OSHA SALT LAKE TECHNICAL CENTER

GUIDELINE FOR DEVELOPING SAMPLING AND ANALYTICAL METHODS

WITH VALIDATION REQUIREMENTS

Version: 1.0

April 28th, 2023

Method Development Team

Industrial Hygiene Chemistry Division
OSHA Salt Lake Technical Center
Sandy UT 84070-6406

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For assistance with accessibility problems in using figures and illustrations presented in this
guideline, please contact the OSHA Salt Lake Technical Center at (801) 233-4900.

Content

Introduction.................................................................................................................................................. 4
1 Preliminary Considerations................................................................................................................... 4
1.1 Target Concentration Selection.................................................................................................... 4
1.2 Sampling...................................................................................................................................... 5
1.2.1 Active Sampling....................................................................................................................... 5
1.2.2 Surface Sampling..................................................................................................................... 5
1.2.3 Bulk Sampling.......................................................................................................................... 6
1.3 Sampler Testing Techniques....................................................................................................... 6
1.3.1 Test Atmosphere...................................................................................................................... 6
1.3.2 Direct Spiking........................................................................................................................... 6
1.4 Internal Standard Selection.......................................................................................................... 7
1.5 Interference.................................................................................................................................. 7
2 Validation Testing.................................................................................................................................. 7
2.1 Limits of Detection and Quantitation............................................................................................ 7
2.2 Analytical Calibration................................................................................................................... 8
2.3 Analytical Recovery..................................................................................................................... 9
2.4 Stability of Prepared Samples.................................................................................................... 10
2.5 Post Sampling Storage.............................................................................................................. 11
2.6 Method Precision....................................................................................................................... 11
2.7 Sampler Capacity....................................................................................................................... 12
2.8 Effect of Humidity....................................................................................................................... 13
2.9 Cassette Wall Wiping Removal Efficiency..................................................................................14
2.10 Sampling and Analytical Interferents..........................................................................................14
2.10.1 Sampling Interferents............................................................................................................. 14
2.10.2 Analytical Interferents............................................................................................................. 15
2.11 Analytical Reproducibility........................................................................................................... 15
2.12 Confirmation Analysis................................................................................................................ 15
3 Estimation of Method Uncertainty....................................................................................................... 16
3.1 Sampling and Storage Uncertainty............................................................................................16
3.1.1 Air Volume Sampled.............................................................................................................. 16
3.1.2 Sampling Efficiency................................................................................................................ 17

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 3.1.3 Post Sampling Storage.......................................................................................................... 18
3.1.4 Sampling Interferents............................................................................................................. 18
3.2 Analytical Uncertainty................................................................................................................ 18
3.2.1 Calibration Standards............................................................................................................. 18
3.2.2 Analytical Recovery................................................................................................................ 18
3.2.3 Stability of Prepared Standards............................................................................................. 19
3.2.4 Method Precision................................................................................................................... 19
3.2.5 Method Bias........................................................................................................................... 20
3.2.6 Effect of Humidity................................................................................................................... 20
3.2.7 Cassette Wall Removal Efficiency.........................................................................................20
3.2.8 Analytical Interferent.............................................................................................................. 20
3.2.9 Instrument Response Drift..................................................................................................... 21
3.3 Combined Standard Uncertainty and Expanded Uncertainty.....................................................21
4 Preparation of Written Method............................................................................................................ 22
References................................................................................................................................................ 23

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Introduction

This guideline provides a uniform and practical means for validating sampling and analytical methods for
workplace exposure monitoring of hazardous chemicals by the Occupational Safety and Health
Administration (OSHA). The overall goal is to provide OSHA with usable sampling and analytical methods
that produce reliable results. It defines required laboratory tests and statistical calculations with
acceptance criteria for determining sampling and analytical parameters. This guideline replaces previous
method validation guidelines used by OSHA1,2,3 and harmonizes validation tests, statistical calculations,
terms, and definitions. This guideline also explains how OSHA will evaluate sources of both sampling and
analytical uncertainty and report a combined uncertainty value for sampling and analysis using the
approach outlined in the International Organization for Standardization (ISO) standard ISO 20581:2016. 4
Unique methods and procedures that do not fit within the framework of this guideline may also be
developed to fulfill the needs of the agency using other appropriate tests and procedures.

Before approval and use, all validated methods will be reviewed by technically competent individuals, and
found to demonstrate that a method is clearly written using standardized language, that terms and
numerical data are correctly used and presented, that consideration for inclusion of method target
analytes in a group that may use the same sampling media and conditions for simultaneous analysis has
been made, and that the validation requirements of this guideline have been met. All traceability
documentation and data associated with a method must be stored in a standardized format and made
accessible during the process of development and review, and for future reference.

1 Preliminary Considerations

Review the literature, regulatory standards, and other appropriate sources of information to determine
how and where the chemical substance to be sampled and analyzed is used in workplaces and how it is
or may be regulated. Review existing or related sampling and analytical methods to identify the potential
to add the chemical substance to an existing or prospective sampling group. Identify other common
chemicals present in those workplaces that could cause sampling and analytical interferences.

Consider the relevant chemical and physical properties in determining appropriate sampling, sample
preparation, and analytical techniques. If possible, incorporate sampling and analysis into a current OSHA
sampling group method so that common samplers, reagents, and instrumentation can be used, or
consider creation of a new sampling group as appropriate. For example, a non-polar analyte that can be
collected on coconut shell charcoal for desorption with carbon disulfide, and that may be analyzed by gas
chromatography with flame-ionization detection should be evaluated for inclusion into OSHA Method
5000.5 For stand-alone methods, explore ways to use common instrumentation configurations aligned
with other methods, and/or eliminate preparation steps as well as hazardous waste generation when this
is compatible with acceptable quality and usefulness for enforcement sampling.

1.1 Target Concentration Selection

Determine the concentration of the chemical substance at which the validation will be performed. This
value, referred to as the target concentration (TC), may be an OSHA Permissible Exposure Limit (PEL),
an American Conference of Governmental Industrial Hygienists (ACGIH) Threshold Limit Value (TLV), or
some other occupational exposure limit (OEL). The method can be validated at more than one
concentration if the chemical substance has multiple exposure limits; for example, an 8-hour time
weighted average (TWA), and one or more of the following: a ceiling, peak, short-term exposure limit

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(STEL), or action level. The equivalent analyte mass collected on a sampler at T C with the proposed
sampling volume (dependent on the sampling time and rate) is referred to as the target mass (TM).

