Arsi UNIVERSITY

DEPARTMENT : FOOD SCIENCE AND POST

HARVEST TECHNOLOGY
Course: Food Analysis (FSPT1074)

BY: Habtamu T.
June 4,2020 G.C
 Chapter one
Introduction to food Analysis
1.1 Introduction
 Food Is products derived from plants or animals
that can be taken in to the body to yield energy and
nutrients for the
 maintenance of life
growth and
repair of tissue.
 Historically, people secured food through two
methods:
 hunting and gathering
 agriculture 2
Today, most of the food consumed by the
world population is supplied by the
food industry
 Sources (origins) of our food are:
 Plant origin
Animal origin

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Food analysis is the evaluation of a food to
determine its conformity to the required
standard or specification

Food analysis also refers to the discipline

dealing with the development, application
and study of analytical procedures for
characterizing the properties of foods
and their constituents

 It is a tool used to ascertain the quality of

food
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 Laboratory safety
1. protective clothing (lab coat ) must be worn at all time
when working in the laboratories

2. Goggles are strongly recommended for your eye

protection

3. As far as possible do not work alone in the lab

4. In case of chemical sling on you wash thoroughly under

running water. an emergency shower is provided for series
contamination.

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5. If a chemical enters your eyes, irrigate the inside of the eye
thoroughly using running water.

6. Do not use your mouth to suck chemicals or any sample into

a pipette

7. Do not heat or mix anything close to the face

8. Do not pour volatile solvents in a beaker in the open

laboratory space but make use of the hood for volatile
substance .
9. Handle acids with special care. When diluting an acid
such as sulfuric acid, add acid to the water and not water
to acid to avoid over heating /explosion
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10. Clean up spilled chemicals immediately

11. Do not use cracked or chipped glassware

12. Handle all equipment with care.

13. Follow the procedure closely.

14. If a chemical is accidentally swallowed,

get medical help immediately.
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 Planning the Experiment

1. Search for relevant information on the assay to be

familiar with the principle and theory of the experiment

2. Prepare reagents and arrange the equipment

3. Prepare food sample for the analysis

4. Follow the procedure of the experiment

5. Record all observation and readings in your note book

immediately

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1.2 Who analysis foods?
 Food analysis is done by scientists working
in all of the major sectors of the food
industry including:

Food Manufacturer
 Ingredients Suppliers
 Analytical Laboratory Services
Government Laboratory
University Research Laboratories

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Reasons for Analyzing Foods
 Why do we need to carry out food analysis?
The various purposes that foods are analyzed
are briefly discussed below.

A. To answer specific question for government regulatory

purpose
B. To ensure safety of food supply

C. To answer specific question for quality control

D. For research and new product development purpose

Evaluating new processes for making food

products
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 A. Government Regulations and Recommendations
Why are Government regulations and recommendations
designed?
1. maintain the general quality of the food supply
2.ensure the food industry provides consumers with foods
that are wholesome and safe,
3. inform consumers about the nutritional composition of
foods so that they can make knowledgeable choices about
their diet,
4. enable fair competition amongst food companies, and
to eliminate economic fraud.

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 Government Departments Responsible for regulating
the composition and quality of foods in USA:
Food and Drug Administration (FDA),
the United States Department of Agriculture (USDA)
the National Marine Fisheries Service (NMFS) and
the Environmental Protection Agency (EPA).
In Ethiopia, EFMHACA (Ethiopian Food, Medicine and Health
Care Administration and Control Authority)
Responsibility of the Departments:
regulating particular sectors of the food industry and
publish documents that contain detailed information about the
regulations and recommendations pertaining to the foods
produced within those sectors.

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Government regulations agencies they have the right to control
i. Standards
ii. Nutritional labeling
iii. Authenticity
iv. Food inspection and grading
i. Standards
 Government agencies have a specified number of
voluntary and mandatory standards concerning the
composition, quality, inspection, and labelling of
specific food products.
1. Mandatory Standards
a) Standards of Identity
 These regulations specify the type and amounts of
ingredients that certain foods must contain.
B. Standards of Quality
 Standards of quality have been defined for certain foods (e.g., canned
fruits and vegetables) to set minimum requirements on the color,
tenderness, mass and freedom from defects.
C. Standards of Fill-of-Container
 These standards state how full a container must be to avoid consumer
deception, as well as specifying how the degree of fill is measured.
Example: canned food
2. Voluntary Standards
Standards of Grade: A number of foods, including meat, dairy products and
eggs, are graded according to their quality
 Eggs have grade AA to B
 Meat can be graded as “prime”, “choice”, “standard” and etc
 In Ethiopia, Ethiopian standard Agency (ESA)
 ii. Nutritional Labeling

 One of the major reasons for introducing these regulations is

that consumers could make informed choices about their diet
 (total calorific value of the food, total fat, saturated fat,
cholesterol, sodium, carbohydrate, dietary fiber, sugars,
protein, vitamins, calcium and iron)
 “The label may also contain information about nutrient content
claims (such as “low fat”, “low sodium” “high fiber” “fat free”
etc)”
iii. Authenticity/ genuine
 Price of foods usually dictated by the quality of the
ingredients that they contain
• Example,
• a packet of premium coffee may claim that the
coffee beans are from Ethiopia
 iv. Food Inspection and Grading

