Wastewater typically contains large numbers of
viruses, bacteria, and protozoans. Additional
microbes are seeded into wastewater during
the secondary treatment process, and are
encouraged to multiply to assist in the
breakdown of organic matter and nutrients.
Even finished effluent from wastewater
treatment plants may contain significant
numbers of microorganisms. The chlorination
or UV light treatment that occurs at the
end of the wastewater treatment process is
intended to control pathogens and reduce
microbial numbers, but does not produce
sterile water. Furthermore, the likely routes
of exposure of utility workers or the general
public to microbes that may have been added
to wastewater accidentally or intentionally is
through inhalation of aerosols and perhaps
limited dermal contact, as opposed to
ingestion. Consequently, there is much less
emphasis placed on screening for microbial
contaminants in wastewater during a suspected
contamination event compared to a drinking
water contamination incident.
Possible exceptions may include microbes such
as the anthrax bacterium, Bacillus anthracis,
whose spores could pose an inhalation risk
if they ended up in the wastewater system.
Various parts of the wastewater collection and
treatment systems generate aerosols that may
potentially impact health via the inhalation
route. Still another situation where the need
may arise to analyze wastewater for the
presence of microbial contaminants might be
if the decision is made by officials to discharge
to or bypass the wastewater treatment plant,
following an intentional or unintentional
biological contamination incident, allowing
elevated numbers of potentially harmful
microbial contaminants to enter natural
waterways if such discharge or bypass is
not otherwise prohibited by CWA Section
301(f), 40 CFR 122.41(m), or another law or
regulation.
Analysis of wastewater for specific bacterial,
viral, or protozoal contaminants is complicated
by high
background levels
of microbes
in wastewater.
Additionally,
efforts to
concentrate
wastewater
samples for
microbial analysis are complicated by the high
solids content of wastewater.
For all of these reasons, an extensive screening
procedure is not recommended at this time
for microbes in wastewater following a
contamination threat or incident. Should the
need for detailed microbial analysis arise, an
attempt may be made to screen wastewater
samples using molecular techniques (e.g.,
Polmerase Chain Reaction - PCR) or
traditional culture methods. In the event that
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select biological agents (such as anthrax spores
or the biotoxins ricin or botulinum toxin) are
believed to be involved in a contamination
incident, samples may be analyzed by the
Centers for Disease Control and Prevention's
laboratory since they are authorized to work
with these microbes.
9 Forensic Implications of Sample
Collection and Analysis
It is important to note that if a contamination
event in wastewater is the result of an
intentional or accidental release, there will
likely be legal ramifications. Any samples
collected and analysis conducted during the
incident response may ultimately be used for
evidentiary purposes. Therefore, sampling
and analytical procedures should be accorded
greater attention to detail.
10 Data Analysis and Reporting
The responsibility of the laboratory during an
emergency does not end with sample analysis.
At a minimum, the lab should report the
results in a timely manner to the recipients
designated by incident command. Additionally,
the laboratory may be asked to assist in
the analysis and interpretation of the data.
The Water Laboratory Alliance - Response
Plan has suggestions for the maintenance
and reporting of data. The following are
some general guidelines for the analysis and
reporting of results:
The laboratory and the client (e.g., the
Utility Incident Commander or the overall
Incident Commander) should agree on the
format and content of the report before
data are released by the lab. In general, the
report should be thorough enough so that
all information is available. However, if
too much detailed information is reported,
the laboratory may confuse the client.
During a suspected contamination
incident, it is important that all
relevant information be managed
through incident command. Therefore,
analytical results should be reported
only to those individuals designated by
incident command, and it will be their
responsibility to subsequently inform
other stakeholders.
In a crisis situation, the laboratory may
be asked to provide tentative results
(sometimes called a rolling report) prior
to complete data review and confirmation.
In this case the lab may need to provide
appropriate caveats regarding the validity
of the data at that stage of the analysis.
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The laboratory should remain available
to assist in the analysis and interpretation
of both preliminary and final results. The
laboratory staff has a unique perspective
regarding the reliability of the methods
and interpretation of results.
11 Summary
The response to the threat of an intentional or
accidental contamination event in wastewater
often necessitates sample collection and
analysis. The analytical response will
begin at a fairly basic level with rapid
testing of wastewater in the field during the
site characterization process. Should the
contamination threat be deemed 'Credible',
definitive analyses will need to be conducted
in one or more laboratories. An important
challenge to labs analyzing such samples is the
potential risk to personnel handling samples
which may contain potentially hazardous
substances. Another challenge is accurately
detecting, identifying, and quantifying one or
more contaminants from the array of thousands
of chemical, microbes, and radionuclides that
could accidentally or intentionally end up in a
wastewater collection or treatment system.
Module 4 discusses safety procedures that
should be employed to protect the analysts.
It also recommends general approaches
that could be used to begin the process of
eliminating possible contaminants and target
the agent that is actually present. In the case
of many contaminants, a variety of both
standardized and exploratory techniques may
need to be utilized.
The Module emphasizes the need for utility,
government, and commercial laboratories to
prepare their own Laboratory Guides, follow
emergency procedures contained in the Water
Laboratory Alliance -Response Plan, and
prepare site-specific analytical approaches
based on the recommendations provided in the
Wastewater Response Protocol Toolbox.
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