1.2 Sampling

1.2.1 Active Sampling

Active sampling methods determine the concentration of an airborne chemical substance by actively
pumping air presumed to contain the substance into a vessel, or through a sampling medium, followed by
prescribed quantitative analytical procedures.

Use an appropriately designed sampling medium, such as a sorbent tube with separate front and back
sections, or a cassette with adequate support for a filter, to allow the production of results with required
quality and certainty. Consider how the analyte will interact with the sampling medium and any
components such as sorbent separators, supports, and cassette walls. For aerosol sampling, select a
sampler that will collect the appropriate size fraction (e.g., respirable) when this is required.

If possible, use a 4-hour sampling time for substances with an 8-hour TWA exposure limit. Select a flow
rate that will provide sufficient time for the analyte to collect, react, or adsorb to the sampling medium.
Flow rates between 50-200 mL/min for sorbent tubes, and 1-2 L/min for filters and OSHA Versatile
Samplers (OVS), are recommended. Optimal sampling time and rate will depend on the lowest mass per
sample reportable, defined as the limit of quantitation (LOQ) in Section 2.1, and the sampler capacity
defined in Section 2.7. If the LOQ for a prepared sample would be ≥ one-tenth the TM value, evaluate the
use of a higher flow rate or longer sampling time. If sampler capacity is exceeded evaluate the use of a
lower flow rate or shorter sampling time.

For substances that have a peak, ceiling, or other short-term exposure limit, optimize the sampling rate to
collect sufficient analyte mass so that the LOQ is ≤ one-half the corresponding TM value for the specific
exposure limit when sampling for a period relevant to the short-term exposure limit.

At each proposed sampling rate, measure the pressure drop across the sampler and confirm a
commercial sampling pump is available that can maintain a constant flow at the measured pressure drop
with adequate flow rate precision and accuracy.

1.2.2 Surface Sampling

Surface sampling is used to determine possible exposure to hazards that may be absorbed through the
skin or be ingested. A potentially contaminated surface is wiped with an appropriate sampling medium to
collect a target analyte.

Assure the wipe medium is large enough to have the capacity to handle the required analyte mass
expected within the area sampled, yet small enough to be practical for preparation in the laboratory. The
sampling medium should allow sampling of a 10×10 cm area, and be durable enough to withstand the
wiping procedure without tearing. Determine if the medium will need a wetting agent or if it can be used
dry. If a wetting agent is used, verify it will not interfere with detection of the target analyte.

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1.2.3 Bulk Sampling

Bulk sampling is not specifically addressed in this guideline. However, bulk sampling should be
addressed when necessary, for example, if required by an expanded standard, or if needed as an
appropriate reference material for performing supporting tests and analyses needed to correctly analyze
samples collected in the field. When specifying special requirements related to shipment of a bulk
material, assure the recommended shipping vessel is compatible chemically and physically, and is gas
tight if the sampled material is expected to contain volatile components.

1.3 Sampler Testing Techniques

1.3.1 Test Atmosphere

The use of a dynamically generated controlled test atmosphere is preferred for evaluating sampling
approaches when air contaminants are encountered in the gas phase. When safety concerns or other
problems prevent the use of a dynamic test atmosphere generation system, consider the use of a static
test atmosphere (e.g., target analyte injected into a gas-sampling bag). All test atmospheres generated
must be non-condensing. Generate test atmospheres using either dry or humid air as specified in Section
2, Validation Tests. Generate humid test atmospheres with an absolute humidity of 15.7 ± 3.0 milligrams
of water per liter of air, and dry test atmospheres with an absolute humidity of 3.92 ± 1.6 milligrams of
water per liter of air. These water concentration values represent 80% and 20% relative humidity
respectively at 22.2 °C. Perform sampling for testing using the proposed sampling time and sampling rate.

1.3.2 Direct Spiking

Use vapor spiking to test air samplers for air contaminants expected to be encountered in the vapor
phase when test atmosphere generation is not possible. This involves injecting the analyte, or the analyte
dissolved in a non-interfering solvent, directly onto a glass wool plug immediately upstream of a sampler.
Air is then drawn through the plug and sampler using the proposed sampling time and rate to move
analyte into the sampler. As a general rule, no more than 20 µL of solution should be used when vapor
spiking.

pike analyte onto media when vapor spiking is not possible. This involves spiking onto the first media
section intended to capture analyte that airflow encounters in an air sampler (e.g., the front section of a
sorbent tube). When using this technique ensure the analyte is loaded only onto the front section of a
sampler that contains more than one section. For samplers that contain a filter supported by a backup
pad, or a filter placed in front of a sorbent bed, be sure to spike in such a way that the analyte is not
immediately transferred to the support pad or sorbent bed. This can be done by removing a filter and
placing it on a suitable support, such as the rim of a small beaker so that only the outside edge of the filter
touches the support. Then carefully deliver liquid to about 5-10 small spots across the face of the filter.
Once the delivery solvent has evaporated, the filter can be placed back into its sampler and have air
drawn through it for testing where this is required.

Also use direct spiking to test wipe sample media that have been prepared using a proposed sampling
technique (e.g., when media is to be moistened before use). Spike the analyte directly onto the wipe
medium to test recovery, or onto a non-porous smooth surface such as a glass plate to test sampling
efficiency and recovery, as specified in Section 2. The testing surface should be the same dimensions as
the sampling area recommended in the method.

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1.4 Internal Standard Selection

Where an internal standard may be used in an analytical process, identify an internal standard that can be
used to correct for sample introduction, matrix interferences, instrument drift, and variable signal noise.
Assure internal standards are compatible with the sampling medium, analytical technique,
instrumentation, and are not typically found in the workplace.

1.5 Interference

The effect of water vapor on sampling and analysis must always be considered. Water vapor effects have
been known to include altered sampler capacity, analytical recovery, and sampler storage stability.
OSHA’s experience is that water will usually have a detrimental effect on sampling and analysis, thus air
sampler testing is mostly done using humid air; however, testing is also done using dry air as in some
cases this has also been shown to be detrimental.