1.routinely analyses the properties of food products

2.ensure that they meet the appropriate laws and regulations.
• thus, accurate analytical techniques required
B. Food Safety
 Analyze foods to ensure that they are safe
• No harmful or toxic substances - HACCP
• A food may be considered to be unsafe because it contains
harmful microorganisms (e.g., Listeria, Salmonella), toxic
chemicals (e.g., pesticides, herbicides) or extraneous matter
(e.g., glass, wood, metal, insect matter)
 In many situations it is important to use analytical techniques
that have a high sensitivity, i.e., that can reliably detect low
levels of harmful material.
C. Food Quality Control
 The food industry is highly competitive and food
manufacturers are continually trying to increase their market-
share and profits.
 To do this they must ensure that their products are of higher
quality, less expensive, and more desirable than their
competitors, whilst ensuring that they are safe and nutritious.
 To insure quality control
 Characterization of raw materials.
 Monitoring of food properties during processing
D. Research and development
 In recent years, there have been significant changes in the
preferences of consumers for foods that are healthier, higher
quality, lower cost and more exotic.
 Individual food manufacturers must respond rapidly to these
changes in order to remain competitive within the food
industry.
 To meet these demands food manufacturers often employ a
number of scientists whose primary objective is to carry out
research that will lead to:
 the development of new products
 analytical techniques here needed to:
1. characterize the overall properties of foods (e.g., color,
texture, flavor, shelf-life etc.),
2. to ascertain the role that each ingredient plays in determining
the overall properties of foods, and
3. to determine how the properties of foods are affected by various
processing conditions (e.g., storage, heating, mixing, freezing).
1.2. Properties to be Analyzed
 Food analysts are interested in obtaining information about a
variety of different characteristics of foods and analytical
procedures are used to provide information about a wide
variety of different characteristics of foods, including their:
A. Composition
B. Structure
C. Physicochemical properties
D. Sensory attributes
 A. Composition
 Chemical Composition: it describes the water, fat, carbohydrate;
protein etc content of a food.
 The composition of a food largely determines its safety, nutrition,
physicochemical properties, quality attributes and sensory
characteristics.
B. Structure
 Two foods that have the same composition can have very different
quality attributes if their constituents are organized differently
• The structure of a food can be examined at different levels:
a) Molecular structure ( 1 – 100 nm). their three-dimensional
structure and their interactions with each other
b) Microscopic structure ( 10 nm – 100 mm). can be observed by
microscopy
Eg: discrete phases (emulsion droplets), fat crystals, protein
aggregates and small air cells
c) Macroscopic structure ( > 100 mm). can be observed by the
unaided human eye,
e.g., sugar granules, large air cells
 Hence, two foods that have the same composition can have
very different quality attributes if their constituents are
organized differently.
 For example, a carton of ice cream taken from a refrigerator
has a pleasant appearance and good taste, but if it is allowed to
melt and then is placed back in the refrigerator its appearance
and texture change dramatically and it would not be acceptable
to a consumer.
 Thus, there has been an adverse influence on its quality, even
though its chemical composition is unchanged, because of an
alteration in the structural organization of the constituents
caused by the melting of ice and fat crystals.
C. Physicochemical Properties
 These properties include:
Optical
 Rheological
 Stability
 Flavor
 Rheological properties: rheology is the study of the flow and
deformation of matter, and texture deals with a consumer’s
perception of rheology.
 Rheology is a factor in food process engineering, shelf-life
testing, and quality control
 The stability of a food is a measure of its ability to resist
changes in its properties over time. These changes may be
chemical, physical or biological in origin. The stability of a
food is a measure of its ability to resist changes in its
properties over time. These changes may be chemical, physical
or biological in origin.
 D. Sensory Attributes

• Ultimately, the quality and desirability of a food product is

determined by its interaction with the sensory organs of
human beings,
• e.g., vision, taste, smell, feel and hearing. Therefore,

• sensory properties of new or improved foods are usually

tested by human beings (acceptable and desirable
properties)

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 Sensory perception of individuals:

often fairly subjective, being influenced by certain factors:

• current trends,

• nutritional education,

• climate,

• age,

• health, and

• social, cultural and religious patterns.

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 Disadvantages of sensory analysis:

1.time consuming

2. expensive to carry out,

3. tests are not objective,

4. it cannot be used on materials that contain poisons or toxins,

and

5. it cannot be used to provide information about the safety,

composition or nutritional value of a food.

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 1.3 Type of sample analyzed
– Raw material

– Process control samples

– Finished product

– Competitor’s samples

– Complaint sample

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 1.4 Steps in Analysis
The success of any analytical method relies on the proper
selection and preparation of the food sample and careful analysis,
calculations and interpretation of the data
A. Select and Prepare Sample
 Sampling is the initial point for sample identification
 In analyzing food samples, all results depend on obtaining a
representative sample and converting the sample to a form
that can be analyzed.
B. Perform the Assay (methods)
 Assay for each analysis for different component or
characteristic or specific type of product is unique.
 Example: Methods to determine fatty acid and amino acid
are different.
C. Calculate and Interpret the Results
 Important to make appropriate calculations to
interpret the data correctly.
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Example: in Calculation of total protein, Choosing the
right factor is important
1.5 Choosing an Analytical Technique:
Most appropriate technique for the specific
application must be chosen.
Basis for the choice of a method:
1.property to be measured,
2.the type of food to be analyzed, and
3. the reason for carrying out the analysis.
Often it is necessary to consult scientific and
technical publications.

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Tabulated Official Methods of Analysis:

1. Association of the Official Analytical Chemists

(AOAC)
2. American Oil Chemists Society (AOCS) and
others

Steps:

a) a particular laboratory develops a new analytical

procedure

b) The method is then tested by a number of independent

laboratories

c) The results of these tests are collated and compared

d) After rigorous testing the procedure may be accepted,

modified or rejected as an official method.
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1.6 criteria for Selecting an Appropriate Technique:

Precision: A measure of the ability to reproduce an answer between

determinations performed by the same scientist (or group of
scientists) using the same equipment and experimental
approach.

Reproducibility: A measure of the ability to reproduce an answer by

scientists using the same experimental approach but in
different laboratories using different equipment.

Accuracy: A measure of how close one can actually measure the true
value of the parameter being measured, e.g., fat content, or
sodium concentration.

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Simplicity of operation: A measure of the ease with which
relatively unskilled workers may carry
out the analysis.

Cost: The total cost of the analysis, including the reagents,

instrumentation and salary of personnel required to
carry it out.

Speed: The time needed to complete the analysis of a single

sample or the number of samples that can be analyzed
in a given time.

Sensitivity: A measure of the lowest concentration of a

component that can be detected by a given
procedure.
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Specificity: A measure of the ability to detect and quantify
specific components within a food material, even
in the presence of other similar components,

e.g., fructose in the presence of sucrose or glucose.

Safety: Many reagents and procedures used in food analysis

are potentially hazardous

e.g. strong acids or bases, toxic chemicals or

flammable materials.

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Destructive/Nondestructive: In some analytical methods
the sample is destroyed during the analysis,
whereas in others it remains intact.

On-line/Off-line: Some analytical methods can be used to

measure the properties of a food during
processing, whereas others can only be
used after the sample has been taken from
the production line

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Official Approval: Various international bodies have
given official approval to methods that have been
comprehensively studied by independent analysts and
shown to be acceptable to the various organizations
involved, e.g., ISO, AOAC, AOCS.

Nature of Food Matrix: The composition, structure

and physical properties of the matrix material
surrounding the analyte often influences the type of
method that can be used to carry out an analysis, e.g.,
whether the matrix is solid or liquid, transparent or
opaque, polar or non-polar.
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 If there are a number of alternative methods available for
measuring a certain property of a food, the choice of a
particular method will depend on which of the above criteria is
most important.

