ASTM E3290-21

Standard Test Method for Establishing Performance of Equipment and Assays for Field Detection of Fentanyl and Fentanyl-Related Compounds

SCOPE
1.1 General:  
1.1.1 This test method provides a procedure for characterizing the performance of field portable fentanyl detection equipment and assays when utilizing the test samples and statistical considerations described in Specification E3243.  
1.1.2 This test method describes sample preparation and analysis protocols to use when characterizing the performance of various types of field fentanyl detection equipment or assays in a laboratory environment including gas chromatography/mass spectrometry (GC/MS), high pressure mass spectrometry (HPMS), ion mobility spectrometry (IMS), Fourier Transform Infrared (FTIR) spectroscopy, Raman spectroscopy, colorimetric assays, and immunoassays.  
1.1.3 The intent of this test method is to analyze samples in a manner that is analogous to how they are analyzed in the field by Federal and State/Local/Tribal/Territorial (SLTT) law enforcement and first responders, but under much more controlled and reproducible conditions than those that would generally be achievable when conducting field testing.  
1.2 Units:  
1.2.1 When creating test sample mixtures, all concentrations are stated as weight/weight percent (mg/mg) for solid sample mixture test samples, and weight/volume (mg/mL) for solid and liquid test samples that are dissolved in a solvent. When creating diluted liquid test samples (for example, for detection of compounds solubilized in solvent prior to analysis), all concentrations are stated as volume/volume percent (for example, µL/mL).  
1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.4 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

General Information

• Status: Published
• Publication Date: 30-Apr-2021
• ICS: 71.040.40 - Chemical analysis
• Technical Committee: E54 - Homeland Security Applications
• Drafting Committee: E54.01 - CBRNE Detection and CBRN Protection

Relations

• Refers: ASTM E3131-17 - Standard Specification for Nucleic Acid-Based Systems for Bacterial Pathogen Screening of Suspicious Visible Powders
• Effective Date: 15-Oct-2017
• Refers: ASTM E2771-11e3 - Standard Terminology for Homeland Security Applications
• Effective Date: 15-Dec-2011
• Refers: ASTM E2771-11 - Standard Terminology for Homeland Security Applications
• Effective Date: 15-Dec-2011
• Refers: ASTM E2771-11e1 - Standard Terminology for Homeland Security Applications
• Effective Date: 15-Dec-2011
• Refers: ASTM E2771-11e2 - Standard Terminology for Homeland Security Applications
• Effective Date: 15-Dec-2011

Overview

ASTM E3290-21: Standard Test Method for Establishing Performance of Equipment and Assays for Field Detection of Fentanyl and Fentanyl-Related Compounds provides a comprehensive protocol for evaluating the performance of field-portable detection equipment and assays. Developed by ASTM International, this standard supports law enforcement and first responders in identifying fentanyl and related synthetic opioids effectively and reliably under controlled, laboratory conditions. By following this method, organizations can ensure their detection systems meet necessary performance metrics and are fit for purpose in real-world applications.

Key Topics

• Performance Characterization: Procedures for systematically assessing detection equipment or assays using defined test samples and statistical approaches described in ASTM E3243.
• Sample Preparation and Analysis: Guidelines for preparing bulk and trace samples, including quantification and mixing protocols for target and non-target compounds, ensuring consistency and reproducibility.
• Applicable Technologies: Coverage of major field detection technologies, such as:
  • Gas Chromatography/Mass Spectrometry (GC/MS)
  • High Pressure Mass Spectrometry (HPMS)
  • Ion Mobility Spectrometry (IMS)
  • Fourier Transform Infrared (FTIR) Spectroscopy
  • Raman Spectroscopy
  • Colorimetric Assays
  • Immunoassays
• Controlled Laboratory Testing: Emphasis on conducting tests under controlled conditions, imitating field analysis while achieving greater reproducibility.
• Handling Interferents: Consideration of common cutting agents and interfering substances to evaluate detector specificity and minimize false positives/negatives.
• Sample Concentration Units: Instructions for expressing concentrations, including weight/weight for solids, weight/volume for liquid samples, and volume/volume for diluted liquids.
• Documentation and Safety: Requirements for thorough documentation of sample preparation, test execution, and adherence to rigorous safety and PPE protocols when handling hazardous opioids.

Applications

ASTM E3290-21 is used by:

• Federal, State, Local, Tribal, and Territorial (SLTT) Law Enforcement: To validate and select detection equipment that meets field requirements for identifying fentanyl and related substances.
• First Responders and HazMat Teams: Providing assurance that portable devices and assays will deliver accurate results during critical incidents involving suspected opioid substances.
• Manufacturers and Supplier Evaluations: Allowing equipment suppliers to benchmark products against a recognized standard for performance claims.
• Laboratory Personnel: Guiding laboratory-based testing of field detection devices under well-controlled conditions to simulate field usage scenarios.
• Regulatory Compliance and Purchasing: Supporting purchasing decisions and compliance with technical regulations and safety best practices.

By implementing this standard, organizations can select and deploy fentanyl detection technologies that have been rigorously tested and are proven to deliver reliable performance, thereby improving public safety and operational effectiveness.

Related Standards

To fully implement and understand ASTM E3290-21, users should consult these related documents:

• ASTM E3243: Specification for Field Detection Equipment and Assays Used for Fentanyl and Fentanyl-Related Compounds (statistical testing framework and sample definitions)
• ASTM E3289: Guide for Using Equipment and Assays for Field Detection of Fentanyl and Fentanyl-Related Compounds (field deployment guidance)
• ASTM E2771: Terminology for Homeland Security Applications
• ISO 17034: General requirements for the competence of reference material producers
• Eurachem/CITAC Guide CG 4: Quantifying Uncertainty in Analytical Measurement

These standards ensure a harmonized approach to testing, quality control, and result interpretation within the broader context of chemical detection and occupational safety.

