Guidance for Industry and FDA

Staff
Assayed and Unassayed Quality
Control Material

Document issued on: June 7, 2007

The draft of this document was released on February 3, 1999.

For questions regarding this document contact Carol Benson, 240-276-0490, email
carol.benson@fda.hhs.gov.

U.S. Department of Health and Human Services

Food and Drug Administration
Center for Devices and Radiological Health
Office of In Vitro Diagnostic Device Evaluation and Safety
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Preface
Public Comment
Written comments and suggestions may be submitted at any time for Agency consideration to the
Division of Dockets Management, Food and Drug Administration, 5630 Fishers Lane, Room 1061,
(HFA-305), Rockville, MD, 20852. Alternatively, electronic comments may be submitted to
http://www.fda.gov/dockets/ecomments. When submitting comments, docket number 98D-1232.
Comments may not be acted upon by the Agency until the document is next revised or updated.

Additional Copies
Additional copies are available from the Internet
http://www.fda.gov/cdrh/oivd/guidance/2231.pdf You may also send an e-mail request to
dsmica@fda.hhs.gov to receive an electronic copy of the guidance or send a fax request to 240-
276-3151 to receive a hard copy. Please use the document number 2231 to identify the guidance
you are requesting.

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Table of Contents
I. Introduction ............................................................................................................................. 4
II.Classification and Identification of QC Materials................................................................... 5
III.
Assayed and Unassayed QC Material for Qualitative and Quantitative Assays..................... 7
IV.Information to include in a 510(k) for Assayed QC Materials ............................................... 8
A. Device Description .......................................................................................................... 8
B. Performance Evaluation and Results ............................................................................... 9
1. Matrix Effects.......................................................................................................... 9
2. Analyte Value Assignment .................................................................................. 10
3. Stability ................................................................................................................. 10
4. Surrogate QC material ..................................................................................... 11
V. Labeling................................................................................................................................. 11

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Guidance for Industry and FDA Staff

Assayed and Unassayed Quality Control

Material

This guidance represents the Food and Drug Administration's (FDA's) current thinking on
this topic. It does not create or confer any rights for or on any person and does not operate to
bind FDA or the public. You can use an alternative approach if the approach satisfies the
requirements of the applicable statutes and regulations. If you want to discuss an alternative
approach, contact the FDA staff responsible for implementing this guidance. If you cannot
identify the appropriate FDA staff, call the appropriate number listed on the title page of this
guidance.

I. Introduction
This guidance document provides recommendations to manufacturers regarding
preparation of premarket notifications and labeling for quality control (QC) material.
These materials are intended to monitor reliability of a test system and help minimize
reporting of incorrect test results. These materials are often the best source of ongoing
feedback that a laboratory has to monitor whether results reported to physicians are
sufficiently reliable. QC materials may be marketed together with a specific test system,
or alternatively, for more general use.

Both assayed and unassayed QC materials are discussed in this guidance document. Both types
of QC materials are subject to the Quality System Regulation (QSR), 21 CFR, Part 820, and
labeling regulation 21 CFR 809.10. However, most types of unassayed QC materials are exempt
from premarket notification (see Section II below for exceptions). Although premarket
notifications are not required for unassayed QC materials, some aspects of this guidance
document such as labeling, stability, and matrix effects are still relevant for these materials.
Assayed QC materials have analyte values1 or ranges assigned by the manufacturer and
presented in the labeling. Unassayed QC materials do not have manufacturer-assigned values or
ranges; rather, only the end user laboratories assign values and ranges. These two categories of
QC materials are further described in Section III. When QC materials are included as a
component of a test system that is itself exempt from premarket notification, the QC materials
also do not need premarket notification. However, as with unassayed QC materials, many of the

1
In this guidance document we use the term values to refer to units of assay output, or other
units from which assay output could be inferred by an end user.

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recommendations in this guidance document for evaluation and labeling still apply.

This guidance addresses external QC material. It is not intended to address internal controls,
process controls (such as control lines on single use devices) or, electronic QC. This guidance
does not address calibrators. A separate guidance is available for calibrators at,
http://www.fda.gov/cdrh/ode/calibrator.pdf. This guidance also does not address QC materials
for genetic tests for heritable diseases or pharmacogenetic testing. These types of materials will
be addressed in other OIVD (Office of In Vitro Diagnostic Device Evaluation and Safety)
guidance documents. This guidance is not intended to apply to QC materials for use with donor
screening tests. These materials are regulated by Center for Biologics Evaluation and Research
(CBER).

