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សាកលវិទ្យាល័យជាតិមានជ័យ
មហាវិទ្យាល័យ កសិកមម និងកកច្នៃអាហារ
មុខវិជាា ការវិភាគអាហារ

LESSON 1: INTRODUCTION
TO FOOD ANALYSIS
NMU – 2023
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Introduction
• Investigations in food science and technology, by food
industry, governmental agencies, or universities, often
require the determination of food composition and
characteristics.
• The characteristics of foods including chemical
compositions, physical properties, sensory properties
must meet regulations and specifications.
• To ensure the quality and safety of the food supply
• Analysis is done of problem samples and competitor
products.
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Introduction
• The nature of sample and the specific reason for analysis
commonly influence the choice of analytical methods.
• Speed, precision, accuracy and ruggedness are the key
factors.
• Validation of the method is necessary to ensure the
usefulness of the method.
• The success of any method relies on the proper selection
and preparation of the food sample, carefully performing
the analysis, and doing the appropriate calculations and
interpretation of the data.
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Introduction
• Official methods are critical in the analysis of food, to
ensure that they meet the legal requirements.
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Trends and Demands

• Consumers demand a wide variety of products that are of
high quality, nutritious, and offer a good value.
• Also, they are concerned about the safety of foods, which
has increased the testing of foods for allergens, pesticide
residues, and products from genetic modification of food
materials.
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Trends and Demands

• Analytical methods must be applied across the entire food
supply chain to achieve the desired final product quality.
• Food companies rely on others to supply high-quality and
safe raw ingredients and packaging materials, therefore,
they have select suppliers, who perform the analytical
tests, to ensure compliance with detailed specifications.
• The results of these analytical tests related to the
specifications are delivered as a Certificate of Analysis
(COA) with the ingredient/raw material.
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Steps in Analysis
• Select and prepare sample: obtain a representative
sample and convert the sample to a form that can be
analyzed.
• Perform the assay: chemicals, reagents, apparatus, and
step-by-step instructions.
• Calculate and interpret the results
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Choice and Validity of Method

• Why need to select a method?
• Numerous methods are available to assay food samples.
• Characteristics of the method
• Objective of the assay: reference, definitive, official, or
primary, rapid secondary or field methods
• Consideration of food composition and characteristics:
performance of many analytical methods is affected by
food matrix
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Choice and Validity of Method

• One must consider certain characteristics of any method,
such as specificity, precision, accuracy, and sensitivity.
• For data to be valid, equipment must be standardized and
appropriately used, and the performance limitations must
be recognized.
• A major consideration for determining method validity is
the analysis of materials used as controls, often referred
to as standard reference materials or check samples.
• Standard reference materials are important tools to
ensure reliable data.
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Choice and Validity of Method

• The choice of method is often made easier by the
availability of official methods.
• Several nonprofit scientific organizations have compiled
and published these methods of analysis for food
products, which have been carefully developed and
standardized.
• AOAC International (1884) is an organization to serve
the analytical methods needs of government regulatory
and research agencies
• And other: AACC International, AOCS
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EVALUATION OF
ANALYTICAL DATA
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Evaluation of Analytical Data

• our effort would be for naught if there were not some way
for us to evaluate the data obtained from assays.
• Several mathematical treatments are available to provide
an idea of how well a particular assay was performed
Measures of Central Tendency
Reliability of Analysis
Curve fitting: Regression Analysis
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Measures of Central Tendency

• To increase accuracy and precision, the analysis of a
sample is usually performed (repeated) several times, at
least three times.
• We usually determine the mean (or average) using all the
values obtained and report the results of the mean.

x
x1  x2  x3    xn

x i

n n
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Measures of Central Tendency

• Example 1
• we measured moisture content of a sample of uncooked
hamburger four times and obtained the following results:
64.53%, 64.45%, 65.10%, and 64.78%.
• Calculate the mean of moisture content
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Measures of Central Tendency

• Another determination is the median, which is the
midpoint or middle number within a group of numbers.
• The median is not used often, because the mean is such
a superior experimental estimator.
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Reliability of Analysis
• Accuracy refers to how close a particular measure is to
the true or correct value.
• The problem in determining accuracy is that most of the
time we are not sure what the true value is.
• A term that is much easier to deal with and determine is
precision. This parameter is a measure of how
reproducible or how close the replicate measurements
become.
• If repetitive testing yields similar results, then the
precision of that test was good, or the precision often is
called error.
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Reliability of Analysis
 18

