Food Analysis-4

SAMPLING AND SAMPLING TECHNIQUES

• Analysis of the properties of a food material depends on the
successful completion of a number of different steps:
▪ planning (identifying the most appropriate analytical procedure),
▪ sample selection,
▪ sample preparation,
▪ performance of analytical procedure,
▪ statistical analysis of measurements, and
▪ data reporting.
 Sample Selection and Sampling Plans
• A food analyst often has to determine the characteristics of a large quantity of food
material, such as the contents of a truck arriving at a factory, a days worth of
production, or the products stored in a warehouse.
• Ideally, the analyst would like to analyze every part of the material to obtain an accurate
measure of the property of interest, but in most cases this is practically impossible.
• Many analytical techniques destroy the food and so there would be nothing left to sell if
it were all analyzed. Another problem is that many analytical techniques are time
consuming, expensive or labor intensive and so it is not economically feasible to
analyze large amounts of material.
• It is therefore normal practice to select a fraction of the whole material for analysis, and
to assume that its properties are representative of the whole material.
• Selection of an appropriate fraction of the whole material is one of the most important
stages of food analysis procedures, and can lead to large errors when not carried out
correctly.
 Populations, Samples and Laboratory Samples
• It is convenient to define some terms used to describe the characteristics of a
material whose properties are going to be analyzed.
• Population: The whole of the material whose properties we are trying to obtain
an estimate of is usually referred to as the population.
• Sample: Only a fraction of the population is usually selected for analysis, which
is referred to as the sample. The sample may be comprised of one or more sub-
samples selected from different regions within the population.
• Laboratory Sample: The sample may be too large to conveniently analyze using
a laboratory procedure and so only a fraction of it is actually used in the final
laboratory analysis. This fraction is usually referred to as the laboratory sample.
• The primary objective of sample selection is to ensure that the properties of the
laboratory sample are representative of the properties of the population,
otherwise erroneous results will be obtained. one must always be aware that
analysis of a limited number of samples can only give an estimate of the true
value of the whole population.
 Sampling Plans
• To ensure that the estimated value obtained from the laboratory sample is a good
representation of the true value of the population it is necessary to develop a sampling plan.
• A sampling plan is a clearly written document that contains precise details that an analyst
uses to decide the sample size, the locations from which the sample should be selected, the
method used to collect the sample, and the method used to preserve them prior to analysis.
• It should also stipulate the required documentation of procedures carried out during the
sampling process.
• For certain products and types of populations sampling plans have already been developed
and documented by various organizations which authorize official methods, e.g., the
Association of Official Analytical Chemists (AOAC).
• The choice of a particular sampling plan depends on
➢the purpose of the analysis,
➢the property to be measured,
➢the nature of the total population and of the individual samples, and
➢the type of analytical technique used to characterize the samples.
 Developing a Sampling Plan
• Different sampling plans have been designed to take into account differences in the types of
samples and populations encountered, the information required and the analytical techniques
used. Some of the features that are commonly specified in official sampling plans are listed
below.
• To ensure that the estimated value obtained from the laboratory sample is a good
representation of the true value of the population, it is necessary to develop a sampling plan.
• A sampling plan should be a clearly written document that contains precise details that an
analyst uses
➢to decide the sample size,
➢locations from which the sample should be selected,
➢method used to collect the sample,
➢the method used to preserve them prior to analysis
➢It should also stipulate the required documentation . of procedures carried out during the
sampling process.
 Sample size
• The size of the sample selected for analysis largely depends on;
➢the expected variations in properties within a population,
➢the seriousness of the outcome if a bad sample is not detected,
➢the cost of analysis, and
➢the type of analytical technique used
• Given this information it is often possible to use statistical techniques to design a
sampling plan that specifies the minimum number of sub-samples that need to be
analyzed to obtain an accurate representation of the population.
• Often the size of the sample is impractically large, and so a process known as
sequential sampling is used. Here sub-samples selected from the population are
examined sequentially until the results are sufficiently definite from a statistical
viewpoint. For example, sub-samples are analyzed until the ratio of good ones to
bad ones falls within some statistically predefined value that enables one to
confidently reject or accept the population.
 Sampling
• Consist of obtaining a larger group ( referred to as the population ). From this
sample, one hopes to obtain an estimate of the true value of the parameters of
interest with sufficient accuracy for the intended purposes.
• Sampling permits a reduction in cost and personnel while allowing information to
be obtained quickly and comprehensively.
• A laboratory sample is anything that is sent to a laboratory for analysis. It can be of
any size or quantity. The anticipated use of the results from the sample will
determine the sampling procedure.
• Populations may be finite or infinite. For finite populations, sampling provides an
estimate of lot. For infinite populations, sampling determines concern at many
points.
 TYPES OF SAMPLING
1. Non-probability sampling: Judgments sampling; Convenience
sampling; Haphazard sampling, Quota sampling.
2. Probability sampling: Simple random sampling; Systematic sampling;
Stratified random sampling; Clustered sampling; Composite sampling.
3. Mixed sampling: Mixed sampling combines random and purposeful
sampling. The lot is subdivided based on purposeful sampling
methods and items from within the group are selected randomly.
 NON-PROBABILITY SAMPLING
• Probability of inclusion of any portion of the whole in the sample is not equal. In these
sampling plans the investigator determines which sample will be selected. Accurate
estimates of the entire population are not possible because sampling error cannot be
determined.
a. Judgments sampling: Depends on the person choosing the sample. Frequently this method
is the only practical and feasible way to obtain a sample. If the investigator is experienced
in sample selection and the limitations in extrapolation of the results are understood, this
method may better represent the true state of the population than the random sampling.
b. Convenience sampling: It is often referred to as ‘chunk sampling’ or ‘grab sampling’ The
first pallet or easiest to get the box is selected. Such a sample is not representative of the
whole. Restricted sampling may be required when the entire population is not accessible.
Results are not representatives of the entire population.
c. Haphazard sampling: Selection of any portion of the sample. It should be avoided.
d. Quota sampling: It is the division of the lot into groups representing various categories. A
specific number of samples are selected from each group by judgment. The sampling plan
is less costly than random sampling but is also less reliable.
 PROBABILITY SAMPLING
• It provides a scientific method for selection of samples according to a statistical plan. The
chance of including each item is known and sampling error may be calculated.
a. Simple random sampling: It requires that the number of units in the population to the
sample is known. Each unit is assigned (in order) a number. A specific quantity of random
numbers is selected between 1 and the total number of units in the lot. Random number
tables may be used. Units are chosen corresponding to the random numbers and evaluated
for the characteristics of interest.
b. Systematic sampling: It is applied when a complete list of sample units is not available.
The first sample unit is selected at random and every nth unit after that is selected. This
plan is used when materials are continuously distributed over time or space. It is most
frequently used in production line sample.
c. Stratified random sampling: Population to be sampled is divided into subgroups such that
units within each group are as homogenous as possible. Group means are as widely
different as possible. Samples are taken randomly from each subgroup. This procedure
provides the most representative cross section of the entire population because no part is
excluded. It is less expensive than simple random sampling.
 PROBABILITY SAMPLING
d. Clustered sampling: Population is divided into subgroups termed ‘clusters’ such
that each subgroup is as similar to all others as possible. Heterogeneity is within the
cluster. This process is more efficient and less costly than simple random sampling for
populations that can be easily divided into homogenous groups. Clusters should be
small and number of sample units from each cluster is about the same.

