Applications of food analysis techniques

1) Quality control
Particularly in food and beverage industries. Analysis techniques helps check out on the raw
materials, quality during processing and quality of products.
2) Official laboratories
Public health labs, KEBS, government chemist. They are also used in this area to give evidence in
court on food quality in case of disputes.
3) Research institution
E.g. food research, agriculture research institutions. They analyze e.g. quality of new hybrid variety,
effect on quality on consumer health due to pesticide and fungicide.
Food Analysis - Need and importance
All food products require analysis at various stages right from reception of raw materials through
production and sometimes even after the product reaches the market. Food is analyzed by government
agencies, food industries and researchers for various needs and reasons.
1.2.1 Food Safety
The first and foremost reason for food analysis is to ensure its safety. A customer first expects his food to
be safe. Food safety testing has become very essential as more and more contamination scares arise.
Melamine in milk products, carbendazim in orange juice, fish tainted with PCBs (polychlorinated
biphenyl), mercury-tainted milk powder and food supplements containing unauthorized food ingredients
are a few of the recent contamination alerts across the world.
A food may be unsafe due to the presence of three different types of hazards namely
i. Biological hazard - harmful microbes (e.g., Listeria, Salmonella)
ii. Chemical hazard - toxic chemicals (e.g., pesticides, herbicides)
iii. Physical hazard - extraneous matter (e.g., glass, wood, metal, insect matter).
1.2.2 Ensuring quality
The food industry sector has become highly competitive as consumers and buyers are becoming more
aware of the importance of safe, high quality products. It has led to companies emphasizing the quality of
their products. In order to do this, a food industry needs to apply analytical methods across the entire food
supply chain, from the raw ingredients to the final product. For example, when a consumer lodges
complaint about a specific characteristic of a product, it is the responsibility of the manufacturer to
perform analysis on the complaint sample and identify the problem pertaining to it. At the same time, it
also helps the manufacturer to rectify the same in future.
1.2.3 Pricing

Page 1 of 25
In many cases, the price of the product is directly linked with the quality. For e.g., a milk processor pays
the producer based on the fat content of the raw milk. The percentage of sugar in the cane determines its
price in sugar industries. There is difference in price of a pack of local coffee beans and that from a
specific part of the world. Also there exists a difference in the price of sugar free sweets from that of the
normal ones. Only analysis helps in confirming that the manufacturer is authentic.
1.2.4 Government Regulations and Recommendations
Food industries are subject to particularly strict regulations on the quality and safety of their products
which is not without reason. If contaminated food reaches the retail market, the consequences can be
serious. With the growing concerns about the food & health safety, the food regulatory authorities have
imposed stringent mandatory norms for the presence of various toxicants, which if present beyond a
prescribed residual level might prove hazardous to human health.
1.2.5 Nutritional labelling
Proper analytical technique have been employed by manufacturers to give accurate information. In 1990,
the US government passed the Nutritional, Labelling and Education Act (NLEA), which revised the
regulations pertaining to the nutritional labelling of foods and made it mandatory for almost all food
products to have standardized nutritional labels. Major reason for introducing these regulations was to
enable the customers to make informed choices about their diet. Nutritional labels states the total calorific
value of the food, as well as total fat, saturated fat, cholesterol, sodium, carbohydrate, dietary fiber, sugar,
proteins, vitamins, calcium and iron. The label may also contain information about the nutrient content
claims such as low fat, low sodium, high fibers, fat free etc. the information provided on the label can be
used by consumers to plan a nutritious and balanced diet, to avoid over consumption of food components
linked with health problems and to encourage greater consumption of foods that are beneficial to health.
1.2.6 Detection of adulteration
Adulteration is defined as the addition or subtraction of any substance to or from food, so that the natural
composition and quality of food substance is affected. In India, about 25-30% of edibles sold in the
market in India are intentionally adulterated. In 2012, a study in India conducted by the Food Safety
Standards Authority of India (FSSAI) across 33 states found that milk in India is adulterated with
detergent, fat and even urea, as well diluted with water. This indicates the role of food analysis in
detecting food adulteration.
1.2.7 Characterization of raw materials
Companies increasingly rely on select suppliers to supply high-quality, safe raw ingredients and
packaging materials. These suppliers perform analytical tests to ensure compliance with
specifications for raw materials. Results of these analytical tests related to the predetermined
specifications are delivered as a Certificate of Analysis (COA) with the raw material. Companies must