---

Keywords: ASTM E3290-21, fentanyl detection, field detection equipment, synthetic opioid detection, portable assays, GC/MS, HPMS, IMS, FTIR, Raman, law enforcement, first responders, sample preparation, laboratory testing, false positives, workplace safety.

Standards Content (Sample)

This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
Designation: E3290 − 21
Standard Test Method for
Establishing Performance of Equipment and Assays for
Field Detection of Fentanyl and Fentanyl-Related
Compounds
This standard is issued under the fixed designation E3290; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
INTRODUCTION
Asaresultofthewidespreaduseofsyntheticopioids,firstrespondersandotherresponsepersonnel
mayencounterfentanylandfentanyl-relatedcompoundswhileworkinginthefield.Toensurethatfirst
responders can identify these compounds in the field, it is critical for end-users to understand the
performance metrics of detection equipment and assays to ensure these products meet the intended
end-user needs. It is also necessary to understand the potential effects on detection systems when
common cutting agents and other interferents, including drugs (for example, heroin), are present in
samples.
The test methods presented in this standard are intended for equipment and assays commonly
employed in the field for the detection of suspected drugs that may contain fentanyl and
fentanyl-related compounds. The test methods presented here are not intended for laboratory
equipment, but rather for testing of field detection equipment in a laboratory setting.The test methods
are also not intended to reflect testing approaches that would be performed in the field (for guidance
on using field detection equipment in the field, the user is referred to the companionASTM standard
Guide E3289). This test method is a companion standard to Specification E3243. The specification
describes a statistical testing approach to quantify performance and defines test sample compositions
and amounts. However, Specification E3243 does not describe in detail sample preparation methods
or specific protocols for conducting testing with different types of instruments and assays. These
methods and protocols are described in detail in this test method.
1. Scope 1.1.3 The intent of this test method is to analyze samples in
amannerthatisanalogoustohowtheyareanalyzedinthefield
1.1 General:
by Federal and State/Local/Tribal/Territorial (SLTT) law en-
1.1.1 This test method provides a procedure for character-
forcement and first responders, but under much more con-
izing the performance of field portable fentanyl detection
trolled and reproducible conditions than those that would
equipment and assays when utilizing the test samples and
generally be achievable when conducting field testing.
statistical considerations described in Specification E3243.
1.1.2 This test method describes sample preparation and 1.2 Units:
analysis protocols to use when characterizing the performance 1.2.1 Whencreatingtestsamplemixtures,allconcentrations
ofvarioustypesoffieldfentanyldetectionequipmentorassays are stated as weight/weight percent (mg/mg) for solid sample
in a laboratory environment including gas chromatography/ mixture test samples, and weight/volume (mg/mL) for solid
mass spectrometry (GC/MS), high pressure mass spectrometry and liquid test samples that are dissolved in a solvent. When
(HPMS), ion mobility spectrometry (IMS), Fourier Transform creating diluted liquid test samples (for example, for detection
Infrared (FTIR) spectroscopy, Raman spectroscopy, colorimet- of compounds solubilized in solvent prior to analysis), all
ric assays, and immunoassays. concentrations are stated as volume/volume percent (for
example, µL/mL).
1.3 This standard does not purport to address all of the
This test method is under the jurisdiction of ASTM Committee E54 on
safety concerns, if any, associated with its use. It is the
Homeland Security Applications and is the direct responsibility of Subcommittee
responsibility of the user of this standard to establish appro-
E54.01 on CBRNE Detection and Decontamination.
priate safety, health, and environmental practices and deter-
Current edition approved May 1, 2021. Published July 2021. DOI: 10.1520/
E3290-21. mine the applicability of regulatory limitations prior to use.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
E3290 − 21
1.4 This international standard was developed in accor- 3.1.5 confidence level, CL, n—probability value associated
dance with internationally recognized principles on standard- with a CI; the percentage of intervals that can be expected to
ization established in the Decision on Principles for the include the true population parameter over time.
Development of International Standards, Guides and Recom-
3.1.6 false negative, n—failure to detect a compound within
mendations issued by the World Trade Organization Technical
a sample when it is present.
Barriers to Trade (TBT) Committee.
3.1.7 false positive, n—detection of a compound within a
sample when it is not present. E3131
2. Referenced Documents
3.1.8 lower confidence bound, LCB, n—lowest value of a
2.1 ASTM Standards:
one-sided CI created using a procedure that, when repeated
E2771Terminology for Homeland Security Applications
many times on distinct data sets generated from the same
E3131Specification for Nucleic Acid-Based Systems for
underlying stochastic process, will include the true measure of
Bacterial Pathogen Screening of Suspicious Visible Pow-
performance a proportion of times equal to the stated probabil-
ders
ity. E3131
E3243Specification for Field Detection Equipment and
3.1.8.1 Discussion—The LCB ensures that the POD attains
Assays Used for Fentanyl and Fentanyl-Related Com-
a satisfactory value for the CL selected and determines the
pounds
minimum number of samples that shall be analyzed.
E3289Guide for Using Equipment and Assays for Field
Detection of Fentanyl and Fentanyl-Related Compounds
3.1.9 measurement process, n—step, or series of systematic
steps, used to detect a material or determine if a system or
2.2 Eurachem Standard:
Eurachem/CITAC Guide CG 4Quantifying Uncertainty in instrument performs as intended. E3131
Analytical Measurement, Third edition (2012)
3.1.10 non-target compound, n—collection of diluents/
