RAJESH NAIR

DIRECTOR CALF-NDDB, ANAND, GUJARAT

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 Coverage
Need of AQC in food testing laboratories

Analytical quality system of food testing laboratories

Relationship between Quality System, Quality Assurance & Quality Control

Variables of data accuracy

Analytical Quality Control (AQC)in food testing laboratories

ISO/IEC 17025:2005: Concepts

Internal quality control checks

External quality control checks

Trend Analysis using control charts

Conclusion
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 Need of AQC in food testing laboratories

• Emerging food safety issues such as presence of microbial pathogens, bacteria, viruses,
environmental toxins, allergens, adulterants, residues of agricultural drugs and toxic
chemicals in food commodities, increased consumer awareness and rapidly growing
international trade in food have stimulated the demand for accurate food safety testing
all over the world.
• Information about the quality of the food commodity can be generated through quality
testing in analytical laboratories.
• Analytical quality control, commonly shortened to AQC refers to all those processes and
procedures designed to ensure that the results of laboratory analysis are consistent,
comparable, accurate and within specified limits of precision.

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 Analytical quality system of food testing laboratories
• Quality is hard to define in a way that is appropriate for all
situations
• Therefore, quality means that the laboratory results meet
the customer’s expectations and are accurate and
defensible
• Analytical quality system of a food testing laboratory
includes Quality assurance which further includes quality
control.
• The meaning of the terms ‘Quality Control’ and ‘Quality
Assurance (QA)’ often vary according to the context
• In practical terms, QA relates to the overall measures taken
by the laboratory to regulate quality
• Whereas quality control describes the individual measures
which relate to the quality of individual samples or batches
of samples.

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Relationship between Quality System, Quality Assurance
& Quality Control

• Quality management works on the

organizational level to implement an overall
Quality quality policy.
System • A quality system refers to the organizational
resources, processes and procedures to
Quality implement quality management, which is
Assurance broader than both Quality Assurance (QA) and
Quality Control (QC).
• QA program is the backbone of the laboratory
quality system.
Quality • QA provides a management tool within the
Control organization.
• QC is a process within the QA program. The
process is to collect evidence that the desired
level of quality is achieved.

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 Variables of data accuracy

Post analytical variables

Analytical variables

Preanalytical variables
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 Analytical Quality Variables

PREANALYTICAL
Analytical
Laboratory facilities Standard material & Equipment
• Shelf life and storage
• Laboratory facilities
• Classes of equipment
• Design of the laboratory • Maintainenance, calibration and repair
• Environmental control Sample collection and handling
• Housekeeping control • Sampling
Staffing and organization • Transportation
• Test material identification
• Management structure
• Control and storage
• Job description Methods of analysis
• Staff training • Selection method
• Staff performance • Validation of analytical methods
• Support personnel • Quality control procedures
• Quality assurance procedures
• Chemicals, reagent and standards

POST ANALYTICAL
• External proficiency testing
• Maintenance of laboratory records
• Statistical Quality Control
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 Analytical Quality Variables
Laboratory facilities
• Facilities must allow Laboratories work to proceed both effectively and safely
• Facilities should reflect the general features of work programme in long term rather than
the specific pattern of current work
Design of the laboratories
• Laboratory should be designed with efficiency in mind
• Even though the final design of the laboratory is made by architects and engineers the
technical hand should be involved in establishing food control laboratories
• Design of the laboratory should meet minimum requirements to perform several
functions such as chemical analysis of foods for proximate, trace metals, additive,
nutrients residues and contaminants and for some basic food microbiology as well
Environmental control
• Adequate control of temperature, humidity, and dust is important for staff comfort,
instrumental performance and safe working (e.g. with flammable solvents)
• Computers need to be protected from strong magnetic fields
• Direct exposure of chemicals and reagents to sunlight and fluorescent light must be
avoided
• Delicate balances and optical instruments may need to be protected from vibrations
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• All these needs have to be documented and identified
 Analytical Quality Variables
Sampling

Analytical data reported must reflect the composition of received samples as a whole
Test material identification

Without clear and unambiguous receipt and identification of each individual sample,
quality assurance can not be maintained

Each test material must be given an identification code, which can uniquely identify that
material and records of its analysis
Control and storage

Deterioration of the test material invalidates the test results

Therefore test material must be stored, so as to ensure their integrity, safety, legality and
stability
Equipment

Each item of equipment used in the course of work must be known to be in the proper
working order

An instrument may be used only by staff with appropriate training

For each instrument, the nature and frequency of performance checks and the person
responsible for should be specified 9
 Analytical Quality Variables
SELECTION OF THE METHOD

Any analytical method used on test materials must be followed in accordance with
intended purpose of analysis and must have quality control procedure associated with it

Reference methods

Official methods

In- house methods

Screening methods

Protocols

VALIDATION OF THE METHOD

Validation is a process of establishing documentary evidence demonstrating that a
procedure, process, or activity carried out in production or testing maintains the desired
level of compliance at all stages.