Page 2 of 25
have means to control and monitor these COAs. With careful control over the quality of raw materials,
less testing is required during processing and on the final product.
Some companies perform their own tests on the raw materials received. If the lot does not meet their
standards then it is rejected. Variations in the properties of raw materials might lead to changes in the
properties of the final product. By analyzing the raw materials it is often possible to predict their
subsequent behavior during processing so that the processing conditions can be altered to produce a final
product with the desired properties. For example, the colour of potato chips depends on the concentration
of reducing sugars in the potatoes used for production. Higher the concentration of reducing sugars,
browner the potato chip. Thus by analytically measuring the concentration of reducing sugars in the
potatoes the frying conditions can be altered to produce the optimum colored potato chip.
1.2.8 Monitoring during processing
Monitoring the food product at certain stages during processing is advantageous as it can help eliminate
any deviations from the actual product quality. If such testing is done during processing, corrective
measures can be taken at the right time which prevents the rejection of the entire lot of products. For e.g.
In the case of biscuits with peanut, it is required to know that the biscuits have correct shape and also if
there is a peanut of required size in the Centre of the biscuit. Biscuits without peanuts and those with
incorrectly located peanuts have to be rejected. Also the specific line which misplaces peanuts has to be
corrected to adjust the peanut’s position. This can be done by image processing and analysis during
processing.
Fig showing the actual picture of the biscuits on the left. The one on the right shows red and green colour
to indicate the rejected and acceptable biscuits.

Also, the taste and crispiness of the biscuits can be found out by sensory and texture analysis. All these
analyses can help monitoring the quality of biscuits during processing.
1.2.9 Final product characterization
Testing the final product will ensure that the product meets the necessary quality standards, labelling
requirements and government regulations.
1.2.10 Research and Development
Various scientists and researchers throughout the world are continuously analyzing food materials for
better understanding of their properties so that improvement can be done during their processing. Their

Page 3 of 25
research is mainly directed towards investigating the structure and interaction of food ingredients, and
how they are affected by changes in temperature, pressure and mechanical agitation. Scientists working
for food companies are usually involved in product development. These scientists help improve existing
products or develop new products.

SAMPLING
 Sampling is the process of selecting a group of individuals from a population to study
them and characterize the population as a whole.
 The population includes all members from a specified group and all possible outcomes
or measurements that are of interest. The exact population will depend on the scope of the
study.
 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.
 The sampling frame is the information that locates and defines the dimensions of the
universe.
 Laboratory Sample. The sample may be too large to analyze using a laboratory
procedure conveniently, so only a fraction is used in the final laboratory analysis. This
fraction is usually referred to as the laboratory sample
A GOOD SAMPLE SHOULD SATISFY THE BELOW CONDITIONS-
1. Representativeness: The sample should be the best representative of the population
under study.
2. Accuracy: Accuracy is defined as the degree to which bias is absent from the sample. An
accurate (unbiased) sample is one that exactly represents the population.
3. Size: A good sample must be adequate in size and reliability.
SAMPLING PLAN
 sampling plan should be a 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 before
analysis.

Page 4 of 25
  It should also stipulate the required documentation of procedures carried out during the
sampling process.
 The choice of a particular sampling plan depends on;
1. the purpose of the analysis
2. the property to be measured
3. the nature of the total population and the individual samples
4. The type of analytical technique used to characterize the samples.
 For certain products and types of populations, sampling plans have already been
developed and documented by various organizations that authorize official
methods, e.g., the Association of Official Analytical Chemists (AOAC).
TYPES OF SAMPLING TECHNIQUES
There are several different sampling techniques available, and they can be subdivided into two
groups-
1. Probability sampling involves random selection, allowing you to make statistical inferences
about the whole group.
There are four types of probability sampling techniques
 Simple random sampling
 Cluster sampling
 Systematic sampling
 Stratified random sampling
2. Non-probability sampling involves non-random selection based on convenience or other
criteria, allowing you to easily collect initial data. There are four types of Non-probability
sampling techniques.
 Convenience sampling
 Judgmental or purposive sampling
 Snowball sampling
 Quota sampling
Choosing Between Probability and Non-Probability Samples
The choice between using a probability or a non-probability approach to sampling depends on a
variety of factors:
1. Objectives and scope of the study

Page 5 of 25
 2. Method of data collection
3. Precision of the results
4. Availability of a sampling frame and resources required to maintain the frame
5. Availability of extra information about the members of the population