2.3 ISO Standard:
cutting agents, dyes/colorants, and other drugs that are not the
ISO 17034General requirements for the competence of
target compound, which in this case is fentanyl or any
reference material producers
fentanyl-related substance.
2.4 OSHA Standards:
3.1.10.1 Discussion—Non-target compounds should not
29 CFR 1910.132OSHA Personal Protective Equipment
give a positive test result for fentanyl or fentanyl-related
Standard
substances;thisisconsideredafalsepositiveresult.Ideally,all
29 CFR 1910.134OSHA Respiratory Protection Standard
non-target compounds should result in negative detection
results for fentanyl and fentanyl-related compounds.
3. Terminology
3.1.11 operator, n—person operating an on-site chemical
3.1 Definitions:
assessment technology.
3.1.1 accuracy, n—closeness of agreement between a test
3.1.11.1 Discussion—This definition differs slightly from
result and the accepted reference value. E2771
that in Specification E3131 as it applies to the chemical
3.1.2 assay, n—quantitative or qualitative test used to deter- detection technology referred to in this test method.
mine the presence or absence of a chemical compound.
3.1.12 precursor, n—chemical compound that is used in the
3.1.3 bulk sample, n—total sample amount (including the
synthesis of fentanyl or fentanyl-related compounds.
sum of target and non-target compounds) that is visible to the
3.1.13 probability of detection, POD, n—proportionofposi-
naked eye.
tive analytical outcomes for a qualitative method for a given
3.1.3.1 Discussion—The amount of sample available for
matrix at a given concentration.
testing bulk samples according to this standard is >1 µg, but
3.1.14 reach-back support, n—service that allows equip-
≤10 mg.
ment users in the field to be in real-time contact with subject
3.1.4 confidence interval, CI, n—range of values created
matter experts (SMEs) to provide advice an assessment of
using a procedure that, when repeated many times, on distinct
analysis results, including, but not limited to identification of
data sets, generated from the same underlying stochastic
one or more substances in an unknown sample.
process, will bracket the true measure of performance, such as
3.1.14.1 Discussion—This service is often available 24 h a
probability of detection (POD), the proportion of times stated.
day/7 days per week via paid subscription through an equip-
E3131
ment (instrument or assay) manufacturer. For example, a field
chemical detection instrument like a Raman spectrometer may
2 be used to scan an unknown sample suspected of containing
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM
narcotics. The user can send the resulting data to reach-back
Standards volume information, refer to the standard’s Document Summary page on
support for review or interpretation. The SMEs providing the
the ASTM website.
3 support can identify poor quality data and recommend settings
Available from the Eurachem organization, https://www.eurachem.org/
for a successful rescan, perform advanced data analysis tech-
index.php/publications/guides/quam.
Available from International Organization for Standardization (ISO), ISO
niques to identify chemicals present in the sample, and quickly
Central Secretariat, Chemin de Blandonnet 8, CP 401, 1214 Vernier, Geneva,
provide decision support capabilities and guidance in response
Switzerland, https://www.iso.org.
to end-user requests and inquiries. Reach-back services are
Available from Occupational Safety and Health Administration (OSHA), 200
Constitution Ave., NW, Washington, DC 20210, http://www.osha.gov. commonly used for aiding in the interpretation of ambiguous
E3290 − 21
data and for confirmation of both positive and negative results 3.2.7 FTIR—Fourier transform infrared (also known as
(especially those with which decisions of consequence may be Fourier transform infrared spectroscopy).
taken).
3.2.8 GC—gas chromatography.
3.1.15 reference material, n—substance sufficiently homog-
3.2.9 GC/MS—gas chromatography/mass spectrometry.
enous and stable with respect to one or more specified
3.2.10 HPMS—high pressure mass spectrometry.
propertiesthathasbeenestablishedtobefitforitsintendeduse
3.2.11 IMS—ion mobility spectrometry.
in the measurement process; properties can be quantitative or
3.2.12 ISO—International Organization for Standardization.
qualitative. ISO 17034
3.2.13 LCB—lower confidence bound.
3.1.16 saturation, n—condition in which the detector re-
sponse no longer increases with increased sample concentra-
3.2.14 MS—mass spectrometry.
tion.
3.2.15 NIOSH—National Institute for Occupational Safety
3.1.16.1 Discussion—This can occur when there is too
and Health.
much sample introduced to an instrument or assay. For optical
3.2.16 OSHA—Occupational Safety and HealthAdministra-
detection instruments, too much exposure to light, including
tion.
ambient light, can saturate the detector.
3.2.17 POD—probability of detection.
3.1.17 sensitivity, n—change in the response of a measuring
3.2.18 PPE—personal protective equipment.
instrument divided by the corresponding change in the stimu-
3.2.19 RSD—relative standard deviation.
lus. Eurachem/CITAC Guide CG 4:2012
3.2.20 SERS—surface enhanced Raman spectroscopy.
3.1.18 specificity/selectivity, n—ability of a measurement
procedure to determine accurately and specifically the analyte 3.2.21 SLTT—state, local, tribal, and territorial.
of interest in the presence of other components in the sample
3.2.22 SME—subject matter expert
matrix under the stated conditions of the test.
3.2.23 TM—test module
Eurachem/CITAC Guide CG 4:2012
3.2.24 US&R—urban search and rescue (which may be
3.1.19 target compound, n—any of a collection of fentanyl
FEMA or state teams).
and fentanyl-related compounds.
4. Summary of Test Method
3.1.20 test module, TM, n—set of samples with particular
characteristics used to establish the detection technology per-
4.1 Test samples containing known concentrations/amounts
formance for a given type of sample.
of target and non-target analyte as defined in Annex A1 of
3.1.20.1 Discussion—TMs include target compound
Specification E3243 are selected, mixed, and prepared for
samplesatthreedifferentpercentsamplecompositions(≥95%,
testing.
10%, and 1%), non-target samples that do not contain
4.2 Instrument or assay start-up, preparation, system