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 Analytical Quality Variables

Initial Validations

Methods must first undergo in-house validation and quality assurance programme which
specify what parameters to include in validation

Validation of a method developed in house must additionally identify factors which are
revealed by an inter laboratory trial

Thus it will include demonstration of the robustness of the method and identification of
any aspect of the method which are critical to its ability to generate accurate data

On going validation
• Some form of ongoing check that the method is continuing to work properly is necessary
• The nature and frequency of these checks need to reflect the complexity of the method,
the degree of skills required, its reliability, track record, number of samples analysed and
the quality of data required
• Usually the checks will take the form of some or all quality control procedures
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 Analytical Quality Control (AQC) in food testing laboratories
Quality Control refers to a measuring process, or to check a result and provide assurance that
all activities are performing within predetermined limits.
Why QC ?
• Safety of the consumer is being ensured through quality testing of food commodity in
analytical laboratories.
What is the goal ?
• To check that the results being produced are fit for purpose.
• Rapid detection of SIGNIFICANT errors
• Provide accurate results in a timely manner
• Cost effective and simple to use
• To identify the source of error

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ISO/IEC 17025:2005: General requirements for the
competence of testing and calibration laboratories
4. Management Requirements 5. Technical Requirements
1. Organization
1. General
2. Management System
2. Personnel
3. Document Control
3. Accommodation & environmental
4. Review of requests, tenders and contracts conditions
5. Subcontracting of tests and calibrations 4. Test & calibration methods &
6. Purchasing services and supplies method validation
7. Service to the customer 5. Equipment
8. Complaints 6. Measurement Traceability
9. Control of nonconforming testing and/or 7. Sampling
calibration work 8. Handling of test & calibration
10. Improvement items
11. Corrective action 9. Assuring the quality of test &
12. Preventive action calibration results (5.9.1)
13. Control of records 10. Reporting the results
14. Internal audits
15. Management reviews 13
 ISO 17025 Clause 5.9.1 says....
• The laboratory shall have quality control procedures for monitoring the validity of tests
and calibrations 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.

• This monitoring shall be planned and reviewed and may include, but not be limited to,
the following

a. Use of CRMs and/or RMs ;

b. Proficiency-testing;
c. Replicate tests;
d. Retesting;
e. Correlation of results for different characteristics

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Quality control checks in daily life

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 Classification of AQC

• Internal Quality Control

• External Quality Control

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 Internal quality control checks
Internal quality control consists of all the procedures undertaken by a laboratory for the
continuous evaluation of its work in order to ensure the consistency of day-to-day results
and their conformity with defined criteria
This may take a variety of forms including the use of

1. Replicate determinations
2. Reagent blank
3. Method blank
4. Solvent blank
5. Positive control
6. Negative control
7. Use of in-house standards
8. Use of internal standards
9. Use of certified reference material
10. Spike recovery
11. Retesting /Blind sample
12. Calibration verification
13. Trend analysis using control charts

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 Replicate determinations

• Conducting repeat analysis of the same sample

• Where results are customarily reported on the basis of single

determination, it should be practice to include in an analytical
batch replicate determinations on the same test material

• Preferably Replicate of the same test material should be

randomised in batch and ideally the analyst should not be aware of
their presence

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 Blank/Reagent Blank/ Method Blank/ Solvent Blank
• A blank or method blank determination is an analysis of a sample without the analyte or
attribute, or an analysis without a sample, i.e. going through all steps of the procedure
with the reagents only
• It is used to determine the contribution of the reagents and the preparative analytical
steps to error in the measurement
• For convenience, some analysts practice "forcing the blank to zero" by adjusting the
instrument. Some instruments even invite or compel analysts to do so
• This is equivalent to subtracting the blank value from the values of the standards before
plotting the calibration graph
• From the standpoint of Quality Control this practice must be discouraged
• This is becoming more and more common practice with modem sophisticated hi-tech
instruments
• Whatever the case, a decision on how to deal with blanks must made for each procedure
and laid down in the SOP concerned

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 Positive control

• Where it is anticipated that the majority of the test portion will not contain significant
amounts of analyte, it is important to analyse test portion to which analyte has been
added and show that the analysis produces expected results

• This approach can be used to check the effectiveness of the screening test or in presence
or absence analysis

