TITLE

Proximate analysis of a given sample of cereal and legume mix.

AIM

To quantitatively estimate the components of a given sample of cereal and legume mix namely;

 Fat

 Moisture

 Protein

 Fibre

 Ash

 Carbohydrate

 Energy content.

INTRODUCTION

Food typically consists of ingredients including carbs, protein, fat, vitamins, moisture, and

mineral stuff (ash). Ancient staple foods that are high in carbohydrates, fiber, and micronutrients

include cereal and legume grains. They offer a decent amount of proteins as well. Protein

efficiency is increased when cereals and legumes are mixed because their necessary amino acid

compositions are complimentary (Young & Pellett, 1994). Both types of whole grains are

essential for the prevention of cardiovascular disease, type 2 diabetes, certain malignancies,

digestive tract illnesses, overweight, and obesity, according to Mellen, 2006, et al (Duranti,

2006; Mellen, Walsh, & Herrington, 2008; Ye, Chacko, Chou, Kugizaki, & Liu, 2012).
When consumed in their unrefined state, cereals are a rich source of essential nutrients; but, after

the removal of the outer layers—the bran and germ—the endosperm is primarily composed of

carbohydrates and lacks the bulk of the other nutrients (Abdulrahman & Omonyi, 2016). This

means that even while grains are packed with nutrients, the refining process lowers the quality of

them. Some have a lot of protein, while others have almost none. While some cereals have a lot

of fat, others have relatively little of it. However, a commonality among all cereals is that they

have a high carbohydrate content and a low water content. Legumes added to cereal products

have nutritional benefits that improve the amino acid balance as well as the products'

processability and physical characteristics.

The elements of a cereal legume mix will be estimated in this investigation. The components of

the cereal legume mix will be identified using the proximate composition analysis of food

samples.

Proximate analysis is the process of estimating a food's essential ingredients using techniques

that enable a reasonably rapid and accurate assessment of different food fractions, typically

without the use of expensive equipment or chemicals. Protein, fat, moisture, ash, and

carbohydrate levels are determined using a range of methods, including extraction, Kjeldahl, and

NIR. The given cereal-legume mix's protein, fat, moisture, ash, and carbohydrate contents were

ascertained using AOAC Methods.

METHODOLOGY:

Determination of Moisture Content - AOAC 930.15

A petri dish that was empty was dried in the oven, chilled, and its weight was recorded. The

material was weighed and distributed equally over the bottom of the petri dish in an amount of

about five grams (5g). The dish was cooked for 4 hours at 105 °C in a forced draft oven. After

cooling the dish and sample in a desiccator, the sample's weight was collected after drying, and

the moisture content was computed as a percentage of moisture lost. The formula used to

determine the moisture content (MC) was

%MC = [Weight of dish + Sample (before)] – Weight of dish + Sample (after)] x 100

Weight of sample

Determination of Crude Fat - AOAC 2003.05

The sample was weighed into a thimble containing non-absorbent cotton and packed with around

5 g of the sample. The sample-containing thimble was set within the Soxhlet equipment'

extractor. Weighing was done with a clean, dried Soxhlet flask and around 200 cc of petroleum

ether, the extraction solvent. To assist in condensing the organic solvent, the Soxhlet apparatus

was set up and cold water was allowed to flow through the condenser. The apparatus was left to

operate for six hours, after which the Soxhlet flask containing the fat extraction was removed and

dried for one hour in an air oven at 105 °C. It was then weighed after cooling in a desiccator.

Using the equation, the fat content was estimated as a percentage of crude fat.

( %FC=( Weight of flask +Oil ) – Weight of empty flask x 100 )

Weight of Sample

Determination of Total Ash content – AOAC 942.05

A porcelain crucible was weighed and its base was labeled with a pencil after it had already been

lit and cooled. The sample was weighed into the crucible at a weight of around 2 g. The crucible

was positioned with tongs in a muffle furnace set at 550 °C and let to burn up for two hours. The

ash was then weighed after cooling in a desiccator to room temperature. The following equation

was used to determine the ash content as a percentage of total ash:

%AC = [Weight of crucible + Sample (before)] – Weight of crucible + Sample (after)] x 100

Weight of sample

Determination of Crude Fibre - AOAC 978.10

2g of the defatted sample (Ws) were placed in a round bottom flask with around 100 mL of

1.25% H2SO4 and allowed to boil for 30 minutes. The material was filtered and rinsed with

warm water following the 30-minute period. 100 mL of 1.25% NaOH was then poured over the

residue after it had been put into the flask. After another 30 minutes of boiling, filtering, and

washing with warm distilled water and 80% alcohol, the remaining material was left behind. The

residue was subsequently dried for two hours at a temperature of around 130 °C in a hot air oven

before cooling in a desiccator for roughly 30 minutes. The dried fiber weight was measured in a

pre-dried crucible (W1), and the sample was burned for two hours at 550°C before cooling.