fentanylorfentanyl-relatedcompounds,andfentanylsynthesis
checks, and calibrations are performed per manufacturer’s
precursors or other compounds used for synthesis. TMs are
instructions and guidance.
given in Annex A1 of Specification E3243.
4.3 Test samples and control samples are analyzed accord-
3.1.21 test sample, n—amount and identity of a particular
ing to manufacturer’s instructions and guidance.
substance (target and non-target compounds) prepared for
4.4 Required documentation is collected.
testing.
4.5 Results are evaluated and recorded.
3.1.22 trace sample, n—total sample amount, including
target and non-target compounds, ≤1µg used for testing.
4.6 Results are reported.
3.1.22.1 Discussion—In this test method, a trace sample is
4.7 In this test method, each detection technology is pre-
intended to represent a sample not visible to the naked eye, a
sented in a separate section (Sections12–19). Sections that
situation commonly encountered by first responders. For prac-
pertain to all detection technologies include Sections5–11
tical purposes, it is assumed that the sizes represented by bulk
and Sections 20 and 21.
and trace samples sufficiently cover the range of situations
encountered in the field where the testing equipment may be
5. Apparatus
used.
5.1 Appropriate personal protective equipment (PPE), in-
3.2 Acronyms: 6
cluding appropriate nitrile gloves.
3.2.1 ATR—attenuated total reflectance.
5.2 Analytical balance capable of weighing to 0.0001g.
3.2.2 CDC—Centers for Disease Control.
5.3 Chemical fume hood suitable for working with organic
3.2.3 CFR—Code of Federal Regulations.
solvents and highly toxic material such as fentanyl in liquid
and solid form.
3.2.4 CI—confidence interval
3.2.5 CL—confidence level.
3.2.6 FEMA—Federal Emergency Management Agency. Seehttps://www.tandfonline.com/doi/full/10.1080/15459624.2020.1784426.
E3290 − 21
5.4 Mixing apparatus and containers. 7. Required Documentation
5.5 Calibrated dispensing devices to accurately and consis-
7.1 Include, at a minimum, enough information in the
tently deliver necessary aliquots between 10.00 mL and
documentation of all chemical compounds used in testing and
1.00µL.
samplepreparationanddilution(forexample,solvents)totrace
the material used to its source, including chemical
5.6 Instrument or assay kit in operational readiness.
manufacturer, catalog or product number or both, lot number,
5.7 For Raman Instruments:
purchase date, and any manufacturer-provided quality
5.7.1 Shroud (for example, black or light blocking cloth, if
information/certificates.
supplied or required by manufacturer).
7.2 Record all calculations (for example, for weighing and
5.8 Reagents and Materials:
diluting).
5.8.1 See Annex A1 of Specification E3243 for a listing of
7.3 Prepare any instrument or assay for testing in accor-
target and non-target chemical compounds needed for testing.
dancewithmanufacturer’sguidance,includinganycalibration,
5.8.2 Ultrapure water.
tuning, or operational checks. Record all protocols used and
5.8.3 Ethanol.
results obtained.
5.8.4 Methanol.
5.8.5 Other solvents as indicated by an instrument or assay
7.4 Record control sample identity and control sample
manufacturer.
results and all TM samples associated with those controls (see
5.8.6 Volumetric flasks.
8.9).
5.8.7 Swabs.
7.5 Save all associated raw data and data analysis/reporting
5.8.8 Alcohol wipes.
outputs provided to the user after testing a TM sample or
control sample.
6. Hazards
6.1 Take all necessary precautions when working with
8. Selecting and Preparing Test Samples
fentanyl and fentanyl-related compounds, semi-synthetic
opioids, other drugs, and other hazardous chemicals contained
8.1 Select applicable TMs and prepare samples containing
in the target, non-target, and precursor test panels. Implement known concentrations and amounts of target and non-target
best practices for working with chemicals, including use of
compounds. See Table 1, Table 2, and Table 3, and Annex A1
proper PPE. Fentanyl and fentanyl-related compounds can be in Specification E3243.
extremely hazardous if appropriate handling and PPE are not
8.2 Prepare bulk samples (>1µg but <10mg) in suitably
adhered to. In general, an exposure of 2 to 3 mg of fentanyl or
sized sealable containers. For individual samples in each TM,
20 µg of carfentanil can be lethal if inhaled, ingested, or
2 mLglass vials are recommended. For Raman measurements,
absorbed through mucous membranes. Dermal exposure is not
flat bottom “33 expansion” glass vials are recommended;
typically associated with significant hazards. Seven grains of
curved and thick bottom glass vials should be avoided.
table salt represent about 1 mg.
8.3 Prepare trace samples starting with bulk sample
6.2 Safety Data Sheets for all chemicals (targets, non-
amountsassuggestedinTable1atthetimeoftestingfollowing
targets, precursors, solvents) should be consulted before use.
manufacturer’s guidance specific to the instrument/assay being
The user of this test method should also be aware of the
evaluated for further sample preparation/dilution (for example,
hazardsassociatedwiththeoperationofthechoseninstruments
solvent dissolution followed by serial dilution).
or assays.
8.3.1 Smaller sample amounts may be used subject to
6.3 Some instruments may contain hazards related to radio-
guidance from the instrument or assay manufacturer, SMEs, or
active materials or lasers and some assays contain hazardous
prior testing knowledge.
chemicals.Consultthemanufacturer-providedusermanualand
8.4 Each sample container in a specific TM should contain
training materials for hazards specific to the instrument or
the same mass of sample within a 10% uncertainty (for
assay being used.
example, 2 mg 6 0.2 mg); the amounts do not need to be the
same for all TMs, as long as the amounts are uniform within a
specific TM.
See https://www.cdc.gov/niosh/topics/fentanyl/risk.html.
TABLE 1 Recommended Bulk Sample Amounts for Different Detection Technologies
Detection Technology Recommended Bulk Amount of Sample per Vial
Colorimetric Assay 1to2mg
FTIR Spectroscopy 1to3mg
Gas Chromatography/Mass Spectrometry (GC/MS) 1to2mg
High-Pressure Mass Spectrometry (HPMS) 1to2mg
Immunoassay 1to2mg
Ion Mobility Spectrometry 1to2mg
Raman Spectroscopy 9 to <10 mg
