Experiment 3

Siddhant Unde
J18IMT650

 Analysis of Wheat Flour

 Determination of damaged starch from whole wheat
flour

1. 1000 Kernel Weight

The 1000-kernel weight (TKW) can be determined from the average weight of 300 individual
kernel replicated using the Perten Single Kernel Characterization System (SKCS 4100). The
instrument weighs each seed to the nearest 0.01 mg and automatically calculates the TKW
based on the average weight of the 300 individual seeds. The average TKW is reported in
grams. The 1000 kernel weight is used as an indicator of wheat milling quality which is
considered better than test weight. In order to enhance the correlation between TKW and
milling extraction rate, TKW can be adjusted to a specific moisture. Higher the TKW greater
the wheat flour extraction

Apparatus - Weighing machine, Wheat kernel Grains.

Procedure - 1.  Multiply the desired plant population per square foot by the 1,000 kernel
weight in grams.

2.  Divide that by the seedling survival rate (in decimal form).

3.  Divide by 10 to get pounds per acre.

An example- 1. Multiply desired plant population (30 plants per square foot) x the 1,000
kernel weight (31g) = 930

2.  Divide by the seeding survival rate (0.91) = 1,022

3.  Divide by 10 = 102 pounds per acre.

2. Bulk Density
The bulk density (g/cm3) of flour is the density measured without the influence of any
compression. The bulk densities of flours ranged from 0.762 g/cc to 0.820 g/cc. The highest
bulk density was observed W55 flour (0.820 g/cc) followed by W70 flour (0.786 g/cc),
W85 flour (0.774 g/cc) and lowest for wheat flour (0.762 g/cc). The present study revealed that
bulk density depends on the particle size and initial moisture content of flours. Bulk density
of composite flour increased with increase in the incorporation of different flours with wheat
flour. It is clear that decreased the proportion of wheat flour increase the bulk density of
composite flours. The high bulk density of flour suggests their suitability for use in food
preparations. On contrast, low bulk density would be an advantage in the formulation of
complementary foods. Highest bulk density of composite flour (W55) suggests its suitability to
be used as thickener in food products and for use in food preparation since it help to reduce
paste thickness which is an important factor in convalescent and child feeding.

Apparatus- Wheat Flour, Cylindrical vessel Weighing Machine, etc.

Procedure - 1. Pass a quantity of powder sufficient to complete the test through a 1.0 mm
sieve, if necessary, to break up agglomerates that may have formed during storage and allow
the obtained sample to flow freely into the measuring vessel until it overflows.

2. Carefully, scrape excess powder from the top of the cup by smoothly moving the edge of
the blade of a spatula perpendicular to and in contact with the top surface of the cup, taking
care to keep the spatula perpendicular to prevent packing or removal of powder from the cup.

3. Remove any material from the side of the cup and determine the mass (M) of the powder
to the nearest 0.1%. Calculate the bulk density (g/ml) using the formula M/V0 in which V0 is
the volume of the cup.

4. Determine the mass (M0) of the powder to the nearest 0.1% by subtraction of the
previously determined mass of the empty measuring vessel.

5. Calculate the bulk density (g/ml) using the formula M0/100 and record the average of three
determinations using three different powder samples.

3. True Density

Flour, wheat weighs 0.593 gram per cubic centimetre or 593 kilogram per cubic meter,
i.e. density of flour, wheat is equal to 593 kg/m³. In Imperial or US customary measurement
system, the density is equal to 37.02 pound per cubic foot [lb/ft³], or 0.3428 ounce per cubic.
In physical properties, it was observed that wheat flour was having the true density (TD) of
 0.75±0.17 g/ml whereas the true density varied from 0.74±0.12 to 0.65±0.42 g/ml in
composite flour.

Density of Grain Wheat (material)
1. Density of Grain Wheat in a few select units of density measurement:
2. Density of Grain Wheat g cm3 = 0.79 g/cm³
3. Density of Grain Wheat g ml = 0.79 g/ml.
4. Density of Grain Wheat g mm3 = 0.00079 g/mm³
5. Density of Grain Wheat kg m3 = 790 kg/m³
6. Density of Grain Wheat lb in3 = 0.029 lb/in³

4. Falling Number –

Falling Number is particularly important in milling wheat. Minimum requirements for FN are

largely determined by industry, where consistency is a primary concern, and is the reason
why most wheat shipments must achieve a FN of 300 sec or higher. Falling numbers is a test
that helps identify the structural integrity of the starch chains. To conduct the test, grind the
grain into meal, add water, mix with a stirrer and add heat for 60 seconds. The falling number
is the number of seconds it takes for the stirrer to fall to the bottom of the test tube. The
longer it takes the stirrer to fall, the higher the quality of starch (longer chains).If the stirrer
falls quickly, that is an indicator alpha amylase enzyme may be at high enough levels to start
the starch degradation process for seed germination. In fact, the falling number of a sample is
very closely related (r2=0.924) to the concentration of alpha amylase. If falling number is
high then enzymes can be removed from the flour and if it is too low enzymes cannot be
removed.