Besides the effect of water, test other potential sampling and analytical interferences as needed.
Interferences can result in reduced sampler capacity, may react with an analyte or a derivatizing agent, or
negatively impact instrument performance.

2 Validation Testing

Validation tests are presented in logical order; however, the order in which tests are completed is not
important. Use calibration materials provided by a manufacturer accredited to ISO 17034:2016 6, where
practical, and traceable to National Institute of Standards and Technology (NIST) or equivalent national or
international standards, where possible. All other reference materials used must be traceable to certified
reference materials where possible. Use reagents of acceptable purity (e.g., reagent grade or better).
Ensure that supplier information (including lot numbers) for reagents, standards, and sampling media, and
expiration dates are captured in the traceability system used to document method development activities.
Evaluate and respect expiration dates for standards and reagents. Validation testing instrumentation must
be properly maintained and must be verified to be performing properly. Optimize analytical parameters so
that analyses may be completed at all required validation levels. Record all needed details regarding
instrument performance and maintenance status, traceability, and testing details to allow evaluation and
re-creation of all tests completed. Ensure that all relevant testing is documented, whether successful or
not, to leave a data trail that may be important for future method development work. Compliance with
these traceability and documentation requirements will result in a digitized data packet for validation work.
Complete all final validation tests of record using the sampling and analytical conditions described in the
final method.

2.1 Limits of Detection and Quantitation

Determine the Limit of Detection (LOD) and Limit of Quantitation (LOQ) for each sample collection
medium using the following procedure:

1. Estimate the background response from analysis of a sample medium blank.

2. Directly spike three separate samples at each of five evenly spaced levels, for a total of fifteen
spiked media samples, with the highest spiking level producing an instrument response about
ten times the background response of the media blank, and the lowest spiking level producing a
response about two times the background response.

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3. Prepare the fifteen spiked samples for analysis, along with three media blanks, and analyze in a
random order.

4. Plot instrument response versus calculated mass per sample and determine the slope ( m ) from
an ordinary least-squares (OLS) linear regression line equation.
5. Calculate the standard error of estimate for the OLS line equation using Equation 1:

S y / x=
√ ∑ ( y i−^y )2
( 1)
n−k

where S y / x is the standard error of estimate; y iis the observed instrument response; ^y is the
calculated response from the line equation; n is the number of samples and blanks (eighteen);
and k is 2 for linear regression.

6. Calculate the LOD using Equation 2:

3.3 × S y/ x
LOD=
m
(2 )

where LOD is the limit of detection in terms of mass per sample; S y / x is the standard error of
estimate; and m is the slope of the OLS line. Equation 2 assumes that the probability of a false
positive and of a false negative detection occurrence are both 5%, each sample will be analyzed
once, and results are not blank corrected.7-11

7. Calculate the LOQ using Equation 3:

10× S y / x
LOQ=
m
(3 )

where LOQ is the limit of quantitation in terms of mass per sample and is set to approximately 3
times the LOD; S y / x is the standard error of estimate; and m is the slope of the OLS line.

2.2 Analytical Calibration

Determine the analytical calibration procedure using the following procedure:

1. Establish a reporting limit (RL) value by selecting a mass that is ≥ LOQ and ≤ 0.1× the TM value.
If the LOQ is greater than one-tenth the TM value, select the LOQ as the RL value.

2. Prepare three separate analytical standards at ten evenly spaced levels, for a total of thirty
standards, covering the RL to at least 2.2× the TM value.

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3. Analyze each of the thirty standards in a random order.

4. As discussed in the National Institute of Standards and Technology/Sematech Handbook of

Statistical Methods “graphical analysis of the residuals is the single most important technique for
determining the need for model refinement or for verifying that the underlying assumptions of the
analysis are met.”12 Plot instrument response versus mass per sample as an OLS regression.
Then, calculate the residuals and plot residuals versus mass per sample. Visually examine both
the OLS regression and residual plots to assess the quality of the regression model and fit. 13,14

5. Perform a Shapiro–Wilk test, at the 95% confidence level, to assess normality of the residuals. 15
If the test leads to rejection of the null hypothesis that residuals are normally distributed (i.e., p-
value < 0.05), examine the use of a weighted least-squares (WLS) fitting technique 16 (e.g., x−1
weighting factor), or transform17 the data and return to Step 4.

6. Perform a Levene test, using means at the 95% confidence level, to assess the equality of
variances of the residuals.18 If the test leads to rejection of the null hypothesis that variances are
equal (i.e., p-value < 0.05), examine the use of a WLS fitting technique or transform the data and
return to Step 4.

7. Perform a lack-of-fit test, at the 95% confidence level, to assess the appropriateness of the
model and fit (e.g., WLS linear regression using a weighting factor of x −1).13,19 If the test leads to
rejection of the null hypothesis that the data fit the model (i.e., p-value < 0.05), examine the use
of a polynomial model (e.g., quadratic, cubic) and return to Step 4.

8. After determining the regression model and fitting technique, establish the analytical calibration
procedure to be used in the method validation test and presented in the method. For example,
“analytical calibration will be performed with five equally spaced standards, over the range of the
RL to 2.2× the TM value using WLS linear regression with a weighting factor of x−1.”

9. Prepare six spiked media samples at the RL and analyze using the proposed analytical
calibration procedure. Calculate percent recovery for each sample, correcting for the analytical
recovery value determined in Section 2.3, and the mean recovery. If the mean recovery of the
spiked samples is not within ±25% of the calculated value, adjust the regression model, fitting
technique, or RL of the proposed analytical calibration procedure and complete this test again.

2.3 Analytical Recovery

Determine analytical recovery for each type of sample collection medium/analyte tested using the
following procedure:

1. Directly spike six samples at each of five mass loading levels (i.e., 0.1, 0.5, 1.0, 1.5, and 2.0× the
TM value) for a total of thirty spiked media samples. The 0.1× TM value is not needed for peak,
ceiling, or other short-term exposure limits. For an OEL that may include multiple compounds,
such as for lead, perform analytical recovery tests using a reasonable subset of the possible
compound types (e.g., one soluble and two or three insoluble compounds that may be
reasonably expected to be found in the workplace).

