FREQUENTLY ASKED QUESTIONS.

This page was last updated April 04, 2005.

Click on any of the following 29 questions to proceed directly to the question.

Q1: Does the CAPE Technologies dioxin kit have regulatory approval?

Q2: Aren’t immunoassays for hazardous wastes a new and untested technology?

Q3: What is the “Triad Approach” to hazardous waste site assessment and
        remediation, and what does Method 4025 have to do with it?

Q4: What do I gain by using Method 4025 instead of Method 8290?

Q5: If I don’t want to use the kit myself, can CAPE Technologies analyze my
       samples for me? 

Q6: What exactly was Method 4025 intended for?

Q7: What matrices can be analyzed by Method 4025?

Q8: The matrix I want to analyze is not listed.  Can I still use the test?

Q9: The EPA web site says Method 4025 is for screening at 500 ppt. 
       
Is that all I can do with this kit?

Q10: How does this test work?

Q11: What kind of result does the kit give?

Q12: How long does it take to get results?

Q13: How many samples can be analyzed in a day?

Q14: Is Method 4025 just for yes/no results or can I generate quantitative data?

Q15: Can I get individual congener values for my samples?

Q16: What compounds does the kit detect?

Q17: What is the detection limit of the kit?

Q18: How reproducible are the kit results?

Q19: How large a sample is needed for immunoassay analysis?

Q20: How much sample cleanup is needed for immunoassay analysis?

Q21:   What kind of facility is needed for this method?  Can I run this test in 
           the field?

Q22: What must I buy to get started and how much will it cost?

Q23: How much does it cost per sample?

Q24: How soon can I get started?

Q25: How complicated is the test protocol?

Q26: How much training is required?

Q27: What if I need help while I am using the kit?

Q28: Why isn't this kit as easy to use as the PCB and other immunoassay kits 
         I have used before?

Q29: Does this test have problems with mass labeled internal standards?


Q1: Does the CAPE
Technologies dioxin kit have regulatory approval?

A1: Yes.  In 2001 the US EPA Office of Solid Waste and Emergency Response
(OSWER) accepted Method 4025 into the SW-846 Compendium of Solid Waste
Methods.  Method  4025 was developed around and is based on the CAPE
Technologies DF1 Dioxin/Furan Immunoassay Kit.  This kit is the only
commercially available immunoassay product accepted by the US EPA for dioxin
analysis.  Method 4025 is posted on the new methods section of the
EPA OSWER SW-846 web site 


Q2: Aren’t immunoassays for hazardous wastes a new and untested
technology?

A2: Definitely not.  The 4000 series of solid waste screening methods  within
SW-846 includes methods such as for PCBs, PAHs, and petroleum fuels.  These
methods were approved by the US EPA in the early 1990s and have been used
widely since then.  The CAPE Technologies DF1 Immunoassay is very similar in
principle
to the other 4000 series immunoassays and has been in commercial use
since 1997, receiving US EPA acceptance in 2001 for use in Method 4025.  CAPE
Technologies Technical Reference
TR-005 (scroll down the document to get to the
beginning of the text) describes the EPA-OSWER program for evaluation and
approval of immunoassay and related methods
in the 4000 series.  The US EPA 
has embraced a concept called the
“TRIAD approach” to hazardous waste site
assessment and remediation, implementation of which explicitly requires
rapid
analytical methods such as the 4000 series immunoassays.



Q3: What is the “Triad Approach” to hazardous waste site assessment and
       remediation, and what does Method 4025 have to do with it?

A3: The “Triad Approach” is a systematic approach to streamlining the handling
of waste sites.  The strategy aims to reduce overall costs and accelerate site
closures by circumventing some of the bottlenecks in the conventional approach
to site closure.
Triad’s three parts are:
          1) systematic and flexible planning;
          2) on-site measurement technologies;
          3) dynamic workplans. 

The US EPA is currently trying to educate hazardous waste site stakeholders 
about the benefits of this approach.  From the
EPA fact sheet on Triad : “The US
EPA supports the
adoption of streamlined approaches to sampling, analysis, and
data management activities conducted during site assessment, characterization, 
and cleanup.  This position reflects the growing
trend towards using smarter,
faster, and better technologies and work strategies.”  The report further states: 
“If used correctly,
innovative rapid-turnaround field analytical and software
tools
coupled with on-site decision making can significantly condense a project’s
overall budget and lifetime, while significantly
increasing the likelihood that the
gathered data will guide transparent decisions.”

