ARBAMINCH UNIVERSITY

SAWLA CAMPUS

GROUP ASSIGNMENT

COURSE NAME: FORTIFICATION FOOD PRODUCTS

GROUP 1

MEMBERS’ NAME ID

1. DAWIT TADESSE RAMIT/568/11

2. GENEMO TOLA RAMIT/862/11
3. HANA MULETA RAMIT/941/11
4. SOLOMON TEKETEL RAMIT/1691/11
5. TEMESGEN BUNARO RAMIT/1507/09
6. YESHIWORK CHEKOL RAMIT/1951/11

FEBRUARY 2023

SAWLA, ETHIOPIA

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Table of Contents
Quality Control System and Monitoring Of Fortified Food Products.............................................1

1 Introduction..............................................................................................................................1

1.1 Food Fortification.............................................................................................................1

1.1.1 Large-Scale Food Fortification..................................................................................1

1.1.2 Bio fortification.........................................................................................................2

1.1.3 Point-of-Use or Home Fortification...........................................................................2

1.2 Health Risks Associated with Consumption of Micronutrient Fortified Foods................2

1.3 Quality control system......................................................................................................3

1.3.1 Food legislation.........................................................................................................5

1.3.2 Internal monitoring....................................................................................................6

1.3.3 External and commercial monitoring........................................................................6

1.3.4 Laboratory analysis....................................................................................................7

1.3.5 Codex and food fortification......................................................................................7

1.3.6 Premix quality............................................................................................................8

1.4 Quality Assurance/Control in Food Processing................................................................9

1.5 HACCP and QACCP........................................................................................................9

1.6 Analysis of Vitamin and Minerals..................................................................................11

1.6.1 Vitamins...................................................................................................................11

1.6.2 Vitamin A................................................................................................................15

1.6.3 Minerals...................................................................................................................16

2 Reference...............................................................................................................................21

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List of figures
Figure 1Monitoring and evaluation system for food fortification Source (Allen et al 2006)..........5
Figure 2 General steps for measuring iron content in food...........................................................17
Figure 3 Comparison b/n two samples with differing iron content...............................................17

List of Tables

Table 1 Dietary Reference Intake (DRI) values of water-soluble vitamins..................................12

Table 2 Dietary Reference Intake (DRI) values of fat-soluble vitamins.......................................13

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 Quality Control System and Monitoring Of Fortified Food Products

1 Introduction
1.1 Food Fortification
Food fortification is defined as the deliberate addition of one or more micronutrients to particular
foods, with the outcome of to increase the intake of these micronutrient(s) in order to prevent a
demonstrated deficiency and provide a health benefit. The concept of adding a product to a
food/beverage to affect a biological function was first recorded in 400 B.C. by the Persian
physician, Melanpus, who suggested adding iron filings to wine to increase soldiers’ “potency.”

In the early 20th century, the industrial fortification of widely-consumed foods developed as a
strategy to prevent and reduce the prevalence of specific nutritional deficiencies of public health
importance in non-emergency as well as emergency contexts; these initiatives emerged thanks to
major discoveries in nutrition science, and advances in the synthesis and mass production of key
vitamins and minerals.

The joint WHO/FAO guidelines on food fortification distinguish three approaches to food
fortification: mass, targeted, and market-driven (Iannotti LL, et al 2006). Mass fortification refers
to the addition of micronutrients to edible products that are consumed regularly by the general
public, such as cereals, oils and vegetable fats, milk, and condiments (Iannotti LL, Bjelakovic G
et al 2012).

Targeted fortification refers to the fortification of foods designed for specific population
subgroups, such as complementary weaning foods for infants, foods for institutional programs
aimed at schoolchildren or preschoolers, and foods used under emergency situations. Market
driven fortification refers to a situation in which a food manufacturer takes the initiative to add
one or more micronutrients to processed foods in order to increase sales and profits. Universal
fortification refers to fortification of foods consumed by animals as well as humans, with
iodization of salt as the main example.

1.1.1 Large-Scale Food Fortification

Industrial or large-scale food fortification (LSFF) is the addition of micronutrients during
processing to commonly consumed foods such as salt, flours, oil, sugar and condiments. LSFF
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programs can be categorized as either mandatory—meaning they are initiated and regulated by
the government—or voluntary where food processors add nutrients to their foods on their own
volition but is still governed by regulatory limits. Mandatory fortification programs are
increasingly common, especially when it comes to fortified flour and iodized salt. Salt iodization
is perhaps the most common form and between 1990 and 2008, the number of households
globally consuming iodized salt rose from 20 percent to 70 percent. Currently, over 130
countries have mandated iodized salt.