FDA's guidance documents, including this guidance, do not establish legally enforceable
responsibilities. Instead, guidances describe the Agency's current thinking on a topic and should be
viewed only as recommendations, unless specific regulatory or statutory requirements are cited. The
use of the word should in Agency guidances means that something is suggested or recommended,
but not required.

Abbreviated 510(k) Submissions

As explained in The New 510(k) Paradigm - Alternate Approaches to Demonstrating
Substantial Equivalence in Premarket Notifications
(http://www.fda.gov/cdrh/ode/parad510.html), a manufacturer may submit either a Traditional
510(k) or an Abbreviated 510(k). An Abbreviated 510(k) provides a means to streamline the
review of data in a 510(k) through a reliance on FDA-recognized consensus standards, special
controls, or FDA guidance documents. Guidance on the content and format for Abbreviated and
Traditional 510(k)s is available at http://www.fda.gov/cdrh/ode/guidance/1567.html. Also, see
Section 514(c)(1)(B) of the Act and the FDA guidance, Use of Standards in Substantial
Equivalence Determinations http://www.fda.gov/cdrh/ode/guidance/1131.html on use of
standards in an abbreviated 510(k).

The Least Burdensome Approach

The issues identified in this guidance document represent those that we believe should be
addressed before your device can be marketed. In developing the guidance, we carefully
considered the relevant statutory criteria for Agency decision-making. We also considered the
burden that may be incurred in your attempt to follow the guidance and address the issues we
have identified. We believe that we have considered the least burdensome approach to resolving
the issues presented in the guidance document. If, however, you believe that there is a less
burdensome way to address the issues, you should follow the procedures outlined in the
document, A Suggested Approach to Resolving Least Burdensome Issues. It is available on
our Center web page at: http://www.fda.gov/cdrh/modact/leastburdensome.html.
.

II. Classification and Identification of QC Materials

The various types of QC materials are identified, and classified, as follows in the CFR:

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a.) 21 CFR 862.1660 Quality control material.

(a) Identification. A quality control material (assayed and unassayed) for clinical
chemistry is a device intended for medical purposes for use in a test system to
estimate test precision and to detect systematic analytical deviations that may arise
from reagent or analytical instrument variation. A quality control material (assayed
and unassayed) may be used for proficiency testing in interlaboratory surveys. This
generic type of device includes controls (assayed and unassayed) for blood gases,
electrolytes, enzymes, multi-analytes (all kinds), single (specified) analytes, or
urinalysis controls.
(b) Classification. Class I (general controls). Except when used in donor
screening tests, unassayed material is exempt from premarket notification
procedures, subject to 21 CFR 862.9.

b.) 21 CFR 862.3280. Clinical toxicology control material.

(a) Identification. A clinical toxicology control material is a device intended to
provide an estimation of the precision of a device test system and to detect and
monitor systematic deviations from accuracy resulting from reagent or instrument
defects. This generic type of device includes various single, and multi-analyte
control materials.
(b) Classification. Class I (general controls). Except when used in donor screening
tests, unassayed material is exempt from premarket notification procedures in subpart
E of part 807 of this chapter subject to Sec. 862.9.

c.) 21 CFR 864.5425. Multipurpose system for in vitro coagulation studies.

(a) Identification. A multipurpose system for in vitro coagulation studies is a device
consisting of one automated or semiautomated instrument and its associated reagents
and controls. The system is used to perform a series of coagulation studies and
coagulation factor assays.
(b) Classification. Class II (performance standards).

d.) 21 CFR 864.8625. Hematology quality control mixture.

(a) Identification. A hematology quality control mixture is a device used to ascertain
the accuracy and precision of manual, semiautomated, and automated determinations
of cell parameters such as white cell count (WBC), red cell count (RBC), platelet
count (PLT), hemoglobin, hematocrit (HCT), mean corpuscular volume (MCV),
mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin
concentration (MCHC).
(b) Classification. Class II (performance standards).

e.) 21 CFR 866.2480. Quality Control Kit for Culture Media.

(a) Identification. A quality control kit for culture media is a device that
consists of paper discs (or other suitable materials), each impregnated with a
specified, freeze-dried, viable microorganism, intended for medical purposes
to determine if a given culture medium is able to support the growth of that

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microorganism. The device aids in the diagnosis of disease caused by

pathogenic microorganisms and also provides epidemiological information on
these diseases.
(b) Classification. Class I (general controls). The device is exempt from the
premarket notification procedures in subpart E of part 807 of this chapter subject to
the limitations in Sec. 866.9.

III. Assayed and Unassayed QC Material for Qualitative

and Quantitative Assays
A. Assayed QC Materials
Assayed QC materials have analyte values specified in the labeling by the manufacturer. These
analyte values and associated ranges are provided as guidelines to the end user; the clinical
laboratory establishes its own ranges based on its own test system and criteria. QC materials for
qualitative tests, as well as quantitative tests may be assayed.