Probability Sampling
Probability sampling is normally preferred when conducting major studies, especially when a
population frame is available, ensuring that we can select and contact each unit in the population.
Probability sampling allows us to quantify the standard error of estimates, confidence intervals to
be formed and hypotheses to be formally tested.
The main disadvantage is Bias in selecting the sample and the costs involved in the survey.
1. Simple random sampling
In Simple Random Sampling, each observation in the population is given an equal probability of
selection, and every possible sample of a given size has the same probability of being selected.
One possible method of selecting a simple random sample is to number each unit on the
sampling frame sequentially and make the selections by generating numbers from a random
number generator.
Simple random sampling can involve the units being selected either with or without replacement.
Replacement sampling allows the units to be selected multiple times whilst without replacement
only allows a unit to be selected once. Without replacement, sampling is the most commonly
used method.
Ex: If a sample of 20 needs to be collected from a population of 100. Assign unique numbers to
population members and randomly select 20 members with a random generator. Train and test
split in ML
Applications
1. Train and test split in machine learning problems
2. Lottery methods
Advantages
1. Minimum sampling bias as the samples are collected randomly
2. Selection of samples is simple as random generators are used
3. The results can be generalized due to representativeness

Page 6 of 25
Disadvantages
1. The potential availability of all respondents can be costly and time consuming
2. Larger sample sizes

2. Systematic sampling
In systematic random sampling, the researcher first randomly picks the first item from the
population. Then, the researcher will select each nth item from the list. The procedure involved
in systematic random sampling is very easy and can be done manually. The results are
representative of the population unless certain characteristics of the population are repeated for
every nth individual.
Steps in selecting a systematic random sample:
1. Calculate the sampling interval (the number of observations in the population divided by
the number of observations needed for the sample)
2. Select a random start between 1 and sampling interval
3. Repeatedly add sampling interval to select subsequent households
Ex: If a sample of 20 needs to be collected from a population of 100. Divide the population into
20 groups with a members of (100/20) = 5. Select a random number from the first group and get
every 5th member from the random number.
Applications
1. Quality Control: The systematic sampling is extensively used in manufacturing industries
for statistical quality control of their products. Here a sample is obtained by taking an
item from the current production stream at regular intervals.
2. In Auditing: In auditing the savings accounts, the most natural way to sample a list of
accounts to check compliance with accounting procedures.
Advantages
1. Cost and time efficient
2. Spreads the sample more evenly over the population
Disadvantages
1. Complete population should be known
2. Sample bias If there are periodic patterns within the dataset

Page 7 of 25
3. Stratified random sampling
In Stratified random sampling, the entire population is divided into multiple non-overlapping,
homogeneous groups (strata) and randomly chosen final members from the various strata for
research. Members in each of these groups should be distinct so that every member of all groups
get equal opportunity to be selected using simple probability.
4. Cluster sampling
Cluster sampling divides the population into multiple clusters for research. Researchers then
select random groups with a simple random or systematic random sampling technique for data
collection and data analysis.
Steps involved in cluster sampling:
1. Create the clusters from the population data
2. Select each cluster as a sampling frame
3. Number each cluster
4. Select the random clusters
After selecting the clusters, either complete clusters will be used for the study or apply the other
sampling methods to pick the sample elements from the clusters.
Ex: A researcher wants to conduct an academic performance of engineering students under a
particular university. He can divide the entire population into multiple engineering colleges
(Which are clusters) and randomly pick up some clusters for the study.
Advantages
1. Saves time and money
2. It is very easy to use from the practical standpoint
3. Larger sample sizes can be used
Disadvantages
1. High sampling error
2. May fail to reflect the diversity in the sampling frame
Non-probability sampling
Non-Probability samples are preferred when accuracy in the results is not important. These are
inexpensive, easy to run and no frame is required. If a non-probability sample is carried out
carefully, then the bias in the results can be reduced.

Page 8 of 25
The main disadvantage of Non-Probability sampling is “dangerous to make inferences about the
whole population.”