1.1.2 Bio fortification

In contrast to LSFF where nutrients are added during post-harvest processing, bio-fortification is
the process by which food crops are grown to improve their nutritional value. Bio fortification
projects mainly concentrate on boosting iron, zinc and provitamin A carotenoid in different food
crops through plant breeding or agronomical (mineral fertilizer); some projects have also bio
fortified with amino acids and protein. Examples of bio fortification projects include iron bio
fortification of rice, beans, maize and sweet potato; zinc bio fortification of wheat, rice, beans,
sweet potato and corn; and Vitamin A bio fortification of sweet potatoes, corn and cassava.

1.1.3 Point-of-Use or Home Fortification

Point-of-use fortification is the addition of vitamins and minerals to food that has been cooked
and is ready to be eaten. Formerly known as “home fortification”, the WHO adopted the term
“point-of-use” in 2012 to reflect the many settings where this type of intervention can take place
such as in schools and refugee camps; and in 2016, recommended point-of-use fortification of
complementary foods with micronutrient powders (MNPs) as a key intervention for improving
micronutrient intake (to improve iron status and reduce anemia in particular) in children aged 6–
24 months.

1.2 Health Risks Associated with Consumption of Micronutrient Fortified Foods

The aim of food fortification is to provide a significant level of the nutrient at regular
consumption of the food vehicle. The levels also need to take into account variations in food
consumption so that the safety of those at the higher end of the scale and impact for those at the
lower end are ensured. They should also consider prorated intakes by young children to ensure
efficacious and safe dosages (WHO 2001).

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An adverse health effect has been defined as any impairment of a physiologically important
function and as any change in morphology, physiology, growth, development or life span of an
organism.

Vitamins and minerals perform specific and vital functions in a variety of body systems and are
crucial for maintaining optimal health [29] and hence, used in food fortification strategies.

However, in a long period consumption of micronutrient fortified foods may cause toxicity due
to the accumulation of these nutrients in human body.

Food fortification may lead to differential exposure to synthetic vitamins. Accumulation of

excess vitamins cause side effects ranging from mild (i.e., nausea, vomiting, and abdominal pain)
to severe toxicity resulting in increased intracranial pressure (Darnton-Hill I, Nalubola R 2002)
liver abnormalities. In addition, there is an evidence that some vitamins (beta-carotene, vitamin
A, and vitamin E) seem to increase mortality. Because Vitamin A is fat soluble, and excess of
this vitamin can be stored in the liver and may result in hypervitaminosis A. From this point of
view, it seems unnecessary to take vitamins every day, although estimated daily average
requirements and the recommended dietary allowances.

Similarly, minerals are of critical importance in the diet, even though they comprise only 4–6%
of the human body. Each element has its individual impact in the structural and functional
integrity of the living cells and organisms. Iron is a potent pro-oxidant and cannot be actively
excreted by humans

1.3 Quality control system

Quality control (QC) is defined as the techniques and assessments used to document compliance
with established technical standards through the use of objective and measurable indicators that
are applicable to products (fortified foods) (WHO/FAO 2006). It is usually conducted by
government, or at least for the government by an appropriate agency. QC focuses on quality
earlier in the process than quality assurance.

For food fortification to have a health impact, it is essential that the fortified food maintain its
expected quality from the time of leaving the factory, during storage, and up to the time of
consumption. Thus, the quality of a fortified food needs to be monitored at key points along the

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distribution system. The aim of this monitoring is to identify points at which the quality of the
fortified food is compromised, such as during production due to improper mixing of the food and
fortification premix or during distribution due to inappropriate storage.

It distinguishes two main categories of monitoring; regulatory monitoring and household and
individual monitoring. Household and individual monitoring is concerned with coverage and
impact assessments and is not discussed in this article. It is important to note that unless
regulatory monitoring is implemented to ensure that a quality product is reaching its consumers,
there is limited value in undertaking coverage or impact assessments.

Regulatory monitoring, or food control, includes all monitoring activities at the production level
(i.e. Factories and packers), as well as monitoring at customs warehouses and retail stores.
External and commercial monitoring is carried out by the relevant regulatory authorities at these
locations. Internal monitoring refers to monitoring by the producers themselves, as part of
quality control and assurance programs. The primary aim of regulatory monitoring in relation to
food fortification is to ensure that the fortified foods meet the nutrient standards set prior to
implementation.

