Standard Guide for Spiking into Aqueous Samples

SIGNIFICANCE AND USE
5.1 Matrix spiking is commonly used to determine the bias under specific analytical conditions, or the applicability of a test method to a particular sample matrix in that context, by determining the extent to which the spiked analyte or component is recovered from the sample matrix under these conditions. Reactions or interactions of the analyte or component of interest with the sample matrix may cause a significant positive or negative effect on recovery and may render the chosen analytical, or monitoring, process ineffectual for that sample matrix.  
5.2 Matrix spiking can also be used to monitor the performance of a laboratory, individual instrument, or analyst as part of a regular quality assurance program. Changes in spike recoveries or recovery limits from the same or similar matrices over time may indicate variations in the quality of analytical results.  
5.3 Spiking can be used to compare the recoveries of like spikes from reagent water samples and natural matrix samples (measured with and without spike) to distinguish between (1) unusual interference and (2) inherent method recovery and instability effects. This guide does not attempt to deal with the statistical significance of differences in spike recoveries from different matrices.  
5.4 Special precautions shall be observed when nonlaboratory personnel perform spiking in the field. It is recommended that all spike preparation work be performed in a laboratory by experienced analysts so that the field operation consists solely of adding a prepared spiking solution to the sample matrix. Training of field personnel and validation of their spiking techniques are necessary to ensure that spikes are added accurately and reproducibly. Duplicate field spikes can be used to document the reproducibility of the technique. When environmentally labile compounds are used as spikes, the spiking solution shall be protected up to the point of use by appropriate means such as chilling, protection from sunlight and o...
SCOPE
1.1 This guide covers the general technique of “spiking” a broad range of materials into aqueous media. This guide will serve the analyst in preparing spiked samples for quality control purposes. Guidance is also provided to aid the analyst in calculating recoveries and interpreting results. It is the responsibility of the analyst to determine whether the procedures and materials described here are appropriate to the task at hand.  
1.2 The procedures in this guide are focused on “matrix spike” preparation, analysis, and interpretation of results. The applicability of these procedures to the preparation of calibration standards, calibration check standards, laboratory control standards, reference materials, and other quality control materials by spiking is incidental. A sample (the matrix) is fortified (spiked) with the analyte of interest for a variety of analytical and quality control purposes. While the spiking of multiple sample portions is discussed, the method of standard additions is not covered.  
1.3 This guide is intended for use in conjunction with the individual analytical test method that provides procedures for analysis of the analyte or component of interest. The test method is used to determine an analyte or component’s background level and, again after spiking, its now elevated level. Each test method typically provides procedures not only for samples, but also for calibration standards or analytical control solutions, or both. These procedures include preparation, handling, storage, preservation, and analysis techniques. These procedures are applicable by extension, using the analyst’s judgement on a case-by-case basis, to spiking solutions, and are not reiterated in this guide. See also Practice E200 for preparation and storage information.  
1.4 These procedures apply only to analytes that are soluble in water at the concentration of the spike plus any background material, or to analytes soluble in...

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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: D5810 − 96 (Reapproved 2021)
Standard Guide for
Spiking into Aqueous Samples
This standard is issued under the fixed designation D5810; 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.
1. Scope a homogeneous solution of analyte and sample. Meaningful
recovery data cannot be obtained if an aqueous solution or
1.1 This guide covers the general technique of “spiking” a
homogenoussuspensionoftheanalyteofinterestinthesample
broad range of materials into aqueous media. This guide will
cannot be attained. These procedures may be applicable to
serve the analyst in preparing spiked samples for quality
microbiologicalpreparationsifthehomogeneityofthesuspen-
control purposes. Guidance is also provided to aid the analyst
sioncanbeadequatelymaintainedthroughoutthecourseofthe
in calculating recoveries and interpreting results. It is the
analysis, for example, by mechanical agitation or stirring.