Detailed information about Triad is available from the EPA at website
 
http://clu-in.com/triad .  Of particular note are two recent articles published by 
staff from the EPA Office of Technology Innovation in the journals
Remediation and Quality Assurance


Q4: What do I gain by using Method 4025 instead of Method 8290?

A4: First, Method 4025 should not be considered a replacement for
Method 8290 (the “conventional” dioxin method), but rather should be viewed
as a complementary method.  Method 4025 does things which Method 8290 
cannot and vice versa.  Both methods are valuable tools to the analytical 
chemist and site
 manager. 
The advantages of Method 4025 include:
     lower cost;
     higher sample throughput;
     faster turnaround time;
     and on site analysis.

Most importantly, Method 4025 can allow you to utilize the
Triad approach
described by the US EPA
(EPA fact sheet on Triad)) in ways that would not be
possible using
only Method 8290.  The greatest benefit to you depends on your
individual situation.  Reduced analytical costs can be very important, but there 
are many other factors to consider

For soil remediation, the decrease in heavy equipment costs because of real time
screening may be vastly greater than the
reduction in analytical costs. 
For site assesment and mapping
, the greatest benefit may be in completing the 
work in a few weeks rather than several months.
For complex sites
, the biggest advantage may be in being able to develop a map
with high enough spatial resolution to show the complexity of dioxin distribution. 
A recent large scale implementation of Method 4025 is described  in CAPE
Technologies Technical Reference
TR-006 . 


Q5: If I don’t want to use the kit myself, can CAPE Technologies analyze my
       samples for me? 

A5: Yes.  As noted in our description of Products & Services and our
Analytical Options Matrix
, we also offer analytical services
based on our 
own
products.  We can also provide rapid turnaround to support rapid on-site
decision
making.   In many cases, results can be reported by phone or email 
the same
day your samples are received in our lab.  
Contact CAPE Technologies directly to discuss specifics.


Q6: What exactly was Method 4025 intended for?

A6: The original design of Method 4025 was to provide semi-quantitative 
screening analysis in soil at 500 ppt, as described in detail in Application Note
AN-007 .  A modification of the sample preparation protocol portion of Method
4025 (called modified Method 4025 or 4025m)
has been validated for quantitative
soil analysis, as described in Application Note
AN-008 .  Other modifications of 
this same
sample preparation protocol expand the scope of the CAPE 
Technologies  DF1 Dioxin/Furan Immunoassay Kit even further.


Q7: What matrices can be analyzed by Method 4025?

A7: At CAPE Technologies, we use Method 4025 and modifications of Method 
4025 to analyze soils, sediments, water, surface wipes, pulverized concrete,
hydrocarbon oils, food oils, feed additives, and a variety of other matrices.  
If you want to use Method 4025 or 4025m in your lab, two Application Notes
describing soil analysis are available, as noted in the previous answer.  
Applications for other matrices have not yet been released for use by customers,
but we will work with you to bring these methods into your lab if you wish.
Contact CAPE Technologies for more detail.


Q8: The matrix I want to analyze is not listed.  Can I still use the test?

A8: Possibly.  If we have no in house application that can be transferred to you, 
we will
work with you to develop the validation data necessary to support the use
of the kit for
your matrix.  
Contact CAPE Technologies  for assistance.


Q9: The EPA web site says Method 4025 is for screening at 500 ppt. 
      
Is that all I can do with this kit?

A9: Definitely not.  The target for the initial EPA Method 4025 validation was
selected by the EPA as a practical level for many sites of interest (and also near
the detection limit of Method 8280,
the low resolution GC-MS method for dioxin). 

The EPA 4000 Series of methods allows kit manufacturers to update methods
without going through the validation process again, as long
as the changes are 
minor and claims of kit performance are supported by reliable data.  
This conforms to the Performance
Based Measurement Systems (PBMS) concept
supported by EPA OSWER
since the early 1990s and summarized in section 6.3 
of CAPE
Technologies Technical Reference
TR-005 .