There are three common ways in which regulatory monitoring or food control management
systems are organized:

In the multiple agency approach, responsibilities for food control are shared between various
government ministries (e.g., health, agriculture, commerce, trade, and industry) or across
government agencies at different levels (central, regional, local). Involvement of multiple
agencies in the food control system might have its challenges, as it takes time to develop and
implement regulations, time to make changes, and time to act and coordinate unless there are
urgent food safety issues.

In the single agency approach, all responsibility for protecting public health and food safety is
combined into a single food control agency with clearly defined terms of reference.

In the integrated agency approach, policy, risk assessment and management, development of
standards and regulations, and coordinating functions are merged in a food safety and control
agency at the national level, although responsibilities for food inspection and enforcement,

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education and training, etc. remain with existing agencies at the national, regional, and local
levels.

Figure 1Monitoring and evaluation system for food fortification Source (Allen et al 2006)

1.3.1 Food legislation

Regulatory monitoring systems for fortified foods are based on national legislation, regulations,
and standards, which establish requirements for foods and ideally also the expected roles and
responsibilities of various stakeholders. The objectives of any food legislation are or should be
the protection of health, protection of consumers’ rights, and facilitation of trade in safe and
healthy food. The development of food legislation has to be based on risk analysis principles.
Food standards can be based either on Codex Alimentarius standards or on alternative
independent, objective, and scientific risk assessments. Legislation has to be developed,
evaluated, and revised in a comprehensive, open, and transparent manner, involving consultation
with the public, the scientific community, and trading partners.

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A legal framework for food fortification has to be underpinned by a Food Law, which is, ideally,
brief but lays out the general principles. This legislation then has to be supported by technical
regulations and standards that are based on the latest available science and/or accepted
international norms. National governments should consider Codex Alimentarius standards when
developing their own national food fortification standards so that they will be based on the best
available science and so that trade is facilitated internationally.

1.3.2 Internal monitoring

Internal monitoring refers to quality control and quality assurance practices as conducted by
industry, i.e. Producers, importers, and packers. Quality control is product-oriented and focuses
on defect identification, whereas quality assurance is process-oriented and focuses on defect
prevention. Internal monitoring may thus be defined as a system to control all parts of the
milling/salt production process to ensure the consistent production of flour/salt that meets both
regulatory and commercial requirements. As such, internal monitoring does not involve only one
test or one check, but is made up of a variety of components and mechanisms, including premix
procurement and storage controls, feeder and dosifier installation, feeder calibration, feed rate
calculations, process controls (such as check weighing of premix addition), record keeping, and
laboratory analysis. It also includes quantitative tests for vitamins and minerals in wheat flour
and iodine in salt (WHO, FAO 2011). Internal monitoring systems tend to be better in larger
production facilities that have the capacity and resources to establish strong procedures and
qualified personnel to undertake them.

1.3.3 External and commercial monitoring

External monitoring refers to the inspection, testing, and auditing activities carried out at
production centers, factories and packers, and importation sites.

Government authorities are responsible for external monitoring, which is implemented to assure
that the production facility is able to produce a quality and safe product that meets national
regulations. Thus, external monitoring must assess whether the facility has adequate raw
materials, equipment, systems, and procedures in place to do this on a continuous basis.

Contrary to popular perception, the objective of a monitoring visit is not to see if a sample
collected on that day conforms to national regulations.

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Rather, as noted above, external monitoring would aim to verify that internal monitoring is
adequate and ongoing. Thus, a typical external monitoring system has the following components:
mill or factory inspections (including review of records), sampling policies and procedures,
laboratory analysis, and enforcement procedures.