Convenience sampling
Convenience sampling is the easiest method of sampling and the participants are selected based
on availability and willingness to participate in the survey. The results are prone to significant
bias as the sample may not be a representative of population.
Applications
1. Surveys conducted in social networking sites and offices
Examples: The polls conducted in Facebook or Youtube. The people who are interested in taking
the survey or polls will attend the survey and the results may not be accurate as the results are
prone to significant bias.
Advantages
1. It is easy to get the sample
2. Low cost and participants are readily available
Disadvantages
1. Can’t generalize the results
2. Possibility of under or over representation of the population
3. Significant bias
Quota sampling
This method is mainly used by market researchers. The researchers divide the survey population
into mutually exclusive subgroups. These subgroups are selected with respect to certain known
features, traits, or interests. Samples from each subgroup are selected by the researcher.
Quota sampling can be divided into two groups-
1. Controlled quota sampling involves introduction of certain restrictions in order to limit
researcher’s choice of samples.
2. Uncontrolled quota sampling resembles convenience sampling method in a way that
researcher is free to choose sample group members
Steps involved in Quota Sampling
1. Divide the population into exclusive sub groups

Page 9 of 25
 2. Identify the proportion of sub groups in the population
3. Select the subjects for each subgroup
4. Ensure the sample is the representative of population
Ex: A painting company wants to do research on one of their products. So the researcher uses the
quota sampling methods to pick up painters, builders, agents and retail painting shop owners.
Advantages
1. Cost effective
2. Doesn’t depend on sampling frames
3. Allows the researchers to sample a subgroup that is of great interest to the study
Disadvantages
1. sample may be overrepresented
2. Unable to calculate the sampling error
3. Great potential for researcher bias and the quality of work may suffer due to researcher
incompetency and/or lack of experience
Judgement (or Purposive) Sampling
In Judgement (or Purposive) Sampling, a researcher relies on his or her judgment when choosing
members of the population to participate in the study. Researchers often believe that they can
obtain a representative sample by using sound judgment, which will result in saving time and
money.
As the researcher’s knowledge is instrumental in creating a sample in this sampling technique,
there are chances that the results obtained will be highly accurate with a minimum margin of
error.
Ex: A broadcasting company wants to research one of the TV shows. The researcher has an idea
of the target audience and he can choose the members of the population to participate in the
study.
Advantages
1. Cost and time effective sampling method
2. Allows researchers to approach their target market directly
3. Almost real-time results
Disadvantages
1. Vulnerability to errors in judgment by researcher

Page 10 of 25
 2. Low level of reliability and high levels of bias
3. Inability to generalize research findings
Snowball sampling
This method is commonly used in social sciences when investigating hard-to-reach groups.
Existing subjects are asked to nominate further subjects known to them, so the sample increases
in size like a rolling snowball. For example, when surveying risk behaviors amongst intravenous
drug users, participants may be asked to nominate other users to be interviewed.
This sampling method involves primary data sources nominating other potential primary data
sources to be used in the research. So the snowball sampling method is based on referrals from
initial subjects to generate additional subjects. Therefore, when applying this sampling method
members of the sample group are recruited via chain referral.
Ex: Individuals with rare diseases. If a drug company is interested in doing research on the
individuals with rare diseases, it may be difficult to find these individuals. So the drug company
can find few individuals to participate in the study and request them to refer the individuals from
their contacts.
Advantages
1. Researchers can reach rare subjects in a particular population
2. Low-cost and easy to implement
3. It doesn’t require a recruitment team to recruit the additional subjects
Disadvantages
1. The sample may not be a representative
2. Sampling bias may occur
3. Because the sample is likely to be biased, it can be hard to draw conclusions about the
larger population with any confidence
Finally,
1. Reducing sampling error is the major goal of any selection technique.
2. A sample should be big enough to answer the research question, but not so big that the
process of sampling becomes uneconomical.
3. In general, the larger the sample, the smaller the sampling error, and the better job you
can do.
4. Decide the appropriate sampling method based on the study or use case.

Page 11 of 25
Sample Identification
Laboratory samples should always be labeled 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,
e) Method used to select the sample.
The analyst should always keep a detailed notebook 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.
PREPARATION OF LABORATORY SAMPLES
Once the selected a sample that represents the properties of the whole population, we must
prepare it for analysis in the laboratory. Preparing a sample for analysis must be done very
carefully to make accurate and precise measurements.

1. 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) 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 the 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. Some 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

Page 12 of 25
(e.g., proteases, cellulases, lipases) or chemical methods (e.g., strong acids, strong bases,
detergents).
2. 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.
Sample preservation
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.
1. 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.
2. 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.
3. 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.