• It is useful for checking the ability to correctly identify test portions containing analyte at
the action level

• It can be used to asses the incidences of false negatives

• This practice can be proved to be more useful in nutritional analysis of a product claimed
to be fortified with the respective nutrients
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 Negative control

• In a reverse situation of positive control where, most test portion contains significant
amount of analyte, it is important to include number of test portions which are
essentially free of the analyte so as to provide confidence that laboratory contamination
levels are acceptably low

• Use of a matrix which is essentially free of the analyte is particularly important in analysis
for trace amounts of additives

• In case of residues and contaminants it is preferred to have reagent blanks, which contain
no test matrix

• The other form of negative control uses test materials known to contain analyte at levels
just below the action level, results from these provide a check on the incidence of false
positive

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 In-house standards
• When a large number of similar analysis are to be conducted over a period of time it is
useful to check the self consistency of the data by use of an in house standard material

• For this a relatively large amount of the matrix containing an appropriate level, it can be
homogenised and kept under conditions where the analyte is stable

• At specified intervals test portions from this in-house standards are included in a normal
analytical batch

• Over a period of time it is possible to see whether the method is giving consistent data
and to obtain an estimate of the long term precision

• A large number of repeat analysis conducted on the in-house standard over a short time
allow the in-house precision (repeatability) of the method to be calculated

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 Internal standards
• Internal standards are similar in analytical behaviour to the compounds of interest, and
not expected to be found in the samples
• The response of the target compound is normalized to the response of the reference
standard. This reference standard is called an internal standard because it is contained
within the aliquot of the sample or sample extract that is actually injected into the
instrumentation
• The ratio of the peak area (or height) of the target compound in the sample or sample
extract to the peak area (or height) of the internal standard in the sample or sample
extract is compared to a similar ratio derived for each calibration standard
• The analyst needs to demonstrate that the measurement of the internal standard is not
affected by target analytes, surrogates or by matrix interferences · This is not as useful for
GC and HPLC methods with non-MS detectors, unless the internal standards could be
separated from target compounds chromatographically
• The retention times of the target compound and the internal standard may be used to
calculate the relative retention time (RRT) of the target compound and can then be used
to compensate for small retention time shifts

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 Use of Certified Reference Material
• These are homogenous test materials in which the analyte has been determined with
great care usually by a number of expert laboratories and preferably using a number of
different analytical techniques
• By analysing appropriate certified reference material from time to time it is possible to
check the accuracy of the results
• The in-house standards demonstrates the precision (consistency) of the data being
produced while CRM demonstrates the accuracy i.e. how near the results are to correct
values
• Since most analytical instrumentation is comparative, it requires a sample of known
composition (reference material) for accurate calibration. These reference materials are
produced under stringent manufacturing procedures and differ from laboratory reagents
in their certification and the traceability of the data provided.
• Quality management system involving laboratory accreditation as per ISO/IEC-17025
requires metrological traceability to Certified Reference Materials (where possible) when
using reference materials for calibration

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 Spike Recovery
• Spike, recovery is an important method for assessing the accuracy of analytical techniques
for particular sample types
• Matrix match standards are ideal for residue testing to get a better recovery
• Spike-and-recovery is used to determine whether analyte detection is affected by a
difference between the diluent, dilution accuracy of calibration standards prepared, the
standard curve and the sample matrix
• Recovery shall be determined in each batch of samples and the recovery factor obtained
for the specific batch shall be used in quantitative determination of an analyte in sample
• Standard recovery criteria are as given in the table, again it depends on respective
regulations.
concentration Accepted recovery range
Less than 1 µg/kg -50 to +20
Between 1 to 10 µg/kg -30 to +10
More than 10 µg/kg -20 to +10
• Calibration curve should include at least five calibration points and the working range
should be clearly described
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 Retesting /Blind sample

• Retesting of retained sample

• The stable parameters to be considered as the storage may

adversely affect the result

• The variation should be recorded and investigated

• One of the commonly followed QC in food labs.

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 Calibration verification
• Calibration is the process of testing and adjusting an instrument, kit, or test system
readout to establish a correlation between the instrument’s measurement of the
substance being tested and the actual concentration of the substance
• Use of CCV standards are recommended in heavy metal testing
• Calibration verification is the periodic confirmation by analysis of a calibration standard
that the instrument performance has not changed significantly from the initial calibration
• Calibration verification should also be performed under the following conditions
1. Whenever major maintenance is performed or a critical component part of an analyser has
been replaced
2. Whenever reagent lots are completely changed (unless it has been stated and shown that
these lot changes do not affect test results, as with manufacturer’s instructions and guidelines
in package inserts and analyser specific manuals)
3. When control values are found to be continually unacceptable, as with shifts and trends in
Shewharts/Levy-Jennings graphs over a period of time

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 AQC checks in Food Microbiology Lab

• Standards & critical consumable checks

• Reference culture quality checks
• Critical consumable quality verification
• Media/Diluent sterility checks
• Spiking samples with reference microbial cultures
• Duplicate analysis of sample for enumeration tests
• Analysis of same sample by different method, personnel, equipment etc.