When QC materials are marketed for a specific assay, or assays, the manufacturer should assign
analyte values of the QC material, relative to the medical decision points associated with that
assay. Therefore, we consider all QC materials recommended by the manufacturer for a specific
assay, (or those included as part of an assay system) to be assayed QC material.

The following are types of labeling information that might render a control an assayed control:

Units are assigned to the control(s) in your labeling (examples of units include, but are
not limited to, IU/mL, copies/mL and mg/mL).

Units are used to describe the preparation of the control(s) in your labeling, such that an
assays expected value might be inferred by the user.

The controls are described in the labeling as for use with one or more specific assays.

External QC materials for qualitative tests marketed to waived test sites should be assayed by the
manufacturer and appropriately labeled, since these types of testing sites do not typically assign
control values. Labeling should include the expected target value, the relationship of this value
to the cut-off point of importance in the assay being controlled, and estimates of performance
around the target value.

B. Unassayed QC Materials
Unassayed QC materials do not have assigned analyte values provided by the manufacturer and
are not linked to specific assays/assay systems by labeling or other means. The end user, rather
than the manufacturer assigns expected results to unassayed QC material. The manufacturer
may indicate whether a specific analyte is present or absent in the QC material preparation
without indicating an expected assay result.

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While manufacturing of a QC material should include some type of analyte level assessment as
part of the production process, for unassayed QC material you should not provide end users with
values from which they might mistakenly infer expected assay results.

The following are examples of types of label information that alone would not render a control
an assayed:

Relative dilutions (e.g., linear dilutions ranging from 1:2 to 1:20) are specified, but the
absolute and/or source concentration is unspecified and it is clear that the user laboratory
is responsible for determining the value of the control.

Relative levels such as high, medium and low are provided, but without any specific
values or concentrations and it is clear that the user laboratory is responsible for
determining the value of the control.

For assays that target infectious microorganisms, specific protein(s) or nucleic acid
sequence(s) that make up the QC material are specified but do not provide information on
likely test performance (strong negative, weak positive, strong positive or other control
values). (This does not include QC material for pharmacogenetic tests, or tests for
heritable markers. These types of QC materials are addressed in the guidance document,
Guidance for Industry and FDA Staff - Class II Special Controls Guidance Document:
Quality Control Material for Cystic Fibrosis Nucleic Acid Assays,
http://www.fda.gov/cdrh/oivd/guidance/1614.pdf)

IV. Information to Include in a 510(k) for Assayed QC Materials

A. Device Description
Your device description in the 510(k) should include the following types of information, as
applicable:

Intended use (including the types of assays and analyzers the material is intended to be
used with).
Information concerning the composition of the QC material, including:
Concentrations of each analyte in each level of QC material.
The relevance of the concentrations chosen (e.g., concentrations represent common
medical decision points, concentrations are chosen to incorporate the range of the
assay and to challenge the cut-off or decision point of clinical importance).
Base matrix (e.g., serum, blood, urine, buffer), and any known matrix effects that
affect assay performance (see definitions in Section IV.B.1 below).
Added components such as stabilizers, preservatives, clarifiers, conjugated materials
etc., and any known matrix effects of these added components that affect assay
performance.
Any other information pertinent to users concerning how the QC material is prepared.
If you market calibrators and control material for a single assay, we recommend that

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you clarify what steps were taken to ensure that these materials will provide
assessments independent of each other. For example, the controls and calibrators
could be prepared from separate lots of serum pools (or other materials).
Analyte source, e.g., human or animal species, synthetic or recombinant. For
recombinant nucleic acid material, you should include the vector, the source of the
cloned nucleic acid region or gene, and specific nucleic acid sequence. For a
microorganism, you should include the strain, and portion of the microorganism, the
media or cell line used for culture.
Safety information, including methods you used to test for infectious agents. When
blood products are used, you should include a certification statement that the
animal/human source components used in the control are safe and that any blood
product derived material has been tested by FDA approved (or equivalently
recognized) assays and found to be negative for the communicable disease agents, as
stated in 21 CFR Part 610. If inactivation methods have been used for infectious
agents, you should describe the methods and the results demonstrating non-
infectivity.

B. Performance Evaluation and Results

1. Matrix Effects
The matrix refers to all components of a material system, except for the analyte2. For QC
materials this generally refers to the base material (e.g., serum, buffer) and added
components such as stabilizers or preservatives. The matrix effect refers to the influence of a
property of the sample, other than the analyte, on the measured value of the analyte.
Viscosity, surface tension, turbidity, ionic strength, and pH are among causes of matrix
effects.