1.3.4 Laboratory analysis

Laboratory analysis plays an important role in regulatory monitoring, both internal and external.
However, the results of laboratory analysis are subject to several sources of variation and do not
provide conclusive evidence of compliance or noncompliance. Over-reliance on laboratory
results can therefore be very misleading and counterproductive to ensuring high-quality fortified
food. Food inspectors will have to consider laboratory analysis in combination with additional
critical information, including information obtained from mill or factory inspections

1.3.5 Codex and food fortification

The following standards, guidelines, and codes of practice adopted by the Codex Alimentarius
Commission should specifically be considered in the development of national legislation
regarding food fortification [4]:

 Codex General Principles for the Addition of Essential Nutrients to Foods (GL 9-1987)
 Codex General Standard for the Labelling of Pre- packaged Foods [Codex Stan 1-1985
(Rev. 1-1991)]
 Codex General Standard for the Labelling and Claims for Pre-packaged Foods for Special
Dietary Uses (Codex Stan 146-1985)
 Codex General Guidelines on Claims (CAC/GL 1-1979 Rev. 1-1991)
 Codex Guidelines on Nutritional Labelling (CAC/ GL 2-1985)
 Recommended International Code of Practice: General Principles of Food Hygiene
[CAC/RCP 1 (Rev.2-1985)]
 Guidelines on the Application of the Hazard Analysis Critical Control Point (HACCP)
System (CAC/GL 18-1993)

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1.3.6 Premix quality
Large-scale food fortification has emerged as one of the most feasible, cost-effective, and
sustainable evidence-based interventions to address population-level vitamin and mineral
deficiencies, estimated to affect hundreds of millions of the world’s population.3 Food
fortification also improves food and nutrition security by providing greater availability of, and
access to, micronutrients necessary for health and well-being. However, in order for fortification
programs to be truly sustainable, they need to be embedded within food systems along with the
inputs and resources required to ensure the quality and safety of fortified foods. This includes
consistent access to quality-assured vitamin and mineral premix.

Manufacturing of premix requires highly sophisticated quality assurance and control measures
and expensive laboratory equipment for product validation and testing. Imported premix
therefore has a competitive advantage on quality due to several reasons:

The imported suppliers have state of the art manufacturing facilities with adequate process
controls, quality testing and stricter standards since they have to comply with several
international regulations as they supply premix globally. The storage and distribution network of
international suppliers is well established to ensure fortificants are stored appropriately with
limited exposure to the environment.

The aim of quality assurance of premix is to ensure that, the factory has enough supply of premix
and are properly labeled, premix meet the specifications established for flour fortification such as
the micronutrients levels declared on the label is the same as the one in the certificate of analysis
and lastly premix are stored under suitable conditions as described on the label by manufacturer
and used on the FIFO basis.

Vitamin and mineral premix is one of the most significant recurring input costs for large scale
food fortification (Guinot P, Jallier V et al 2012). Accessing suppliers, volatile prices for premix,
lack of quality assurance and monitoring of delivered product and lack of funds to purchase
premix are major factors which affect the procuring of adequate quality premix (Guinot P, Jallier
V et al 2012).

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1.4 Quality Assurance/Control in Food Processing
The maintenance of a well-functioning Quality Assurance (QA) program is essential if a
consistent product is to result which meets all required standards. Such a program should be
based on Hazard Analysis and Quality Analysis Critical Control Point (HACCP and QACCP)
systems. HACCP and QACCP are more proactive than traditional approaches to QA/QC
activities. The establishment of such programs is the responsibility of QA personnel but the
execution of it involves everyone in the company. To avoid ambiguity regarding responsibility
for any QA function, it is important to assign specific HACCP/QACCP accountabilities to
responsible persons and groups (Corlett, 1992).

A QA program must consider all activities impacting upon product quality, from raw materials
and ingredients used to product handling through distribution channels all the way to the final
consumer. In respect of this, Wilson (1991) has outlined the following required components of a
QA system:

i. Raw material control - standard specifications must be adopted for all ingredients which
must then be inspected to ensure conformity;
ii. Process control - all chemical, physical and microbiological hazards as well as quality
factors must be identified, critical control points (CCP) must be established, monitored
and a record made of any action taken;
iii. Finished product control - this requires that the finished product be unadulterated,
properly labelled and that the integrity of the finished be protected from the environment.

1.5 HACCP and QACCP

All food production activities must be monitored and controlled within the framework of an
effective QA program. The addition of nutrients to a food for the purpose of fortification adds to
the control points which have to be considered. Poor manufacturing control leading to
excessively high levels of nutrients in the finished product could have important health
implications for the consumer if intake of the nutrient reaches the toxic dose. Conversely, low
levels of nutrients in the finished product could render it nutritionally ineffective. This could also
have serious health implications if the target population in the fortification program was at high
nutritional risk. Poor manufacturing control could also lead to other quality defects related to
interactions of added nutrients with other components of the system.
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The following steps in the implementation of a quality assurance program in the production of a
fortified food have been outlined by Wilson (1991):