Based on this policy, we have provided data using the DF1 Dioxin/Furan
Immunoassay Kit with a more rigorous cleanup.  These data, in Application Note
AN-008 , demonstrate quantitative correlation between Methods 4025m and 8290
in the low to mid-ppt range. 

Variations of this cleanup, which is a simpler and much faster version of that
used for Method 8290, have been used at CAPE Technologies
to analyze a wide
variety of sample matrices, in some cases as low as 0.5 ppt in solids.  PBMS
dictates that the analyst provide QA data to validate performance claims.  
Such QA is a routine part of our analytical services.  In addition, our kit pricing
and cost per sample estimates are based on the assumption that 40% of each
immunoassay
kit will be used for standards and QA samples.


Q10: How does this test work?

A10: Several helpful schematic diagrams are given in the section on
Immunoassay Background
.  These provide background
information on
immunoassay
in general, plus a schematic depiction
of the DF1 test.  A succinct
text summary of the
principles behind the DF1 kit is given in Section C of the kit
insert
-- scroll down to Section C of 
IN-DF1.


Q11: What kind of result does the kit give?

A11: The kit is designed to correlate with the toxic equivalent concentration 
(TEQ)
contributed by polychlorinated dibenzodioxins and dibenzofurans
(PCDD/Fs).  The DF1
Dioxin/Furan Immunoassay correlates with TEQ because
response correlates with
toxicity of the individual congeners.  Dioxin/furan 
analysis by Method 8290 arrives
at the same TEQ number, which is the value
most commonly
used for regulatory purposes.


Q12: How long does it take to get results?

A12: For samples analyzed in your lab, sample preparation time is the key 
variable and
may vary among applications.  Same day results
are possible for the
fastest sample
preparation methods, while other applications may require 24 hours.
Consult your
specific Application
Note or Contact CAPE Technologies for more
specific information.


The flexibility of the DF1 Dioxin/Furan Immunoassay also allows the design of
methods
suited to specific situations.  For example, an engineering firm testing a
pilot scale
dioxin destruction system requested a same day method for  screening
hydrocarbon oils
at 5 ppb.  By modification of both cleanup and immunoassay
protocol, they were able to
generate results in 4 hours.  For certain samples,
coupling of the immunoassay to
faster extraction methods, such as supercritical
fluid extraction or
accelerated solvent extraction, could reduce the analytical 
cycle time
significantly.


For samples analyzed by CAPE Technologies, standard turnaround is 10 
business days from receipt of samples.  Rush turnaround (50% premium) is
5 business days from receipt of samples.  Extra rush turnaround (100% 
premium) is 2 business days from receipt of samples.  Reports are typically
delivered by email, with hard copy backup if desired.


Q13: How many samples can be analyzed in a day?

A13: Batch processing capability is an important part of the DF1 kit.  Sample
throughput
depends on analyst skill and experience, as well as the sample
preparation protocol used
and the number of quality
assurance samples.

The customer cited in Technical Reference
TR-006 analyzed
as many as 40
samples per day (24 hour turnaround time), based on about 12 person-hours per
day.  Their total for the project was 1100 samples in about 8 weeks, based on 
4 batches per week.  They used no automated equipment and performed Method
4025 as described in Application Note
AN-007 in their own laboratory.

The study summarized in Application Note AN-008 utilized the split approach 
under Option 2 of the
Analytical Options Matrix .  The rate per day was 10-15
samples, broken down as follows.  The customer laboratory extracted 234 
samples and reduced the extracts to paraffin oil, requiring about 5 person days.
These samples were
shipped to CAPE Technologies, put through the coupled
column cleanup of
AN-008 , and analyzed by immunoassay, along with all the QA
samples noted in Table 1 of
AN-008 .  The extract cleanup
and analysis, including
dilutions for quantitation of high samples, required about 4 person-weeks.


Q14: Is Method 4025 just for yes/no results or can I generate quantitative data?

A14: Method 4025 was designed for making semi-quantitative decisions, but it is
possible to develop quantitative results using Method 4025m, as described in
AN-008 . 
For accurate absolute quantitation, you must use a set of 4025m-8290 comparison
data to verify the EIA calibration.  You must also include sufficient QA samples 
to determine that extraction recoveries and cleanup recoveries are acceptable.