responsibility of the analyst to determine whether the proce-
dures and materials described here are appropriate to the task 1.5 Matrix spiking may be performed in the field or in the
at hand. laboratory,dependingonwhichpartoftheanalyticalprocessis
to be tested. Field spiking tests the recovery of the overall
1.2 The procedures in this guide are focused on “matrix
process, including preservation and shipping of the sample.
spike” preparation, analysis, and interpretation of results. The
Laboratoryspikingteststhelaboratoryprocessonly.Spikingof
applicability of these procedures to the preparation of calibra-
sample extracts, concentrates, or dilutions will test only that
tion standards, calibration check standards, laboratory control
portion of the process subsequent to addition of the spike.
standards, reference materials, and other quality control mate-
rials by spiking is incidental.Asample (the matrix) is fortified 1.6 The values stated in SI units are to be regarded as
(spiked) with the analyte of interest for a variety of analytical standard. No other units of measurement are included in this
and quality control purposes. While the spiking of multiple standard.
sample portions is discussed, the method of standard additions
1.7 This standard does not purport to address all of the
is not covered.
safety concerns, if any, associated with its use. It is the
responsibility of the user of this standard to establish appro-
1.3 This guide is intended for use in conjunction with the
priate safety, health, and environmental practices and deter-
individual analytical test method that provides procedures for
mine the applicability of regulatory limitations prior to use.
analysis of the analyte or component of interest. The test
1.8 This international standard was developed in accor-
method is used to determine an analyte or component’s
dance with internationally recognized principles on standard-
background level and, again after spiking, its now elevated
ization established in the Decision on Principles for the
level. Each test method typically provides procedures not only
Development of International Standards, Guides and Recom-
for samples, but also for calibration standards or analytical
mendations issued by the World Trade Organization Technical
control solutions, or both. These procedures include
Barriers to Trade (TBT) Committee.
preparation, handling, storage, preservation, and analysis tech-
niques.Theseproceduresareapplicablebyextension,usingthe
2. Referenced Documents
analyst’s judgement on a case-by-case basis, to spiking
solutions, and are not reiterated in this guide. See also Practice
2.1 ASTM Standards:
E200 for preparation and storage information.
D1129Terminology Relating to Water
D1193Specification for Reagent Water
1.4 Theseproceduresapplyonlytoanalytesthataresoluble
D3694Practices for Preparation of Sample Containers and
in water at the concentration of the spike plus any background
for Preservation of Organic Constituents
material, or to analytes soluble in a solvent that is itself
D3856Guide for Management Systems in Laboratories
water-soluble. The system used in the later case must result in
Engaged in Analysis of Water
D4375Practice for Basic Statistics in Committee D19 on
This guide is under the jurisdiction ofASTM Committee D19 on Water and is
thedirectresponsibilityofSubcommitteeD19.02onQualitySystems,Specification,
and Statistics. For referenced ASTM standards, visit the ASTM website, www.astm.org, or
CurrenteditionapprovedJuly1,2021.PublishedJuly2021.Originallyapproved contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
in 1996. Last previous edition approved in 2015 as D5810–96 (2015). DOI: Standards volume information, refer to the standard’s Document Summary page on
10.1520/D5810-96R21. the ASTM website.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
D5810 − 96 (2021)
Water (Withdrawn 2018) 5.3 Spiking can be used to compare the recoveries of like
E200Practice for Preparation, Standardization, and Storage spikes from reagent water samples and natural matrix samples
of Standard and Reagent Solutions for ChemicalAnalysis (measured with and without spike) to distinguish between (1)
unusual interference and (2) inherent method recovery and
3. Terminology
instability effects. This guide does not attempt to deal with the
3.1 Definitions:
statistical significance of differences in spike recoveries from
3.1.1 For definitions of terms used in this standard, refer to
different matrices.
Terminology D1129.