e. Composite sampling: It is commonly used for flour, seeds and other items in bags.
It is also useful for solid samples in bulk. Two or more random samples are combined
to give one sample for analysis. This procedure averages differences within the
population.
 Operating Characteristic (OC) Curves
• Operating Characteristic (OC) curves are used
extensively in acceptance sampling.
• The Operating Characteristic (OC) curve shows the
probability of acceptance, Pa, for any level of lot
quality.
• On the horizontal axis is the quality characteristic. This
OC curve enables you to evaluate the probability of
acceptance for any true lot quality level-on a what-if
basis.
• This way, you can design sampling plans that perform
the way you want.
1. If the lot quality is 0.093 fraction defective, then the
probability of acceptance, Pa, is 0.05.
2. If the lot quality is 0.018 fraction defective, then the
probability of acceptance, Pa, is 0.95.
Requirements of Good Sampling Methods
• Good sampling techniques and good sampling practices.
• Inspection of the lot before sampling.
• Use of suitable sampling devices for the particular commodity and type of sample
desired.
• Use of suitable containers to hold the sample.
• Maintenance of the integrity of the sample and associated records.
• Use of adequate precautions in preserving, packing and delivery of the sample to the lab
in a timely manner.
• Provision of appropriate storage conditions for the sample both prior to and following
analysis.
• Cost versus benefits analysis, and
• A review of program objectives and regularity requirements, are to be assessed and
brought together in a sampling plan that serves as a guide to management, as well as to
operating personnel as a firm plan to achieve quality in sampling.
 SAMPLE LOCATION
1. Homogeneous versus heterogeneous population:
• The ideal population would be exactly the same at every location. Such a population
would be homogeneous. Sampling from a homogeneous population is simple.
Unfortunately in the real world such populations are rare. Most populations are
heterogeneous.
• Results obtained with samples taken from heterogeneous population will depend on
the location of sampling. The food material within the sample selected from the
population is usually heterogeneous, i.e., its properties vary from one location to
another. Sample heterogeneity may either be caused by in the properties of different
units within the sample (inter-unit variation) e. g. a box of oranges, some of good
quality and some of bad quality. Variations within the individual units in the sample
(intra-unit variation). E. g. individual orange, whose skin has different properties
than its flesh.
 SAMPLE LOCATION
2. Manual versus continuous sampling:
• Manual sampling is done by humans. Regardless of the process, it is imperative that
the unit being sample be as homogenous as possible prior to sampling. For liquids in
small containers, this is achieved by shaking prior to sampling. For liquids in silos,
aeration maintains a homogenous unit.
• For grains in rail cars samples are probed from several points at random and a
composite sample prepared to represent the whole. Granular or powdered samples
may be taken with the aid of triers or probes that are inserted into the material. The
solid products may be sampled by cutting representative portions from specific
areas
• .Continuous sampling is performed by mechanical sampling devices.
 Sample collection.
• Sample selection may either be carried out manually by a human being
or by specialized mechanical sampling devices.