Reliability of Analysis
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Reliability of Analysis
• The most commonly used statistical evaluation of the
precision of analytical data is the standard deviation.
• The standard deviation (SD) measures the spread of the
experimental values and gives a good indication of how
close the values are to each other.

  i
( x x ) 2

n
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Reliability of Analysis
• In case of number of replicates is small (≤30), n is
replaced by n–1 term
• The standard deviation (SD) is calculated by

SD 
 i
( x x ) 2

n 1
( x1  x ) 2  ( x2  x ) 2  ...  ( xn  x ) 2

n 1
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Reliability of Analysis
• Example 2
• Determine standard deviation in Example 1.
• 64.53%, 64.45%, 65.10%, 64.78%.
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Reliability of Analysis
• One way to get a feel for the standard deviation is to
calculate what is called the coefficient of variation (CV),
also known as the relative standard deviation.

SD
CV   100%
x
• The small CV value indicates the high level of precision or
reproducibility of the replicates.
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Reliability of Analysis
• Example 3
• Determine CV in the above Example.
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Reliability of Analysis
• If the true value is not known, we can calculate only
Precision.
• If we know the true value, we can determine the accuracy.
• One term that can be calculated is the absolute error,
which is simply the difference between the experimental
value and the true value.
E abs  x  T
• A more useful measurement of error is relative error.

x T
E rel 
T
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Reliability of Analysis
• Example 4
• Using moisture data of uncooked hamburger, calculate
absolute and relative error if the true moisture content of
the sample is 65.05%.
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Source of Errors
• it is unreasonable to expect an analytical technique to be
entirely free of error.
• The best is that the variation is small.
• Systematic or determinate error produces results that
consistently deviate from the expected value in one
direction or the other (b).
• Random or indeterminate errors are always present in
any analytical measurement.
• Blunders are easy to eliminate, since they are so
obvious; the results are not close to an expected value.
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Sensitivity and Limit of Detection

• Sensitivity relates to the magnitude of change of a
measuring device (instrument) with changes in compound
concentration.
• Limit of detection (LOD) is the lowest possible increment
that we can detect with sssome degree of confidence
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Curve Fitting
• Curve fitting is a generic term used to describe the
relationship and evaluation between two variables.
• The x-axis data are called the independent variable,
while the y-axis data, the dependent variable, may have
error.
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Curve Fitting
• We can mathematically determine the best fit of the line
by using linear regression.
• Line Equation: y = ax + b

a
 ( x  x )( y  y )
i i

 (x  x) i
2

b  y  ax

r
 ( x  x )( y  y )
i i

 (x  x)  ( y  y)
i
2
i
2
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Curve Fitting
• Example 5
• Using the data, calculate the concentration of a sample
with an area of 4000 at 272 nm.
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SAMPLING AND SAMPLE