5.4 Special precautions shall be observed when nonlabora-
3.2 Definitions of Terms Specific to This Standard:
tory personnel perform spiking in the field. It is recommended
3.2.1 matrix spike, n—the quantity (mass) of a component
thatallspikepreparationworkbeperformedinalaboratoryby
(analyte) of interest that is added to a sample (matrix) in order
experienced analysts so that the field operation consists solely
to test the bias as measured by recovery (of that component
of adding a prepared spiking solution to the sample matrix.
under specific analytical conditions) and reported as percent
Training of field personnel and validation of their spiking
recovery (P).
techniques are necessary to ensure that spikes are added
accuratelyandreproducibly.Duplicatefieldspikescanbeused
3.2.2 spike, v—the addition of a known amount of an
to document the reproducibility of the technique. When envi-
analyte of known identity to a measured volume of a sample
ronmentally labile compounds are used as spikes, the spiking
(from a specific matrix) to determine the efficiency with which
solutionshallbeprotecteduptothepointofusebyappropriate
the added analyte can be “recovered” from (measured in) that
meanssuchaschilling,protectionfromsunlightandoxygen,or
matrix by the analytical system after exposure to a specific
chemical preservation.
portionofananalyticalprocess.Matrixspikingisaprocessfor
accomplishing this. The precision and bias estimates from
NOTE1—Anyfieldspikedsample,ifknowntothelaboratory,shouldbe
several trials under specific analytical conditions represent the
labeledasafieldspikeinthefinalresultsreport.Also,wheneverpossible,
field spiking of volatile compounds should be avoided.
measurement efficiency with which the analyte may be deter-
mined under these conditions.
5.5 It is often tacitly assumed that an analyte component is
recovered from samples to approximately the same extent that
3.2.3 spikingsolution,n—thesolutioninwhichoneormore
a spike of the same analyte is recovered from a spiked sample.
spikes are dissolved (along with any necessary preservatives).
One reason that this assumption may be incorrect is that the
This solution acts as a carrier to provide ease of measurement
spike may not be bound up in the sample (for example, with
and more rapid and thorough mixing of the spike into the
suspendedmatter)inthesamewaythatthenaturallyoccurring
sample, as compared to adding the spike as a pure compound.
analyte is bound in the sample. The spike may therefore be
4. Summary of Guide
recovered from the sample differently than the background
4.1 This guide describes a technique for the addition of a level of the analyte. It is not good practice to correct analytical
known amount of an analyte to an aqueous sample.Appropri- data using spike recoveries for this reason, as well as the fact
ate concentrations of the spike relative to the original concen- that bias corrections can add variability. However, spike
recovery information should be reported along with related
tration in the sample are discussed. Applications of the tech-
nique and aids in the interpretation of results obtained are sample analysis results.
described.
5.6 This guide is also applicable to the use of spikes for
quantification by the method of standard additions and to the
5. Significance and Use
addition of surrogates and internal standards.
5.1 Matrix spiking is commonly used to determine the bias
under specific analytical conditions, or the applicability of a
6. Apparatus
test method to a particular sample matrix in that context, by
6.1 Pipetters—Plunger-actuated pipetters, to dispense small
determining the extent to which the spiked analyte or compo-
volumesofspikesolutions.Thesemustbecalibratedandtested
nent is recovered from the sample matrix under these condi-
carefully for repeatability before use.
tions. Reactions or interactions of the analyte or component of
interestwiththesamplematrixmaycauseasignificantpositive
6.2 Volumetric Transfer Pipets—Class A, used to deliver
or negative effect on recovery and may render the chosen
knownvolumesofsampleandtoaddlargervolumesofspiking
analytical, or monitoring, process ineffectual for that sample
solutions.
matrix.
6.3 Volumetric Flasks—Class A volumetric flasks may be
5.2 Matrix spiking can also be used to monitor the perfor-
used to measure known volumes of sample.
mance of a laboratory, individual instrument, or analyst as part
6.4 Balance—Ananalytical(0.1-mg),semimicro(0.01-mg),
of a regular quality assurance program. Changes in spike
or micro (0.001-mg) balance.
recoveriesorrecoverylimitsfromthesameorsimilarmatrices
over time may indicate variations in the quality of analytical
7. Reagents
results.