i. Product specifications - All specifications for fortificants, food vehicle and any other
ingredients must be documented as well as acceptable deviations of these. These include
specification of particle size, color, potency, and level of fortification as well as any other
requirement which might be deemed necessary.
ii. Product safety assessment - This involves an assessment of microbiological, chemical
and physical hazards for all ingredients and the finished product
iii. Product analysis - Sampling and testing procedures for all ingredients and the finished
product must be explicitly stated.
iv. Determination of critical and quality control points - Based on firsthand knowledge of the
total process (including the plant facility, equipment and environment) stages at which
inadequate control could lead to unacceptable health risk or adversely affect product
quality are identified. The system of controls and actions to be taken at each control point
are documented.
v. Recall system - A mechanism must be put in place whereby product can be recalled if
such action becomes necessary.
vi. QA audit - Periodic checks are necessary to verify that the QA system is effective and
product quality is maintained up to the ultimate consumer.
vii. Feedback mechanism - Response to consumers and other relevant groups to correct any
deficiencies discovered.
viii. Documentation of QA system - Details of the QA program used in the production of the
fortified food must be readily available to relevant individuals and organizations.

Shortcomings of many fortification programs in the past have been due to failure to establish an
adequate quality assurance program. Evaluation of the fortification of sugar with vitamin A in
Guatemala showed that only 30% of samples tested were fortified at levels within the legal limits
(Nestle, 1993). A study of iodine content in iodized salt samples obtained from several plants in
India also provided an example of the need for greater control in processing.

In the determination of critical and other control points for any process accurate flow diagrams
outlining the total process have been used. The construction of an accurate flow diagram for any

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given process requires firsthand knowledge of the processing facility and its environs so that all
factors which might be expected to impact on product safety could be identified. Annex 1
includes a list of common critical control points and examples of monitoring procedures in the
processing of selected fortified foods, which are intended to demonstrate the main points of a
quality assurance program.

I. Use of HACCP serves to improve food safety control and should be applied on that basis;
II. The elaboration of food safety policies by government and international agencies should
use risk analysis as the basis for establishing food safety priorities and for focusing
inspection resources. These policies should be implemented through national strategic
plans;
III. In the post Uruguay Round of GATT, the Codex Alimentarius Commission should
recognize the importance of its role in harmonizing and establishing food standards,
guidelines and recommendations particularly as it relates to safety of food in international
trade. Codex should develop a strategic plan which will include a strengthening of the
scientific basis for risk analysis, equivalence and the elaboration of its standards,
guidelines and other recommendations and should include specific instructions to the
Codex Committees on incorporating HACCP.

1.6 Analysis of Vitamin and Minerals

1.6.1 Vitamins
The concept of a vitamin was developed in the early part of last century. The development of
chemical and instrumental methods for the separation, identification and quantitative analysis of
vitamins has become extremely important to the food industry and to academic and
governmental institutions, to access the nutritional value of food and food products for human
health and nutritional labeling.

1.6.1.1 Chemistry of vitamins

Vitamins are a group of organic substances that are required in the diet of humans and animals
for normal growth, maintenance of life and normal reproduction. Vitamins act as catalysts; very
often either the vitamins themselves are coenzymes, or they form integral parts of coenzymes. A
substance that functions as a vitamin for one species does not necessarily function as a vitamin

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for another species. The vitamins differ in structure, and there is no chemical grouping common
to them all.

Water-soluble vitamins

Many vitamins work together to regulate several processes within the body. A lack of vitamins
or a diet that does not provide adequate amounts of certain vitamins can upset the body’s internal
balance, or block one or more metabolic reactions.

Table 1 Dietary Reference Intake (DRI) values of water-soluble vitamins

Fat-soluble vitamins

Vitamin A is a generic term for a large number of related compounds. Retinol (an alcohol) and
retinal (an aldehyde) are often referred to as preformed vitamin A. Retinal can be converted by
the body to retinoic acid, the form of vitamin A known to affect gene transcription. Retinol,
retinal, retinoic acid and related compounds are known as retinoid. Beta-carotene and other
carotenoids that can be converted by the body into retinol are referred to as provitamin a
carotenoids.

