Te ch ni cal N ote 7 0 8

Automate System Suitability Testing with

Chromatography Software
Frank Tontala, Thermo Fisher Scientific, Germering, Germany

Key Words
Chromeleon Chromatography Data System, (CDS), System Suitability
Testing, Chromatographic Suitability, SST

Goal
To illustrate and define the tools used by the Chromeleon Chromatography
Data System (CDS) to perform Automated System Suitability Testing

Introduction
System Suitability Testing (SST) is commonly used by
laboratories to ensure that the complete analytical system
(including instrument, reagents, columns and analysts) is
suitable for the intended application.
The general goal of system suitability testing is to monitor
The United States Pharmacopeia (USP) Chromatography chromatographic results to ensure chromatographic
General Chapter states: suitability (e.g. by testing tailing factor, column efficiency
and resolution of critical peak pairs) and consistent
“System Suitability Tests are an integral part of gas and
system performance (e.g. by using replicate injections
liquid chromatographic methods. They are used to verify
of test standards).
that the resolution and reproducibility of the chromato-
graphic system are adequate for the analysis to be done. SST calculations are usually performed on standards
The tests are based on the concept that the equipment, before any samples are analyzed to ensure that potentially
electronics, analytical operations and samples to be valuable samples are not injected into an unsuitable
analyzed constitute an integral system that can be system. In a manual environment, this can require user
evaluated as such.” interaction with every batch of samples and result in
considerable throughput delays. Furthermore, if calcula-
tions are done manually, there is the potential for error
and incorrect pass or fail results.

To overcome these problems, the Thermo Scientific

Dionex Chromeleon 7 Chromatography Data System
includes automated System Suitability Testing as part of
sequence acquisition and processing. Using Chromeleon™,
all SST calculations are automatically performed by
the software.
2 Based on user-specified criteria, Chromeleon can These specifications will vary depending on the actual
determine if the system is suitable and, if desired, even conditions of the analytical method, but provide a good
stop an analytical run if any of the tests fail. starting point. The FDA also recommends that at least 5
injections be made to determine repeatability. Commonly,
System Suitability Testing is not limited to laboratories
all other tests are checked against these 5 injections as
required to comply with guidelines set by the FDA and
well, providing more confidence that the system is suitable
other regulatory bodies. Any laboratory can benefit
for performing the analysis.
from the ability of Chromeleon 7 to perform automated
SST on hundreds of different types of calculated results. System Suitability Functionality in
Possible test criteria include ensuring sample concentra- Chromeleon 7
tions are within expected ranges, setting limits for detector Setting up Automated System Suitability Testing in
signal-to-noise, limiting the acceptable levels of impurities Chromeleon 7 is easy. A wizard guides the user through
and much more. selecting the test criteria, calculations are performed
automatically and predefined report templates simplify
FDA Guidelines on System Suitability Testing
reporting the results.
The FDA (CDER) 1 guidelines on Validation of
Chromatographic Methods recommend the following What Parameters Can Be Tested?
tests and specifications: Any value that Chromeleon can calculate can be used in
automated system suitability testing. This includes the key
Table 1: FDA Recommended SST Specifications values recommended by the FDA, as well as many others
such as relative retention time, area %, peak width and
Test Specification concentration.
Capacity Factor (k’) k’ > 2
Visualizing Tests and Results
RSD ≤ 1% Figure 1 shows the Chromeleon Chromatography Studio
Precision/Injection Repeatability (RSD)
(n ≥ 5 is desirable)
displaying a chromatogram, processing method (middle
Resolution of Analyte peak from closest pane) and computed results (bottom pane).
Rs > 2
peak (Rs)
The processing method has been configured to perform
Tailing Factor (T) T≤2
the five system suitability tests recommended by the FDA.
Number of Theoretical Plates (N) N > 2000 For example, Test 1 checks that the resolution of the
acetophenone peak versus the next peak is > 2. When
acquiring a sequence of samples using this processing

Figure 1. A Chromeleon processing method is shown above, configured with FDA recommended suitability tests
method, the “Fail-Action” column 3
directs that if the test fails to meet
this criterion, then sequence
acquisition will be halted.