7.1 Purity of Reagents—At a minimum, reagent grade
chemicals shall be used in all spike preparations. Reagents of
The last approved version of this historical standard is referenced on
www.astm.org. thehighestavailablepurityshallbeusedforspikeanalytesand
D5810 − 96 (2021)
demonstrated to be free of interfering substances for the longer periods than dilute solutions. Alternatively, prepare
subsequent tests to be performed. If possible, a primary spike or spiking solution fresh for each batch of samples.
standard grade shall be used. Unless otherwise indicated, it is
8. Sampling
intended that all reagents conform to the specifications of the
Committee onAnalytical Reagents of theAmerican Chemical
8.1 Althoughsamplingmethodologyisbeyondthescopeof
Society. Other grades may be used, provided that the reagent this guide, a properly split or duplicate sample is of utmost
isofsufficientlyhighpuritytopermititsusewithoutadversely
importance to the successful measurement of spike recovery.
affecting the bias and precision of subsequent determinations. This is especially critical in samples containing suspended
Purchasedspikingsolutionsshallbedemonstratedtobefreeof
sediment or volatile components.
substancesthatwouldinterferewithsubsequentanalysesbeing
8.2 Sample containers shall be selected and prepared, and
performed, and the supplier’s stated concentration shall be
samples shall be preserved in accordance with Practices
verified by analysis prior to use. Compensatory errors associ-
D3694.
atedwithself-referencingshouldbepreventedbyusingspiking
solutions of a standard originating from a source, when
9. Procedure
available, different from that of the routine method calibration
9.1 Use relevant good laboratory practices in accordance
standards.
with Guide D3856 and Practice E200.
7.2 Purity of Water—Unless otherwise indicated, references
9.2 Performananalysisonatleastoneportionofthesample
to water shall be understood to mean reagent water as defined
to estimate the concentration of the component(s) of interest.
by the individual test method to be used to analyze a sample
9.3 Use the result of this analysis to determine the appro-
after spiking. If more than one test method is to be used, the
priate amount of spike and spiking solution to be added to the
minimum criteria of each test method must be met. If test
sample. If this is not possible (such as when spiking in the
method reagent water specifications are not available, refer-
field),estimatetheconcentrationsofthecomponentsofinterest
ences to water shall be understood to mean reagent water as
based on prior knowledge of the sample source.
defined by Type I of Specification D1193 and demonstrated to
9.3.1 To be of maximum value for quantification of the
be free of interfering substances for the test(s) being per-
analyte(s) or for the evaluation of method accuracy, the
formed.
concentration in the spiked sample should be at least double,
7.3 Solvents—Spectroscopic, high-pressure liquid chroma-
butideallynotoverfivetimes,theconcentrationoftheanalyte
tography(HPLC),orultrapuregrademethanolispreferablefor
in the unspiked sample, as long as the total analyte concentra-
use as a solvent for relatively water-insoluble components in
tion can be brought within the test method’s dynamic range.
most trace-organic analyses. Other water-soluble solvents may
Spike concentrations below this range lead to highly variable
be useful as solvents for certain analytes. Most inorganic
spike recoveries, as described in Section 11. Higher spike
spiking solutions are prepared in water or dilute aqueous acid
concentrationsmaymasktheeffectthatrealinterferences,such
solution. Solvents shall be checked before use by analysis for
as matrix effects, are having on the analyte at its background
interfering substances.
levels,leadingtoover-optimisticestimatesofanalyterecovery.
9.3.2 If the spiked component is not present in the sample,
7.4 Spiking Solutions—Spiking solutions of each analyte of
but is added only to validate the recovery of an analytical
interest are prepared individually or in combination, either
method, the concentration after
...

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