Surface Enhanced Raman Spectroscopy (SERS) 1to2mg
E3290 − 21
8.5 When preparing TM samples record all sample prepa- 8.7.5.2 Standard solutions are prepared by dissolving each
ration calculations, mixing calculations, and test data used to individual chemical component of the mixture sample in an
qualify accurately prepared mixtures. appropriate volume of solvent (5% or less volumetric uncer-
tainty) to produce a “parent” solution. Parent solutions are
8.6 Weigh materials with a balance that can measure≤2mg
diluted prior to quantitative analysis such that the concentra-
with a maximum 10% uncertainty.
tions or signal intensities of the compounds are similar (within
8.7 Preparation of Multicomponent TM Samples (TM2,
a factor of two) to those expected for the five aliquots of the
TM3, and TM4):
mixture.
8.7.1 Determine how much total mass of each chemical is
8.7.5.3 Calculate the relative standard deviation (RSD) for
necessary for the desired mixtures. Calculate the exact mass of
reported values of each component among the five samples.
each chemical needed to achieve the specified percent compo-
The RSD is calculated for each component by dividing the
sition for a given mixture and prepare enough mixture so that
standard deviation of the relative concentration or relative
the minimum amount of any one component in the sample
intensity of the five samples by its mean and multiplying the
mixture is not less than 1 to 2 mg.
result by 100%. The error in the mean concentration for each
8.7.2 Aminimaldegreeofsampleprocessingisnecessaryto
component is determined using the standard solution’s relative
provide results that can be compared among different testing
concentration or relative signal intensity. Both error and RSD
events. Prior to weighing, all solids should be processed
values of 20% or less indicate that adequate mixing was
through a mesh-60 sieve. Retain all particles smaller than
achieved.
mesh-60 for testing. If necessary, use a mortar and pestle with
8.7.5.4 If the mixture is acceptable for use in a TM, weigh
minimal grinding to reduce the particle sizes sufficiently to
out material into 2 mL vials following the guidelines of Table
pass through the mesh-60 sieve. Do not overgrind as this will
1 for the instrument or assay to be tested.
result in sample physical properties that do not reflect real-
8.8 Analysis and Reporting of Control Samples:
world illicit drug samples.
8.8.1 Throughout the course of testing, analyze available
8.7.3 Weigh out each chemical to within 10% of the
positiveandnegativecontrolsamplesusingatestingprocedure
calculated (desired) mass and combine in an appropriate
recommended by the manufacturer to demonstrate that the
mixing container. Ensure that the balance has a measurement
instrument or assay is operating correctly, and that the testing
uncertaintyof10%orlessforthesmallestmasstobeweighed.
environment is free from contamination. Test the specific
The smallest or minimum weight for a balance is determined
control samples recommended by the manufacturer (note that
by multiplying the balance’s repeatability by two and dividing
insomecasesthemanufacturer’sinstructionsmayonlycallfor
theresultbythedesiredmeasurementuncertainty,expressedin
onetypeofcontrolsample–eitherpositiveornegative;follow
decimal form. For example, the minimum weight to obtain at
manufacturer’s guidelines).
least 10% uncertainty for a balance with a reported repeatabil-
8.8.2 Test positive and negative control samples at least
ity of 0.04 mg is (0.04 mg × 2)/0.1 or 0.8 mg.
onceper10testsamplesanalyzedoratleastonceeachdaythat
8.7.4 Mix the combined chemicals using an appropriate
tests are performed, whichever is greater. Follow manufactur-
mixer and mixing settings. Handle the final mixed sample
er’s guidance.
with utmost care to avoid unnecessary shaking or jarring that
8.8.3 Do not include results from control samples in any
may result in the separation of mixture components.
calculation of LCB or POD.
8.7.5 Verifythesamplemixtureissufficientlyhomogeneous
8.8.4 Stoptestingiffailureswithcontrols,including,butnot
by analyzing five subsamples using a quantitative analytical
limited to, indication of contamination occur. Testing shall
technique such as solvent dissolution followed by liquid
remain on hold until a cause for the failure, or failures, has
chromatography with ultraviolet or mass spectrometric detec-
been found and corrected. Invalidate all TM sample testing
tion (LC/UV or LC/MS). Other techniques are acceptable.
results associated with the failed controls. Repeat the invali-
Collect five samples of at least 2 mg each from different
dated tests, but only after the cause of the failure has been
locations in the final mixed sample. This can be accomplished
identified and corrected, and the instrument/assay has been
by removing the mixture from the mixing container and
cleared for performance following the appropriate manufactur-
placing it onto a clean surface (for example, a metal plate,
er’s protocols.
silicon wafer, or weighing paper). Dissolve each of the five
aliquots in an appropriate volume of solvent (5% or less
8.9 Analysis and Reporting of Test Samples:
volumetric uncertainty) and analyze using one or more ana-
8.9.1 Depending on the number of samples in each TM
lytical methods that can determine absolute or relative concen-
chosen for testing (31, 14, or 4), testing may cease after a
trations (or signal intensities) for all mixed components.
certain number of failures for that TM as the minimum
8.7.5.1 The quantitative values obtained from the analysis
acceptable performance level of 0.85/80% LCB/CLwould not
should be compared to values obtained from standard solu-
be attainable (see Section 5 in Specification E3243). However,
tions.
testing may continue until all the samples in the selected TM
have been tested if desired. In this case, testing of all samples
LabRamI,ResodynAcousticMixers,Butte,MT,USA.Settings:75Gfor5min
planned for allows for obtaining a more precise estimate of
mixing time. Use 2 mLGC vials with 500µLinsert. The make, model and settings
performance even though it will be below 0.85/80% LCB/CL.
are provided as examples that were tested and shown to produce well-mixed
8.9.2 For instruments that have reach-back support, two test
multicomponent samples. Other products that can be shown to produce well-mixed