Page 13 of 25
 4. 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.
Physicochemical Properties
 The physiochemical properties of foods (rheological, optical, stability, flavor) ultimately
determine their perceived quality, sensory attributes and behavior during production,
storage and consumption.
 The optical properties of foods are determined by the way that they interact with
electromagnetic radiation in the visible region of the spectrum, e.g., absorption,
scattering, transmission and reflection of light. For example, full fat milk has a whiter
appearance than skim milk because a greater fraction of the light incident upon the
surface of full fat milk is scattered due to the presence of the fat droplets.
 Rheological properties of foods are determined by how the shape of the food changes, or
how the food flows, in response to some applied force. For example, margarine should
be spreadable when it comes out of a refrigerator, but it must not be so soft that it
collapses under its own weight when it is left on a table.
 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.
Chemical stability refers to the change in the type of molecules present in a food with
time due to chemical or biochemical reactions, e.g., fat rancidity or non-enzymatic
browning. Physical stability refers to the change in the spatial distribution of the
molecules present in food with time due to the movement of molecules from one
location to another, e.g., droplet creaming in milk.
Biological stability refers to the change in the number of microorganisms present in a
food with time, e.g., bacterial or fungal growth.

 Foods must therefore be carefully designed so that they have the required
physicochemical properties over the range of environmental conditions that they will
experience during processing, storage and consumption, e.g., variations in temperature or

Page 14 of 25
mechanical stress. Consequently, analytical techniques are needed to test foods to ensure
that they have the appropriate physicochemical properties.

Page 15 of 25
  The flavor of a food is determined by how certain molecules in the food interact with
receptors in the mouth (taste) and nose (smell) of human beings. The perceived flavor of
a food product depends on the type and concentration of flavor constituents within it, the
nature of the food matrix, as well as how quickly the flavor molecules can move from
the food to the sensors in the mouth and nose. Analytically, the flavor of a food is made
of resistant materials.

Sensory Evaluation

 Sensory evaluation is the evaluation of the sensory properties of food (appearance,

flavour, texture).
 It can be carried out by the following steps:
 Look at the food and describe the overall appearance.
 Smell the food and describe its aroma.
 Cut the food and feel its texture.
 Chew the food and describe the taste and mouth feel.

The sensory properties of food are:

1. Appearance: Involves the sense of sight and can be described in terms of colur, size, shape,
consistency and crumb. Example; golden brown, small, round, smooth.
2. Texture: involves the senses of sight and touch.
Mouthfeel is the sensation when we bite and chew food.
Different cooking methods produce different textures Example: soft, chewy, crispy, sticky,
grainy, coarse etc.
3. Flavor: is produced by a combination of taste and aroma.
Taste: the taste buds on tongue enables to detect the different tastes of food- examples; sweet,
spicy, brand, buttery, nutty.
Aroma; involves the sense of smell- detected by the nose. Example: fragrant, burnt, fishy, fruity.
Conducting Sensory Evaluation
A panel of judges is selected. They should be unbiased for tasting.
Physical, psychological and environmental conditions should be maintained as these affect one’s
judgement.

Page 16 of 25
The sampling has to be done homogenously.
Preference Test
Judging should be done in individual booths.
This assures independent judgment and communication between panel members should not be
allowed except for consultation with the panel leader on any point of doubt.
The best time of day for sensory testing is morning 10.00 am to 12 noon and 3 to 5 pm.
The size of the panel is usually 50 to 100 people to avoid any experimental error.
Judgement should be done quickly, but not hurriedly.
They are valuable in developing new foods and in evaluating quality.
These tests are designed to provide information on selected characteristics and to indicate
preference or acceptability of products.
Environment for conducting sensory test
 Separate sensory booths should be provided so that judges do not interact with each other
except when preparing profiles because judges work together to develop vocabulary needed to
describe food samples.
 Controlled air and lighting so that food is correctly visible and booth is free from odours
other than the sample. Temperature should be comfortable and non-smoking zone observed.
 Small sinks should be provided for spitting out samples or rinsing ones mouth.
Sample preparation and presentation
 Samples should be of identical size.
 They should be in identical shape or from identical portions e.g. edge of cake from one
sample and centre slice from another sample should not be taken.
 It should be served at the customary temperature example. e.g. Soup should be served
hot.
 Sample plates should be marked.
 Plates or containers used should be of identical size and colour.
 Necessary cutlery, glass of water should be provided at room temperature.
 Only a limited number of samples should be evaluated at a time to avoid fatigue and for
efficient judging.