For either semi-quantitative or quantitative analysis, CAPE Technologies  
provides
a downloadable Excel spreadsheet
tool, Calculation Module C . 
This tool performs
sigmoid curve fitting and calculation of
sample 
concentrations, so there is no need for
you to purchase additional
software.


Q15: Can I get individual congener values for my samples?

A15: No. The test result is a single value which is the sum of responses to all the
individual congeners.



Q16: What compounds does the kit detect?

A16: Essentially only toxic dioxin and furan congeners.  Detection is structure
based, as noted in Answer 10 above.  Specificity data for individual congeners are
given in Table 2 of the Kit Insert
-- scroll to Table 2 in pdf document
IN-DF1. 
Strong
recognition by the DF1 Dioxin/Furan Immunoassay generally requires 
both the dioxin/furan core structure
and the 2378- chlorination pattern.  For
example, test response to non-toxic PCDD/Fs
is greatly reduced because of 
their deviation from this chlorination pattern. 
Recognition
of the PCBs most similar to 2378-TCDD (PCBs 77 and 126) is 
very limited because the core structure to which the chlorines are attached
is not
a dioxin or furan. 
When both core structure and chlorination
pattern are changed, as for PCB 153,
there
is no detectable recognition.


Additionally, the carbon column based cleanup procedure described in
Application Note
AN-008 retains only the tetra and higher dioxin and furan
congeners,
discarding the trichloro- and lower dioxins and furans, as well as
the PCB’s.



Q17: What is the detection limit of the kit?

A17: The detection limit is 4 pg of 2378-TCDD, which is sufficient for ppt to ppb
level analysis.  The best accuracy and precision are obtained with 10-20 pg of
2378-TCDD.  More complete response data are given in
Table 1 of the Kit Insert
(scroll to Table 1 in IN-DF1 ) and in graphic form as the starting values in
Calculation Module C .
 
Detection
limits  in the original sample will vary by matrix, the matrix load 
(amount
of sample introduced into the immunoassay), and the degree of sample
cleanup used.
Low to mid pg/g levels for soils are
straightforward, as shown by Application 
Note
AN-008 .  A simple
and minor modification of that protocol allows for soil
analysis
below 10 pg/g.  Some food matrices are done easily at levels a
s low as
0.5 pg/g.  Clean water with low particulate loads, such as finished drinking water,
can be analyzed
as low as 30 pg/L using a 2 to 4 L sample.


Q18: How reproducible are the kit results?

A18: Table 1 of the Kit Insert (scroll down to Table 1 of IN-DF1) of the kit 
insert
includes standard deviations for each data point which indicate excellent
reproducibility.
  The data tables in Application Notes AN-007  and AN-008
provide quantitative accuracy and precision data from analysis of real soil 
samples. 



Q19: How large a sample is needed for immunoassay analysis?

A19: Solid sample sizes are generally 5 g, but the actual analysis may require 
only
a small fraction of this or possibly more than this, depending on the target
sensitivity.
 
Section
s I-2 and I-3 (scroll down to Sections I-2 and I-3 of the Kit Insert
IN-DF1 ) describe how to determine the amount of sample to use for your
application.



Q20: How much sample cleanup is needed for immunoassay analysis?

A20: Because of the intrinsic specificity of the immunoassay, less cleanup is
required than
for Method 8290, but the exact amount depends primarily on the
target sensitivity.  Generally, decision levels at or above 1 ppb TEQ can be done
with only a simple liquid
oxidation of the extract such as in Application Note 

AN-007
.  Lower decision levels can be reached using a simple one step coupled
column cleanup such as in Application Note
AN-008 .


Q21:   What kind of facility is needed for this method? Can I run this test in 
           the field?

A21:  Electricity and a temperature controlled workspace are required, but 
running water is not.  If your lab (fixed or field) has the equipment noted in the
following answer, you will be able to
perform the test.



Q22: What must I buy to get started and how much will it cost?

A22: The minimal equipment required to perform the immunoassay procedure
is described in Equipment List
EL-001 and costs less than $2000 total. 
The equipment which
may be required to perform the sample preparation
is
described in Equipment List
EL-002 . It is best to look at specific Application 
Notes, such as
AN-007 and AN-008, which contain lists of required supplies and
equipment. 