PREPARATION
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Sampling Procedures
• In most cases quality attributes are measured on small
portions of material that are taken periodically from
continuous processes or on a certain number of small
portions taken from a lot.
• The small portions taken for analysis are referred to as
samples, and the entire is called a population.
• The process of taking samples from a population is called
sampling.
• The correct sample is critical to obtain the accurate
estimation of the population.
• The analysis can be obtained accurately, quickly, and with
less expense and time by sampling.
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Sampling Procedures
• The first step in any sampling procedure is to clearly
define the population.
• A sample of a lot must be used to make inferences about
that particular lot; conclusions cannot be extended to
other lots in the warehouse.
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Sampling Procedures
• After sampling, a series of stepwise procedures – from
sample preparation, laboratory analysis, data processing,
and interpretation – are conducted.
• In each step, there is a potential for error, so the final
result depends on the cumulative errors at each stage.
• The reliability of sampling is dependent more on the
sample size than on the population size.
• The larger the sample size the more reliable the sampling.
• However, sample size is limited by time, cost, sampling
methods, analysis, and data processing.
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Sampling Procedures
• The International Union of Pure and Applied
Chemistry (IUPAC) defines a sampling plan as: “A
predetermined procedure for the selection, withdrawal,
preservation, transportation, and preparation of the
portions to be removed from a lot as samples”
• A sampling plan should be selected on the basis of the
sampling objective, the study population, the statistical
unit, the sample selection criteria, and the analysis
procedures.
• Depending on the purpose of the sampling plan, samples
are taken at different points of the food production system,
and the sampling plan may vary.
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Sampling Procedures
Factors that affect the choice of sampling plan:
• Purpose of the inspection: accept or reject; determine the
variability
• Nature of the product: homogenous or heterogeneous,
cost of sample
• Nature of test method: critical or minor; destructive; cost
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Acceptance Sampling
• Acceptance sampling is a procedure that is used to
determine if a shipment of products has enough quality to
be accepted.
• Acceptance sampling can be performed by the food
processor before receiving materials from a supplier, or by
a buyer.
• Lot acceptance sampling plans may fall into the following
categories: single, double, multiple, sequential, and
skip plans.
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Acceptance Sampling
• In single sampling plans, the decision of accepting or
rejecting a lot is based just on one sample of items taken
at random.
• Multiple sampling plans are extensions of double
sampling plans for which more than two samples are
drawn to reach a conclusion.
• The ultimate extension of multiple sampling is sequential
sampling.
• In skip lot sampling only a fraction of the submitted lots
is inspected.
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Manual Sampling
• To obtain a manual sample the sampler must attempt to
take a “random sample” to avoid bias in the sampling
method.
• the sample must be taken from a number of locations
within the population to ensure that it is representative of
the whole population.
• For liquids, this can be done by shaking prior to sampling.
Liquids may be sampled by pipetting, pumping, or
dipping.
• when sampling grain, samples are obtained by probing
from several points at random.
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Problems in Sampling
• Sampling bias may compromise reliability.
• Errors also may be introduced by not understanding the
population distribution and selection of an inappropriate
sampling plan .
Unreliable data also can be obtained a poor sample
storage resulting in sample degradation.
• Samples should be stored in a container that protects the
sample from moisture and other environmental factors
(heat, light, air…)
• How to protect our samples ?!!!!!
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Problems in Sampling
• Store in opaque glass for light-sensitive samples
• Store under nitrogen or inert gas for oxygen-sensitive
samples
• Refrigeration or refreezing for unstable samples.
• Use of preservatives, if applicable
•
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Problems in Sampling
• Mislabeling of samples causes mistaken sample
identification.
• For example, plastic bags that are to be stored in ice
water should be marked with water-insoluble ink .
• The container must be sealed to protect against
tampering, eg. an official or legal sample
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Preparation of Samples
1) Size Reduction: If the particle size or mass of the
sample is too large, it must be reduced in particle size.
The samples are thus homogenized to ensure negligible
differences between each portion.
2) Grinding is important both for sample preparation
prior to analysis and for food ingredient processing. To
homogenize moist samples, bowl cutters, meat mincers,
tissue grinders, mortars and pestles, or blenders are
used, while mortars and pestles and mills are best for
dry samples.
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Preparation of Samples
3) Enzymatic inactivation: Enzyme activity therefore must
be eliminated or controlled since it may degrade the food
components being analyzed. Heat denaturation, freeze
storage (–20C or lower), changing pH, salting out.
4) Lipid oxidation protection: Unsaturated lipids are
sensitive to oxidative degradation and should be protected
by storing under nitrogen or vacuum. Antioxidant, cold
storage.
5) Microbial growth and contamination: Microorganisms are
present in almost all foods and can alter the sample
composition. Freezing, drying, and chemical preservatives
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Summary
• Food quality is monitored at various processing stages
but 100% inspection is rarely possible
• To obtain a representative sample for analysis, sampling
and sample reduction methods must be developed and
implemented.
• The selection of the sampling procedure is determined by
the purpose of the inspection, the food product, the test
method, and the characteristics of the population.
• Increasing the sample size will generally increase the
reliability of the analytical results.
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Summary
• Sampling is a vital process, as it is often the most variable
step in the entire analytical procedure
• There is no sampling plan that is risk free.
• The consumer risk is the probability of accepting a poor
quality product, while the vendor risk is the probability of
rejecting an acceptable product.
• Sampling methods may be manual or continuous.
• Ideally, the sampling method should be statistically sound.
• Each sample must be clearly marked for identification and
preserved during storage until completion of the analysis.
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Summary
• Often, only a portion of the sample is used for analysis,
and sample size reduction must ensure that the portion
analyzed is representative of both the sample and
population.
• Sample preparation and storage should account for
factors that may cause sample changes.
• Samples can be preserved by limiting enzyme activity,
preventing lipid oxidation, and inhibiting microbial
growth/contamination.