Page 17 of 25
Members of the Panel
 The judge should neither be too hungry or too well fed.
 Smoking, chewing gum or nibbling snacks 20 minutes prior to the test should not be
permitted.
 The judges should be healthy and not suffering from a cold as this will affect their sense
of taste and smell.
Microbiological methods for Food Analysis
Microbiological analysis of food and food products is important as growth of microbes occur in
food contaminated with pathogenic bacteria and fungi which critically affects the quality of food.
These pathogenic microorganisms has hazardous effects on human health. Since microbiological
parameters have been established as responsible factors for food safety, hence, microbiological
evaluation for food safety are key subject to current area. Major objectives of microbiological
analysis of food are to evaluate the presence or absence of different microbes recommended for
analysis and determining their nature in terms of pathogenicity and toxicity. It’s also important to
know about different categories of levels of contamination. Ingredients and water used for food
preparation should also be evaluated for microbial testing in addition to testing of the final
product. Quality of water and ingredients determine the quality of the food product. Various
microbiological methods for: total bacterial count, total coliforms count, total yeast and mold
count, enumeration of E. coli, S. aureus, Bacillus cereus.
Explain texture evaluation in food analysis (6 mks)

Page 18 of 25
Determination of ash

Ash refers to the inorganic residue remaining after either ignition or complete oxidation of
organic matter in a foodstuff.
Two major types of ashing are used:
a. Dry ashing, primarily for proximate composition and for some types of specific
mineral analyses;
b. Wet ashing (oxidation), as a preparation for the analysis of certain minerals.
Microwave systems now are available for both dry and wet ashing, to speed up the
processes. Most dry samples (i.e., whole grain, cereals, dried vegetables) need no
preparation, while fresh vegetables need to be dried prior to ashing.
High-fat products such as meats may need to be dried and fat extracted before ashing. The
ash content of foods can be expressed on either a wet weight (as is) or on a dry weight
basis.
Definitions
Dry ashing refers to the use of a muffle furnace capable of maintaining temperatures of 500–
600◦C. Water and volatiles are vaporized, and organic substances are burned in the presence of
oxygen in air to CO2 and oxides of N2. Most minerals are converted to oxides, sulfates,
phosphates, chlorides, and silicates. Elements such as Fe, Se, Pb, and Hg may partially volatilize
with this procedure, so other methods must be used if ashing is a preliminary step for specific
elemental analysis.
Wet ashing is a procedure for oxidizing organic substances by using acids and oxidizing agents
or their combinations. Minerals are solubilized without volatilization. Wet ashing often is
preferable to dry ashing as a preparation for specific elemental analysis. Wet ashing often uses a
combination of acids and requires a special perchloric acid hood if that acid is used.
Importance of Ash in Food Analysis
a. It is a part of proximate analysis for nutritional evaluation. Ashing is the first step in preparing a food
sample for specific elemental analysis. Because certain foods are high in particular minerals, ash content
becomes important. One can usually expect a constant elemental content from the ash of animal products,
but that from plant sources is variable.
Ash Contents in Foods
The ash content of most fresh foods rarely is greater than 5%. Pure oils and fats generally contain
little or no ash; products such as cured bacon may contain 6% ash, and dried beef may be as high
as 11.6% (wet weight basis).

Page 19 of 25
Fats, oils, and shortenings vary from 0.0 to 4.1% ash, while dairy products vary from 0.5 to
5.1%.
Fruits, fruit juice, and melons contain 0.2–0.6% ash.
Dried fruits are higher (2.4–3.5%).
Flours and meals vary from 0.3 to 1.4% ash.
Pure starch contains 0.3% and wheat germ 4.3% ash.
METHODS
Sample Preparation
It cannot be overemphasized that the small sample used for ash, or other determinations, needs to
be very carefully chosen so that it represents the original materials. A 2–10-g sample generally is
used for ash determination.
For that purpose, milling, grinding, and the like probably will not alter the ash content much;
however, if this ash is a preparatory step for specific mineral analyses, contamination by
microelements is of potential concern. Remember, most grinders and mincers are of steel
construction. Repeated use of glassware can be a source of contaminants as well. The water
source used in dilutions also may contain contaminants of some microelements. Distilled-
deionized water always should be used.
Dry Ashing
Principles and Instrumentation
Dry ashing is incineration at high temperature (525℃ or higher). Incineration is accomplished with a muffle
furnace. All crucibles should be marked for identification. Marks on crucibles with a felt-tip marking pen will
disappear during ashing in a muffle furnace. Laboratory inks scribed with a steel pin are available commercially.
Crucibles also may be etched with a diamond point and marked with a 0.5M solution of FeCl3, in 20% HCl. An iron
nail dissolved in concentrated HC1 forms brown goo that is a satisfactory marker. The crucibles should be fired and
cleaned prior to use.
The advantages of conventional dry ashing are that it is a safe method, it requires no added reagents or blank
subtraction, and little attention is needed once ignition begins. Usually a large number of crucibles can be handled at
once, and the resultant ash can be used additionally in other analyses for most individual elements, acid-insoluble
ash, and water-soluble and insoluble ash.
Disadvantages
Time required (12–18 h or overnight)
Expensive equipment.
There will be a loss of the volatile elements and interactions between mineral components and crucibles.
Procedures