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Table 2 Dietary Reference Intake (DRI) values of fat-soluble vitamins

1.6.1.2 Extraction and purification methods

Extraction and purification of vitamins in foods, whether they are inherently present or added
during food manufacture, is a challenging task due to the sensitivity of these compounds towards
light, oxygen, heat and ph. The first step in the analysis is extraction from the food sample,
which is done with a combination of either strong acid, or alkali, or solvent, with heating. This is
necessary because the measurement techniques cannot be used for solid food substances.

The next step is enzyme treatment to release any of the water-soluble vitamins bound to proteins
and other components in the food matrix. The conventional method for the isolation of fat-
soluble vitamins from a simple food matrix includes solvent extraction, using solvents such as
hexane, ethanol, methanol, and tetrahydrofuran and petroleum ether. Complex food matrices are
usually saponified prior to extraction in order to disrupt the matrix and degrade triacylglycerol to
glycerol and produce soaps of the free fatty acids.

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1.6.1.3 High performance liquid chromatography (HPLC)
High performance liquid chromatography (HPLC) is a form of column chromatography used
frequently in biochemistry and analytical chemistry. It is sometimes referred to as high-pressure
liquid chromatography. High performance liquid chromatography combined with a UV–Vis
detector is the most common method for identification and quantification of fat- and water-
soluble vitamins in foods. Several HPLC methods have been presented for the determination of
vitamins in fortified food products and supplements.

The high performance liquid chromatography method used to determine vitamins and their
isomers has high sensitivity, good selectivity and the ability of simultaneous multicomponent
determination.

1.6.1.4 Gas chromatography

Gas chromatography (GC) procedures were developed for water- and fat-soluble vitamins before
the advent of HPLC practices. GC can be used to determine the homologs of some vitamins and
their isomers.

1.6.1.5 Capillary electrophoresis

Capillary electrophoresis (CE) is a fairly recent analytical technique that allows the rapid and
efficient separation of sample components based on differences in their electrophoretic abilities
as they migrate or move through narrow bore capillary tubes. In spite of considerable
developments, CE still suffers from problems of poor peak shape, lack of sensitivity and poor
robustness.

1.6.1.6 Microbiological methods

Microbiological assay is applicable only to the B vitamins. The rate of growth of a species of
microorganism that requires a vitamin is measured in growth media that contain various known
quantities of a foodstuff preparation containing unknown amounts of the vitamin. This method
uses test microorganisms such as bacteria, protozoa or yeast, where growth is proportional to the
presence of a specific vitamin. These microorganisms are cultivated in the presence of known
quantities of the studied vitamin and in the presence of a food extract where the vitamin needs to
be evaluated.

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1.6.2 Vitamin A
Vitamin A is the generic name applied to a group of fat soluble compounds that have the
biological activity of all-trans-retinol and includes: retinol (alcohol), retinal (aldehyde), retinoic
acid (carboxylic acid), and pro-vitamin A carotenoids such as ß-carotene. Retinol and its related
compounds consist of four isoprenoid units joined head to tail, and contain five conjugated
double bonds.

Pro-vitamin A carotenoids are found in fruits and vegetables, as well as in egg yolk, and they are
transformed in the organism to all-trans-retinol. Retinol is referred to as preformed vitamin A
and it is found in liver, milk, butter, cheese, eggs and fish liver oils, mainly esterified with fatty
acids.

Retinyl palmitate and retinyl acetate are the two main retinyl esters used in food fortification.
The vitamin A compound for flour fortification is usually embedded into a protecting matrix that
contains starches and antioxidants to provide stability and dispersibility in water.

Qualitative method for determining vitamin A in fortified wheat flour

Vitamin A (retinyl palmitate) used for fortifying flours is extracted into organic solvents after
mixing the flour with water and 2-propanol. The organic solution containing vitamin A is then
reacted with chromogenic solutions to produce a blue solution. A new proposed modification to
extend the life of the blue color involves the addition of florisil to the vitamin A extract before
addition of the chromogenic solution. This adsorbent (florisil) adsorbs the vitamin A, and the
blue complex developed by the reaction with the chromogenic reagent takes place in this solid
matrix. When the concentration of vitamin A is high enough (above 1 mg/kg), the blue color
lasts for a few minutes before changing to a light brown reddish color. Because the results are
not reproducible at low concentrations of vitamin A, the use of florisil is not recommended,
unless the concentration of vitamin A is above 1mg/kg.