• Manual sampling may involve simply picking a sample from a conveyor

belt or a truck, or using special cups or containers to collect samples
from a tank or sack.

• The manner in which samples are selected is usually specified in

sampling plans.
 Preparation of Laboratory Samples
• Once we have selected a sample that represents the properties of the
whole population, we must prepare it for analysis in the laboratory.

• The preparation of a sample for analysis must be done very carefully in

order to make accurate and precise measurements.
 Making Samples Homogeneous
• The food material within the sample selected from the population is usually heterogeneous,
i.e., its properties vary from one location to another. Sample heterogeneity may either be
caused by variations in the properties of different units within the sample (inter-unit
variation) and/or it may be caused by variations within the individual units in the sample
(intra-unit variation).
• The units in the sample could be apples, potatoes, bottles of ketchup, containers of milk etc.
An example of inter-unit variation would be a box of oranges, some of good quality and
some of bad quality. An example of intra-unit variation would be an individual orange,
whose skin has different properties than its flesh. For this reason it is usually necessary to
make samples homogeneous before they are analyzed, otherwise it would be difficult to
select a representative laboratory sample from the sample.
• A number of mechanical devices have been developed for homogenizing foods, and the
type used depends on the properties of the food being analyzed (e.g., solid, semi-solid,
liquid). Homogenization can be achieved using mechanical devices (e.g., grinders, mixers,
slicers, blenders), enzymatic methods (e.g., proteases, cellulases, lipases) or chemical
methods (e.g., strong acids, strong bases, detergents).
 Reducing Sample Size
• Once the sample has been made homogeneous, a small more manageable portion is
selected for analysis.