Page 20 of 25
AOAC International has several dry ashing procedures (e.g., AOAC Methods 900.02 A or B,
920.117, 923.03) for certain individual foodstuffs. The general procedure includes the following
steps:
1. Weigh a 5–10-g sample into a tared crucible. Predry if the sample is very moist.
2. Place crucibles in a cool muffle furnace. Use tongs, gloves, and protective eyewear if
the muffle furnace is warm.
3. Ignite 12–18 h (or overnight) at about 550◦C.
4. Turn off muffle furnace and wait to open it until the temperature has dropped to at
least 250◦C, preferably lower. Open door carefully to avoid losing ash that may be
fluffy.
5. Using safety tongs, quickly transfer crucibles to a desiccator with a porcelain plate
and desiccant. Cover crucibles, close desiccator, and allow crucibles to cool prior to
weighing.
The ash content is calculated as follows:
% ash (dry basis)
= (wt after ashing−tare wt of crucible)/(original sample wt×dry matter coefficient)
× 100
Wet Ashing
Principle, Materials, and Applications
Wet ashing is sometimes called wet oxidation or wet digestion. Its primary use is preparation
for specific mineral analysis and metallic poisons. Often, analytical testing laboratories use only
wet ashing in preparing samples for certain mineral analyses (e.g., Fe, Cu, Zn, P).
Advantages
 Minerals will usually stay in solution, and there is little or no loss from volatilization
because of the lower temperature.
 The oxidation time is short and requires a hood, hot plate, and long tongs, plus safety
equipment.
Disadvantages
 It takes virtually constant operator attention
 Corrosive reagents are necessary, and
 Only small numbers of samples can be handled at any one time.

Page 21 of 25
  If wet digestion utilizes perchloric acid, all work needs to be carried out in an expensive
special fume hood called a perchloric acid hood.
Unfortunately, a single acid used in wet ashing does not give complete and rapid oxidation of
organic material, so a mixture of acids often is used. Combinations of the following acid
solutions are used most often: (1) nitric acid, (2) sulfuric acid-hydrogen peroxide, and (3)
perchloric acid. Different combinations are recommended for different types of samples.
The nitric–perchloric combination is generally faster than the sulfuric–nitric procedure. While
wet digestion with perchloric acid is an AOAC procedure (e.g., AOAC Method 975.03), many
analytical laboratories avoid if possible the use of perchloric acid in wet ashing and instead use a
combination of nitric acid with either sulfuric acid, hydrogen peroxide, or hydrochloric acid.
Wet oxidation with perchloric acid is extremely dangerous since the perchloric acid tends to
explode. The perchloric acid hood that must be used has wash-down capabilities and does not
contain plastic or glycerol-base caulking compounds.
Procedures
The following is a wet ash procedure using concentrated nitric and sulfuric acids
1. Accurately weigh a dried, ground 1-g sample in a 125-ml Erlenmeyer flask (previously acid
washed and dried).
2. Prepare a blank of 3 ml of H2SO4 and 5 ml of HNO3, to be treated like the samples. (Blank is
to be run with every set of samples.)
3. Add 3 ml of H2SO4 followed by 5 ml of HNO3 to the sample in the flask.
4. Heat the sample on a hot plate at ca. 200◦C (boiling). Brown-yellow fumes will be observed.
5. Once the brown-yellow fumes cease and white fumes from decomposing H 2SO4 are observed,
the sample will become darker. Remove the flask from the hot plate. Do not allow the flask to
cool to room temperature.
6. Slowly add 3–5 ml of HNO3.
7. Put the flask back on the hot plate and allow the HNO3 to boil off. Proceed to the next step
when all the HNO3 is removed and the color is clear to straw yellow. If the solution is still dark
in color, add another 3–5 ml of HNO3 and boil. Repeat the process until the solution is clear to
straw yellow.

