INTERNAL QUALITY CONTROL

in the ANALYTICAL LABORATORY

 INTERNAL QUALITY CONTROL

The need for quality control in an analytical laboratory

According to ISO/IEC 17025

 The laboratory shall have quality control procedures

for monitoring the validity of tests undertaken.

 The resulting data shall be recorded in such a way

that trends are detectable and, where practicable,
statistical techniques shall be applied to the
reviewing of the results.
According to ISO/IEC 17025

 The monitoring shall include e.g., regular use of

certified reference materials and/or internal quality
control using secondary reference materials.

 Quality control data shall be analysed and, where

they are found to be outside pre-defined criteria,
planned action shall be taken to correct the
problem and to prevent incorrect results from being
reported.
QUALITY ASSURANCE TOOLS
 Planned quality management
 Established procedures for:
 Training and development of staff
 Validation (equipment and method)
 Various routines in the daily work
 Internal/ external control measures
 Handling of the measurement uncertainty
 Internal audits
 Systematic recording (e.g., forms, system, archiving)
Important Elements of a Quality System

Codex Alimentarius Recommendations for the Assessment

of Testing Laboratories for Export and Import

 Participation in PT Schemes
 Use of reference materials as analytical/laboratory
quality assurance measures
 Use of internal quality control (IQC) procedures
 Use of validated methods of analysis
 Accreditation to ISO/IEC 17025:2005
PROFICIENCY TESTING

• Proficiency Testing Schemes can be regarded as a

routine, but relatively infrequent, check on analytical
errors.
• The main beneficial effect of PTs is that of
encouraging participants to install effective quality
control systems.
• It has been shown that laboratories with effective
IQC systems perform better in a PT Scheme.

• Without the support of a well-developed IQC system,

the value of participation in PT is negligible.
IUPAC Harmonized Guidelines for IQC, 1995
Benefits from Participation in PT
Schemes
• Identifying measurement problems
• Comparing methods or procedures
• Improving performance
• Educating staff
• Exchange of information wit the PT Provider
• Instilling confidence in Staff, Management, and Users
of laboratory services

Eurachem, 2011. Use and Interpretation of PT Schemes

REFERENCE MATERIAL (RM)

Material or substance whose property values are

sufficiently homogeneous and well-established to be
used for quality control

• Certified Reference Materials

• Standard Reference Materials
• Secondary Reference Materials
(e.g., PTRMs)
• In-house Reference Materials
Uses of RMs

• Method Validation and Measurement

Uncertainty

• Verification of the Correct Use of Method

• Calibration

• Quality Assurance and Quality Control

(QA/QC)
Suitability of RMs

• Measurand
• Concentration range/ Level
• Matrix match and potential interferences
• Sample size
• Homogeneity and Stability
• MU
 What is Internal Quality Control (IQC)?
One of a number of concerted Determined by a comparison of
measures to ensure that the data the accuracy achieved in a
produced in the laboratory are fit laboratory at a given time with a
for their intended purpose. required level of accuracy.

Routine practical procedures that enable

the analytical chemist to accept a result
as fit for purpose, or reject the results
and repeat the analysis (monitoring of
the analytical system)

Fitness for purpose: degree to which data produced by a measurement

process enables a user to make technically & administratively correct
decisions for a stated purpose
IUPAC Harmonized Guidelines for IQC, 1995
 Basic Approach to IQC

Analysis of control materials alongside the test

materials under examination.

Interpretation of control data must be based on

documented data, objective criteria and statistical
principles wherever possible.

IUPAC Harmonized Guidelines for IQC, 1995

 Basic Approach to IQC

Results are viewed primarily as indicators of

performance of the analytical system.

Analytical system: range of circumstances that contribute to the quality

of analytical data , including equipment, reagents, procedures, test
materials, personnel, environment and quality assurance measures.

Secondarily, as a guide to the errors associated

with
individual test results.

IUPAC Harmonized Guidelines for IQC, 1995

 INTERNAL QUALITY CONTROL

 Internal quality control at the chemical analytical

laboratory involves a continuous, critical evaluation of the
laboratory’s own analytical methods and working
routines.

 The control encompasses the analytical process starting

with the sample entering the laboratory and ending with
the analytical report.

Nordtest Report TR 569, Internal Quality Controll, 4th ed., 2011

 LABORATORY WITH QUALITY CONTROL
MANAGEMENT

Responsibility Documentation Customer Relations Deliveries Follow-up

Sampl Sample Testing/ Data Reporting Test

e handling Calibration handling results Report

Staff Qualifications Proced./Methods Traceability IQC

RELIABILITY/ DATA QUALITY!!!
TECHNICAL COMPETENCE
 METHOD VALIDATION IS VITAL IN QA!!!