For example, accuracy and use of an official method may be

the most important criteria in a government laboratory which
checks the validity of compositional or nutritional claims on
food products,

Whereas speed and the ability to make nondestructive

measurements may be more important for routine quality
control in a factory where a large number of samples have to
be analyzed rapidly.

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Sampling and Sample
Preparation

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 1. Introduction
What is research?

• “Scientific research is systematic,

controlled, empirical, and critical
investigation of natural phenomena guided
by theory and hypotheses about the
presumed relations among such
phenomena.”
– Kerlinger, 1986

• Research is an organized and systematic

way of finding answers to questions

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 Definition of some terms
Population

 The entire lot or production from which the sample is taken

is called a population

The group to which you want to generalize your findings.

The larger group you are representing with your sample.

The larger group to which your results will apply

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Sample

 The small portions taken from the population for analysis

are referred to as samples

A subset of the population being studied from which data is

actually collected

A good sample accurately represents all kinds of

elements/members in proportion to their presence in the
population.

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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

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 2.1 Sampling

 The process of obtaining a portion sample that is

representative of the whole is referred to as sampling

The two primary objectives of sampling are:

To estimate the average value of a characteristic
To determine if the average value meets the
specifications defined in the sampling plan

Sampling purposes vary widely among different food industries;

however, the most important categories include the following
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1. Nutritional labeling
2. Detection of contaminants and foreign matter
3. Statistical process control (Quality Assurance)
4. Acceptance of raw materials, ingredients, or
products (Acceptance Sampling)
5. Release of lots of finished product
6. Detection of adulterations
7. Microbiological safety
8. Authenticity of food ingredients, etc. 43
The first step in any sampling procedure is to clearly define
the population with respect to production lot, a day’s
production, the contents of a warehouse

Once sampling is conducted, a series of stepwise procedures

needed to obtain data from the samples are:

1. Select and prepare samples

2. Perform the assay (laboratory analysis)
3. Data processing, and interpretation

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2.2 Sampling Plans
–Sampling plan is “
A predetermined procedure for the
selection, withdrawal, preservation,
transportation, and preparation of the
portions to be removed from a lot as
samples”

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”
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 For certain products and types of populations sampling plans

have already been

developed and documented by various

organizations which authorize official methods
The choice of a particular sampling plan is depends on
the purpose of the analysis
the property to be measured
the nature of the total population and of the individual samples
the type of analytical technique used to characterize the samples.

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2.3 Factors Affecting Choice of Sampling Plans
Factors to be considered
Purpose of the inspection
Nature of the product
Nature of the test method
Nature of the population being
Investigated
Questions
Is it to accept or reject the lot?
Is it to measure the average quality of the lot?
Is it to determine the variability of the product?
Is it homogeneous or heterogeneous?
What is the unit size?
How consistently have past populations met
specifications?
What is the cost of the material being sampled?
Is the test critical or minor?
Will someone become sick or die if the population fails
to pass the test?
Is the test destructive or nondestructive?
How much does the test cost to complete?
Is the lot large but uniform?
Does the lot consist of smaller, easily identifiable sub
lots?
What is the distribution of the units within the
population?
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 2.3.1 SAMPLING PROCEDURES

 The reliability of analytical data is compromised if

sampling is not done properly.

As shown in Table 2-1, the use of the data to be obtained

will determine the sampling procedure.

Details for the sampling of specific food products are

described in the Official Methods of Analysis of AOAC
International and in the Code of Federal Regulations (CFR).

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 The AOAC Method 925.08 describes the method for sampling

flour from sacks.

The number of sacks to be sampled is determined by the
square
root of the number of sacks in the lot.
 The sacks to be sampled are chosen according to their
exposure.
 The samples that are more frequently exposed are sampled
more often than samples that are exposed less.
 Sampling is done by drawing a core from a corner at the top
of the sack diagonally to the center. 49
The sampling instrument is a cylindrical, polished trier with a
pointed end.
 It is 13 mm in diameter with a slit at least one third of the
circumference of the trier.
 A second sample is taken from the opposite corner in a similar
manner.
The cores are stored for analysis in a clean, dry, airtight
container
that has been opened near the lot to be sampled.
The container should be sealed immediately after the sample is
added.
A separate container is used for each sack.
Additional details regarding the container and the procedure
also are described below.

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 Title 21 CFR specifies the sampling procedures required to
ensure that specific foods conform to the standard of identity.
 In the case of canned fruits, 21 CFR 145.3 defines a sample unit
as “container, a portion of the contents of the container, or a
composite mixture of product from small containers that is
sufficient for the testing of a single unit”
 Furthermore, a sampling plan is specified for containers of
specific net weights.
 The container size is determined by the size of the lot.
 A specific number of containers must be filled for sampling of
each lot size.
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 The lotis rejected if the number of defective units exceeds the
acceptable limit.
 For example, out of a lot containing 48,001–84,000 units, each
weighing 1 kg or less, 48 samples should be selected.
 If six or more of these units fail to conform to the attribute of
interest the lot will be rejected.
 Based on statistical confidence intervals, this sampling plan will
reject 95% of the defective lots examined, that is, 5% consumer
risk
 The discussion below describes general considerations to take
into
account when obtaining a sample for analysis
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1. Homogeneous vs. Heterogeneous Populations

Homogeneous population is an ideal and would be

uniform throughout and identical at all locations

sample can be taken from any location and the analytical

data obtained will be representative of the whole

 However, this occurs rarely, as even in an apparently

uniform product, such as sugar syrup, suspended particles
and sediments in a few places may render the population
heterogeneous
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Heterogeneous, the location within a population
where a sample is taken will affect the subsequent data
obtained

However, sampling plans and sample preparation can

make the

sample representative of the population or

take heterogeneity into account in some other way

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2. Manual vs. Continuous Sampling

Manual sampling
Attempt to take a “random sample” to avoid human bias
The sample must be taken from a number of locations
within the population
Shake prior to sample for liquids in small containers
 when sampling grain from a rail car, mixing is
impossible and samples are obtained by probing from
several points at random within the rail car

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 Such manual sampling of granular or powdered material is
usually achieved with triers or probes that are inserted into the
population at several locations.
 Errors may occur in sampling as rounded particles may flow into
the sampling compartments more easily than angular ones.