2. Set the spiked samples aside for one hour to assure complete analyte sorption.

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3. Prepare four control standard replicates, without media, corresponding to each equivalent
concentration that would be obtained from processing samples with no preparation loss at the
same levels as in paragraph 1, for a total of twenty standards if five mass loading values are
used. Use the same equipment (e.g., syringe), technique, reagents, working standards, and
spiking volumes to prepare the control standards as used to prepare the directly spiked samples.

4. Prepare the spiked samples for analysis. Analyze control standards and samples at each level,
grouping each level together and evenly spacing the samples between the corresponding control
standards.

5. Calculate percent recovery for each sample using Equation 4:

Is
Ri= ×100
I std
(4 )

where Ri is the recovery of an individual spiked sample, in percent; I s is the instrument response for
the spiked sample; and I std is the mean response signal of the four control standards at the
equivalent level.

6. Calculate the mean recovery and variance for each of the five levels tested.

7. Apply a Dixon Q test for possible mean recovery outlier values, and a Cochran C test for within-
level variance outlier values across the five levels tested (both at the 95% confidence level). 20 A
mean recovery outlier, or a difference in variance between levels, is usually an indicator of a
poor analytical practice, such as using a 50-µL syringe to spike 1.00 µL of analyte.

8. Calculate the analytical recovery (RA) as the mean of the 30 individual sample recoveries. The
RA must be within 75 to 125%; however, a value between 95% and 105% is preferred, and the
mean recovery of each level should be within ±5% of the RA value.

9. Apply a two-tailed Student’s t-test at the 95% confidence level to determine if the bias of the
analytical recovery is significant compared to the reference value of 100%. If bias is significant,
sample results should be corrected, or include the bias as an uncertainty component of
analytical recovery.

2.4 Stability of Prepared Samples

Determine the stability of prepared samples for each type of sample collection media tested using the
following procedure:

1. Directly spike six samples at the T M value. Set the resulting spiked media samples aside for one
hour to assure complete analyte sorption. (Note: The 1.0× T M value samples prepared for
Section 2.3, can be used for this stability determination.)

2. Prepare four control standards, without media, corresponding to the equivalent concentration
that would be obtained from processing samples loaded at the T M value with no preparation loss.

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 Use the same equipment (e.g., syringe), technique, reagents, working standards, and spiking
volumes to prepare the control standards as used to prepare the spiked samples.

3. Prepare the spiked samples for analysis and immediately analyze. Analyze control standards
and samples, evenly spacing the samples between the corresponding control standards.

4. Reanalyze the same prepared samples again approximately one, two, and three days after the
initial analysis using four freshly prepared control standards for each analysis.

5. Calculate the recovery of each spiked sample using Equation 4. Plot percent recovery versus
days of storage and obtain an OLS linear regression line equation. Stability of prepared samples
(∆ ps) is calculated as the absolute difference between the final and initial test day recoveries
calculated using the line equation. If recovery changes by more than 5%, provide appropriate
directions for timely analysis in the method (e.g., could include requirements to perform analysis
within 24 hours, change storage conditions, or recap).

2.5 Post Sampling Storage

Determine the post sampling storage limitations for each type of sample collection medium/analyte
combination tested using the following procedure:

1. Collect eighteen air samples at the T C from a humid test atmosphere at the recommended
sampling time and with the recommended sampling rate, or for analytes where test atmosphere
generation is not possible, spike samples at the T M value followed by drawing humid air through
the samplers for the recommended sampling time at the recommended sampling rate. For wipe
samples, prepare eighteen samples by spiking the analyte onto the sampling medium as it is to
be used in the field (e.g., moistened).

2. Prepare and analyze three samples on the day of preparation (Day 0) using the analytical
calibration procedure determined in paragraph 8 of Section 2.2.

3. Store the remaining fifteen samples on a bench-top at room temperature.

4. Prepare and analyze three samples every third or fourth day, using freshly prepared calibration
standards, so that the duration of the entire storage test is about eighteen days.

5. Calculate the recovery for each sample, but do not correct sample results for analytical recovery
using the mean recovery value determined in Section 2.3. Plot percent recovery versus days of
storage and obtain an OLS linear regression line equation. Calculate the final and initial test day
recoveries using the line equation, then calculate post sampling storage stability ( ∆ ss) as the
absolute difference between the final and initial test day recoveries using the OLS lineequatiion
to obtain these. Recovery after eighteen days must be ≥75% and ∆ ssmust be ≤10%.

6. Perform a refrigerated post sampling storage stability test if recovery from room temperature
storage is ≤75% or ∆ ss is >10%. Freezer storage can also be tested if necessary.

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2.6 Method Precision

Determine method precision for each type of sample collection medium tested using the following
procedure:

1. Collect six air samples at each of five levels (i.e., 0.1, 0.5, 1.0, 1.5, and 2.0× the T C) from a
humid test atmosphere using the recommended sampling time and sampling rate, or for analytes
where test atmosphere generation is not possible, spike six samples at each of five levels (i.e.,
0.1, 0.5, 1.0, 1.5, and 2.0× the T M value) followed by drawing humid air through the samplers for
the recommended sampling time at the recommended sampling rate. The 0.1× TM can be
excluded for peak, ceiling, or other short-term exposure limits.

For wipe samples, collect six samples at each of five levels (i.e., 0.1, 0.5, 1.0, 1.5, and 2.0× the
TM value) by spiking the analyte onto a surface, and then wiping the surface with the sampling
medium using the proposed wiping technique. Use a surface that is extremely smooth and non-
porous such as a glass plate. A suggested technique is to draw a 10 cm × 10 cm square with a
marker on a glass plate and then spike the analyte on the reverse side. The analyte need not be
uniformly distributed within the confines of the test area. If the analyte is delivered onto the
surface in solution, allow any solvent used to evaporate before proceeding. If the analyte is
volatile proceed as rapidly as possible.

2. Prepare and analyze samples at each level as an independent application of the proposed
method, including use of new calibration standards prepared using the analytical calibration
procedure determined in paragraph 8 of Section 2.2.

3. Calculate percent recovery for each sample, correcting for analytical recovery using the mean
recovery value determined in Section 2.3.

4. Calculate the mean recovery and variance for each of the five levels tested.

5. Apply a Dixon Q test for possible mean recovery outlier values, and a Cochran C test for within-
level variance outliers across the five levels tested (both at the 95% confidence level). 20 A
statistical difference in variance between levels, or a mean recovery outlier, should be
investigated to determine if the cause is due to the testing procedure or method performance.