multicomponent samples when following the protocol in 8.7 may also be used. results are obtained (see Section 21):
E3290 − 21
8.9.2.1 The direct readout result, and handheld probe. Sampling directly from solids may require
8.9.2.2 The result provided by reach-back. additional accessories or software methods.
8.9.3 If 4 samples are chosen to be tested in an applicable
12.8.1 Bulk Sample Testing in GC/MS Mode:
TM, testing may cease when a direct readout result failsAND
12.8.1.1 Dissolve the sample in manufacturer-
when any result provided by reach-back fails;
recommended solvent to a concentration of 1 to 10 mg/mL
8.9.4 If 14 samples are chosen to be tested in an applicable
(1 to 10 µg/µL; weight/volume) using volumetric glassware or
TM, testing may cease when a single direct readout result fails
pipettes that provide a volumetric uncertainty of 5% or better,
AND when a result provided by reach-back fails;
and an analytical balance that has a measurement uncertainty
8.9.5 If 31 samples are chosen to be tested in an applicable
of 10% or better (never measure less than the minimum
TM, testing may cease when more than three direct readout
manufacturer-recommended weight for a balance). Follow
results fail AND when more than three results provided by
manufacturer’s recommendations for further sample handling
reach-back fail.
(for example, shake to mix and particulate settling) and further
dilution,ifnecessary,soasnottointroduceaconcentrationthat
9. Preparation of Apparatus
will saturate the instrument.
9.1 Prepare instruments or assays according to manufactur-
12.8.1.2 Once further sample handling and dilution has
er’s guidance, including considerations of temperature, ambi-
occurred, inject 1µL of solution into instrument using a
entlightandbrightness,humidity,dust,andotherenvironmen-
standard5-µLGCsyringeifthereisnootherspecificguidance.
tal factors.
12.8.2 Bulk Sample Testing in Vapor Detection Mode:
12.8.2.1 For high vapor pressure liquids and solids, survey
9.2 Ensure analytical balance and pipettes are calibrated.
mode can be used for quick identification using a sampling
9.3 Ensure glassware and any weighing accessories (for
wand or handheld probe and passing the probe over an open
example, spatula) are clean.
vial of test sample, taking care not to saturate the instrument.
12.8.2.2 If sampling in vapor mode, slowly approach
10. Calibration and Standardization
sample with wand or probe until sufficient signal is observed.
10.1 Ensure each instrument is calibrated and not in need of
12.8.2.3 Sampling directly from low vapor pressure solids
service per manufacturer’s instructions.
may require additional accessories or software methods. Note
10.2 Ensureassaysarenotexpiredandpackagingorstorage
that the intent of this test method is to analyze samples in a
has not been compromised.
manner that is analogous to how they are analyzed in the field
by Federal and SLTT law enforcement and first responders.
11. Conditioning
Some sampling approaches, while available, may not reflect
11.1 Consult user manuals for any instrument or test-
how law enforcement and first responders actually use this
specific setup features.
equipment,inwhichcasetheyshouldnotbeused.Consultwith
SMEs and equipment end-users.
12. Gas Chromatography/Mass Spectrometry (GC/MS)
12.8.2.4 Perform scan according to manufacturer’s
Test Procedure
instructions, appropriate for sample type and identification
12.1 Following procedures outlined in Section 8, choose to
mode.
test bulk amounts of test sample (>1 µg but <10 mg, note that
12.8.3 Trace Sample Testing in GC/MS Mode:
samples may need to be diluted to avoid saturating the
12.8.3.1 Dissolve the sample in manufacturer-
detector) or trace amounts of test sample (≤1 µg target
recommended solvent to a concentration of 0.1 to 1 mg/mL
compound), choose the test module (TM1, TM2, TM3, TM4,
(0.1 to 1 µg/µL; weight/volume) using volumetric glassware or
andTM5),selectthenumberofsamplestobetested(31,14,or
pipettes that provide a volumetric uncertainty of 5% or better,
4), and choose the number of different sample types to test (14
and an analytical balance that has a measurement uncertainty
or 4; for clarification, see 4.13 in Specification E3243).
of 10% or better (never measure less than the minimum
12.2 Refer to user manual and training materials for the manufacturer-recommended weight for a balance). Shake to
mix.
specific instrument being tested.
12.8.3.2 1µLofthissolutionisinjected(nomore),resulting
12.3 Power on instrument and allow it to perform boot
intherequiredintroductionofnomorethan1µgofsampleper
sequence,includingallowingittoreachoperatingtemperature.
the definition of trace sample. Alternative concentrations and
12.4 Log in and perform system checks as required.
volumes are acceptable (for example, 2 µL injection of a
0.5µg⁄µL solution) as long as no more than 1 µg of sample is
12.5 Install appropriate vapor or trace sampling accessory
for sample to be tested. introduced.Introductionof<1µgofsampleisacceptable.Note
that for TM2 (10% target compound), 1 µg of sample equates
12.6 Ensure that target compounds are present in the library
to 100 ng of target compound and for TM3 (1% target
to be used for analysis.
compound), 1µg of sample equates to 10 ng of target com-
12.7 Don appropriate PPE, including powder-free nitrile
pound.
gloves.
12.8.3.3 Perform scan according to manufacturer’s
12.8 AnalysisofbulksamplescanbedoneinGC/MSmode instructions, appropriate for sample type and identification
or using just MS for vapor detection using a sampling wand or mode.
E3290 − 21
12.9 Data Inspection and Analysis: of 10% or better (never measure less than the minimum
12.9.1 For TM1, TM2, and TM3, if the target compound manufacturer-recommended weight for a balance). Follow
cannot be identified as fentanyl or a fentanyl-related manufacturer’s recommendations for further sample handling
compound, the analysis of the test sample has failed (see 8.9). anddilution,ifnecessary,soasnottointroduceaconcentration
Retest only if an error has been made by the testing entity or that will saturate the instrument.
the instrument has malfunctioned based on the failure of 13.9.1.2 Spike a manufacturer-recommended volume (typi-
control samples (see 8.8). cally 1 to 2 µL) of the solubilized dilute sample onto an