 Similarly, hygroscopic materials flow more readily into the

sampling devices than do nonhygroscopic materials.

 Horizontal core samples have been found to contain a larger

proportion of small-sized particles than vertical ones
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Continuous sampling
 is performed mechanically.
 less prone to human bias than manual sampling.

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2.4 Sampling by Attributes and Sampling by Variables
Sampling plans are designed for examination of either attributes
or variables:
 Attribute sampling
is performed to decide on the acceptability of a
population based on whether the sample possesses a
certain characteristic or not
 dichotomous form (Data with two possible
alternatives)
Ex: Present / absent
Eg Presence of Clostridium botulinum) contamination
in
canned foods
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Variable sampling
is performed to estimate quantitatively the
amount of a substance (e.g. moisture content,
etc.) or a characteristic (e.g., color) on a
continuous scale
The estimate obtained from the sample is
compared with an acceptable value
 In general, variable sampling requires smaller sample
size than attribute sampling

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2.4.1 Acceptance sampling
 Acceptance sampling is a procedure that serves a very
specific role: to determine if a shipment of products or
ingredients has enough quality to be accepted.

 Acceptance sampling can be performed by the food

processor before receiving a lot of materials from a supplier,
or by a buyer who is evaluating the processor’s output

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 Lot acceptance sampling plans that may be used for
evaluation of attributes or variables, or a combination of
both, fall into the following categories:
•Single sampling plans
Allow accept/reject decision to be made by inspection
of one sample of a specific size
 If the number of defects does not exceed a specified acceptance
number (C), the consumer accepts the entire lot.
 Any defects found in the sample are either repaired or returned to the
producer.
 If the number of defects in the sample is greater than C, the consumer
subjects the entire lot to 100 percent inspection or rejects the entire
lot
and returns it to the producer
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•Double sampling plans
 management specifies two sample sizes (n 1 and n 2 )
and two acceptance numbers (C 1 and C 2 ).
 If the quality of the lot is very good or very bad, the
consumer can make a decision to accept or reject the lot
on the basis of the first sample, which is smaller than in
the single-sampling plan.
To use the plan, the consumer takes a random sample of
size n 1 . If the number of defects is less than or equal to
C 1 , the consumer accepts the lot.
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If the number of defects is greater than C 2 , the consumer
rejects the lot.
If the number of defects is between C1 –C 2 , the
consumer
takes a second sample of size n 2.
If the combined number of defects in the two samples is
less
than or equal to C 2 , the consumer accepts the lot.
Otherwise, it is rejected.
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64
•Multiple sampling plans
The amount of sampling depending on the overall lot
quality
Reject low quality lots and accept high quality lots
quickly

65
 The analyst plots the total number of defectives against the
cumulative sample size, and if the number of defectives is less
than
a certain acceptance number (C 1 ), the consumer accepts the
lot.

 If the number is greater than another acceptance number (C 2 ),

the consumer rejects the lot.

 If the number is somewhere between the two, another item is

66
inspected.
67
There are two types of risks in acceptance sampling:

1 Consumers’ risk: This is the chance that a poor quality lot is

passed as good the risk of accepting a substandard
lot/population.
• This should be as low as possible, usually 5%.
• The actual level depends on the consequence of failure are very
severe, then the risk should be minimized further, may be < 1%.
Will someone get sick and/ or die if we fail to detect
a defect that should have been rejected?
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2. Vendors’ risk: This is the chance of rejecting a good lot.

 This is usually set at 5- 10%. Sampling plan should protect

both the consumer and vendor.

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 acceptable quality level (AQL) The quality level desired by

the consumer.

 The producer of the item strives to achieve the

AQL, which typically is written into a contract or
purchase order.

For example, a contract might call for a quality level

not to exceed one defective unit in 10,000, or an
AQL of 0.0001.

70
producer’s risk (ά ) The risk that the sampling plan will fail
to verify an acceptable lot’s quality and, thus, reject it (a
type I error).
 lot tolerance proportion defective (LTPD) The worst level
of quality that the consumer can tolerate.
 consumer’s risk ( β) The probability of accepting a lot
with LTPD quality (a type II error)

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 2.5 Statistical considerations

2.5.1 Sampling techniques

 Sampling techniques are the processes by which the

subset of the population from which you will collect data

are chosen.
 There are two general types of sampling techniques:
1. Non-Probability Sampling
2. Probability Sampling
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1. Non –probability sampling.
The process of selecting a sample from a population without
using (statistical) probability theory.

NOTE: In Non –probability sampling each element/member

of the population does not have an equal chance of being
included in the sample, and
 the researcher cannot estimate the error caused by not
collecting data from all elements/members of the population.

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 Types of Non –probability sampling

1. Judgment sampling
Is solely at the discretion of the sampler and therefore
is highly dependent on the person taking the sample

 This method is used when it is the only practical way

of obtaining the sample
It may result in a better estimate of the population than
random sampling if sampling is done by an experienced

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2. Convenience sampling
 Is performed when ease of sampling is the key factor.
The first pallet in a lot or the sample that is most
accessible is selected

This is also called “chunk sampling” or “grab

sampling”
Although this sampling requires little effort,
the sample obtained will not be
representative of the population, and therefore
is not recommended
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3. Restricted sampling
may be unavoidable when the entire population is not
accessible
This is the case if sampling from a loaded boxcar, but
the sample will not be representative of the population
4. Quota sampling
 is the division of a lot into groups representing various
categories, and samples are then taken from each group
 This sampling method is less expensive than random
sampling but also is less reliable

76
2. Probability sampling

 A probability sampling scheme is one in which every unit in

the population has a chance (greater than zero) of being
selected in the sample, and this probability can be accurately
determined.

 In probability sampling, the sampling error can be estimated

and the chance of item being selected does not depend on the
inspector but on standardized statistical procedures.

77
Types of probability sampling

1. Simple random sampling: Applicable when population is small,

homogeneous & readily available.
 All subsets of the frame are given an equal probability.

 Each element of the frame thus has an equal probability of

selection.
 It provides for greatest number of possible samples. This is done
by assigning a number to each unit in the sampling frame.
.
78
 A table of random number or lottery system is used to
determine which units are to be selected.

Disadvantages

If sampling frame large, this method impracticable. Minority

subgroups of interest in population may not be present in
sample in sufficient numbers for study

79
2. Systematic sampling
 is used when the unit list is not available and when the
unit are evenly distributed in time or space like in a
production line.

The first unit is randomly selected and then every nth is

selected.

