HALABJA TECHNICAL COLLEGE OF APPLIED SCIENCES

Food science & quality control

High diploma study

Scientific report

Protein Analysis in Food: Assessing

Nutritional Value and Quality

Submitted by:
Pshtiwan L. N.

pshtiwan.noori.tcas@spu.edu.iq

Submitted to:
Dr. muhammed S. rasheed
muhammed.rasheed@spu.edu.iq
Content:

 Introduction
 Protein structure
 Amino Acid Profiling
 Protein Composition in Common Food Sources
 The function of dietary protein in human health
 Protein Digestibility
 Protein Extraction
 Methods of Protein Analysis
 Conclusion
 References
Introduction

Proteins are nitrogen-containing substances which can be fashioned through amino acids.
They function the primary structural issue of muscle and different tissues inside the body.
In addition, they're used to produce hormones, enzymes and hemoglobin. Proteins can also
be used as electricity; but, they're no longer the number one preference as an power
source. [15].

Throughout diverse food processing procedures, food proteins can also go through various
chemical alterations. Despite the extensive intake of processed ingredients globally, there is
nevertheless a whole lot to find out regarding the perfect nature of those modifications.
This task stems partially from the complicated person of the adjustments worried and
partially from the analytical difficulties imposed by using the difficult nature of the meals
matrix. The complexities are similarly heightened by using the shortage of chemically
primarily based analytical checks that accurately gauge amino acid availability in biologically
meaningful phrases [1].

To examine meals proteins efficiently, it's miles essential to isolate the protein in a form
devoid of materials that might disrupt the analytical take a look at. This calls for using an
extraction approach that minimizes analytical artifacts [2].

In the realm of ingredients, severa elements can impact protein nice past simply amino acid
composition and inherent digestibility. Intrinsic factors encompass the protein's supply,
whether or not the protein possesses antinutritional homes, and the processing and garage
records of the food. Additionally, there are extrinsic factors unrelated to the protein itself,
together with the health fame of the person consuming the food [3].

There is a commonplace false impression that a food product created from uncooked
ingredients that meet all dietary protein necessities will routinely continue to be a
nutritionally adequate supply until
Protein structure

The form of a protein could be very important to its feature. To apprehend how a protein
gets its final shape or conformation, we need to apprehend the four tiers of protein
structure: primary, secondary, tertiary, and quaternary [20].

1. Primary structure:
The handiest degree of protein structure, number one structure, is definitely the
collection of amino acids in a polypeptide chain.

1. Secondary structure:
The subsequent degree in protein structure, called secondary shape, pertains to
compact folded formations going on locally inside a polypeptide, arising from
interactions a few of the backbone atoms [18].
2. Tertiary stracture:

The complete 3-dimensional association of a polypeptide is termed its tertiary shape.

This structural configuration typically arises from the interactions among the R groups of
the amino acids constituting the protein [18].

3. Quaternary stracture:
Numerous proteins include a solitary polypeptide chain and showcase only the 3
aforementioned degrees of shape. Nevertheless, sure proteins are composed of a
couple of polypeptide chains, known as subunits. The amalgamation of those
subunits contributes to the formation of the protein's quaternary structure.
Amino Acid Profiling

The human body abounds with considerable molecules referred to as biomolecules. Among
these, pivotal molecules encompass proteins, carbohydrates, lipids, and nucleic acids,
which feature as biopolymers featuring a recurring unit called the monomer. "Mono"
signifies one, and "poly" signifies many, leading to the formation of a polymer while severa
monomers hyperlink collectively. Each polymer is characterised via its distinct type of
monomer. Understanding the various monomer gadgets and the mechanisms through
which they be part of together is important in comprehending the construction of those
expansive molecules. In this regard, amino acids can be considered a monomer [18].

Proteins represent highly intricate polymers composed of 21 different amino acids,

interconnected through substituted amide bonds. At the elemental level, proteins, on a
weight-to-weight basis, consist of approximately 50–55% carbon, 6–7% hydrogen, 20–23%
oxygen, 12–19% nitrogen, and 0.2–3.0% sulfur.

α-Amino acids are the basic structural units of proteins. These amino acids consist of a α-
carbon atom covalently attached to a hydrogen atom, an amino group, a carboxyl group,
and a side chain R group [9].