12.9.2 For TM4, if the result indicates the presence of appropriate dry swipe using a standard 5-µL GC syringe.
fentanyl or a fentanyl-related compound, the analysis of the
13.9.1.3 Perform scan according to manufacturer’s
testsamplehasfailed(see8.9).Retestonlyifanerrorhasbeen instructions, appropriate for sample type and identification
made by the testing entity or the instrument has malfunctioned
mode.
based on the failure of control samples (see 8.8). 13.9.2 Bulk sample testing using vapor detection mode:
12.9.3 ForTM5, if the identification of the test sample does
13.9.2.1 For high vapor pressure liquids and solids, vapor
not indicate the presence of the precursor-related compound,
detection mode can be used by passing the instrument’s vapor
the analysis of the test sample has failed.
inlet over an open vial of test sample, taking care not to
12.9.4 If the instrument is working below optimal
saturate the instrument.
conditions, maintenance procedures or recalibration may be
13.9.2.2 If sampling in vapor mode, slowly approach
required. Consult manufacturer if necessary, to troubleshoot.
sample with instrument until sufficient signal is observed.
Conduct all testing under homogeneous instrument conditions.
13.9.2.3 Perform scan according to manufacturer’s
Maintenance or recalibration procedures may affect instrument
instructions, appropriate for sample type and identification
performance and effectively result in different testing condi-
mode.
tions for samples tested before and after the procedure occurs,
13.9.3 Trace sample testing in thermal desorption mode:
which may invalidate the results.
13.9.3.1 Dissolve the sample in manufacturer-
recommendedsolventtoaconcentrationof1mg/mL(1µg/µL)
13. High-Pressure Mass Spectrometry (HPMS) Test
using volumetric glassware or pipettes that provide a volumet-
Procedure
ric uncertainty of 5% or better, and an analytical balance that
13.1 Following procedures outlined in Section 8, choose to
has a measurement uncertainty of 10% or better (never
test bulk amounts of test sample (>1 µg but <10 mg; note that
measure less than the minimum manufacturer-recommended
samples may need to be diluted to avoid saturating the
weight for a balance).
detector) or trace amounts of test sample (≤1 µg target
13.9.3.2 Dilute the 1 µg/µL solution by a factor of 4 (5%
compound), choose the test module (TM1, TM2, TM3, TM4,
uncertainty or better) to prepare a 0.25 µg/µL dilution. Spike
andTM5),selectthenumberofsamplestobetested(31,14,or
4µLof the diluted sample onto an appropriate dry swipe using
4), and choose the number of different sample types to test (14
a standard 5-µL GC syringe. Spiking smaller volumes onto a
or 4; for clarification, see 4.13 in Specification E3243).
swipe or use of different dilutions is acceptable as long as no
more than 1 µg of total sample amount is introduced. Note that
13.2 Refertousermanualandtrainingmaterialsforspecific
for TM2 (10% target compound), 1 µg of sample equates to
instrument being tested.
100 ng of target compound and for TM3 (1 % target
13.3 Power on instrument and allow it to perform boot
compound), 1 µg of sample equates to 10 ng of target
sequence,includingallowingittoreachoperatingtemperature.
compound.
13.4 Log in and perform any required system checks as
13.9.3.3 Allowtheswipetodryforatleast30s,oruntildry,
required.
and then analyze.
13.9.3.4 Final test samples can be generated through mul-
13.5 Install appropriate vapor or dry swipe accessory for
tipleroundsofswabtoswabserialdilutiontoreachappropriate
sample to be tested.
sample concentration if necessary.
13.6 Log in and perform system checks as required.
13.9.3.5 Perform scan according to manufacturer’s
13.7 Ensure that target compounds are present in the library
instructions, appropriate for sample type and identification
to be used for analysis.
mode.
13.9.4 Reviewscanresultsandinterpretaccordingtomanu-
13.8 Don appropriate PPE, including powder-free nitrile
facturer’s guidance.
gloves.
13.10 Data Inspection and Analysis:
13.9 Analysis of bulk samples can be done in thermal
13.10.1 For TM1, TM2, and TM3, if the target compound
desorption mode (requires solubilization of sample and appli-
cation to a swipe) or in the vapor detection “sniffer” mode. cannot be identified as fentanyl or a fentanyl-related
compound, the analysis of the test sample has failed (see 8.9).
13.9.1 Bulk sample testing in thermal desorption mode:
13.9.1.1 Dissolve the sample in manufacturer- Retest only if an error has been made by the testing entity or
the instrument has malfunctioned based on the failure of
recommended solvent to a concentration of 1 to 10 mg/mL (1
to 10 µg/µL; weight/volume) using volumetric glassware or control samples (see 8.8).
pipettes that provide a volumetric uncertainty of 5% or better, 13.10.2 For TM4, if the result indicates the presence of
and an analytical balance that has a measurement uncertainty fentanyl or a fentanyl-related compound, the analysis of the
E3290 − 21
testsamplehasfailed(see8.9).Retestonlyifanerrorhasbeen horizontal). Note that spiking volumes larger than 2 µL can
made by the testing entity or the instrument has malfunctioned lead to undesirable wicking; spiking a volume of 1 µLis ideal.
based on the failure of control samples (see 8.8).
14.8.1.3 Perform scan according to manufacturer’s
13.10.3 For TM5, if the identification of the test sample instructions, appropriate for sample type and identification
does not indicate the presence of the precursor-related mode.
compound, the analysis of the test sample has failed.
14.8.2 Bulk Sample Testing using Vapor Detection Mode:
13.10.4 If the instrument is working below optimal
14.8.2.1 For high vapor pressure liquids and solids, vapor
conditions, maintenance procedures or recalibration may be
detection mode can be used by passing the instrument’s vapor
required. Consult manufacturer if necessary, to troubleshoot.
inlet over an open vial of test sample, taking care not to
Conduct all testing under homogeneous instrument conditions.
saturatetheinstrument.Notethatopioidsandmanyotherillicit
Maintenance or recalibration procedures may affect instrument
drugs do not have an adequate vapor pressure to allow
performance and effectively result in different testing condi-
detection. However, many solvents that are used in the synthe-