80
Advantages:
 Sample easy to select, suitable sampling frame can
be identified easily and sample evenly spread over
entire reference population
Disadvantages:
 Sample may be biased if hidden periodicity in
population coincides with that of selection.
 Difficult to assess precision of estimate from
one survey.

81
3. Stratified sampling

involves dividing the population (size N ) into a

certain number of mutually exclusive homogeneous
subgroups (size N1, N2, N3, etc.) and
then applying random or another sampling
technique to each subgroup
is used when subpopulations of similar characteristics
can be observed within the whole population

82
Stratified sampling is used when sub-populations of
similar characteristics can be observed within the
whole population.
An example of stratified sampling would be a company
that produces tomato juice in different plants.
If we need to study the residual activity of
polygalacturonase in tomato juice we can stratify on
production plants and take samples on each plant.

83
4. Cluster sampling
 Entails dividing the population into subgroups, or
clusters, and then selecting randomly only a certain
number of clusters for analysis

In stratified sampling sample are taken from every

single subgroup, while
in cluster sampling only some randomly
selected clusters are sampled
The clusters selected for sampling may be either
totally inspected or sub sampled for analysis
84
The main difference between cluster sampling and
stratified sampling is that in the latter samples are taken
from every single subgroup, while in cluster sampling
only some randomly selected clusters are sampled.
The clusters selected for sampling may be either
totally inspected or sub sampled for analysis.
This sampling method is more efficient and less
expensive than simple random sampling, if populations
can be divided into clusters. Going back to the tomato
juice example, when using cluster sampling we would
consider all processing plants, but we would select
randomly just a few for the purpose of the study.

85
5. Composite sampling

is used to obtain samples from bagged products

such as flour, seeds, and larger items in bulk

Small aliquots are taken from different bags, or

containers, and combined in a simple sample (the
composite sample) that is used for analysis

86
 2.6 SAMPLING PROCEDURES

 The reliability of analytical data is compromised if

sampling is not done properly.

As shown in Table 2-1, the use of the data to be obtained

will determine the sampling procedure.

Details for the sampling of specific food products are

described in the Official Methods of Analysis of AOAC
International and in the Code of Federal Regulations (CFR).

87
 The AOAC Method 925.08 describes the method for sampling

flour from sacks.

The number of sacks to be sampled is determined by the
square
root of the number of sacks in the lot.
 The sacks to be sampled are chosen according to their
exposure.
 The samples that are more frequently exposed are sampled
more often than samples that are exposed less.
 Sampling is done by drawing a core from a corner at the top
of the sack diagonally to the center. 88
The sampling instrument is a cylindrical, polished trier with a
pointed end.
 It is 13 mm in diameter with a slit at least one third of the
circumference of the trier.
 A second sample is taken from the opposite corner in a similar
manner.
The cores are stored for analysis in a clean, dry, airtight
container
that has been opened near the lot to be sampled.
The container should be sealed immediately after the sample is
added.
A separate container is used for each sack.
Additional details regarding the container and the procedure
also are described below.

89
 Title 21 CFR specifies the sampling procedures required to
ensure that specific foods conform to the standard of identity.
 In the case of canned fruits, 21 CFR 145.3 defines a sample unit
as “container, a portion of the contents of the container, or a
composite mixture of product from small containers that is
sufficient for the testing of a single unit”
 Furthermore, a sampling plan is specified for containers of
specific net weights.
 The container size is determined by the size of the lot.
 A specific number of containers must be filled for sampling of
each lot size.
90
 The lotis rejected if the number of defective units exceeds the
acceptable limit.
 For example, out of a lot containing 48,001–84,000 units, each
weighing 1 kg or less, 48 samples should be selected.
 If six or more of these units fail to conform to the attribute of
interest the lot will be rejected.
 Based on statistical confidence intervals, this sampling plan will
reject 95% of the defective lots examined, that is, 5% consumer
risk
 The discussion below describes general considerations to take
into
account when obtaining a sample for analysis
91
1. Homogeneous vs. Heterogeneous Populations

Homogeneous population is an ideal and would be

uniform throughout and identical at all locations

sample can be taken from any location and the analytical

data obtained will be representative of the whole

 However, this occurs rarely, as even in an apparently

uniform product, such as sugar syrup, suspended particles
and sediments in a few places may render the population
heterogeneous
92
Heterogeneous, the location within a population
where a sample is taken will affect the subsequent data
obtained

However, sampling plans and sample preparation can

make the

sample representative of the population or

take heterogeneity into account in some other way

93
2. Manual vs. Continuous Sampling

Manual sampling
Attempt to take a “random sample” to avoid human bias
The sample must be taken from a number of locations
within the population
Shake prior to sample for liquids in small containers
 when sampling grain from a rail car, mixing is
impossible and samples are obtained by probing from
several points at random within the rail car

94
 Such manual sampling of granular or powdered material is
usually achieved with triers or probes that are inserted into the
population at several locations.
 Errors may occur in sampling as rounded particles may flow into
the sampling compartments more easily than angular ones.

 Similarly, hygroscopic materials flow more readily into the

sampling devices than do nonhygroscopic materials.

 Horizontal core samples have been found to contain a larger

proportion of small-sized particles than vertical ones
95
Continuous sampling
 is performed mechanically.
 less prone to human bias than manual sampling.

96
2.7 SAMPLE PREPARATION

The aim is to make sure that the sample represent the whole
portion.

It is necessary to determine whether the analysis is for entire

food or for the edible portion only.

Changes during preparation and contamination should be

minimized.

Chances of contamination should minimized by using clean

and sanitized utensils and minimizing manual handling.

97
 Chapter Two
Principle and application of analytical techniques

3.1 Introduction

 Instrumental methods of chemical analysis have become the principal means

of obtaining information in diverse areas of science and technology.

The speed, high sensitivity, low limits of detection, simultaneous detection

capabilities, and automated operation of modern instruments, when compared
to classical methods of analysis, have created this predominance.

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  An instrument for chemical analysis converts information about the physical
or chemical characteristics of the analyte to information that can be
manipulated and interpreted by human.

FIGURE Block diagram showing the overall process of an instrumental measurement.