Cost of equipment for sample preparation will vary according to the application, 
but is
still quite modest and may already be covered by your present equipment.
If you choose
option 2 or 3 from the
Analytical Options Matrix  (CAPE
Technologies does some or all
of the work), these requirements will be reduced
accordingly.

The two sizes of immunoassay kit starter package kit described in  
Products & Services
  are available to all first time customers at reduced prices.
The
large starter package (
described in document ML-DF1-ST-B )  provides
enough immunoassay materials for training, with enough left over to analyze
30 
to 50 samples.  The total expenditure for 
equipment and the first kit can be as l
ittle
as the cost of analyzing 4 or 5 samples by EPA Method 8290.



Q23: How much does it cost per sample?

A23: It varies, but is always a small fraction of the cost of Method 8290. 
The
Analytical Options Matrix gives approximate prices for three different 
options: 
- if you analyze the samples;
- if CAPE
Technologies analyzes the samples; 
- and if the work is split.

Exact costs may vary by geographic region, sample cleanup requirements, and
other factors such as the proportion of immunoassay kit materials used for 
quality assurance.



Q24: How soon can I get started?

A24: If you are sending your samples to CAPE Technologies for us to analyze, 
we may
be able to provide results the same day we receive the samples.  If timing
is critical it
is best to
contact us directly to discuss specific details.

If you are doing the analysis, CAPE Technologies can generally ship kits within 
two business days, often the same or next day.  One to four weeks of lead time
may be needed for the required photometer equipment (
see
  EL-001 ), which is
purchased from
an outside vendor.  This potential delay can be avoided by  
renting a photometer from CAPE Technologies.



Q25: How complicated is the test protocol?

A25: Not very complicated.  Our most widely used sample preparation methods
are described in detail
and summary form in AN-007 and AN-008 .  The protocol
for the immunoassay is summarized in the
Protocol Summary (within document
PS-DF1) and
described in detail in the Section J of the kit insert (scroll down to
Section J of the
document IN-DF1 ).


Q26: How much training is required?

A26: Not much.  Some new users with non-immunoassay analytical experience
have
mastered the technique with only telephone assistance.  Most new users can
begin
producing useful data within the first 1 to 3 runs (one half or less of a large
kit).  Training
at your facility or ours is possible at additional cost.  

Note: If you have PowerPoint software on your computer, you can view the online
 
AN-008 training presentation.   This PowerPoint presentation is 7.7 MB and will
take approximately one to two minutes to download for those who have broad-band
connection to the Internet.  Please note that some browsers will allow download
only, in which case download the presentation, and then play it with your 
PowerPoint software.


For more information  Contact CAPE Technologies 


Q27: What if I need help while I am using the kit?

A27: If the answer to your problem or question can not be found in our
extensive
web site
material or printed literature, contact us by
email or other means . 
CAPE Technologies does not just manufacture kits.  We have extensive 
experience in
applications development and technical service, including kit
troubleshooting.  Our final
product is customer satisfaction, and we will provide
whatever technical assistance is
required to help you
achieve your analytical goal.


Q28: Why isn't this kit as easy to use as the PCB and other immunoassay kits
         I have used before?

A28: For the same reason that the average PCB lab will not perform a dioxin
analysis.  Conventional dioxin analysis (
Method 8290) is far more expensive and
difficult than
PCB analysis by Method 8082
.  This is partly because the 
chemistry
of dioxin analysis is intrinsically more difficult.  A major portion of 
this difficulty is
due to the fact that sensitivity targets are typically several 
orders of magnitude
lower than for PCB and other analytes.  This drives matrix
concentration factors to extreme levels and forces the
analyst to apply some
form of cleanup to sample
extracts.
  However, the proportional advantages of
Method 4025  in speed and cost relative to Method 8290 are about the
same as
 for PCB analysis by Method
4020 vs. Method 8082. 



Q29: Does this test have problems with mass labeled internal standards?

A29: Yes and no.  The immunoassay is based on structural recognition rather 
than on the mass separation used by Method 8290.  Therefore, mass labeled
internal standards, which are structurally identical to native PCDD/Fs, can 
interfere with the immunoassay.  The most efficient and economically 
advantageous use of the DF1 kit avoids conventional mass labeled internal
standards.  However, it is possible to create
immunoassay compatible internal
standards by slightly modifying existing standard
mixtures.  
For further information,
contact CAPE Technologies .