The “Result” column in the method

panel shows the post acquisition
results of each test. Note that the
result for the %RSD test is “Not
Executed,” because the displayed
injection is only the second line of the
sequence, meaning that an insufficient
number of samples (2 out of 5) have
been analyzed. Note also that the
Theoretical Plates test has failed,
since the computed value (1278) is
less than the test criterion (2000)

Reporting Results
Chromeleon comes with a compre-
hensive set of default reports,
which can be used as they are, or
can be easily customized to meet
the specialized needs of the lab.
Predefined report tables that include
system suitability related information Figure 2. An example of an SST report is shown above.
can be inserted in any report. Figure 2
shows a customized system suitability
injection report that includes the SST
results, along with an overall pass/fail
determination.

System Suitability Testing

During Acquisition
During sequence acquisition, Chrome-
leon automatically computes system
suitability results at the completion of
each injection. The Automated System
Suitability Testing can be configured
such that Chromeleon can stop the
sequence acquisition if a test fails.
This ensures that samples are not
injected onto a system that is not
suitable for sample analysis, and that
valuable samples and reagents are
not wasted. Figure 3. This table depicts an Instrument Audit Report showing a sequence abort.

All acquisition-related system

suitability events are recorded in the
instrument audit trail.

Figure 3 shows the details logged in

the audit trail describing the tests
performed, the results obtained and
the actions performed.
4 Other Uses for Automated
System Suitability Test
Calculations
The System Suitability Testing
functionality in Chromeleon is not
limited to the system performance
values cited by the FDA. Any
chromatographic value that Chrome-
leon can calculate can be part of the
test criteria. This means that Auto-
mated System Suitability Testing may
be used for other purposes, such as
testing product quality.

Comparing Results Against

Specifications
In this example, tablets containing an
Figure 4. Chromeleon SST may be used to test Tablet Purity and % Label Strength
active ingredient are analyzed for
% Label Strength (%LS), which is
defined as the amount of the active
ingredient in the tablet as a percent-
age of the expected amount. The
published label strength for the tablet
is 50 mg ± 5%. In addition, the total
amount of impurities (% Area of all
other peaks in the chromatogram)
must be less than 2%.

To implement these criteria, two tests

are defined in Chromeleon. The first
test checks that the %LS value for
the active ingredient is between 95
and 105. The second test checks that
the %Area of the active peak is
greater than or equal to 98% (which
is equivalent to testing that all the
other peaks comprise less than 2%
of the total peak area).

Figure 4 shows the Chromeleon

Figure 5. Chromeleon control charts can be used to monito theoretical plates over time.
Chromatography Studio displaying
a chromatogram, the processing
lead to higher data quality and • The analyst is not trained
method with the 2 SST entries
greater laboratory productivity. sufficiently on either the analytical
(middle panel) and the computed
method or the instrument.
results (bottom panel). Common reasons for inconsistent lab
performance include: Figure 5 shows a control chart created
Using Control Charts to Monitor automatically using the built-in query
Lab Performance and Data • The instrument is not suitable for and interactive charting tools of
Quality the analytical method. For exam- Chromeleon. This chart displays the
When Chromeleon System Suitability ple, instruments injecting samples theoretical plates obtained on a
Tests are used routinely, trends in onto a heated column that do not system over time. In this example,
SST results can be easily monitored. pre-heat the mobile phase can though the system suitability checks
Chromeleon includes tools that sometimes produce lower theoreti- routinely passed, clearly there was
facilitate generating control charts cal plates. a period of sub-optimal chromato-
to monitor chromatographic results • The instrument needs to be graphic performance. Clicking on any
over time. These charts can be used serviced. For example, the detector data point in the chart will open the
to identify trends or anomalies in lamp may be nearing the end of its source data, allowing further investi-
the performance of systems, methods rated life and need replacement. gation of the underlying cause.
or users. Once such information is
• The analytical method is not
available, it is possible to determine
optimized, resulting in marginal
the causes of the failure and appro-
performance.
priate corrective action, which will
Building an Automated 5
System Suitability Test in
Chromeleon
Chromeleon provides a wizard (the
SST Wizard) to help users easily
specify automatic system suitability
tests. This wizard guides users
through each step, as follows:

SST Wizard Screen 1 – Selecting

the Test Parameter
On the first page of the wizard, the
user selects the parameter to be tested
(See Figure 6). In addition to the
commonly used system performance
parameters, any chromatographic or
calculated result may be specified,
using the <Custom Test> entry.

SST Wizard Screen 2 –

Specifying the Applicable Figure 6. The System Suitability Test is selected.
Injections
On the next wizard page, the user
defines the injections to which the
test will be applied. (Figure 7). For
example, system suitability tests are
not generally performed on blank
injections (unless assessing signal
noise) or samples. Rather, the tests
are typically applied to the five or
more specific System Suitability
Test injections.

Alternately, injections may be defined

by Injection Type or virtually any
other property of the injection or its
position in the sequence.

Figure 7. The set of injections for the test is specified.

6 SST Wizard Screen 3 – Defining
the Test Criteria
The Evaluation page (Figure 8)
defines how the pass/fail result will be
determined. For tests on a single
injection (Figure 9), the user simply
specifies the operator and the
threshold value. For tests that are
based on multiple injections (e.g.
%RSD of replicates) the type of
statistical calculation and the
required injection types and grouping
of replicates are also specified. This
automatically delays test evaluation
until the required number of
injections have been processed.

SST Wizard Screen 4 –

Specifying the Peak and Channel
Conditions Figure 8. Evaluation criteria may be specified for a single-injection test.
On the Peak/Channel page (Figure 10),
the user defines the component(s)
and channel to which the test will be
applied. If the name of the peak is
known, it can be selected directly.
Otherwise, the peak can be selected
by its position in the chromatogram
(peak number) or by its properties (for
example, the peak with the highest area
or with the lowest width). Alternately,
it is possible to have the test performed
on all peaks.

Figure 9. Evaluation criteria may be specified for a multiple-injection test.

Figure 10. Tests may be applied to a specific peak and channel.

SST Wizard Screen 5 – Defining
Failure Actions
The final wizard page (Figure 11)

Te ch ni cal N ote 1 0 0 6
allows the user to define what test
result should be reported if the test
cannot be evaluated and, during
sequence acquisition, what action
Chromeleon should take if the
test fails.

Completed Wizard
Upon completion of the Wizard, the
new test is added to the SST table of
the method. Double clicking on any
item in the test row will reopen the
appropriate wizard page (Figure 12).

Figure 11. Conditional actions for the test results may be specified.

Figure 12. The new test is added to the SST table.

Conclusion • Reduced sample loss. Chromeleon • The functionality can also be used
Chromeleon 7 offers many significant can automatically stop sequences to generate control charts to
advantages for System Suitability as soon as it detects a System monitor system performance in the
Testing: Suitability failure. This prevents laboratory and quickly identify
samples from being injected on to a systems that continually produce
• Improved reliability of System
system that is not suitable for the high levels of failure.
Suitability Tests results. Chrome-
analytical method and eliminates • By automating some of the most
leon performs all calculations,
sample loss. repetitive and time consuming
eliminating any errors that can be
caused by manual calculation steps. • The automated SST functionality steps found in laboratories, the
can be extended to perform other Automated System Suitability Test
• Faster generation of System
types of tests, such as comparing functionality saves time and money
Suitability Test results. As soon as
sample results against and increases laboratory
the peak is detected, Chromeleon
specifications. productivity.
will automatically recalculate the
system suitability results. This References
increases throughput by reducing 1. Center for Drug Evaluation and
delays created by having to Research (CDER) Reviewer
manually recalculate the results. Guidance Validation of
Chromatographic Methods,
November 1994.

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