• This is usually referred to as a laboratory sample, and ideally it will have properties
which are representative of the population from which it was originally selected.

• Sampling plans often define the method for reducing the size of a sample in order
to obtain reliable and repeatable results.
 Preventing Changes in Sample
• Once we have selected our sample we have to ensure that it does not
undergo any significant changes in its properties from the moment of
sampling to the time when the actual analysis is carried out, e.g.,
enzymatic, chemical, microbial or physical changes.

• There are a number of ways these changes can be prevented.

 Enzymatic Inactivation
• Many foods contain active enzymes they can cause changes in the
properties of the food prior to analysis, e.g., proteases, cellulases,
lipases, etc.
• If the action of one of these enzymes alters the characteristics of the
compound being analyzed then it will lead to erroneous data and it
should therefore be inactivated or eliminated.
• Freezing, drying, heat treatment and chemical preservatives (or a
combination) are often used to control enzyme activity, with the method
used depending on the type of food being analyzed and the purpose of
the analysis.
 Lipid Protection
• Unsaturated lipids may be altered by various oxidation reactions.
• Exposure to light, elevated temperatures, oxygen or pro-oxidants can
increase the rate at which these reactions proceed.
• Consequently, it is usually necessary to store samples that have high
unsaturated lipid contents under nitrogen or some other inert gas, in
dark rooms or covered bottles and in refrigerated temperatures.
• Providing that they do not interfere with the analysis antioxidants may
be added to retard oxidation.
 Microbial Growth and Contamination
• Microorganisms are present naturally in many foods and if they are not
controlled they can alter the composition of the sample to be analyzed.

• Freezing, drying, heat treatment and chemical preservatives (or a

combination) are often used to control the growth of microbes in foods.
 Physical Changes
• A number of physical changes may occur in a sample, e.g., water may
be lost due to evaporation or gained due to condensation; fat or ice may
melt or crystallize; structural properties may be disturbed.

• Physical changes can be minimized by controlling the temperature of

the sample, and the forces that it experiences.
 SAMPLE IDENTIFICATION
• Laboratory samples should always be labelled carefully so that if any problem
develops its origin can easily be identified. The information used to identify a
sample includes:
a. Sample description,
b. Time sample was taken,
c. Location sample was taken from,
d. Person who took the sample, and
e. Method used to select the sample.
• The analyst should always keep a detailed notebook clearly documenting the sample
selection and preparation procedures performed and recording the results of any
analytical procedures carried out on each sample.
• Each sample should be marked with a code on its label that can be correlated to the
notebook. Thus if any problem arises, it can easily be identified.
 Data Analysis and Reporting
• Food analysis usually involves making a number of repeated
measurements on the same sample to provide confidence that the
analysis was carried out correctly and to obtain a best estimate of the
value being measured and a statistical indication of the reliability of the
value.