Method Validation accreditation

has become an
intrinsic part of QA… proficiency testing
IQC

Performance
BUT IT CANNOT Characteristics
STAND ALONE!!! validation

Fitness for purpose accuracy

analytical measurement
system uncertainty
analytical
result
• method protocol
• type of matrix interpretation
• concentration range of analyte
 Internal control of analytical methods
Current monitoring that the analytical methods applied in the
laboratory are constantly giving reliable results…

 important follow-up upon the validation of the methods

 is the actual method still fit-for-purpose just as after the
validation
o or may some systematic error have appeared?
o or have the reproducibility grown bigger over time?
 maybe time to reconsider method performance
 often supplemented by control of single parameters in the
method process (e.g., balances, temperatures, instrument
 Statistical Quality Control
 Why statistical?

 Why not just measure some control sample from time to

time and check the result if OK or NOT OK?
 Statistical Quality Control
 The actual measurement of the control will NEVER give
exactly the expected value
 How big a deviation can be allowed?
 How often should such checks be made?

 The statistically based quality control gives optimal

information from every single control measurement (sample)
- in the light of the awareness of the commonly expected
measurement uncertainty on results from the actual method
Statistical Control
 Means of internal control
Various activities assuring that the laboratory currently
produces reliable results through checks of the analytical
process as a whole – and/ or various (critical) elements in
the analytical process:
 replicate analysis of samples
- checking for precision (note inhomogeneity of
sample)
 analysis of quality control samples
- monitoring method/ laboratory performance over
time
 analysis of blanks
- identifying contamination problems
 Control Materials in IQC

Blank determinations

 Reagent blank – detects purity of reagents and

contamination of the analytical system e.g.,
glassware, environment
 Blank reference material – zero concentration of the
analyte (field blank)
 An inconsistent set of blanks may suggest sporadic
contamination - reject results

IUPAC Harmonized Guidelines for IQC, 1995

 Control Materials in IQC

Spiking and recovery checks

 Traceability is important! Confirmation of identity &
check on purity must be made
Metrological traceability -property of a measurement result
whereby the result can be related to a reference through a
documented unbroken chain of calibrations, each contributing to
the measurement uncertainty. International Vocabulary of
Metrology (VIM) 2012

 calibration standards and spikes must not be traceable

to the same stock solution or the analyst

IUPAC Harmonized Guidelines for IQC, 1995

 Acceptable limits for recovery as a function
of measurand concentration
Active Analyte Ratio Unit Mean Recovery,
Ingredient, % %

100 1 98-102
≥ 10 10-1 98-102
≥1 10-2 97-103
≥ 0.1 10-3 95-105
0.01 10-4 90-107
0.001 10-5 80-110
0.0001 10-6 1 ppm; 1 ug/g 80-110
0.00001 10-7 100 ppb; 0.1 ug/g 80-110
0.000001 10-8 10 ppb; 0.01 ug/g 60-115
0.0000001 10-9 1 ppb ; 1 ug/kg 40-120

AOAC International 2012. AOAC Official Methods of Analysis 2012 Appendix F

 Expected precision (repeatability) as a
function
of analyte concentration
Analyte, % Analyte ratio Unit RSD, %
100 1 100% 1.3
10 10-1 10% 1.9
1 10-2 1% 2.7
0.1 10-3 0.1% 3.7
0.01 10-4 100 ppm (mg/kg) 5.3
0.001 10-5 10 ppm (mg/kg) 7.3
0.0001 10-6 1 ppm (mg/kg) 11
0.00001 10-7 100 ppb (ug/kg) 15
0.000001 10-8 10 ppb (ug/kg) 21
0.0000001 10-9 1 ppb (ug/kg) 30

AOAC International 2012. AOAC Official Methods of Analysis 2012 Appendix F

 INTERNAL QUALITY CONTROL

Statistical tools needed for dealing with quality control

Analytical variation

 Variation among results of repeated

measurements of the same measurand
 This variation follows a certain pattern –
statistical distribution
Samples

Randomly selected elements from a population

- or a subset of some kind of material (e.g., fruit, meat,
water, etc.)
- or a single control sample in a Quality Control Program
Note!!!
Be aware of the origin of the samples in the treatment of
the results of the testing.
- within sample variation vs. between sample variation
- variation of results may be influenced by inhomogeneity
or lack of stability
Example

Example of laboratory internal quality control values for a

solution containing 60,0 μg/l of zinc.