Page 22 of 25
8. While on the hot plate, reduce the volume appropriately to allow for ease of final transfer.
Allow the sample to cool to room temperature, then quantitatively transfer the sample to an
appropriately sized volumetric flask.
9. Dilute the sample to volume with ultrapure water, and mix well. Dilute further, as appropriate,
for the specific type of mineral being analyzed.
Fat analysis
Definitions
Lipids, proteins, and carbohydrates constitute the principal structural components of foods. Lipids are a group of
substances that, in general, are soluble in ether, chloroform, or other organic solvents but are sparingly soluble in
water.
Fats generally refer to those lipids that are solid at room temperature and oils generally refer to those lipids that are
liquid at room temperature. While there may not be an exact scientific definition, the US Food and Drug
Administration (FDA) has established a regulatory definition for nutrition labeling purposes. The
FDA has defined total fat as the sum of fatty acids from C 4 to C24, calculated as triglycerides. This definition
provides a clear path for resolution of any nutrition labeling disputes.
General Classification
The general classification of lipids that follows is useful to differentiate lipids in foods.
Simple Lipids
Ester of fatty acids with alcohol:
• Fats: Esters of fatty acids with glycerol – triacylglycerols
• Waxes: Esters of fatty acids with long-chain alcohols other than glycerols (e.g., myricyl
palmitate, cetyl palmitate, vitamin A esters, and vitamin D esters).
Compound Lipids
Compounds containing groups in addition to an ester of a fatty acid with an alcohol:
• Phospholipids: Glycerol esters of fatty acids, phosphoric acids, and other groups containing
nitrogen (e.g., phosphatidyl choline, phosphatidyl serine, phosphatidyl ethanolamine, and
phosphatidyl inositol)
• Cerebrosides: Compounds containing fatty acids, a carbohydrate, and a nitrogen moiety (e.g.,
galactocerebroside and glucocerebroside)
• Sphingolipids: Compounds containing fatty acids, a nitrogen moiety, and phosphoryl group
(e.g., sphingomyelins)
Derived Lipids

Page 23 of 25
Derived lipids are substances derived from neutral lipids or compound lipids. They have the
general properties of lipids – examples are fatty acids, long chain alcohols, sterols, fat-soluble
vitamins, and hydrocarbons.
Importance of Analysis
1. An accurate and precise quantitative and qualitative analysis of lipids in foods is
important for accurate nutritional labeling.
2. Determination of whether the food meets the standard of identity, and
3. To ensure that the product meets manufacturing specifications.
Inaccuracies in the analysis may prove costly for manufacturers and could result in a product of
undesirable quality and functionality.
By definition, lipids are soluble in organic solvents and insoluble in water.
Therefore, water insolubility is the essential analytical property used as the
basis for the separation of lipids from proteins, water, and carbohydrates in
foods. Glycolipids are soluble in alcohols and have a low solubility in hexane.
In contrast, triacylglycerols are soluble in hexane and petroleum ether, which
are nonpolar solvents. The wide range of relative hydrophobicity of different
lipids makes the selection of a single universal solvent impossible for lipid
extraction of foods. Some lipids in foods are components of complex
lipoproteins and oligosaccharides; therefore, successful extraction requires
that bonds between lipids and proteins or carbohydrates be broken so that
the lipids can be freed and solubilized in the extracting organic solvents.
Sample Preparation
The validity of the fat analysis of a food depends on proper sampling and preservation of the
sample before the analysis. The sample preparation for lipid analysis depends on the type of food
and the type and nature of lipids in the food. The extraction method for lipids in liquid milk is
generally different from that for lipids in solid soybeans. Therefore, there is no single standard
method for the extraction of all kinds of lipids in different foods. Several preparatory steps are
common in lipid analysis. These act to aid in extraction by removal of water, reduction of
particle size, or separation of the lipid from bound proteins and/or carbohydrates.
Pre-drying Sample

Page 24 of 25
Lipids cannot be effectively extracted with ethyl ether from moist food because the solvent
cannot easily penetrate the moist food tissues due to the hydrophobicity of the solvents used or
the hydroscopic nature of the solvents. The ether, which is hygroscopic, becomes saturated with
water and inefficient for lipid extraction. Drying the sample at elevated temperatures is
undesirable because some lipids become bound to proteins and carbohydrates, and bound lipids
are not easily

Page 25 of 25