The characteristics of amino acids are contingent upon the precise nature of the R
organization. Amino acids exhibit covalent properties, forming water-soluble crystalline
substances with improved melting points comparable to salts. The salt-like structure of
amino acids is called zwitterions, In AA transfer of a proton from the acid group (-COOH) to
an amino group (-NH2), the resultant formed compound is called zwitterions. A zwitterion
molecule encompasses both tremendous and negative expenses, yet the general net rate
remains zero. Amino acids function fundamental building blocks for proteins, with twenty-
one acknowledged amino acids identified in living organisms. Out of twenty-one amino
acids, few are essential amino acids, while some of them are non-essential amino acids
[18].
essential, non-essential, and functional amino acids (AA) have traditionally been based on
dietary requirements for nitrogen balance or growth. Essential amino acids (EAA) are those
whose carbon skeletons cannot be synthesized or are inadequately synthesized by the body
relative to its needs, necessitating their provision from the diet to meet optimal
requirements. Conditionally essential amino acids can generally be synthesized adequately
by the organism but must be supplied from the diet under conditions where utilization
rates exceed synthesis rates to meet optimal needs. It is important to consider functional
requirements, such as those related to reproduction and disease prevention, when
classifying amino acids as essential or conditionally essential. Non-essential amino acids
(NEAA) are those that the body can synthesize de novo in sufficient amounts to meet
optimal requirements. It is crucial to note that all 20 protein amino acids and their
metabolites are necessary for normal cell physiology and function. An abnormal
metabolism of any amino acid can disrupt overall body homeostasis, hinder growth and
development, and may even lead to death [19].
Protein Composition in Common Food Sources

Prominent assets of protein encompass animals, flowers, and microorganisms. Animal

sourced protein appears to be the most prominent among the three major sources in terms
of nutritional quality.

Amino acids function the essential constructing blocks of proteins in each plants and
animals. Protein-rich meals encompass eggs, meat, dairy products, poultry, seafood, beans,
peas, processed soy merchandise, nuts, and seeds. All of these foods contain relatively high
levels of protein (e.g., > 40% on dry matter basis). In contrast, most staple foods of plant
origin (except for legumes) have protein content < 15% (dry matter basis). Minerals such as
iron and zinc, along with vitamins like B6, B12, and A, are vital nutrients for both animals
and humans and need to be included in the diet. With the exclusion of vitamin C, animal
tissues serve as reliable sources for most micronutrients. Unlike animals, plants have the
capability to synthesize all vitamins and/or vitamin precursors, including β-carotene, a
precursor of vitamin A. However, vitamin B12 is an exception, being found exclusively in
animal-source foods.

Elevated consumption of animal protein has been associated with a heightened risk of type
2 diabetes, cardiovascular disease, colorectal cancer, and premature mortality, whereas
plant proteins exhibit notable protective effects [10]. Urgent action is required to transition
towards a more plant-based diet, driven by considerations of both environmental
sustainability and public health.

It is essential to consider that the body's utilization of dietary proteins involves a series of
processes: digestion in the gastrointestinal tract, absorption of digestion products (amino
acids, dipeptides, and tripeptides) through enterocytes (absorptive epithelial cells of the
small intestine) into the portal vein, the metabolic transformation of amino acids and small
peptides in a cell- and tissue-specific manner, and the synthesis of tissue proteins from
amino acids in the blood and cells. Therefore, both plant and animal agriculture play a
crucial role in supplying dietary proteins for human consumption [12].

The assessment of a protein's effectiveness entails comparing each its pleasant and
digestibility. Quality refers to the availability of amino acids that it supplies, and digestibility
considers how the protein is best utilized [15].

Proteins from animal sources (a.r.e. eggs, milk, meat, fish, and poultry) provide the highest
quality rating of food sources [16].

When vegetable proteins are combined to make sure the inclusion of all important amino
acids, they function an exceptional protein source, frequently main to a lower inside the
intake of saturated fat and cholesterol. Common sources of those well-balanced vegetable
proteins consist of legumes, nuts, and soy [17].

The function of dietary protein in human health

Proteins exhibit diverse functionalities, including the ability to form networks, create gels,
develop films, retain water, absorb fat, foam, emulsify, and dissolve under varying pH
conditions. These functionalities serve as key indices for measuring protein performance,
and they have found applications in a range of food products such as soups, beverages,
whipped cream, extruded and baked goods.

Various factors contribute to the quality of proteins in foods, extending beyond amino acid
composition and intrinsic digestibility. Intrinsic factors include the protein source and
whether the protein possesses antinutritional properties. Additionally, the processing and
storage history of the food play a crucial role in determining protein quality. Extrinsic
factors, such as the health status of the individual consuming the food, also influence the
overall protein quality [1].

dietary protein serves as the primary source of amino acids required for protein synthesis in
humans. Proteins in the body play crucial roles in various functions, including:

1. forming cell and extracellular structures.