Nordtest Report TR 569, Internal Quality Controll, 4th ed., 2011

Basis for most statistical calculations

• Repeated measurement of the same item – giving a

spread of different values
• The frequency of same values is counted – and charted
(histogram)
 Basis for most statistical calculations

Most (ideally) analytical data behave as in NORMAL

DISTRIBUTION
 Basic estimation for normal distribution

 Estimate the mean,  Variance,

s2

 Estimate for the real  Relative standard deviation

standard deviation, (RSD),

%RSD = %CV,

Nordtest Report TR 569, Internal Quality Controll, 4th ed., 2011

Distribution of average values

 When the results are normally distributed, the mean value x is

defined by the position of the maximum of the curve.
 The shape of the curve is determined by the spread of the single
results, expressed by the standard deviation, s.
 Distribution of average values

 For normal distribution,

calculate a theoretical
spread of the results
around the mean value.
 About 95 % of all results
will be located within the
mean value ± 2sd.
 About 99.7 % of the
results are located within ±
3sd.
 These properties are
applied in the construction
of the control charts.
Basis for the Control Charts

Nordtest Report TR 569, Internal Quality Controll, 4th ed., 2011

Outliers

Single values, which with a given probability, does not

belong to the normally distributed set of data.

Decide whether,
 A certain value is an outlier, and therefore should
be discarded before calculation of average and
spread for the set of data, or
 The value is just an extreme one (given the
number of data), which must be included in the
data set, and therefore influences the calculation
of the mean and standard deviation.
Decisions about Outliers

 Grubb’s Test
• Primarily a test of between-laboratory variability
• i.e., testing the average from the different participants

 Cochran’s Test
• A test of the within-laboratory variabilities
• i.e., testing the standard deviations for the repeated tests
from the different participants
Deciding on potential outliers

 Either
the Grubb’s or Cochran’s Test is applied to identify
stragglers or outliers:
 If the test statistic is less than or equal to its 5% critical
value, the item tested is accepted as correct.
 If the test statistic is greater that its 5% critical value and
less than or equal to its 1% critical value, the item tested
is called a straggler.
 If the test statistic is greater than its 1% critical value, the
item is called a statistical outlier.
Deciding on potential outliers

 Investigatewhether the stragglers and/or statistical

outliers can be explained by some technical error, such
as:
 A slip in performing the measurement,
 An error in computation,
 A simple clerical error in transcribing a test result,
 An analysis of the wrong sample.

 When any stragglers and/or statistical outliers remain

unexplained, the stragglers are retained as correct items
and the statistical outliers are discarded, unless the
statistician for good reason decides to retain them.
Grubb’s Test

 Comparisonof the difference between the extreme value

and the mean with the spread of data

│Suspicious value – x │
G=
s

 Compare with the critical values on a 1% or 5%

confidence interval from a table.
 INTERNAL QUALITY CONTROL

The basic concepts in statistically-based quality

control- and the establishment of control charts
Analytical work under statistical control

 Prerequisites for applying statistical control:

 The variation in results over time is only caused by a
constant influence of random errors
 This variation in analytical results is following the normal
distribution with a given mean and constant variance
 The laboratory has not made any deliberate changes in their
performance of the method since implementation (after
validation)

 If these prerequisites are fulfilled:

 Any major changes in a control result, or sequence of control
results, in comparison with the given mean may then be
caused by something extra-ordinary, i.e., the system may be
Control Charting

 What is control chart(ing)?

 A visual presentation of the actual “Quality” of the
analytical process
 Measurements of a control material with known content
 Subsequent plotting of the values in a diagram

 What are its uses?

 To evaluate the development of the analytical process over
time
 To monitor systematic problems within the system (e.g.,
trending)
The basic Shewhart Control Chart

 Invented by Shewhart in 1924 intended for process industry

 Continuous graphical plot of control values
The Shewhart Control Chart with LIMITS
Limits!!!