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1.6.3 Minerals
Minerals are inorganic species or substances that are of neither animal nor vegetable origin (i.e.,
natural compounds generally not containing a C, H, O, or N skeleton). Inorganic elements
constitute the major part of dry ash that remains after ignition of organic matter, and
consequently dry-ashing techniques are still the primary way to determine total minerals in
foodstuffs. Minerals are nutritional elements known as the main cellular and structural building
materials taking part in osmotic and acid/base regulation [e.g., calcium (Ca), potassium (K),
sodium (Na), phosphorus (P), magnesium (Mg), and chlorine (Cl)]. Trace elements are typically
known as electrolytes, enzymes, and hormones, constituents for which the biological roles are
known for nutritive adding value and health impact e.g., iodine (I), iron (Fe), copper (Cu),
chromium (Cr), molybdenum (Mo), selenium (Se), manganese (Mn), zinc (Zn), and cobalt (Co)].

Mineral analysis in food samples generally requires a preliminary step of sample preparation,
including a dissolution or homogenization step (e.g., blending, mixing, grinding, or slurry
preparation) of the laboratory sample, then a collection of a representative test portion, and
finally decomposition through acid- or alkaline-assisted hot digestion with or without high
pressure.

1.6.3.1 Iron (Fe)

Iron is the fourth most abundant element on earth, but iron deficiency in humans is one of the
most widespread nutritional problems in the world, because the human intestine has reduced
absorption to most iron compounds. In solid form, metal exists as a malleable metal, which
readily oxidizes in moist air.

A general scheme of the steps involved in the different techniques for measuring iron in food is
presented in the figure. Sample is digested to destroy organic matter and reduce complex
molecules to their elements, using either wet or dry digestion. A solution from the digested
product is prepared with diluted acid and then the iron content is measured using Atomic
Absorption (AA), Induced-Coupled Plasma (ICP) or visible spectrophotometry. The visible
spectrophotometry technique needs reduction of ferric ion to the ferrous form in order to use
chromogenic methods that have larger analytical sensitivity than those specific for ferric iron.

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Figure 2 General steps for measuring iron content in food

Qualitative method to determine iron in wheat flour (Spot test for determining added iron)

Principle

Ferric iron, in an acidic medium, reacts with a solution of potassium thiocyanate (KSCN) to form
an insoluble red pigment. Other types of iron, such as ferrous iron and elemental iron can also
react in a similar manner once they are oxidized to the ferric form using hydrogen peroxide.

Interpretation

Unfortified samples of wheat flour might show a reddish coloration, but not well defined red
spots. Number and density of spots might be associated to the iron level in the sample. The more
red spots appear, the higher the concentration of iron in the sample.

Figure 3 Comparison b/n two samples with differing iron content

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1.6.3.2 Zinc
Zinc is an essential trace element for animals and man.' It is a cofactor for a large number of
enzymes in virtually all metabolic pathways, is a component of transcription activation factors
the zinc fingers, and is involved in neurotransmitter function, immune activity and the action of
hormones such as insulin.

Many different analytical techniques have been used to measure zinc in biological samples.
Analytical techniques need to be sensitive, reproducible and accurate, and resistant to
contaminating interference (Reimold EW, Besch DJ 1978).

Flame atomic absorption spectrometry (FAAS)

FAAS is mainly used to determine the concentration of metals in solution in parts per million
(ppm) or parts per billion (ppb) ranges. The metal ions are nebulized as a fine spray into a high
temperature flame where they are reduced to their atoms and subsequently absorb light from an
element specific hollow cathode lamp.

Plasma or serum cannot be analyzed without some initial pretreatment. The protein content is
responsible for a viscosity effect, causing a slower aspiration rate compared with aqueous
calibration solutions. The effect of this interference is removed from the sample either by
dilution with water or protein precipitation with trichloroacetic acid.

The effective working range for the determination of zinc by FAAS is around 1-10 J-l mol/L
(0'65-6'5 mg/L) (Andrew Taylor 1996).

Electro thermal atomic absorption spectrometry (ETAAS)

This technique of analysis involves introducing a small amount of and then resistively heating a
tube of graphite which becomes the atom cell. Very low concentrations of zinc can be measured
by ETAAS, consequently avoidance of contamination from, for example laboratory equipment,
is essential. Furthermore, because of the high sensitivity of ETAAS, many biological specimens
require very large dilutions in order to obtain accurate readings (Andrew Taylor 1996).

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The basic principle is; a sample is introduced into the atom cell where it is desolated and then
atomized. The analyte atoms so formed then quantitatively absorb light in a way that is
proportional to the concentration of the analyte in the cell.