Example:
Spectrophotometry
• Instrument: spectrophotometer Transducer: photocell
• Stimulus: monochromatic light energy Data: electrical current
• Analytical response: light absorption Data processor: current meter

In spectrophotometry, the analytical instrument is spectrophotometer:

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 Mono- Light Photocell Electrical Meter
chromatic absorption current scale
light Current
energy meter

Transducer (specific): device that converts non-electrical to electrical data

Detector (general): device that indicates change in environment

Instrumental Methods for the Analysis of Environmental Samples

Spectroscopic Methods
Utilize electromagnetic radiation (light) to extract chemical information from
the analyte
Atomic Absorption (AA) Spectroscopy
Molecular UV/Vis Spectroscopy
Inductively Coupled Plasma (ICP) Spectroscopy
Chromatographic Methods
Separations techniques based on differential migration of solutes or analytes
in mobile and stationary phase .
Gas Chromatography (GC)
6/11/2020
High Performance Liquid Chromatography (HPLC) 100
How do we know which of these instrumental methods to use?
It depends on a lot of factors, including the nature of the analyte, the type of
information (qualitative or quantitative) desired, the amount of sample available, etc.
Use the following as general guidelines when selecting an instrumental method (It
does not apply to every type of analysis)

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 3.1.1 Atomic Spectroscopy and Molecular Spectroscopy
Spectrometric methods are a large group of analytical methods that are based on
atomic and molecular spectroscopy.
Spectroscopy deals with the interactions of various types of radiation (mainly
electromagnetic radiation) with matter.
Spectrometry and spectrometric methods refer to the measurement of the
intensity of radiation with a photoelectric transducer or other types of electronic
device.
Electromagnetic Radiation: Kind of energy with wave character that can be
characterized by using wavelength (), frequency (), velocity and amplitude.
Electromagnetic radiation is a stream of discrete particles or wave packets of
energy called photons.

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 The Electromagnetic Spectrum

6/11/2020 103
Spectroscopic methods are classified according to the wavelengths or frequencies
that are important for analytical purpose.

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 3.1.2 Interactions of Electromagnetic Radiation
When radiation passes through a sample certain frequency may be selectively
removed by absorption, a process in which electromagnetic energy
is transferred to the atoms, ions or molecules.
Absorption promotes these particles from ground state to one or more higher
energy states.
The lowest energy state of an atom or molecule is called ground state. Higher
energy states are termed excited states.
Generally at room temperature, chemical species are in their ground state.
Absorption of Radiation: Absorption occurs only if :-
a. There is an interaction between the electromagnetic radiation and the
material.
b. The energy of the electromagnetic radiation must exactly corresponds to
the energy of transition in the molecule.

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 Atomic Absorption: The passage of polychromatic ultraviolet or visible radiation
through a medium that consists of mono atomic particles results the absorption of
a few well-defined frequency. Such spectra is very simple due to the small number
of possible energy states for the absorbing particles.

Monochromatic light: radiation of one discrete wavelength

Polychromatic light: radiation of wide distribution of wavelength

 Absorbance (A) is the relative amount of light absorbed by the

sample and is related to transmittance (T).

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Emission of Radiation: Electromagnetic radiation is produced when excited particle
(atoms, ions, or molecules) relax to lower energy levels by giving up their
excess energy as photons. Radiation from an excited source is characterized by
means of an emission spectrum.
Generally There are Three major types of atomic spectroscopy :

Absorption – light of a wavelength characteristic of the element of interest

radiates through the atom vapor. The atoms absorb some of the light. The
amount absorbed is measured.
Emission – sample is heated to excitation/ionization of the sample atoms.
Excited and ionized atoms decay to a lower energy state through emission.
Intensity of the light emitted is measured.
Fluorescence – a short wavelength is absorbed by the sample atoms, a longer
wavelength (lower energy) radiation characteristic of the element is emitted and
measured

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 3.2 Atomic absorption Spectroscopy
Atomic Absorption Spectroscopy (AAS)
– commonly used for elemental analysis
– expose sample to flame or high-temperature
– it is not molecular spectroscopy techniques
It can be sub divided into AAS and AES

Atomic Absorption (AA) is based on the principle that a ground state atom is
capable of absorbing light of the same characteristic wavelength as it would
emit if excited to a higher energy level.
Compound Heat Atoms
Spectra of atoms consist of sharp lines.
Each element has a characteristic spectrum.
The amount of radiation being absorbed is measured and is directly related to the
amount of analyte present by Beer- Lambert Law A= εcl
6/11/2020 110
Measure absorbance or emission of the atomic vapor.
Atomic spectroscopy deals with atoms.
Fe2+ and Fe3+ will not be distinguished.

Comparison of the atomic

emission spectrum of silicon
compared with the molecular
absorption spectrum of ethanal.
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 INTRODUCTION
3.2 Atomic absorption spectrometry (AAS) is an
analytical technique that measures the
concentrations of elements.
• Atomic absorption spectroscopy is a quantitative
method of analysis that is applicable to many metals
and a few nonmetals.
 Minerals are inorganic elements that originated in the
earth and cannot be made in the body.

 They play important roles in variously bodily function

and are necessary to sustain life and maintain optimal
health and thus are essential nutrients
2020-06-11 Atomic Absorption Spectrometry (AAS) 112
 Most of the mineral in the human diet come directly from
plant and water or indirectly from animal foods.
 However the mineral content of water and plant foods varies
geographically because of variation in the mineral content
of soil from region to region.
 They are micronutrients (needed by the body in small
quantities)
In food, minerals are classified as:
1. Major minerals
2. Minor (trace) minerals
3. Toxic (heavy) minerals
 1. Major minerals: are minerals present and needed by
the body relatively in large amounts. Or Major
minerals are those that are required in the amounts
> 100 mg /per day
• Include calcium, chloride, magnesium, phosphorus,
potassium, sodium, and sulfur
2. Minor /Traceminerals: are present and needed by
the body relatively in small amounts. Or Trace
minerals are those that are required in the amounts
< 100 mg /per day

• Include iron, zinc, iodine, selenium, copper, manganese,

fluoride, chromium, molybdenum, arsenic, nickel, silicon,
boron and cobalt

2020-06-11 Atomic Absorption Spectrometry (AAS) 114

 Trace elements :
Minerals that are nutritionally essential for a healthy life.
Examples are ( Iron, Manganese, Copper and Zinc).