 Evaluation of results over time in relation to some limits

 Gives the probability of any exceeding of the limits could
have happened randomly

 Typical sets of limits

 WARNING limits – exceeding results from these limits is
relatively normal IF it doesn’t happen too often – so the
situation must be observed.
 CONTROL limits – a single exceeding result is rather
unlikely (but possible) – so normally action must be taken
(ACTION limits)
Various types of Shewhart Charts

• Single measurements:
 X-Chart
• More measurements per sample:
 X(bar) chart – average of the repeated measurements
 R Chart – range of repeated measurements (xmax – xmin)
 S-kort chart – spread of repeated measurements
• Used in combination:
 x(bar) – R chart
 x(bar) – s chart
 Control Charts

- X-Chart Most important in

chemical analysis
- R-Chart IQC

Nordtest Report TR 569, Internal Quality Controll, 4th ed., 2011

 Control Charts
X-Chart

Mean Value Determination

can be used to monitor the combination of random and systematic
effects for control values, based on single results or a mean of
multiple analyses
• bias may be monitored using a reference material as control
sample by comparing the mean control value over time with the
reference value.
An X-Chart has a central line, upper & lower warning limits (WL)
and upper and lower action limits (AL)

Nordtest Report TR 569, Internal Quality Controll, 4th ed., 2011

 Control Charts

X-Chart: Mean Value Determination

Nordtest Report TR 569, Internal Quality Controll, 4th ed., 2011

 Control Charts
X-Chart

Blank value chart

• special application of an X chart

• provides information about contamination of reagents and state
of the measurement system
• concentration or measured signal may be plotted
• negative and positive control values shall be plotted

Nordtest Report TR 569, Internal Quality Controll, 4th ed., 2011

 Control Charts

X-Chart: Blank Value Determination

Nordtest Report TR 569, Internal Quality Controll, 4th ed., 2011

 Control Charts
X-Chart

Recovery Chart

• Analytical process may be tested for matrix influences by

determining the recovery of spiked additions of standards

• Recovery rate of 100% should be the central line

Nordtest Report TR 569, Internal Quality Controll, 4th ed., 2011

 Control Charts
Range Chart

 Serves the purpose of repeatability control

 Range:
 difference between the largest and smallest single
result for two or more separate samples

 For practical applications in analytical labs, use

duplicate determination (of samples to be analyzed) in
each analysis series

Nordtest Report TR 569: Internal Quality Control Handbook , 4th ed. 2011
 Control Charts
Range Chart
• The range for duplicates
(highest – lowest value) was
used for plotting.
• The mean range for the
same 60 analytical runs was
used as the central line
• The repeatability SD
calculated from the mean
range was used to establish
control limits by multiplication
with factors

Nordtest Report TR 569: Internal Quality Control Handbook , 4th ed. 2011
 Control Charts
Range Chart
Factors for estimation of standard deviation from mean range, calculation of central
line, warning & action limits for construction of R-charts

Nordtest Report TR 569: Internal Quality Control Handbook , 4th ed. 2011
 X-Chart and Range Chart

Nordtest Report TR 569: Internal Quality Control Handbook , 4th ed. 2011
 Control Charts
Setting the Control Limits
X-chart R-chart
Statistical control limits Statistical control limits
• control limits based on control • control limits based on control
sample sample
• From a long time period, e.g. a • From a long time period, mean
year, the SD, s, is calculated from range is calculated, e.g. for
control values duplicate:
• central line: mean value s=mean range/1.128
• warning limits: +2s and -2s • central line: mean range
• action limits: +3s and -3s • upper warning limit: +2.83s
• upper action limit: +3.69s

Nordtest Report TR 569: Internal Quality Control Handbook , 4th ed. 2011
 Control Charts

Advanced control charting:

 CUSUM Chart
 J chart (Royal Society of Chemistry Analytical
Methods Committee Technical Brief AMC TB 12-2003)
 Moving Average (MA)
 Exponentially weighted moving average (EWMA)
chart
 Control Charts

Recommendations:

 Start of QC: preliminary limits from 25 control values or from

method validation results

 For stable QC samples, reliable limits can be based on a one-

year period w/ at least 60 control values

Nordtest Report TR 569: Internal Quality Control Handbook , 4th ed. 2011
 Interpretation of QC

Possible Cases:

1. The method is in control.

2. The method is in control but a long-term evaluation

shows the method is out of statistical control.

3. The method is out of control.

Nordtest Report TR 569: Internal Quality Control Handbook , 4th ed. 2011
 Interpretation of QC

Out-of-control cases
 experience of analyst is important to do remedial actions
 error in the analysis of test samples
 normal action is to do more (at least 2) control analyses;

- if within warning limits: routine samples can be reanalyzed

- if outside warning limits: routine analysis is stopped, find

cause(s) of error

Nordtest Report TR 569: Internal Quality Control Handbook , 4th ed. 2011
 Frequency of Control Analyses (Nordtest)

 As a minimum, 1 control sample in each analytical run must be

analysed; or at least at the beginning & before end of run

 Depends on the stability of the measuring instrument

 If there are errors from calibration drift, control samples need to

be higher

 All measurements after the last approved sample in QC may

have to be reanalyzed

 Balance between the cost of control and the cost of repeating

analyses
 Minimum QC Procedures for a Batch of Analysis
(Determination of trace amounts of analytes in foods and pharmaceuticals,
Horwitz)

1) One QC check standard (instrument check solution).