2.7 Sampler Capacity

Determine sampler capacity for each type of air sample collection medium/analyte combination tested
using the following procedure:

1. Assemble three test systems with each consisting of two samplers connected in series. If a
sampler contains a back section, it should be removed from the first sampler.

2. Sample at the recommended sampling rate through each system in parallel from a humid test
atmosphere containing the analyte at 2.0× the T C for 1.2× the proposed sampling time.
Beginning at 50 to 80% of the recommended sampling time replace the second sampler in each
of the three sample test systems at short time intervals. The time intervals will vary with the
sampling medium, analyte, and sampling time; however, intervals of 15-30 minutes are
preferred. Shorter intervals will lead to more samples requiring analysis and the potential for

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 recovery of insufficient mass for analysis but will also provide more precise data on breakthrough
time at the specified flow rate. These factors should be considered, and some experimentation
may be needed to empirically determine an optimized approach for this.

3. Prepare and analyze the resulting samples using the analytical calibration procedure determined
in paragraph 8 of Section 2.2.

4. If analyte is not detected on the back samplers after 1.2× the proposed sampling time, report
“capacity is not exceeded.” If analyte is detected on the back samplers, determine if the
cumulative analyte mass found on the second samplers is ≥5% of the mass that corresponds to
that calculated to have been present in the sampling volume. The 5% breakthrough volume can
be determined by plotting percent breakthrough versus sampled air volume, fitting the data with
a curve, and using regression to determine the volume corresponding to 5% breakthrough. The
recommended sampling time for a given sampling rate should be 80% of the calculated 5%
breakthrough time determined by the 5% breakthrough volume at a constant sampling rate.

5. Repeat this test by sampling dry air if the absence of water is expected to decrease sampler
capacity.
Sampler capacity can also be determined by monitoring the downstream effluent of a sampler with an
instrument, such as a total hydrocarbon analyzer or infrared spectroscopy instrument, after the response
to the upstream concentration has been established.

When it is not possible to use a dynamically generated controlled test atmosphere, determine sampler
capacity for each type of sample collection medium/analyte combination tested using the following
procedure:

1. Assemble three sampler test systems with two samplers connected in series and no back
section (if applicable) in the front sampler, as described above.

2. Spike each test system with 2.0× the T M value and then draw humid air through each for 1.2× the
proposed sampling time at the recommend sampling rate.

3. Prepare and analyze samples using the analytical calibration procedure determined in paragraph
8 of Section 2.2.

4. Breakthrough will be determined to have occurred if the average mass found on the three back
samplers is ≥5% of the total mass spiked.

2.8 Effect of Humidity

Determine the effect of humidity for each type of sample collection medium/analyte combination tested
using the following procedure:

1. Using dry air, prepare six samples at 2.0× the T C in the same manner used to prepare the
method precision samples in Section 2.6.

2. Prepare and analyze samples using the analytical calibration procedure determined in paragraph
8 of Section 2.2.

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3. Calculate the percent recovery for each sample, correcting for analytical recovery using the
mean recovery value determined in Section 2.3, and the mean recovery value obtained for these
test samples.

4. Calculate the effect of humidity (∆ h) as the absolute difference between the mean dry recovery
and the mean humid recovery taken from the 2.0× T C method precision test described in Section
2.6. If ∆ h is >10%, modify the sampling or analytical procedure if possible and repeat the test.

2.9 Cassette Wall Wiping Removal Efficiency

When a cassette is used in sampling, determine the efficiency of a proposed cassette interior wall wiping
removal technique using the following procedure:

1. Spike the interior wall of six sampler cassettes at the T M value. The analyte need not be
uniformly distributed within the cassette. If the analyte is delivered onto the cassette wall in
solution, allow any solvent used to evaporate before proceeding. Also, if a solvent is used
ensure that it does not react with the cassette material.

2. Wipe the interior of each cassette separately with an individual medium to be tested.

3. Prepare four control standards corresponding to the equivalent concentration that would be
obtained from processing samples loaded at the T M value with no preparation loss. Use the
same equipment (e.g., syringe), technique, reagents, working standards, and spiking volumes to
prepare the analytical standards as used to spike the cassettes.

4. Prepare the media used to wipe the spiked surfaces for analysis. Analyze the control standards
and samples at each level, grouping each level together and evenly spacing the wipe samples
between the control standards. Calculate the recovery of each spiked sample using Equation 4
and the mean recovery. Do not correct for analytical recovery using the mean recovery value
determined in Section 2.3.

5. Calculate the cassette wall wiping removal efficiency (∆ cw) as the absolute difference between
the mean recovery of the 1.0× test for analytical recovery, using the mean recovery value
determined in Section 2.3, and the mean cassette wall wiping recovery. If ∆ cw is >10%,
determine if removal efficiency can be increased with the use of a second wipe. The ∆ cwvalue
must be ≤25%.

2.10 Sampling and Analytical Interferents

Sampling and analytical interferents selected for testing should be based, in part, on interferents possibly
present in workplaces similar to those in which the method may be used. For testing, set the suspected
interferent concentration at an appropriate level, such as its PEL, TLV, or level anticipated to be found in
the workplace

2.10.1 Sampling Interferents

Determine the effect of a suspected sampling interferent by using the following procedure:

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1. Prepare six samples at 1.0× the TC in the same manner as used to prepare the method precision
samples described in Section 2.6 but include the potential interferent. For example, if a test
atmosphere is used in Section 2.6, generate a test atmosphere containing both the analyte and
potential interferent.

2. Prepare and analyze samples using the analytical calibration procedure determined in paragraph
8 of Section 2.2.

3. Calculate the recovery of each sample, correcting for analytical recovery using the mean
recovery value determined in Section 2.3, and the mean recovery value obtained from these test
samples.

4. Calculate the effect of the sampling interferent (∆ si) as the absolute difference between the mean
corrected recovery value for the interferent samples and the mean recovery taken from the 1.0×
the TC method precision test in Section 2.6. If ∆ si is >10%, modify the sampling or analytical
procedure if possible and repeat the test.

2.10.2 Analytical Interferents

Determine the effect of a suspected analytical interferent by using the following procedure:

1. Spike twelve samples at 1.0× the T C, six with analyte only and the other six with both the analyte
and potential interferent.