tions for samples tested before and after the procedure occurs,
sis of opioids do have a measurable vapor pressure.
which may invalidate the results.
14.8.2.2 If sampling in vapor mode, slowly approach
sample with instrument until sufficient signal is observed.
14. Ion Mobility Spectrometry (IMS) Test Procedure
14.8.2.3 Perform scan according to manufacturer’s
instructions, appropriate for sample type and identification
14.1 Following procedures outlined in Section 8, choose to
mode.
test bulk amounts of test sample (>1 µg but <10 mg, note that
14.8.3 Trace Sample Testing in Thermal Desorption Mode:
samples may need to be diluted to avoid saturating the
detector) or trace amounts of test sample (≤1 µg target 14.8.3.1 Dissolve the sample in manufacturer-
compound), choose the test module (TM1, TM2, TM3, TM4, recommendedsolventtoaconcentrationof1mg/mL(1µg/µL)
andTM5),selectthenumberofsamplestobetested(31,14,or
using volumetric glassware or pipettes that provide a volumet-
4), and choose the number of different sample types to test (14 ric uncertainty of 5% or better, and an analytical balance that
or 4; for clarification, see 4.13 in Specification E3243).
has a measurement uncertainty of 10% or better (never
measure less than the minimum manufacturer-recommended
14.2 Refertousermanualandtrainingmaterialsforspecific
weight for a balance).
instrument being tested.
14.8.3.2 Dilute the 1 µg/µL solution by a factor of 4 (5%
14.3 Power on instrument and allow it to perform boot
uncertainty or better) to prepare a 0.25 µg/µL dilution. Spike
sequence,includingallowingittoreachoperatingtemperature.
4µLof the diluted sample onto an appropriate dry swipe using
a standard 5-µL GC syringe. Spiking smaller volumes onto a
14.4 Perform any required system checks prior to sampling.
swipe or use of different dilutions is acceptable as long as no
14.5 Follow instrument-specific guidance for cleaning and
more than 1 µg of total sample amount is introduced. Note that
installation of accessories.
for TM2 (10% target compound), 1 µg of sample equates to
14.6 Ensure that target compounds are present in the library 100 ng of target compound and for TM3 (1 % target
to be used for analysis. compound), 1 µg equates to 10 ng of target compound.
14.8.3.3 Allowtheswipetodryforatleast30s,oruntildry,
14.7 Don appropriate PPE, including powder-free nitrile
and then analyze.
gloves.
14.8.3.4 Final test samples can be generated through mul-
14.8 Analysis of bulk samples can be done in thermal
tipleroundsofswabtoswabserialdilutiontoreachappropriate
desorption mode (requires solubilization of sample and appli-
sample concentration if necessary.
cation to a swipe) or in the vapor detection “sniffer” mode.
14.8.3.5 Perform scan according to manufacturer’s
14.8.1 Bulk Sample Testing in Thermal Desorption Mode:
instructions, appropriate for sample type and identification
14.8.1.1 Dissolve the sample in manufacturer-
mode.
recommended solvent to a concentration of 1 to 10 mg/mL (1
14.8.4 Reviewscanresultsandinterpretaccordingtomanu-
to 10 µg/µL; weight/volume) using volumetric glassware or
facturer’s guidance.
pipettes that provide a volumetric uncertainty of 5% or better,
14.9 Data Inspection and Analysis:
and an analytical balance that has a measurement uncertainty
of 10% or better (never measure less than the minimum 14.9.1 For TM1, TM2, and TM3, if the target compound
manufacturer-recommended weight for a balance). Follow cannot be identified as fentanyl or a fentanyl-related
manufacturer recommendations for further sample handling compound, the analysis of the test sample has failed (see 8.9).
anddilution,ifnecessary,soasnottointroduceaconcentration Retest only if an error has been made by the testing entity or
that will saturate the instrument. the instrument has malfunctioned based on the failure of
control samples (see 8.8).
14.8.1.2 Spike a manufacturer-recommended volume (typi-
cally 1 to 2 µL) of the solubilized dilute sample onto an 14.9.2 For TM4, if the result indicates the presence of
appropriate dry swipe using a standard 5-µL GC syringe. The fentanyl or a fentanyl-related compound, the analysis of the
swipe should be held off of a surface and positioned at a testsamplehasfailed(see8.9).Retestonlyifanerrorhasbeen
45-degree angle when spiking occurs so as to minimize made by the testing entity or the instrument has malfunctioned
wicking (that is, the swipe should not be lying flat or based on the failure of control samples (see 8.8).
E3290 − 21
14.9.3 ForTM5, if the identification of the test sample does 15.12 Fluorescence is a naturally occurring chemical prop-
not indicate the presence of the precursor-related compound, ertythatcausessomemoleculestoemitlight.Fluorescencecan
the analysis of the test sample has failed. be expected from colored substances and certain organic
14.9.4 If the instrument is working below optimal compoundsthatmayappearinoneormoreoftheTMsamples.
conditions, maintenance procedures or recalibration may be The effect will be masking of the Raman signal, which may
required. Consult manufacturer if necessary, to troubleshoot. make it difficult or impossible to identify any compounds
Conduct all testing under homogeneous instrument conditions. present. Lower the laser power to reduce the amount of
Maintenance or recalibration procedures may affect instrument fluorescencefromatestsampleifnecessary.Ifaninstrumentis
performance and effectively result in different testing condi- unable to acquire a scan within five minutes, consider the
tions for samples tested before and after the procedure occurs, analysis of that test as a fail.
which may invalidate the results.
15.13 Because end-users of Raman spectrometers often
measure a sample multiple times, each sample should be
15. Raman Spectroscopy Test Procedure
measured three times in different locations on the sample. In
15.1 Following procedures outlined in Section 8, choose to point and shoot mode, the vial can be positioned at slightly
test bulk amounts of test sample (>1 µg but <10 mg; note that
differentlocations.Forvialadaptors,thevialcanberotatedfor
Raman does not have the sensitivity to detect trace sample different measurements. If any one of the three measurements
amounts),choosethetestmodule(TM1,TM2,TM3,TM4,and
produces a correc
...