2020-06-11 Atomic Absorption Spectrometry (AAS) 115

 3.Heavy Metals (toxic metals)
•The term heavy metal refers to any metallic chemical element
that has a relatively high density and is toxic or poisonous
at low concentrations.
•They have a specific gravity that is at least 5 times the specific
gravity of water.
• Example: Arsenic 5.7,Cadmium 8.65 , Iron 7.9, Lead 11.34
Mercury 13.546

2020-06-11 Atomic Absorption Spectrometry (AAS) 116

Commonly encountered toxic heavy
metals

 Arsenic  Cadmium

 Iron
 Lead

 Mercury  Aluminum

2020-06-11 Atomic Absorption Spectrometry (AAS) 117

 Elements that are detectable by AAS

2020-06-11 Atomic Absorption Spectrometry (AAS) 118

 INTRODUCTION cont’d

• Atomic-absorption spectroscopy (AAS) uses the

absorption of light to measure the concentration of
gas- phase atoms.

2020-06-11 Atomic Absorption Spectrometry (AAS) 119

 INTRODUCTION cont’d

• Flame AAS is predominantly a single-element

technique that uses a flame to generate ground-state
atoms.
• When light is passed through this atom cloud,
specific wavelengths of light are absorbed that are
characteristic of the presence of the metal of
interest.

2020-06-11 Atomic Absorption Spectrometry (AAS) 120

 Introduction
• Atomic absorption spectrophotometer is an
instrument used to determine the
concentration of element in electrolyte

• The technique was introduced in 1953 by

Walsh in Australia

• Atomic absorption spectroscopy quantifies the

absorption of ground state atoms in the
gaseous state
 Contd.
• The atoms absorb ultraviolet or visible light and make
transitions to higher electronic energy level

• The analyte concentration is determined from the amount of

absorption

WHY AAS?
the technique is element selective and.

it provides analytical sensitivities to a very small concentration

level i.e. at the parts-per-million (ppm) level
Basic components of AAS
Atomic absorption spectrometers have 4 principal
components,

A light source ( usually a hollow cathode

lamp )

An atom cell ( atomizer )

A monochromator

A detector , and read out device .

 Schematic Diagram of an Atomic
Absorption Spectrometer

Light source Detector and

atomizer
(hollow cathode monochromator readout device
Lamp )
 Contd.
Schematic Diagram of an Atomic
Absorption Spectrometer
Atomic Absorption Spectrophotometer
How can we obtain the data?
The intensity of the light
coming through the cathode
lamp is measured
The higher the concentration of
the metal that is being observe
in the sample the greater the
absorbance.
The intensity of the light is then
again measured and compared
to the first result. = absorbance
The light can then be absorbed by
the atoms from the sample that
has been vaporized in the flame.
This wavelength can then promote
the electrons to a higher energy
level = excited state
 Contd.
1. Light Source
• commonly there are two types of light sources used in atomic absorption
1. hollow cathode lamp and

2. the electrodeless discharge lamp.

Hollow cathode lamp

 it contain the tungsten anode, and cylindrical hollow cathode Lamp of

made of the element to be determined

 These are sealed in a glass tube filled with an inert gas (neon or argon)

 Each element has its own unique lamp which must be used for that analysis
Hollow cathode Lamp

e
 Contd.
• 1st ionization of some gas atoms take place by
applying a potential difference between the anode
and the cathode

• 2nd The gaseous ions bombard the cathode and

eject metal atoms from the cathode in a process
called ‘sputtering’

• finally Sputtered metal atoms are then excited to

an emission state ,emit radiation, through a kinetic
energy transfer by impact with fill gas ions
 Contd.

2. Atomizer
• Elements to be analyzed needs to be in atomic
sate

• Atomization- is the separation of particles into

individual molecules, and breaking molecules
into atoms
• This is done by exposing the analyte to high
temperatures in a flame or graphite furnace
•
 Contd.
Flam Atomizer
• Converts analyte into free atoms in the form of
vapor phase free atoms.

• flam atomizer contain burner and a nebulizer in the

system

• nebulizer – disperse and convert the liquid sample

into tiny droplets, which can be readily broken down
in the flame
 Burner/Nebulizer

A - Burner
B - Nebulizer
C - Rotational Adjust Knob, D – Horizontal
Adjust Knob, E – Vertical Adjus Knob
 Contd.
3- Monochromators
• It is used to separate out all of the thousands of
lines

• select the specific wavelength of light which is

absorbed by the sample, and to exclude other

• The selection of the specific light allows the

determination of the selected element in the
presence of others
 Contd.
4. Detector and Read out Device

• photomultiplier tube of the detector convert

the light signal into an electrical signal
proportional to the light intensity

• Then the electric signal could be displayed for

readout , or further fed into a data station.
 Working principle of AAS
The sample is vaporized by aspiration of solution into a flame
or evaporation from electrically heated surface
(temperature range: 1800 – 3100 0K)

At this condition where the individual atoms co-exist, a beam of

light is passed through them
 The atoms will absorb in the visible and ultraviolet region
resulting in changes in electronic structure (excited state)
 Working principle of AAS
 So, the resultant light beam coming out of the sample will be

missing the light in the corresponding wave length,

which is a measure of the characteristics of the sample

Based on absorbance quantified concentration will be

computed using standard curve
 Calibration curve
A calibration curve is used to determine the unknown
concentration of an element in a solution

The instrument is calibrated using several solutions of

known concentrations

The absorbance of each known solution is measured and then

a calibration curve of concentration vs. absorbance is plotted

The sample solution is fed into the instrument, and the absorbance
of the element in this solution is measured
The unknown concentration of the element is then calculated
 How do we analyze the data?
• By comparing the light intensity that has passed through the
sample (refer to previous diagram) with that of the same light
after it has passed through a blank, the absorbance is measured.

• The absorbance of different standard solutions of a compound of

the element are also measured and a calibration curve is
constructed.

• Absorbance is plotted against concentration. We then use the

calibration curve to determine the unknown concentration.
 Example of a Calibration curve
• An AAS was used to determine the concentration of lead ions (in ppm)
in fish. The AAS was set up with a lamp that emitted light with a
wavelength that is absorbed by lead atoms. The AAS was calibrated
using different solutions containing known concentrations of lead ions.
The graph on the next slide shows the variation of absorbance with the
concentration of lead.