2. Prepare and analyze samples using the analytical calibration procedure determined in paragraph
8 of Section 2.2.

3. Calculate the recovery for each sample, correcting for analytical recovery using the mean
recovery value determined in Section 2.3. Calculate the mean corrected recovery of the six
samples with potential interferent and the six without.

4. Calculate the effect of the potential analytical interferent ( ∆ ai) as the absolute difference between
the mean corrected recovery values for the samples with and without interferent. If ∆ ai is >10%,
modify the sampling or analytical procedure if possible and repeat the test.

2.11 Analytical Reproducibility

Determine if the analytical procedure is reproducible using the following procedure:

1. Prepare six samples at 1.0× the TC for each sample collection medium/analyte combination in
the same manner used to prepare the method precision samples described in Section 2.6.

2. Submit the samples to the SLTC Production Chemistry Team for analysis along with a complete
draft copy of the proposed method. The analyst assigned these sample will prepare and analyze
them relying solely on the draft method for guidance. If possible, the analyst should use different
analytical equipment and ancillary support equipment to prepare and analyze the samples than
were used to gather method validation data. All sample results must fall within the calculated
expanded uncertainty bounds determined in Section 3.3. If the bounds are exceeded, steps must

15 of 26
 be taken to determine and eliminate the cause (e.g., lack of clarity in the method instructions
provided in the draft copy), followed by a repeat of the test.

2.12 Confirmation Analysis

When a non-orthogonal analytical technique is used, determine alternate analytical conditions to aid in
confirming the identity or purity of the analyte (or relevant derivative). Mass spectrometry can often
provide conclusive identification when combined with another method such as gas chromatography and
should be considered first when possible. Peak response ratios and analysis with alternate detectors may
also provide confirmation.

3 Estimation of Method Uncertainty

For each sampler/analyte combination and target concentration (T C), calculate the relevant sampling and
analytical uncertainty components as described below in Sections 3.1 and 3.2, and then calculate the
combined standard uncertainty and expanded uncertainty as described in Section 3.3. For an analyte
sampled as both a vapor and particulate, where each phase is collected on a separate sampling medium
(e.g., filter preceding a sorbent tube), calculate combined standard uncertainties for each medium and
then calculate a combined standard uncertainty for the sampling procedure.21

3.1 Sampling and Storage Uncertainty

3.1.1 Air Volume Sampled

3.1.1.1 Flow Rate Measurement

Calculate the uncertainty associated with the flow rate measurement of the flow meter, using Equation 5:

CV fr
u fr =
√n
(5)

where u fr is the percent relative standard uncertainty of the flow rate measurement, and CV fr is the flow
rate coefficient of variation (expressed as percent), of n flow measurements. Determine CV fr from
reproducibility measurements using the flow meter with a representative sampler inline at the
recommended flow rate. Take at least three measurements (n = 3).

3.1.1.2 Flow Rate Calibration

Calculate the uncertainty associated with the calibration of the flow meter, assuming a rectangular
probability distribution, using Equation 6:

∆ fc
u fc=
√3
(6 )

16 of 26
where u fc is the percent relative standard uncertainty of the flow meter calibration, and ∆ fc is the
manufacturer’s reported tolerance specification expressed as a percent value.

3.1.1.3 Pump Flow Stability

Calculate the uncertainty associated with the flow stability of the sampling pump, assuming a rectangular
probability distribution, using Equation 7:

∆fs
u fs=
√3
( 7)

where u fs is the percent relative standard uncertainty of the sampling pump flow stability, and ∆ fs is the
flow stability of the sampling pump expressed as percent, assuming the flow rate does not change by
more than ±5% or ±3 mL/min, whichever is greater, during the sampling period.

3.1.1.4 Sampling Time

Calculate the uncertainty associated with sampling time, assuming a triangular probability distribution,
using Equation 8:

( Bst ×t−1
st )
u st = × 100
√6
( 8)

where u st is the percent relative standard uncertainty of the sampling time, t st is the sampling time in min,
and Bst is the sum of maximum bias at the beginning and end of the sampling period (e.g., when
sampling to nearest minute Bst = 0.5 min + 0.5 min = 1 min).

3.1.2 Sampling Efficiency

3.1.2.1 Gas and Vapor Sampling

The relative standard uncertainty associated with gas and vapor sampling efficiency (u se) is negligible
when sampling volume is limited to ensure sampler capacity is not exceeded as determined in Section
2.7.

3.1.2.2 Aerosol Sampling

Components associated with the relative standard uncertainty of aerosol sampling efficiency depend on
the type of sampler and can include the following:

 deviation from the sampling convention (e.g., inhalable, respirable);

 variability between samplers;
 sampler test system calibration;
 sampled concentration.

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These uncertainty components are described in ISO 21832:2020 and estimate values are provided. 22
Calculate the relative standard uncertainty for the sampling efficiency ( u se) by propagation of errors using
the appropriate uncertainty components.

3.1.2.3 Wipe Sampling

Calculate the uncertainty associated with wipe sampling efficiency, assuming a rectangular probability
distribution, using Equation 9:

55 %
u se=
√3
( 9)

where u se is the percent relative standard uncertainty of the sampling efficiency, and 55% is the
calculated difference between 100% and 45%.a

3.1.3 Post Sampling Storage

Calculate the uncertainty associated with post-sampling storage stability, assuming a rectangular
probability distribution, using Equation 10:
∆ss
u ss =
√3
( 10 )
where u ss is the percent relative standard uncertainty of post sampling storage, and ∆ ssis the storage loss
or gain calculated in Section 2.5.

3.1.4 Sampling Interferents

Calculate the uncertainty associated with a sampling interferent, assuming a rectangular probability
distribution, using Equation 11:

∆si
u si=
√3
( 11 )

Where u siis the percent relative standard uncertainty of the effect of a sampling interferent, and ∆ siis the
effect of a sampling interferent (expressed as percent) calculated in Section 2.10.1.

a
It has been demonstrated that across a range of surface types a repeat wipe of a surface, on average, will remove
55% of the contaminant concentration of the initial wipe.23 Assuming the ratio of 0.55 remains constant for further
repeat wipes, the average amount of contaminant removed from the surface by the initial wipe can be determined
S=a1 / ( 1−r ). Setting Sto 1 as the total amount of contaminant
using the equation for an infinite geometric series:
on the surface, r to 0.55 as the common ratio between repeat wipes, and solving for a 1as the amount of contaminant
removed by the initial wipe gives 0.45 (45%).