When the crucible and ash (W2) were cooled, the following was calculated:

%CF = ([Weight of crucible + Sample (before)] – Weight of crucible + Sample (after)] x 100)

Weight of Sample
%CF=W 2 – W 1 x 100/Ws

Determination of Crude Protein - AOAC 2001.11

A digestion tube was filled with around 1 g of the material, and two catalyst tablets and 15 mL of

concentrated H2SO4 were placed over the sample (Kjeltabs). The sample was subsequently

digested in the Kjedahl digester for roughly 2 hours at a temperature of 420 °C. Following

digestion, 10 ml of the digest was removed and diluted to a level of 100 ml in a volumetric flask.

200 cc of distilled water and 50 mL of 40% NaOH were added to 20 ml of the diluted digest.

With the water circulation pump of the condenser turned on, the Kjeltec distillation setup was

placed at a temperature of 150–200 °C. In a conical flask filled with 30 mL of 4% boric acid, two

to three drops of mixed indicator were added. The flask was then attached to the distillation

equipment. The distillate was collected into a conical flask and titrated against 0.1 N HCl in an

amount between 100 and 150 ml (standardized solution). The following equations were used to

compute the percentage of crude protein as well as the percentage of protein content relative to a

blank:

Kjeldahl Nitrogen %

%N = [(Titre of sample – Titre of blank) x N(HCl) x 14.01] x 100

Weight of sample x 10

Crude Protein %

% Crude Protein=% Kjeldahl N x F

Where N = Normality of standardized HCl

14.01 = Molecular weight of Nitrogen

10 = Factor to convert mg/g to percent

Titre = Volume of standardized acid used to titrate a test

F = Conversion factor to Protein

(5.70 for wheat, 6.38 for dairy products and 6.25 for other feeds)

Carbohydrate and energy determination

The carbohydrate content of the sample was determined by difference.

The carbohydrate content was calculated as indicated below:

Carbohydrate (%) = 100 – (% moisture + % fat + % protein + % ash + % fibre)

Calculation of Energy Content

The Atwater factor was used to calculate the energy content of the sample.

The energy content was calculated as indicated below:

Total energy (kcal/100 g) = [(% available carbohydrates × 4) + (% protein × 4) + (% fat × 9)]

RESULTS AND DISCUSSION

Table 1. Proximate composition of given sample of cereal-legume mix.

Sample %Moistu %Fat %Protein %Fibre %Ash %Carboh Energy

ID re y-drate (kCal/100

g)

A 7.37 8.05 19.19 4.26 0.16 60.96 393.05

7.72 8.01 19.19 4.24 0.17 60.67 391.58

Avg ± 7.55 ± 8.03 ± 19.19 ± 0 4.25 ± 0.165 ± 60.82 ± 392.32 ±

Std. 0.175 0.02 0.01 0.005 0.145 0.735

Error

There was no difference between the outcomes for the cereal-legume combination and those

attained by Walles and Moges, other than a much higher ash content and a noticeably lower fiber

content. The larger proportion of legumes in the cereal-legume flour mixture may be to blame for

the elevated ash level given that legumes are known to have a high mineral content. Formulas for

babies should have less fiber because they digest food at a slower rate than adults. The higher

fibre levels in the mixture that was studied may be explained by the fact that the cereal-sample

combination was not created with infants in mind. According to research by (Milkesa, 2020),

malted sorghum-soy composite flour biscuits had a fat level of 9.26% for malted sorghum with

40% soy flour blend and 2.49% for 100% malted sorghum flour. This demonstrates that the fat

content of the composite biscuit was greatly raised by the addition of soy flour, and his fat

content values were comparable to those found for the cereal-legume combination (8.03%).