2. catalyzing enzyme-driven reactions.
3. participating in gene expression.
4. mediating actions of hormones.
5. facilitating muscle contraction.
6. regulating osmotic balance.
7. providing protection against oxidative stress, infections, and bleeding.
8. overseeing metabolic processes.
9. participating in the storage and transport of nutrients such as long-chain fatty acids,
iron, vitamin A, and zinc, as well as oxygen [13].

Consequently, dietary protein intake significantly influences the status of other nutrients in
the body, and ensuring adequate protein nutrition is essential for optimal human growth
and health.

In order for proteins to be utilized by the frame, they have to be metabolized into their only
form, that's amino acids. Twenty amino acids were diagnosed as crucial for human increase
and metabolism. Twelve of these amino acids (11 in kids) are categorized as nonessential,
signifying that the frame can synthesize them and that they do no longer necessarily need
to be acquired through the food regimen. The final amino acids can not be synthesized in
the frame and are classified as vital, indicating that they ought to be acquired via nutritional
intake. The absence of any of these amino acids will compromise the ability of tissue to
grow, be repaired or be maintained [15].

Protein Digestibility

To make amino acids available for cell growth, repair, or protein synthesis, proteins need to
undergo digestion. Despite the uniformity of peptide bonds linking amino acids, various
peptidases are required to break these bonds due to the distinct nature of the side-chains
adjacent to them. Unlike a single lipase or amylase that can break down fats into fatty acids
or starch into glucose, a singular type of peptidase cannot completely cleave a protein chain
into its constituent amino acids. Multiple peptidases act on the proteins in our diet, each
targeting bonds near specific side-chains on the amino acids. Additionally, other enzymes
work on bonds at the ends of the peptide chain, sequentially removing individual amino
acids.

In the stomach, protein hydrolysis occurs through the influence of the hydrochloric acid
secreted in that environment. This acid not only denatures the protein but also facilitates
the breakdown of peptides. Moreover, the acid serves to activate the enzyme pepsinogen,
transforming it into its active form known as pepsin. Pepsin is capable of targeting various
peptide bonds, enabling the fragmentation of the extended protein chain into a series of
shorter polypeptide chains [7].

While most naturally occurring proteins are inherently highly digestible, some feeds may
experience impaired digestibility due to factors such as a dense protein structure or the
presence of anti-nutritional constituents like protease inhibitors, lectins, and tannins.
Controlled heating can mitigate many of these effects; however, excessive heating or
inappropriate chemical treatment during processing can reduce digestibility by adversely
altering physical structure and chemical composition. The availability of lysine and other
amino acids is particularly sensitive to processing conditions, as elevated temperatures can
lead to the formation of Maillard products and other amino acid complexes or products [8].

Conversely, for ruminant feeds, significant efforts have been directed towards the
possibility of reducing the availability of dietary protein for the microflora in the rumen
through chemical and physical treatments while still maintaining high amino acid
digestibility in the small intestine [8].
Protein Extraction

Considerable efforts have been invested in extracting proteins from diverse sources for
various food applications. Currently, proteins from two primary sources, namely milk
proteins and soy protein, are extensively utilized in the food industry, serving either as a
general nutrient supply or as functional ingredients. Milk proteins, such as whey proteins or
caseins, are arguably the most commonly employed proteins in a wide range of food
products. On the non-animal side, soy protein is gaining increasing prominence in food
products due to its health benefits and distinctive physico-chemical properties. Researchers
have also explored proteins from other sources for their functionality and potential
applications in food [5].

Analyzing food proteins is not necessarily a straightforward process. This complexity arises
in part from the heterogeneous nature of foods, comprising a variety of nutrients like lipids,
carbohydrates, and various micronutrients. The composition, structure, or matrix of the
food, along with interactions among different nutrients, may hinder protein accessibility,
potentially leading to an underestimation of the protein content. Furthermore, various
methods are grounded in different analytical principles, determining protein content either
directly or indirectly.

Direct protein determination involves calculating protein content based on the analysis of
amino acid residues. On the other hand, indirect protein determination can be deduced, for
example, by determining nitrogen content or through chemical reactions with functional
groups within the protein. An additional factor contributing to potential inaccuracies in
protein content determination is the process of protein extraction. Certain methods
necessitate a degree of protein extraction before analysis, and as a result, the extraction
yields can impact the final results [6].

