United States
  Environmental Protection
Wastewater Response Protocol
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
  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
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
                        Wastewater Response Protocol Toolbox


                                     Planning and Preparat
                            ion )
                                          Threat Warning
                                      Initial Threat Evaluation



         Immediate Operational
           Response Actions
                                       Site Characterization
                                          and Sampling
                                           Public Health
                                         Response Actions
                                            Is Incident
                                    Remediation and Recovery
                                Wastewater Response Protocol Toolbox

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
                        Wastewater Response Protocol Toolbox


   contamination involving chemical, biological
   and radiochemical contaminants. For more
   information, visit http://www.epa.gov/erln/

   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.

   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/

                     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.
Wastewater Response Protocol Toolbox

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
Chemistry Labs
Microbiology Labs
  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.

                         Wastewater Response Protocol Toolbox

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

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://
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/

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
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
                        Wastewater Response Protocol Toolbox

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

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_

Additionally, there is health and safety
suggestions contained in various government
publications including Biosafety in
                         Wastewater Response Protocol Toolbox


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.

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

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
                         Wastewater Response Protocol Toolbox

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
4 Analytical Approach for
  Unidentified  Contaminants in

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

  • 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.
                        Wastewater Response Protocol Toolbox

   • 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

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.
                        Wastewater Response Protocol Toolbox

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

                        Wastewater Response Protocol Toolbox

  Table 4-1: Sources of Standardized Methods

             Water Contaminant
             Information Tool
              EPA SW-846 methods
              40 CFR Parts 136 and
   Method Index
                       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
                       Promulgated list of
                       defensible methods widely
                       accepted in the analytical
                       community for water and
On-line database containing
chemical, microbiological,
biological, toxicity, and
physical methods for
                        US EPA Office
                        of Water
                        US EPA Office
                        of Solid Waste
                        US EPA Office
                        of Resource
                        and Recovery
                        and US EPA
                        Office of Water
                                                          US Geological
                                                          Survey and
                                                          US EPA
      How to obtain
http //www.epa.gov/wcit
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

                      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
                         Wastewater Response Protocol Toolbox

  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
Trace metals (inorganic)
Total mercury
(inorganic, includes

Purge-and-trap PID/ELCD
Purge-and-trap GC/MS
Solid-phase extraction GC/
graphite furnace AA
Cold vapor AA

Wet chemistry
Gross alpha, gross beta,
gross gamma
802 1B
747 1B

901 2 A





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
                         Wastewater Response Protocol Toolbox

            Table 4-3: Analyte Lists Corresponding to Table 4-2
1 1,1,2-TetrachlorQethane
1 . 1 .2,2-Tetrachloroethane
1,1 ,2-Trichloroethane
1 ,1-Dichloropropene
1 ,2,3-Trichlorobenzene
1 ,2,4-TrichIorobenzene
1 ,2-Dibrorno-3-chtoropropane
1 ,2"Dibromoeth3ne
1 ,2-Dichlorobenzene
1 ,2-Dichloroethane
1 ,2-Dichloropfopane
1 ,3,5-Trimethylbenzene
1 ,4-Oichlorobenzene
2.2-Dichloro propane
2- Nitro propane
Ally I chloride
Bromo benzene
Bromochlorome thane
2,2' 3'.4,6-Pentachlorobiphenyl
Azinphos methyl
Benz(a ) a nth ra cene





Wastewater Response Protocol Toolbox

Table 4-3 (cont.): Analyte Lists Corresponding to Table 4-2
Butyl chloride
Carbon disulfide
Carbon tetrachloride
Cis- 1 ,2-Dichtoroethene
Di ethyl elder
Ethyl methaerylate
Ethy tbenzene
Melhanol (solvent)
Methyt acryiate
Methyl methacrylate
Methyl tert-butyl ether
Methylene chloride
Demeton (mixed isomers)
Dibenz(a , h)anthracene
Diethyl phthalate
Dimethyl phthalate
Di-n-butyl phthalate
Endosulfan I
Endosulfao II
Endosulfan sulfate
Endrin aldehyde
Perth ion
                         Wastewater Response Protocol Toolbox

          Table 4-3 (cont.): Analyte Lists Corresponding to Table 4-2
p-lsopropy (toluene
trans-1 ,2-Dichloroethene
trans-1 ,3-Dichloropropene
trans-1 ,4-Dichloro-2-butene
Vinyl chloride

Heptachlor epoxide
(Isomer B)
lndeno{1 ,2,3-cd)pyrene
Methyl parathion
Wastewater Response Protocol Toolbox

6 Expanded Screening for

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

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
Chemical Warfare Agents
GC, GC/MS, HPLC, LC/MS, Immunoassay test kits
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
                        Wastewater Response Protocol Toolbox

6.1 Expanded Screening for Organic
   Compounds - Sample Preparation

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

Regardless of the detector system employed,
there are a number of widely used sample
preparation techniques. These include the

Large Volume Liquid/Liquid Extraction

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
Figure 4-4. Lab Personnel Using Syringe to
Inject GC.

Micro Liquid-Liquid Extraction

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.
                        Wastewater Response Protocol Toolbox

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

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

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.

                        Wastewater Response Protocol Toolbox

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

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.
                        Wastewater Response Protocol Toolbox

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

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.


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
6.3 Expanded Screening for Inorganic

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.
                        Wastewater Response Protocol Toolbox

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

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

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
                        Wastewater Response Protocol Toolbox

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

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
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
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

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.
                         Wastewater Response Protocol Toolbox

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/
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

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
                        Wastewater Response Protocol Toolbox

                   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
                    Screening results ''T
                       positive?  j.-''
           Additional analysis for
             radionuclides not
                  Preform analysis for
                 specific radionuclides
            Figure 4-5: Protocol for Basic Radionuclide Screening
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

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
                          Wastewater Response Protocol Toolbox


    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
Wastewater Response Protocol Toolbox

    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

Analysis of wastewater for specific bacterial,
viral, or protozoal contaminants is complicated
by high
background levels
of microbes
in wastewater.
efforts to
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
                        Wastewater Response Protocol Toolbox

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.
                        Wastewater Response Protocol Toolbox

   • 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.

                         Wastewater Response Protocol Toolbox

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