Ideally, QC materials simulate the composition of patient samples as closely as possible in

order to minimize matrix effects and correctly reflect the expected performance with patient
samples. Use of matrices such as human serum, urine, or whole blood-derived samples may
help accomplish this. However, in efforts to reduce cost or increase convenience or safety,
modifications might be made to the matrices. For example, animal and synthetic matrices
are used to protect laboratory personnel from exposure to human infectious agents.
Preservatives, stabilizing agents, antimicrobials, and clarifying agents may also be added to
enhance the ease of use and stability of a QC material. Manufacturing processes, (e.g.,
lyophilization or inactivation) may significantly alter the physical, chemical, or biological
properties of the QC material. Although these various matrices and additives at times have
benefits, these deviations from human samples may compromise the QC materials ability to
sufficiently reflect performance of the assay for natural human samples. We consider this
a potential risk to health since QC material is the best source of ongoing feedback a
laboratory has to monitor whether results reported to physicians are sufficiently accurate.
Therefore, we recommend that you evaluate matrix effects of your QC material relative to
the intended use human samples and describe relevant findings in the package insert.

2
See ISO (International Standards Organization) harmonized terminology database.

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Another related issue is that as a result of differences in matrices, QC materials might differ
from patient samples in terms of preparatory steps required for the assay (e.g., dilution,
extraction, centrifugation or other pre-treatment). We recommend that, whenever feasible,
you design the QC material so that it may monitor performance of preparatory steps as well
as the operational steps of the assay itself. If this is not the case, you should indicate to users
in the package insert which operational steps of the assay the QC material might not be able
to monitor.

The degree of evaluation that is appropriate depends on the amount of information generally
known about the specific matrix and how well its biochemical properties are understood. In
some cases the evaluation can be accomplished by spiking the analyte(s) into the QC
material matrix and in parallel into the intended use patient samples across the range of the
assay. You should compare results to determine (1) bias of the QC material matrix relative
to the natural sample (2) differences in precision between the QC material and the sample
and (3) differences in tolerance to factors that could affect assay performance (e.g., stability,
reagent deterioration). When relevant for your QC material, you should include results of
such testing in your package insert.
In some cases you may find it helpful to refer to the Clinical and Laboratory Standards
Institute (CLSI) guideline, EP-14-A2, Evaluation of Matrix Effects for further guidelines,
especially if your QC material is intended for a specific test system.

2. Analyte Value Assignment

You should describe the materials and protocols you used to determine the analyte value and
range you specify in the package insert. You should include the following:

Specific system(s) with which testing was performed (assay and instrument).
The number of replicates, runs and instruments, and time frame of the evaluation.
The statistical analyses used to establish the range.
Results for each analyte level, including coefficients of variation and standard
deviations, with confidence intervals.

You should indicate your acceptance criteria for difference between the nominal (target)
value and the mean values and ranges you determine.

3. Stability
You should describe the process by which you determined stability, in both opened and
closed forms, of the QC material. The term "closed" refers to closed shelf life stability;
"opened" refers to the opened conditions, including reconstituted or on-board conditions.
You should describe the studies performed to determine stability, including:

The conditions, (e.g., temperature, reconstitution conditions) under which the QC

material was stored during the stability evaluation.

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The methods for measuring stability, including the type of instrument and/or assay
used, the parameters measured (e.g., recovery of concentration or activity).
The manufacturers criteria for stability (e.g., 5% recovery based on a specific
method at the expiration date).
Results demonstrating that the manufacturers criteria for stability were met.

You should evaluate QC material stability separately from assay reagent stability, so that
reagent instability will not confound the evaluation.

Manufacturers should conduct real-time stability testing on an ongoing basis. Accelerated

stability testing can sometimes be used in a 510(k) to supplement real-time stability testing,
if real-time testing to support the expiration date is not yet complete. If accelerated stability
testing is used in this way, you should outline your testing conditions in the 510(k). You
should also provide information (e.g., literature) to support that the model you used to
evaluate the accelerated stability results is appropriate for your particular analyte.

4. Surrogate QC material
When a QC material is different in composition from the analyte it is intended to monitor
(e.g., a plasmid containing DNA versus infectious organism), there may be differences
between results obtained with the QC material and those obtained with intended use
samples (as there may be for matrix effects discussed above). In such cases, you should
perform testing on actual clinical samples run in parallel with the QC material to verify
that the QC material is as sensitive as actual patient samples to anticipated analytical
variables. These may include variables such as temperature variations, reagent
deterioration, or pipetting or sample transfer errors. Analogous to evaluation of matrix
effects, this testing helps assure that the same factors that affect a patient diagnostic test
result would have a similar effect on the result obtained with QC material, and could,
thereby, alert the user to analytical error.