Numerous analytical methods have been developed over the years, but only a limited
number of them are commonly employed. The rationale behind the choice of a particular
method in many studies is often inadequately explained. This lack of clarity in method
selection may be attributed to various factors, such as adherence to tradition (utilizing
established analytical procedures in laboratories), absence of appropriate analytical
infrastructure, or the significant economic costs associated with certain methods.
Methods of Protein Analysis

The examination of proteins holds significant economic and societal importance. The
market value of key agricultural commodities, such as cereal grains, legumes, flour,
oilseeds, milk, and livestock feeds, is influenced in part by their protein content. Precise
protein quantitative analysis is essential for quality control and is a prerequisite for
accurate food labeling. Consumers are attracted to proteins from diverse sources based on
their aesthetic appeal, and adherence to religious dietary restrictions involves excluding
specific protein sources. The range of proteins consumed is crucial for addressing food
allergies. The identification of undeclared protein additives and substitutions poses a
growing challenge. Proteins exhibit varying nutritional quality in supporting dietary needs.
In essence, protein analysis has legal, nutritional, health, safety, and economic ramifications
for the food industry [14].

Over the years, a multitude of analytical methods for protein determination has been
developed. However, only a select few are commonly employed, and the rationale behind
the choice of a specific method in many studies is often inadequately explained. Several
factors contribute to this, such as adherence to tradition (employing established analytical
procedures in laboratories), insufficient analytical infrastructure, or the high economic costs
associated with certain methods.

1. The Kjeldahl method:

Johann Kjeldahl delivered a technique in 1883 for figuring out protein content that
involves digesting a meals sample with a robust acid to release nitrogen, that's then
measured through the best titration method. The protein content material of the
meals is calculated based at the awareness of nitrogen. Although the simple
principles of the Kjeldahl technique have endured, contemporary adaptations have
been included to improve efficiency and accuracy in the size procedure.
2. Dumas method:

A modernized version of the Dumas method has emerged as an automated

instrumental technique for swiftly determining protein concentrations in food
samples. Originating from a methodology introduced by scientist Dumas more than a
century and a half ago, this approach is gaining prominence and challenging the
Kjeldahl method as the preferred analysis standard for certain food items, primarily
due to its speed.

3. Methods using UV-visible spectroscopy:

protein concentration assessment using UV-visible spectroscopy. These methods rely
on proteins' light absorption or scattering properties, involving calibration curves
with known concentrations. The sample's absorbance at the same wavelength is
measured, and protein concentration is determined by referencing the calibration
curve, with variations based on specific chemical groups.
4. Biuret Method:
When cupric ions (Cu2+) interact with peptide bonds under alkaline conditions, a
violet-purplish color is generated. The commercially available biuret reagent,
encompassing all necessary chemicals for the analysis, is mixed with a protein
solution. After allowing the mixture to stand for 15-30 minutes, the absorbance is
measured at 540 nm. One significant advantage of this method is its immunity to
interference from substances absorbing at lower wavelengths.

5. Turbimetric method:
Protein molecules, typically soluble in solution, can be induced to precipitate by
introducing specific chemicals, such as trichloroacetic acid. This precipitation results
in the solution becoming turbid. Consequently, the protein concentration can be
ascertained by quantifying the extent of turbidity in the solution.
 6. The Lowry method:

The Lowry method involves the combined action of the biuret reagent and the Folin-
Ciocalteau phenol reagent, with the latter reacting specifically with tyrosine and
tryptophan residues in proteins. This chemical interaction produces a bluish color,
measurable between 500 to 750 nm. A minor peak at 500 nm is suitable for high protein
concentrations, while a prominent peak at 750 nm is used for low protein
concentrations. The Lowry method is particularly sensitive to lower protein
concentrations compared to the biuret method.

Conclusion

In summary, this document delves into the intricate global of proteins, emphasizing their
pivotal roles inside the human body as fundamental structural components, contributors to
hormonal and enzymatic functions, and resources of energy. As proteins go through
numerous chemical alterations at some point of meals processing, the know-how of those
adjustments is critical. The file explores the challenges posed via the complex nature of
these changes, analytical problems, and the shortage of particular exams for assessing
amino acid availability.

To examine meals proteins successfully, proper isolation thru extraction techniques that
decrease analytical artifacts is important. Factors influencing protein excellent extend
beyond amino acid composition, encompassing intrinsic and extrinsic elements inclusive of
antinutritional properties, processing, storage, and the purchaser's fitness status.

The misconception that uncooked substances meeting nutritional protein necessities

ensure a nutritionally ok final product is dispelled. Various processing situations
considerably modify the structure of meals proteins as they have interaction with different
components, impacting the dietary attributes of the meals.

The report emphasizes the important importance of protein evaluation in the food industry,
wherein correct determination holds felony, dietary, fitness, protection, and financial
implications. It highlights the want for clarity in technique choice, acknowledging the
various factors influencing alternatives in analytical strategies.
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