Additionally, Mikesa found crude fiber values ranging from 2.56% to 3.46%, which, despite
being a little lower, are comparable to those found (4.26%). Carbohydrate content was lower

than the cereal-legume mixture and ranged from 47.08% to 59.95% depending on the amount of

soy flour substituted. However, the ash level was much higher (2.8%) than the figure (0.165%)

reported for the cereal-leaven mix. The following ranges of proximate composition for the

various blends were determined by (Walle and Moges, 2017) when optimizing a cereal-legume

blend ratio to improve the nutritional quality and functional features of a complementary food.

These ranges are shown in table 2 below.

Table 2: Range of proximate composition of various cereal-legume ratio blends (Walle

and Moges, 2017)

%Moistur %Fat %Protein %Fibre %Ash %Carbohy- Energy

e drate (kCal/100g)

6.51-6.58 5.80-8.22 14.64-16.50 2.01-2.61 2.04-2.27 66.43 -71.00 394.77 - 405.76

Except for a much higher ash content and a significantly lower fiber content, there was no

difference between the results for the cereal-legume combination and those obtained by Walles

and Moges. Given that legumes are known to have a high mineral content, the higher

concentration of legumes in the cereal-legume flour mixture may be to blame for the increased

ash level. Since babies have lower levels of digestion, formulations for infants should have less

fiber. Given that the cereal-sample combination was not specifically designed for newborns, this

could account for the greater fibre levels in the mixture that was examined. As a result, the

measurements made for the given sample of cereal and legume mix's fat, moisture, protein, fiber,

ash, carbohydrate, and calorie contents were equivalent to those reported in the literature.
CONCLUSION

The fat, moisture, protein, fiber, ash, carbohydrate, and energy contents of the provided sample

of a cereal-legume mix were effectively determined. Given that the cereal-legume mixture has a

sufficient quantity of all the necessary components, it may be said that it has good nutritional

value. As a result, the cereal-legume mixture helps to reduce malnutrition by reducing the

amount of protein in diets and serving as a complete meal.

EXPERIMENT (SPECTROPHOTOMETER ANALYSIS)

Title: Determination of total phenolic content using uvvis spectrophotometer

Introduction

Spectrophotometry is the measurement of light intensity as a light beam passes through a sample

solution to determine how much light is reflected by a chemical substance. The underlying idea

is that each molecule has a specific wavelength spectrum where light is either absorbed or

emitted.

Beer-Lambert According to law, absorbance is influenced by the solute's molar absorptivity, or

absorption coefficient, (ϵ), and the length of the light path (l). A tool used in spectrophotometric

measurements is the spectrophotometer. Choosing a blank cuvette, inserting it into the

spectrophotometer, and finally shutting the lid constitute the mode of operation. The instruments

now read 0.00000A when you click on 0 ABS 100% transmittance. Measure the absorbance

between 300 and 700 nm or at the targeted wavelength using a solution whose concentration is

known.

In the food industry spectrophotometry is used in Quality Control and monitoring of beverage

production where many foods are colored using federally specified and approved food dyes.

Aims/Objectives of the experiment:

To determine the concentration of phenolic content in dried Prekese mix

MATERIAL AND METHODS

Spectrophotometer

Cuvettes

Falcon tube

Test tube

Pipette

Gallic acid

Dried Prekese mix

Distilled water

Vortex mixer

Volumetric flask

Na2CO3

Using Gallic as the reference material, the total phenolic content (TPC) of the dried Prekese mix

sample was calculated using the Folin-Ciocalteau reagent method.

50 mg of the unidentified sample were weighed into a falcon tube to prepare the sample. The

weighted sample was vortexed for two minutes after being dissolved in 50ml of distilled water

up to the 50ml mark. After that, the sample was let to stand for 15 minutes to allow any

undissolved materials to settle.

Stock solution of Gallic acid of concentration 10mg/100ml (0.1mg/ml) was prepared from which

serial dilutions was carried out to obtain concentrations of (0.002, 0.004, 0.008, 0.016, and

0.032)mg/mL
From the respective dilutions and unknown extracts, 1ml of each was pipetted into well labelled

test tubes. To each tube 20ul of Folin-Ciocalteau was added followed with 1000ul of 20%

Na2CO3. The resultant mixture was allowed to incubate at room temperature for 30 minutes.

The blank consisted of 50ml distilled water, 20ul Folin-Ciocalteau reagent, 1000ul of 20%

Na2CO3.

The absorbances of the dilutions and unknown samples were measured and recorded with the

UVVIS spectrophotometer at 760nm.

Results and Discussion

Table. Results showing standard concentrations of Gallic acid solution, Unknown Sample and

their corresponding absorbance readings.