V. Labeling
You should refer to 21 CFR 809.10 for labeling requirements. The following recommendations
are meant to help you apply these requirements to QC materials. Many of the items described in
this section are applicable for both assayed and unassayed QC materials. Items relating to
analyte levels, or specific assays or systems, for which QC materials are intended, are applicable
only for assayed QC materials.

Intended Use
The intended use section should define the QC material and its use. For both assayed and
unassayed QC material, you should include the following:
The analyte(s).
Whether the material is assayed or unassayed.
Whether the material is intended for quantitative or for qualitative systems or assays.

For assayed material you should also include:

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The analyte levels the material is intended to monitor (e.g., assay cutoff, decision points,
limits of assay range, as appropriate).
The system, instrument, or type of test the QC material is intended to be used with.

If the QC material monitors specific aspects of the assay, but not the entire process (e.g., all steps
after pre-treatment, instrument linearity), you should clarify this in the intended use.

Reagents
For assayed QC materials you should include all analytes and levels (if not already included in
the intended use). Inclusion of analyte levels is important even for qualitative assays since it is
important for users to know if the QC material will challenge the cutoff of their particular assay.
You should express analyte concentrations using whichever parameter is most relevant for the
analyte and test system (e.g., weight, activity).

For both assayed and unassayed material you should describe the analyte source (e.g., from
human or animal species, synthetic, or purified chemicals). For recombinant nucleic acid
material, you should include the vector, the source of the cloned nucleic acid region or gene and
specific nucleic acid sequence. For a microorganism, you should include the strain, and if
applicable, the portion of the microorganism (gene, antigen, etc.). You should describe the
media or cell line used for culture.

When applicable for your QC material, you should include donor characterization for the source
material to the extent that it is relevant for clinical use of the QC material.

You should describe the matrix, including the base material (serum, buffer, etc.), stabilizers,
preservatives, or clarifiers added. If any preservatives or other materials require special handling
by the laboratory, you should indicate this to the user.

You should describe inactivation methods used for potentially infectious material in your QC
material. When blood products are used, you should include a certification statement that the
animal/human source components used in the control are safe and that any blood product derived
material has been tested by FDA approved (or equivalently recognized) assays and found to be
negative for the communicable disease agents as stated in 21 CFR Part 610.

Other than the analyte levels, all items above should typically be included in the labeling for
both assayed and unassayed QC material. Labeling for unassayed materials should not include
analyte values since description of values is likely to be interpreted by the user to mean that
these values have been assigned and validated by the manufacturer. For unassayed materials, we
also discourage description of the analyte in relation to reference materials such as WHO, since
this is likely to be interpreted by the user to mean that the QC material is traceable to the
reference material and that the manufacturer has assigned the analyte value.

Operating Instructions
For both assayed and unassayed QC materials, you should include handling and storage
instructions. You should describe stability (expiration dating) under the opened and closed
storage conditions you recommend to users. We recommend that you include a description of

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acceptance criteria for determining stability (e.g., +/- x% recovery using a given assay).

Warnings
For both assayed and unassayed QC materials, you should include applicable warnings
concerning handling (e.g., warnings concerning handling of communicable disease agents, or
chemical components that require special handling).

Assigned target values and ranges

For assayed QC materials only, you should describe how target values and ranges were
established, including instruments or methodologies used for testing, the number of observations,
instruments, laboratories, and any other relevant conditions of testing. For each analyte at each
level, you should include results of the statistical evaluation including the mean(s) and standards
deviation(s), with confidence intervals.

Frequency for QC
You should include a statement that QC materials should be used in accordance with local,
state, and/or federal regulations or accreditation requirements.

Performance characteristics
Where appropriate you should include results of testing for matrix effects and/or effects of
surrogate QC material. You should describe:

Any significant matrix bias along with a brief description of how the bias was
determined.
Any significant difference between the QC material and typical patient samples in terms
of conditions known to cause analytical error.

Limitations
For assayed QC material you should state that the ranges given are intended only as guidelines
and that laboratories should determine the ranges based on their own test system and tolerance
limits.

For unassayed materials you should indicate that values are not assigned by the manufacturer
and that each laboratory should establish its own analyte values and ranges.

You should include a description of any assay conditions that the QC material may not monitor
because of matrix effects or surrogate QC material (e.g., pre-treatment steps, instability under
certain conditions) if these have not already been described in the intended use.

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