A 2.0g sample of the fish was ground up and heated on a hot plate with
10 ml of nitric acid. This mixture was filtered and then sprayed into the
flame of the AAS. The absorbance reading was 6.0. Determine the
concentration of lead ions in the fish.
Graph:
 Application of AAS

• For analysis of food for nutritional content,

contamination , toxicity , and for other
quality assurance

• Clinical analysis- Analyzing metals in

biological fluids such as blood

• Environmental analysis : Monitoring our

environment
 Types of samples for analysis by AAS

Food
 Pharmaceutical
supplements
product

 Standards Entire plant

or part of it

 Cosmetics  Mixture of
known & unknown
herb
2020-06-11 Atomic Absorption Spectrometry (AAS) 144
Limitation of the instrument

• It requires skilled personnel

• The method is a time-consuming, involving

overnight dry ashing

• The AAS instrument has high capital and

maintenance costs
Figure Calibration curve of zinc
T has a range of 0 to 1, %T has a range of 0 to 100%

6/11/2020 147
6/11/2020 148
 For purpose of chemical analysis

Absorbance is directly proportional to:

1. concentration, c, of absorbing species in the sample (A c)
2. path length of light, b, through the sample (A b)
Beer’s law
A = bc
The previous equation is the heart of spectrophotometry as applied to analytical
chemistry, it is called Beer-Lambert law or simply Beer’s law

 Concentration of the analyte is given in unit mol/L (M)

 The path length, b, in cm
 , is called the molar absorptivity or molar absorption coefficient
“Absorbance of 1 M solution measured in a cell of 1 cm path length”
A 1
   L mol 1cm 1  M 1cm 1
bc mol
cm
L
 , is characteristic for each substance at a particular wavelength, . 149
 1
A=bc
certain 

Transmittance, T
constant b
Absorbance, A

One analyte

Slope 0.5

0
Concentration Concentration

Beer’s law is a relation between absorbance and concentration which is a straight

line passes by origin at constant path length, b, and at certain wavelength, .

6/11/2020 150
 ADVANTAGES & DISADVANTAGES:
• Precise and very sensitive
• accurate results can be obtained.
• Moderately expensive
• Can only process one element at a time.
• Slower than ICP-AES
• Can only identify limited types of elements
 Limitation

• It requires skilled personnel

• The method is a time-consuming, involving
overnight dry ashing
• The AAS instrument has high capital and
maintenance costs
 A life-saving technique
• Canada: AAS was used to determine unsafe levels of lead
in children who was lives nearby a lead smelter.
• Japan: From 1932 to 1968, AAS was used to identify the
reason why over 3,000 residents who lives near the
Minimata Bay started showing neurogical problems and
pregnant women starts giving birth to impaired children.
Scientist starts taking samples and performing AAS
process; AAS results shows a very high concentration of
mercury in their blood. This result on stopping the
company, Chisso corporation who dumped
approximately 27 tones of mercury in the bay.
 Safety Precautions
 Exhaust System: AAS flames produce large amounts of heat & the
resultant fumes & vapours may be toxic.
 Gas Cylinders: should be located outside of the laboratory in a
cool well-ventilated area.
 Flammable Solvents: The combination of flame & solvent is a
hazardous situation. Always use a solvent with the highest
flashpoint consistent with the analysis being conducted. Use
covered containers & the smallest practical volume.
 Burners: Keep burners clear & do not allow them to block.
 UV Radiation: Hazardous UV radiation is emitted by flames, hollow
cathode lamps, analytical furnaces. Never look directly at any of
these. Operate the AAS with the door or flame shield closed and
wear appropriate safety glasses.
Gas chromatography
 Introduction
• Chromatography is the term used to describe a separation technique
in which a mobile phase carrying a mixture is caused to move in
contact with a selectively absorbent stationary phase

• The two main chromatographic are GC and HPLC

• In GC we have both mobile and stationary phase

• The comparison of retention times gives GC its analytical usefulness.

 Gas Chromatography Basics

• Gas Liquid Chromatography (GLC)

• Mobile phase does not interact with analyte

• Separation occurs by interaction of analyte differentially

with stationary phase and temperature

• GC preferred method, only applicable to volatile

substances
 Basic Components of Gas
Chromatography

• Carrier Gas
• Sample Inlet
• Column
– Ovens
– Detectors
– Data acquisition system
Basic Components
 Gas carrier
• Carrier gas is the means to move constituents of a sample through the
column and yet the choice of possible gases is restricted

• The choice of a practical carrier gas is simple: nitrogen or helium.

• The gases nitrogen or helium come from cylinder or bottled gas

supply

• the carrier gas has to be cleaned over molecular sieve beds (to reduce
moisture).
 Sample Inlet

• Gas samples can be injected into the column using gas-tight syringes
or using rotary gas switching valves that offer enormous flexibility for
GC instruments

• Sample is loaded into a loop and then, with a change in the valve
position, is swept into the column under flow of the gas source
Injection
 Column
• General types of column in GC
• Packed column
• Capillary column

• Capillary columns are much more effective that

packed columns
 oven

• Most gas chromatographs are equipped with ovens to keep the

column at temperatures from 40 to 350 oC

• Resistive wire coil that radiates into the inner volume of the oven
 Detector
• Detectors that are used in GC are
• FID, ECD, TCD, PID

• The FID relies upon the formation of gaseous ions from organic
molecules combusted in a hydrogen–air flame

• ECD is especially sensitive to and selective for compounds containing

Cl2, O2, and other electronegative element
Flame Ionization Detector
Electron Capture Detector (ECD)
Working Principles of Gas Chromatography
• Once GC has separated a mixture, the components can be
identified using known retention times
• Retention time variation depending on:

- The nature of and the interactions between the

solute/analyte and the stationary and mobile phases.
- The flow rate of the carrier gas
- The temperature of the column
- The length and diameter of the column
 Interpretation of data

• Generally chromatographic data is presented as a graph of detector

response (y-axis) against retention time (x-axis), which is called a
chromatogram.

• The area under a peak is proportional to the amount of analyte

present in the chromatogram.
• By calculating the area of the peak using the mathematical function of
integration, the concentration of an analyte in the original sample can
be determined
 Application of Gas Chromatograph

• Gas chromatography is widely used in food analysis,

especially for

• Trace analysis
• Determination of volatiles (e.g. fatty acids)
• Identification of sources of adulteration
 Limitation of Gas Chromatography

• Limited to volatile samples.

• Not suitable for samples that degrade at
elevated temperatures (thermally labile).

• Requires MS detector for analyte structural

elucidation (characterization).

• Most Non-MS Detectors are destructive.

 Conclusion and Recommendation
• GC is an affordable technique which provides
great versatility and really high resolution for
the analysis of adulterants in food.
• Complex mixtures of volatile compounds can be
directly analyzed by GC, but there is also a
number of interesting derivatives for the study
of non-volatile polar molecules.
• When coupled to spectroscopic techniques,
especially to mass spectrometry, it affords
additional structural information for
compound identification.