Concentration (mg/ml) Absorbance at 760nm

0.002 0.09689

0.004 0.16535

0.008 0.39180

0.016 0.74497

0.032 0.83303

Unknown 0.21676

From the linear equation

Absorbance vrs concentration caliberation curve
0.9
f(x) = 25.0147244623656 x + 0.136225416666667
0.8 R² = 0.841185112404674
0.7
0.6
ance

0.5
 0.3

A
0.2
0.1
0
0 0.005 0.01 0.015 0.02 0.025 0.03 0.035
y = 25.015x + 0.1362 Concentration (mg/ml)

y = 0.21676 m= 25.015

0.21676 = 25.015x + 0.1362

25.015x = 0.21676 - 0.1362

x = 0.08056 / 25.015

x = 0.00322mg/ml

The concentration obtained for the sample from the linear equation on the calibration curve

is 0.00322mg/ml

A mass of 50mg of the unknown sample was weighed into 50ml of distilled water in preparation of

the sample.

0.00322mg = 1ml

T mg = 50ml

T mg = (0.00322mg x 50ml) /1ml

T mg = 0.161mg

The sample contains 0.161mg/g of total phenol

Precaution

To ensure that correct readings are taken at the specified wavelengths, the machine's calibration

status was checked before usage.

DISCUSSION

Based on the outcomes, it is evident that the absorbance and concentration of the standard

solution are, respectively, 0.09689 and 0.02 mg/ml, and that the concentration of the standard

solution% is, similarly, 0.83303 and 0.032 mg/ml. This information demonstrates that the

absorbance value increases with concentration. The total phenolic concentration (Gallic acid

equivalent) is calculated from the presented curve to be 0.00322 mg/ml with an absorbance of

0.21676. With the linear equation y = 25.015x + 0.1362, the experiment's r 2 result is 0.8412. The

antioxidant properties of food are due to their phenolic content. These antioxidants are in charge

of squelching the free radicals that can cause cancer.

CONCLUSION

In conclusion, Prekese contains antioxidants, which serve to reduce free radicals and so reduce

the likelihood of cancer.

EXPERIMENT (MICROBIAL ANALYSIS Content)

Aim: Determination of presence of microbial contamination herbal tea

INTRODUCTION

Bacteria, fungus, archaea, and protists are all types of microorganisms. Viruses and prions are

not considered microbes. The creation of oxygen, symbiotic interactions, evolution, and

decomposition are just a few of the crucial functions that microorganisms play in ecosystems.

Dead animal or plant materials is broken down into simpler molecules during decomposition.

Each microbe has particular adaptive traits and qualities that allow it to perform its duties as a

helpful organism or as the cause of a food-borne disease. Each microorganism's distinctive traits

and adaptations act as its identifying fingerprints.

Cocci, or spherical cells, bacilli, or cylindrical or rod-shaped cells, and spiral or curved forms are

three categories into which the shapes of significant bacteria are categorized.

Molds are multicellular filamentous fungus that can be easily identified by their fuzzy or cottony

appearance when they grow on food.

Microbiology is necessary for all aspects of food production, processing, preservation, and

storage. Microbes like bacteria, molds, and yeasts are utilized to produce food and food

ingredients including wine, beer, bakery goods, and dairy products.

On the other hand, the spread and contaminating of spoilage and harmful bacteria is regarded to

be one of the main causes of food loss in the modern day. During shipping or in other locations

where food is grown, prepared, or stored, hazardous or unclean food practices can lead to

foodborne microbial contamination. The most frequent cause of food poisoning outbreaks is

microbial contamination. Aseptic procedures, sample dilution, appropriate growth media, and

culturing conditions are employed to identify the presence of the various species and amounts of

these pathogenic and food spoilage bacteria in food both qualitatively and quantitatively.

Aim:

To assess the quality of food sample using microbiological culture techniques in a quantitative

measure

Materials and Methods:

Quantitative Assay

Pour plating and Spread plating culture techniques were used in quantitative assessment of the

sample of the herbal tea extract.

Serial Dilution:

To reduce the microbial population and bring it into workable range, the sample was diluted by

dissolving 5ml of the food sample in 45ml of sterile peptone water (0.1%) and subsequently

diluting to the fourth series (10-4).