18 of 26
3.2 Analytical Uncertainty

3.2.1 Calibration Standards

Determine the uncertainty components associated with the concentration of the calibration standards.
These components depend on how the calibration standards are made and can include the following:

 purity of the starting material (e.g., purity >99.0%);

 uncertainty associated with measuring the starting material (e.g., weighing, pipetting);
 uncertainty associated with diluting;
 laboratory temperature.

Calculate the relative standard uncertainty for the calibration standards ( ucs ) by propagation of errors
using the appropriate uncertainty components. Examples of the calculation of ucs can be found in the
Eurachem guide “Quantifying Uncertainty in Analytical Measurement.24

3.2.2 Analytical Recovery

Calculate the uncertainty associated with sample results corrected for analytical recovery using Equation
12:

√( )
2
CV R
uar = A

√n
( 12 )

where uar is the percent relative standard uncertainty of the analytical recovery, CV R is the coefficient of
A

variation of the thirty analytical recovery samples analyzed per Section 2.3 (expressed as a percent
value), n is thirty.

Calculate the uncertainty associated with sample results not corrected for analytical recovery using
Equation 13:

√( ) (√ )
2 2
BR CV R
uar = A
+ A

√3 n
( 13 )

where BR A is the absolute difference between the mean percent recovery of the thirty analytical recovery
samples analyzed per Section 2.6, and the calculated 100% recovery value. If multiple compounds were
tested (e.g., one soluble and two insoluble) calculate uar for each compound and use the largest uar value
to calculate the combined standard uncertainty in Section 3.3.

3.2.3 Stability of Prepared Standards

Calculate the uncertainty associated with the stability of prepared samples, assuming a rectangular
probability distribution, using Equation 14:

19 of 26
 ∆ ps
u ps=
√3
( 14 )

where u ps is the percent relative standard uncertainty of the stability of prepared samples processed and
maintained as required by the method, and ∆ ps is the storage loss or gain calculated in Section 2.4.

3.2.4 Method Precision

Calculate the uncertainty associated with method precision as described in Section C.6.2 of ISO/DIS
22065:201825, using Equation 15:

√
ump = ( CV m ) 2+ 1− ( 1n )(CV
( 15 )
pl
2
)

where ump is the percent relative standard uncertainty of method precision, CV mis the coefficient of
variation for the means of the five levels tested in Section 2.6 (expressed as percent), n is the
number of replicate samples tested per level (six); CV plis the pooled coefficient of variation of the
five levels tested in Section 2.6 (expressed as percent), calculated using Equation 16:

√
2 2 2
( CV 1 ) + ( CV 2 ) + ⋯ + ( CV 5 )
CV pl =
5
( 16 )

where CV 1 through CV 5are the coefficients of variation of the five levels tested expressed as
percent values.

3.2.5 Method Bias

Calculate the uncertainty associated with method bias using Equation 17:

√( )( )
2 2
B mb CV mb 2
umb = + + ( urc )
√ 3 √ n
( 17 )

where umb is the percent relative standard uncertainty of the method bias, Bmb is the absolute difference
between the mean percent recovery of the thirty method precision samples analyzed per Section 2.6, and
the calculated 100% recovery value; CV mbis the percent coefficient of variation of the recovery of the
thirty samples analyzed per Section 2.6, n is thirty, and urc is the percent relative standard uncertainty of
the reference concentration sampled or mass spiked. For gas and vapor dynamic test atmosphere
generation use an estimated urc value of 3% as suggested in ISO/DIS 22065:2018.25

20 of 26
3.2.6 Effect of Humidity

Calculate the uncertainty associated with humidity effect, assuming a rectangular probability distribution,
using Equation 18:

∆h
uh =
√3
( 18 )

where uh is the percent relative standard uncertainty of the effect of humidity, and ∆ h is the effect of
humidity difference calculated in Section 2.8, expressed as a percent value.

3.2.7 Cassette Wall Removal Efficiency

When a cassette is used in sampling, calculate the uncertainty associated with cassette wall wiping
removal efficiency, assuming a rectangular probability distribution, using Equation 19:

∆cw
ucw =
√3
( 19 )

where ucw is the percent relative standard uncertainty of analyte removal from cassette walls, and ∆ cwis
the percent cassette wall wiping removal efficiency calculated in Section 2.9, expressed as a percent
value.

3.2.8 Analytical Interferent

Calculate the uncertainty associated with an analytical interferent, assuming a rectangular probability
distribution, using Equation 20:

∆ ai
uai =
√3
( 20 )

where uai is the percent relative standard uncertainty of the effect of an analytical interferent, and ∆ ai is
the of an analytical interferent calculated in Section 2.10.2, expressed as a percent value.

3.2.9 Instrument Response Drift

Calculate the uncertainty associated with instrument response drift, assuming a rectangular probability
distribution, using Equation 21:

d max
udr =
√3
( 21 )

21 of 26
where udr is the percent relative standard uncertainty of the instrument response drift, and d max is the
percent maximum allowed instrument response drift of a continuing calibration standard.

3.3 Combined Standard Uncertainty and Expanded Uncertainty

Calculate the sampling and storage uncertainty using Equation 22:

√∑ (
ns
2
u s= us )
i
i=1
( 22 )

where u s is the percent relative standard sampling and storage uncertainty, u s is the ith sampling
i

uncertainty component expressed as percent calculated in Section 3.1, and n s is the total number of
relevant sampling uncertainty components addressed in Section 3.1.

Calculate the analytical uncertainty using Equation 23:

√∑ (
na
2
ua = ua )
i
i=1
( 23 )

where ua is the percent relative standard analytical uncertainty, ua is the i th sampling uncertainty
i

component expressed as percent, calculated in Section 3.2, and n a is the total number of relevant analytical
uncertainty components calculated in Section 3.2.

Calculate the combined uncertainty using Equation 24:

√ 2
u= ( u s ) + ( u a )
2

( 24 )

where u is the combined percent relative standard uncertainty for sampling and analysis, u s is the percent
relative standard sampling uncertainty, and ua is the percent relative standard analytical uncertainty.