Inductively coupled plasma atomic emission spectrometry (ICP-AES)

Is a type of emission spectroscopy that uses an inductively coupled plasma to produce exited
atoms and ions that emit EMR at wavelengths characteristic of a particular element. The sample
to be analyzed is introduced as a mist. The most important interferences are due to the matrix.
Viscosity of the sample influences the rate of nebulization or blocks the capillary tube; this
interference is removed by dilution or by destruction of the organic components in the sample.

The lowest concentrations of zinc which can be detected with most ICP-AES instruments are
similar, or slightly higher than those possible with FAAS. Significant dilution is required for
efficient nebulization, application of the technique to the analysis of biological fluids has been
limited. ICP-AES has the advantage of multi-element analysis, for applications where several
trace elements require to be measured in serum or urine (Andrew Taylor 1996).

1.6.3.3 Iodine
The iodine content of iodated salt samples is measured using the iodometric titration method.
The method consists of preparing the reagent solutions, which may last for variable periods of
time, and then using these reagents in the titration procedure. Usually, salt samples of 10 g each
are dissolved in a measured amount of water for the titration analysis. However, in the case of
coarse salt and in salt containing impurities, a bigger sample weight of 50 g salt will yield more
accurate results.

Description of the reaction

The reaction mechanism includes two steps:

 Liberation of free iodine from salt: The addition of H2S4O liberates free iodine from
the iodate in the salt sample. Excess KI is added to help solubilize the free iodine, which
is quite insoluble in pure water under normal conditions.
 Titration of free iodine with thiosulfate: Free iodine is consumed by sodium thiosulfate
in the titration step. The amount of thiosulfate used is proportional to the amount of free

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 iodine liberated from the salt. Starch is added as an external (indirect) indicator of this
reaction and reacts with free iodine to produce a blue color. When added towards the end
of titration (i.e. when only a trace amount of free iodine is left) the loss of blue color, or
end-point, which occurs with further titration, indicates that all remaining free iodine has
been consumed by thiosulfate

Calculation

Mg/kg (ppm) iodine = titration volume in ml x 21.15 x Normality of sodium thiosulfate x

1000 / salt sample weight in g

Titration method for determining salt iodide content

While the use of potassium iodide (KI) is not common for salt fortification in many developing
countries, the following technique is used for analyzing salt iodized with KI.

Liberation of iodine: Bromine water oxidizes iodide ions to free iodine. Sodium sulfite and
phenol are added to destroy excess bromine so that no further oxidation of I- can occur before KI
solution is added.

Titration: The titration reaction with thiosulfate is the same as that described in the iodometric
titration method for iodated salt mentioned earlier.

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 2 Reference
Allen L, de Benoist B, Dary O, Hurrell RF, eds. Guidelines on food fortification with
micronutrients. Geneva:World Health Organization, 2006.

Andrew Taylor Measurement of zinc in clinical samples 1996

Bjelakovic G, Nikolova D, Gluud LL, Simonetti RG, Gluud C (2012) Antioxidant supplements
for prevention of mortality in healthy participants and patients with various diseases.
Cochrane Database Systematic Reviews.

Darnton-Hill I, Nalubola R (2002) Fortification strategies to meet micronutrient needs: successes

and failures. Proceedings of the Nutrition Society 61(2): 231-241 Geneva.

Guinot P, Jallier V, Blasi A, Guyondet C, Van Ameringen M, (2012): An innovative approach

for improving access to quality vitamin and mineral premix in fortification initiative.

Iannotti LL, Tielsch JM, Black MM, Black RE (2006) Iron supplementation in early childhood:
health benefits and risks. The American journal of clinical nutrition 84(6): 1261-1276.

Reimold EW, Besch DJ. Detection and elimination of contaminations interfering with the
determination of zinc in plasma. Clin Chern 1978; 24: 675-80

Report of the joint WHO, UNICEF, FFI, GAIN, MI, and World Bank meeting on regulatory
monitoring of salt and flour fortification programmes in Asia. Manila, Philippines, 27–
29 September 2011 (in press).

WHO/FAO, Guidelines on Food Fortification with Micronutrients, L.H. Allen, et al., Editors.

World Health Organization (2001) Iron deficiency anemia. Assessment, prevention, and control.
A guide for program managers, p.114.

World Health Organization/Food and Agriculture Organization of the United Nations; 2006

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