• A variety of statistical techniques are available that enable us to obtain

this information about the laboratory sample from multiple
measurements.
 Measure of Central Tendency of Data
• The most commonly used parameter for representing the overall properties of a number of
measurements is the mean. The mean is the best experimental estimate of the value that can be
obtained from the measurements. It does not necessarily have to correspond to the true value of
the parameter one is trying to measure.
• There may be some form of systematic error in our analytical method that means that the
measured value is not the same as the true value.
• Accuracy refers to how closely the measured value agrees with the true value. The problem
with determining the accuracy is that the true value of the parameter being measured is often
not known.
• Nevertheless, it is sometimes possible to purchase or prepare standards that have known
properties and analyze these standards using the same analytical technique as used for the
unknown food samples.
• The absolute error Eabs, which is the difference between the true value (Xtrue) and the
measured value (Xi), can then be determined: Eabs = (Xi - Xtrue).
 Measure of Spread of Data
• The spread of the data is a measurement of how closely together repeated
measurements are to each other.
• The standard deviation is the most commonly used measure of the spread of
experimental measurements. This is determined by assuming that the experimental
measurements vary randomly about the mean, so that they can be represented by a
normal distribution.
• The standard deviation SD of a set of experimental measurements is given by the
following equation:
• Measured values within the specified range:
1. SD means 68% values within range (x - SD) to (x + SD)
2. SD means 95% values within range (x - 2SD) to (x + 2SD)
3. SD means >99% values within range (x - 3SD) to (x + 3SD)
 SOURCES OF ERROR
• There are three common sources of error in any analytical technique:
1. Personal Errors (Blunders): These occur when the analytical test is not carried out correctly: the
wrong chemical reagent or equipment might have been used; some of the sample may have
been spilt; a volume or mass may have been recorded incorrectly; etc. It is partly for this reason
that analytical measurements should be repeated a number of times using freshly prepared
laboratory samples. Blunders are usually easy to identify and can be eliminated by carrying out
the analytical method again more carefully.
2. Random Errors: These produce data that vary in a non-reproducible fashion from one
measurement to the next e.g., instrumental noise.
3. Systematic Errors: A systematic error produces results that consistently deviate from the true
answer in some systematic way, e.g., measurements may always be 10% too high. For example,
a nominally 100 cm3 pipette may always deliver 101 cm3 instead of the correct value.
• To make accurate and precise measurements it is important when designing and setting up an
analytical procedure to identify the various sources of error and to minimize their effects. Often,
one particular step will be the largest source of error, and the best improvement in accuracy or
precision can be achieved by minimizing the error in this step.
 STANDARD CURVES: REGRESSION ANALYSIS
• When carrying out certain analytical procedures it is necessary to prepare standard
curves that are used to determine some property of an unknown material.
• A series of calibration experiments is carried out using samples with known
properties and a standard curve is plotted from this data. For example, a series of
protein solutions with known concentration of protein could be prepared and their
absorbance of electromagnetic radiation at 280 nm could be measured using a UV-
visible spectrophotometer.
• For dilute protein solutions there is a linear relationship between absorbance and
protein concentration:
• A best-fit line is drawn through the date using regression analysis, which has a
gradient of a and a y-intercept of b. The concentration of protein in an unknown
sample can then be determined by measuring its absorbance: x = (y-b)/a, where in
this example x is the protein concentration and y is the absorbance. How well the
straight-line fits the experimental data is expressed by the correlation coefficient
r2, which has a value between 0 and 1. The closer the value is to 1 the better the fit
between the straight line and the experimental values: r2 = 1 is a perfect fit. Most
modern calculators and spreadsheet programs have routines that can be used to
automatically determine the regression coefficient, the slope and the intercept of a
set of data.
 STANDARD CURVES: REGRESSION ANALYSIS
Rejecting Data:
• When carrying out an experimental analytical procedure it will
sometimes be observed that one of the measured values is very
different from all of the other values, e.g., as the result of a blunder
in the analytical procedure. Occasionally, this value may be treated
as being incorrect, and it can be rejected. There are certain rules
based on statistics that allow us to decide whether a particular point
can be rejected or not. A test called the Q-test is commonly used to
decide whether an experimental value can be rejected or not.
• Here XBAD is the questionable value, XNEXT is the next closet
value to XBAD, XHIGH is the highest value of the data set and
XLOW is the lowest value of the data set. If the Q-value is higher
than the value given in a Q-test table for the number of samples
being analyzed then it can be rejected:
• For example, if five measurements were carried out and one
measurement was very different from the rest (e.g., 20,22,25,50,21),
having a Q-value of 0.84, then it could be safely rejected (because it
is higher than the value of 0.64 given in the Q-test table for five
observations).
Thanks