Calculate the expanded uncertainty using Equation 25:

U =k ×u
( 25 )

where U is the percent expanded uncertainty of the sampling and analysis procedure, u is the combined
percent relative standard uncertainty for sampling and analysis; and k is the coverage factor. For a two-
sided 95% confidence interval use a coverage factor of 2.

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4 Preparation of Written Method

Include the following sections in OSHA sampling and analytical methods:

 Cover Page
 Introduction
 Sampling Procedure
 Analytical Procedure
 Validation and Estimation of Measurement Uncertainty
 References

Include the following information on the cover page: method number, version number, OSHA PEL, type(s)
of PEL (e.g., “general industry,” “construction,” and/or “shipyard” as applicable), and other appropriate
OEL values, CAS No., brief description of sampling and analytical procedure, recommended sampling
time and rate, LOQ, RL, uncertainty, any special requirements (e.g., cold shipping), status, and release
date. For new methods assign a version number beginning with 1.0. Increment the version number by 0.1
for small changes, such as changes to formatting, and increment by 1.0 for significant changes. When a
method is revised, list the revision date on the cover page and describe the changes in the Introduction.
All methods with a reported combined uncertainty will be considered fully validated.

In the Introduction section, include relevant historical information regarding previous methods and
procedures used or tested by OSHA, along with any informative information from the literature. In the
Sampling Procedure and Analytical Procedure sections, describe the materials and procedures used for
performing sampling and analysis. In the Validation section, describe the results from the validation tests
performed and include a description of the testing procedures. For the Estimation of Measurement
Uncertainty, list the sampling and analytical uncertainty component values. Include comments to ensure
clarity on how values were determined, the combined uncertainty value, and the expanded uncertainty
value with the coverage factor.

Present experimentally derived data with appropriate significant figures. Report percentages to one
decimal place, unless the value is less than 1%, then report two or more decimal places as technically
appropriate. Report LOQ, RL, combined uncertainty, and expanded uncertainty using two significant
figures.

23 of 26
References

24 of 26
1
. Eide, M.; Simmons, M.; Hendricks, W. Validation Guidelines for Air Sampling Methods Utilizing
Chromatographic Analysis, 2010. United States Department of Labor, Occupational Safety & Health
Administration Web site. http://www.osha.gov (accessed March 2021).

2
. Eide, M.; Giles, P.; Simmons, M.; Hendricks, W. Evaluation Guidelines for Air Sampling Methods Utilizing
Spectroscopic Analysis, 2005. United States Department of Labor, Occupational Safety & Health
Administration Web site. https://www.osha.gov (accessed March 2021).

3
. Lawrence, R. Evaluation Guidelines for Surface Sampling Methods, 2001. United States Department of
Labor, Occupational Safety & Health Administration Web site. https://www.osha.gov (accessed March
2021).

4
. ISO 20581:2016, Workplace air – General requirements for performance of procedures for the
measurements of chemical agents.

5
. Pearce, D. Organic Vapor Sampling Group 1 (OVSG-1), CS 2 Extracted Analytes Collected on Coconut
Charcoal Sorbent Tubes (OSHA Method 5000), 2020. United States Department of Labor, Occupational
Safety & Health Administration Web site. https://www.osha.gov (accessed March 2021).

6
. ISO 17034:2016, General requirements for the competence of reference material producers.

7
. Burkhart, A. J. General procedures for limit of detection calculations in the industrial hygiene chemistry
laboratory. Appl. Ind. Hyg. 1986, 1, 153-155.

12
. Graphical Residual Analysis - Initial Model. NIST/SEMATECH e-Handbook of Statistical Methods. United
States Department of Commerce, National Institute of Standards and Technology Web site. https://www.
itl.nist.gov/div898/handbook/pmd/section6/pmd614.htm (accessed March 2021).

13
. Raposo, F. Evaluation of analytical calibration based on least-squares linear regression for instrumental
techniques: A tutorial review. TrAC Trends in Analytical Chemistry 2016, 77, 167-185.

14
. Ellison, S. L. R.; Barwick, V. J. Duguid Farrant, T. J. Regression. Practical Statistics for the Analytical
Scientist: A Bench Guide, 2nd Edition; RCS Publishing: Cambridge UK, 2009; pp 97-98.

15
. Anderson-Darling and Shapiro-Wilk tests. NIST/SEMATECH e-Handbook of Statistical Methods. United
States Department of Commerce, National Institute of Standards and Technology Web site.
https://www.itl.nist.gov/div898/handbook/prc/section2/prc213.htm (accessed March 2021).
16
. Almeida, A. M.; Castel-Branco, A. C.; Falcao, A. C. Linear regression for calibration lines revisited: weighting
schemes for bioanalytical methods. J. Chromatogr. B 2002, 774(2), 215-222.

17
. Transformations to Improve Fit and Equalize Variances. NIST/SEMATECH e-Handbook of Statistical
Methods. United States Department of Commerce, National Institute of Standards and Technology Web
site. https://www.itl.nist.gov/div898/handbook/pmd/section6/pmd624.htm (accessed March 2021).

18
. Levene Test for Equality of Variances. NIST/SEMATECH e-Handbook of Statistical Methods. United States
Department of Commerce, National Institute of Standards and Technology Web site.
https://www.itl.nist.gov/div898/handbook/eda/section3/eda35a.htm (accessed March 2021).

19
. Analytical Methods Committee. Is my calibration linear? Analyst 1994, 119(11), 2363-2366.
20
. Ellison, S. L. R.; Barwick, V. J.; Duguid Farrant, T. J. Outliers in Analytical Data. Practical Statistics for the
Analytical Scientist: A Bench Guide, 2nd Edition; RCS Publishing: Cambridge UK, 2009; pp 49-53.

21
. EN 13936:2014, Workplace exposure – Procedures for measuring a chemical agent present as a mixture of
airborne particles and vapour – Requirements and test methods.

22
. ISO 21832:2020, Workplace air – Metals and metalloids in airborne particles – Requirements for evaluation of
measuring procedures.
24
. Ellison, S. L. R. and Williams, A. (Eds). Eurachem/CITAC guide: Quantifying Uncertainty in Analytical
Measurement, Third edition, (2012). Available from www.eurachem.org.

25
. ISO/DIS 22065:2018, Workplace air - Procedures for measuring gases and vapours using pumped
samplers - Requirements and test methods.