Pour Plating:

Media prepared according to manufacturer’s directions: 1L of distilled water with 17.5g of

dehydrated Plate Count agar heated and autoclaved for 15 minutes at 1210C. Plate count agar

(PCA), which was made by dissolving 4.5g of agar powder in 250ml of distilled water and

sterilizing it at 1210C for 15 minutes to produce sterile molten agar that was then brought to cool

at 400C, was used for the pour plate method. Pipetting 1ml of the dilution series into pre-labeled

Petri dishes carrying the appropriate date, media, sample code, and personnel code was done. To

ensure a complete mixing of the inoculum and media and to achieve a homogenous mixture and

uniform distribution, around 30ml of molten agar was added and gently swelled. The plates were

allowed to stand and settle for 30 minutes before being incubated at 370°C for 24 hours. The

plates were examined for visible colonies, which were counted, and documented with the aid of

the colony counter.

Spread Plating:

The total plate count (TPC) and total Coliform count (TCC) of the sample were calculated using

the spread plate method. Plate count agar (PCA) and Mannitol salt agar (MSA) plates were made

by combining, respectively, 4.5g of PCA and 12.5g of MSA in 250ml of distilled water. The

agars were first sterilized at 1210C for 15 minutes, after which they were dispensed into sterile

Petri plates and let to stand and set for 30 minutes. Aliquots of 100 l of each dilution series were
pipetted into dishes of PCA and MAS that had already been labeled, and they were dispersed

using a sterile spreading rod. Incubation was done at 37 0C for 24 hours and observed visible

colonies were counted and recorded with the help of a colony counter.

Qualitative Assay

The objective of the qualitative assay was to identify Staphylococcus aureus in the food sample.

On mannitol salt agar, this was done using the spread plate approach (MSA). Agar plates of

MSA were covered with 100 l aliquots of the first dilution (10-1) and incubated at 370C for 24

hours. Colonies that had established were observed and duly noted.

Diluent: 0.1 % Buffered Peptone Water (0.1g in 100 mL)

Media: Plate Count Agar (PCA) or PCA supplemented with 1g/L skim powdered milk for dairy

products
Sterile Petri dishes

Sterile media bottle

Sterile Sample containers containing 0.1% PW (9*mass/volume of sample)

Dilution tubes containing 9 ml 0.1% PW

Pipette and sterile pipette tips

Laboratory incubator

METHOD:

Media prepared according to manufacturer’s directions: 17.5g of dehydrated Plate Count

agar in 1L of distilled water, heated and autoclaved at 1210C for 15 mins.

Results and Discussion

Quantitative Assay

The colony counts recorded for the TPC, TCC and S. Aureus are presented in Table below

Test Media 10-1 10-2 10-3 10-4

TPC Plate count agar TFTC (<30) TFTC (<30) TFTC (<30) TFTC (<30)

TCC VRBLA TFTC (<10) TFTC (<10)

S. aureus Manitose salt agar TFTC (<10) TFTC (<10)

The initial dilution (10-1)'s total plate count revealed colony numbers of fewer than 30 cfu. The

same was shown for additional dilutions up to the fourth dilution (10-4). Each of the two carried

out dilutions had a cfu of less than 10 according to the total coliform count. CFU counts for the

qualitative S. aureus test were less than 10, although cfu growth suggests the presence of S.

aureus.

Precaution

Ensure you put all the protective clothing before working in a microbiology lab.

Conclusion

In conclusion, product contains microbial count that is below the maximum permissible standard
CHROMATOGRAPHY

According to Coskun (2016), chromatography is a vital biophysical method that makes it

possible to separate, identify, and purify the constituents of a mixture for qualitative and

quantitative examination. The purpose of chromatography is to extract or separate the solute

from a mixture. The solubility, molecular weight, and ionic charge of the solute and the solvent,

among other physical and chemical characteristics, are used in this separation. Thus,

homogeneous mixes can be successfully separated using chromatography.

What is taken out and separated from the solvent is the solute. It includes passing a mixture that

has been dissolved in a "mobile phase" through a stationary phase, which uses differential

partitioning between the mobile and stationary phases to separate the analyte to be measured

from other molecules in the mixture. Small variations in the partition coefficient of a chemical

lead to unequal retention on the stationary phase, which alters the separation.

The effectiveness of this separation is further influenced by the molecular properties of affinity,

partition, and liquid-solid adsorption. Varied molecules and solutes have different affinities for

the stationary phase or the mobile phase depending on the variations. The mixture advances

along the stationary phase with the help of the mobile phase, but is distinguished by this

difference as it does so. (1997, Porath). Harris (2004) noted that while some molecules of the

mixture move slowly through the chromatographic system and spend more time in the stationary

phase, other molecules move quickly through the mobile phase and are eluted more quickly.