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 External quality control
Laboratories may wish for, or indeed may be required to have, an external check on their
performance. For this, they need to participate in Proficiency testing (PT) schemes.
Example of EQC includes
1. Inter laboratory comparisons
2. Proficiency Testing

Inter laboratory comparison

• It is inter comparison of measurement results of a laboratory
• Successful participation in an inter laboratory comparison is one of the necessary
requirements of quality control of data generated from food testing laboratories when PT
is not available.

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 External quality control
Proficiency testing
• External proficiency testing is an independent external assessment of the correctness of
the results and provides an impartial test of analytical quality: which can be done through
an inter laboratory proficiency testing
• Usually it involves presenting the analyst with the test material and requesting specific
information about its properties such as moisture, fat, protein, ash, fatty acid composition,
toxic metal concentration, pesticide/drug residues etc.
• Ideally the analyst should not be aware that the it is proficiency test, but often this will not
be feasible
• A single bad result from bad testing may prompt actions to rectify the matter, but a single
good result in no way guarantees that all results are of quality achieved on the proficiency
testing
• The proficiency test therefore needs to be conducted on quite regular basis

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 Trend Analysis using control charts

• The control chart, also known as the process-behaviour chart is one of the seven basic
tools of Statistical Quality Control
• A control chart is a graph of test results with respect to time or sequence of
measurements, with limits drawn within which results are expected to lie when the
analysis is in a state of “statistical control”.
• A procedure is in statistical control when results consistently fall within established
control limits.
• It reveals trends that in themselves do not yet result in a rejection of analytical results,
but it will give early warnings of issues or problems that are developing.
• The accuracy of an analysis or manufacturing process is monitored using one or more of
the following control charts:
 Shewhart control chart
 Moving average control chart
 CUSUM control chart

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 Shewhart Control Charts
• Value of the control sample/ QC standard for each batch is plotted on the y axis.
• The batch number (or measurement number) is plotted on the x axis.
• If the analysis (or process) is working satisfactorily the points should be randomly
distributed about the target value and between the upper warning limit (UWL) and lower
warning limit (LWL). Warning limits at the target value±2s (where s is the standard
deviation of a large number of replicate analyses) and the action limits at the target value
±3s

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 Moving average Control Chart
• The average results of a given number of batches is plotted, instead of plotting the result
for each individual batch.
• Each point plotted will be the average of that batch, plus a given number of batches
immediately before it.
• Advantage : This smoothens the data making trends easier to see.
• Disadvantage: This delays the appearance of changes affecting the analysis.
• The warning limits are the target value±(2s/√n) and the action limits are the target value
±(3s/√n), where n is the number of batches used to calculate the moving average.

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 CUSUM Control Chart
• This chart shows up trends more quickly than a Shewhart chart.
• In this chart, the difference between each QC value and the target value (QC result −
target value) is calculated, taking note of the sign.
• The differences are added and plotted against batch number.
• If the analysis is working satisfactorily the points will wander about 0 (the x axis).
• If there is a problem with the analysis or process, the points will move away from the x
axis, up or down.

Target value

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 Trend-Out of Control

• The warning limits can be expected to be breached once in 20 occasions, therefore no

action may be required for a single breach when all other points lie within the warning
limits and are randomly distributed around the target mean.
• More fall outside 3s (outside the upper or lower control limits)
• Two or more consecutive values fall outside 2s (outside the upper or lower warning
limits) on the same side of the mean
• A series of seven or eight consecutive values fall all above or all below the mean
• An increasing or decreasing trend is detected.

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 Conclusion

• Laboratory accreditation does not guarantee that the laboratory is of high quality, but it
helps give customers confidence that the lab is capable of producing quality data.
• Implementation of the AQC requires resources and investment, so there is a significant
cost to quality.
• By applying systematic and continuous QC tools in routine testing, a lab can achieve the
accuracy in their reported results.
• The new trend includes software programs to implement QC activities and statistical
analysis.
• The responsibility of QC does not lie with only the laboratory management or QC /QA
department personnel.
• Each individual working in the lab is responsible for the implementation of QC
techniques. QC is a collective responsibility.

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Thank You For Your Kind Attention