If the result obtained on the check standard differs from the agreed
value by more than ±10%, then all samples in the batch should be
reanalysed
A batch of samples is generally taken to consist of about 10 to 20
samples in a short time interval

2) One reagent or method blank

Where samples containing relatively high concentrations of analyte
are analysed, additional blank determinations should be carried out
immediately after them to eliminate carryovers

NATA Technical Note #23: Guidelines for QC in the Analytical Laboratory- October 1995
 Minimum QC Procedures for a Batch of Analysis
(Determination of trace amounts of analytes in foods and pharmaceuticals,
Horwitz)

3) One duplicate sample analysis pair

4) One spiked sample
– must be prepared from a standard stock solution which is different
from a calibration stock standard
– Should have an analyte concentration within the range of samples
being analyzed.

Note:
 Check samples, reagent blanks, duplicate pair analyses and spiked
samples should be distributed in a random manner in successive
groups of samples analyzed, but with each of them appearing
within each group of samples.

NATA Technical Note #23: Guidelines for QC in the Analytical Laboratory- October 1995
Example:
Group 1: Group 2:
Calibration standards Check sample
(lowest to highest) or spiked sample
Reagent blank Reagent blank
Sample 1 Sample 6
Sample 2 Sample 7
Check sample, or spiked sample A calibration standard
Sample 3 Sample 8
Sample 4 Sample 9
Sample 2 (duplicate analysis) Sample 7 (duplicate analysis)
Reagent blank Reagent blank
Sample 5 Sample 10
Reference material
A calibration standard
NATA Technical Note #23: Guidelines for QC in the Analytical Laboratory- October 1995
Remember!
• Inferences based on IQC results are applicable
only to test materials that fall within the scope of
analytical method validation.

• With professional experience and diligence, IQC is

the principal recourse available for ensuring
appropriate data quality.

• For an IQC program to succeed, there must be a

genuine commitment to quality; must be a part of
total quality management system.

Harmonized Guidelines for Internal Quality Control in Analytical Chemistry Laboratories, IUPAC, 1995.
RECOMMENDED REFERENCES:

ISO Guide 80:2014 – Guidance for the in-house

preparation of quality control materials (QCMs)
ISO 7870-1:2014. Control charts -- Part 1: General
guidelines. (www.iso.org)
ISO 7870-2:2013. Control charts -- Part 1: Shewhart
control charts. (www.iso.org)
ISO 7870-3:2012. Control charts -- Part 3: Acceptance
control charts. (www.iso.org)
ISO 7870-4:2011. Control charts -- Part 4: Cumulative
sum charts. (www.iso.org)
ISO 7873:1993. Control charts for arithmetic average
with warning limits. (www.iso.org)
ISO/TS 13530:2009. Water quality -- Guidance on
analytical quality control for chemical and
physicochemical water analysis. (www.iso.org)
RECOMMENDED REFERENCES:

• M. J. Gardner, Quality control techniques for chemical

analysis: some current shortcomings and possible future
developments, Accred. Qual. Assur., 2007, 12, 653-657
(www.link.springer.com)

• M. Thompson, R. Wood, Harmonized guidelines for internal

quality control in analytical chemistry laboratories, Pure Appl.
Chem., 1995, 67, 49-56 (www.iupac.org)
• Internal Quality ConTROLL, Handbook for chemical
laboratories, 4th Edition, Nordtest Report TR569, 2012
(www.nordtest.info)
RECOMMENDED REFERENCES:

• AMC Technical Briefs,

RSC,(www.rsc.org/Membership/Networking/InterestGroups/
Analytical/AMC/TechnicalBriefs.asp):
• AMC TB 51-2012, Quality control of routine sampling in
chemical analysis
• AMC TB 49-2011, Sporadic blunders
• AMC TB 46-2010, Internal quality control in routine analysis
• AMC TB 12-2003, The J-chart: a simple plot that combines
the capabilities of Shewhart and cusum charts, for use in
analytical quality control
• AMC TB 09-2001, A simple fitness-for-purpose control chart
based on duplicate results obtained from routine test
materials
“Quality begins on the inside...
and then works its way
out.”
“Proficiency is not aBob Moawad
one-time or
occasional compliance to a standard.
It is a consistent, systemic
demonstration of technical
competence.”
RSVP
Thank you for your
attention!