Three essential elements—stationary phase, mobile phase, and separated molecules—are used in

the process of chromatography.

The Stationary phase

This phase is always made up of a solid phase or a liquid layer that has been adsorbed onto a

solid support. As the stationary phase, Silica or Alumina are frequently utilized. A slurry

composed of the adsorbent and an appropriate solvent—typically a non-polar solvent—is mixed

together to build up the column. The sample won't run evenly in the column if the slurry isn't

thoroughly churned to remove all air bubbles. The column is subsequently filled with the

prepared slurry. Additionally, sand is required at the top to help load the sample.

The Mobile phase

This phase can be either liquid or gas-based. The components of the mixture are separated at

different rates by adsorbing to the stationary phase when utilizing chromatography. The mobile

phase, which can be either a liquid or a gas, is moved down a chromatographic column.

How Separation Occurs

Due to their varying rates of transit through the stationary phase, the mixture's constituent parts

separate from one another. The characteristics and selection of the specific mobile and stationary

phases determine which components move faster or slower and how they are separated. The

relative "speed" at which components travel through a chromatographic system is referred to as

retention time.

Steps of Chromatography

There are several types of chromatography and each type has steps that must be followed to be

able to achieve the separation of the components of a mixture for quantitative and qualitative

analysis. Generally, these steps may be followed:

 1. Sample Collection

The Sample is prepared for chromatographic analysis.

2. Sample injection or introduction

The sample is then introduced to the chromatographic system for separation.

3. Sample Separation

Separation of the components of the mixture occurs depending on the characteristics and choice

of the particular mobile and stationary phases.

4. Sample Detection.

The eluted solute is cleaned up and quantified. And if it is connected to a mass spectrometer, the

spectrum is read to determine the substance present.

Types of chromatography

• Column chromatography

• Ion-exchange chromatography

• Gel-permeation (molecular sieve) chromatography

• Affinity chromatography

• Paper chromatography

• Thin-layer chromatography

• Gas chromatography
• Dye-ligand chromatography

• Hydrophobic interaction chromatography

• Pseudo-affinity chromatography

• •High-pressure liquid chromatography (HPLC)

Areas of Application of Chromatography in the Food & Beverage Industry

Chromatography is a useful technique for scientists working in food laboratories. Additionally, it

is employed in medical, pharmaceutical, and clinical laboratories. In the food sector, it is applied

to:

1. Identify the nutrients in a specific food and provide precise information.

Chromatography is used to separate mixtures and test for pollutants or additives in food, which is

how you may find out what's in it.

3. Examining the food's nutritional value: Businesses check their goods for nutrients including

proteins, vitamins, preservatives, and other ingredients. They can check the nutritional value of

ssstheir products utilizing chromatography.

4. Product Development: Additional testing is necessary to assure the safety of food firms'

products as they work to improve their offerings by using various additives and preservatives
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1. Abdulrahman, W. and Omoniyi, A. (2016) ‘Proximate analysis and mineral compositions

of different cereals available in gwagwalada market, FCT, Abuja, Nigeria’,

Researchgate.Net, 3(2), pp. 50–55.

2. Duranti, M. (2006). Grain legume proteins and nutraceutical properties. Fitoterapia, 67–

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3. Mellen, P. B., Walsh, T. F., & Herrington, D. M. (2008). Whole grain intake and

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Diseases, 283–290

4. Ye, E. Q., Chacko, S. A., Chou, E. L., Kugizaki, M., & Liu, S. (2012). Greater whole-

grain 1012 intake is associated with lower risk of type 2 diabetes, cardiovascular disease,

and weight gain. The Journal of Nutrition, 1304–1313.

5. Coskun, O. (2016). Separation techniques: Chromatography. Northern Clinics of

Istanbul, 3(2), 156-160. https://doi.org/10.14744/nci.2016.32757

6. Porath J. From gel filtration to adsorptive size exclusion. J Protein Chem. 1997;16:463–8

7. Harris DC. Exploring chemical analysis. 3rd ed. WH. Freeman&Co; 2004.

8. GenTech Scientific blogpost, 2020. https://gentechscientific.com/applications-of-

chromatography-in-the-food-industry/ accessed on 27th October, 2022