Apparatus- Test tube, Wheat Flour, Stopwatch etc.

Procedure - 1: A moisture test is done on a wheat sample that has been selected and ground.

2: 7-gram ground sub-sample, based on a 14% moisture basis, is used for the falling number
test.

3: Distilled water is added to the ground sample in a falling number test tube.

4: The ground wheat and water mixture is thoroughly shaken, forming a slurry.

5: A stirrer is placed in each falling number tube.

6: Tubes containing the slurry are immersed in the boiling-water bath of the falling number
apparatus. The slurry is stirred with the stirrer for 60 seconds then the stirrer is allowed to
drop by its own weight through the ground wheat and water slurry.

7: The total time in seconds it takes the stirrer to reach the bottom including the 60-second
stirring time is the falling number result, which reflects the sprout damage in the sample. The
falling number reading is then recorded.
5. Sedimentation –
The sedimentation test is a physicochemical test that helps provides information on the
baking quality of wheat flour. It is based on the suspension of flour in a dilute alcohol and
acid solution which causes the flour particles to sediment. The sedimentation test is also used
to grade different classes or types of wheat according to protein quantity and quality. It’s also
a screening tool for the development of new hard wheat genotypes by plant breeders. Unlike
other baking quality tests, the sedimentation test does not assess the rheological properties of
wheat flour-based doughs. Instead, it’s similar to the Solvent Retention Capacity (SRC). The
sedimentation test is based on the density and swelling capacity of gluten-forming proteins in
a flour-water slurry containing a lactic acid–isopropanol solution that has been stained with
bromophenol. Gluten proteins from strong flours tend to display higher rates of hydration
(faster water absorption) and higher hydration capacity (absorb more water) than those of
weak (softer) flours. Sedimentation value is expressed as volume in mL of settled gluten and
is dependent on the quantity as well as quality (strength) of the gluten in the flour sample. As
the amount of water absorbed increases, the compactness of gluten particles decreases due to
reduced density of the gluten particles, therefore sink or sediment more slowly.

Apparatus/ Chemical-Glass stoppered graduated cylinder (100 mL), Chronometer, 100-

mesh sieve with collection pan (for flour preparation), Wheat flours. Isopropyl alcohol 99–
100% pure, Lactic acid stock solution, Methylene blue, Bromophenol blue

Procedure

1. Determine the moisture content of flour being studied. This value will be used to correct
results based on a 14% moisture basis in case different moisture content.
2. Place 10 g flour on the 100-mesh sieve with collection pan. Sift the flour manually for
about 2 min to allow flour particles to become looser. Collect flour in the collection pan
when finished.
3. Weigh 4 g of flour. Place flour into the 100 mL graduated cylinder, using a powder
funnel.
4. Add 50 mL of distilled (or reverse osmosis) water to the graduated cylinder. Run test in
duplicate.
5. Place stopper on the graduated cylinder and mix the flour-water slurry by shaking
vigorously for 30 seconds.
6. Allow slurry to stand for 5 minutes.
7. Add 2 to 3 drops of methylene blue or bromophenol blue indicator into the cylinder, then
add 25 mL of lactic acid / isopropyl alcohol solution (12.5 mL isopropyl and 12.5 mL
lactic acid) with a buret.
8. Gently mix the flour-water slurry with the liquid reagents by inverting and returning the
cylinder to the upright position 10 to 12 times.
9. Immediately after mixing, place the cylinder in an upright position and start timing with a
chronometer.
10. Allow slurry to stand for 5 minutes.
11. At end of 5 min, read volume in mL (estimating tenths of mL) of sediment in cylinder.
The volume in millilitres is the sedimentation value of the flour.
12. Multiply the sedimentation value by the appropriate flour moisture factor detailed in
AACCI Method 56-61.02.
6. Alcoholic Acidity –

Alcoholic acidity is defined as mg of H2SO4 required for 100 g of the sample to have the
same alcohol soluble acids. Grains or their milled products on storage undergo physical, as
well as, chemical changes. Acid phosphates, amino acids and free fatty acids of flours, under
certain conditions increase considerably due to the enzymatic hydrolysis of phytin, protein
and fat, respectively. Milled products deteriorate faster than their parent grains. The amino
acids and acid phosphates are soluble in strong alcohol. The free fatty acids are insoluble in
water but are soluble in fat solvents or in strong alcohol. For this reason, the acidity in flours
is expressed as either fat acidity, wherein, benzene is used as a fat solvent or it is expressed as
alcoholic acidity. Higher alcoholic acidity is an indicator of higher acidity of the germ oil in
the flour. As per FSS Regulations, alcoholic acidity in wheat flour shall not be more than
0.18 per cent by weight; as per Indian Standard, it shall not be more than 0.1 per cent.

Apparatus/Chemicals - Sample of Wheat flour, conical flask, weighing balance, Pipette,

Standard NaOH solution (approximately 0.05 N), Phenolphthalein, Neutral ethyl alcohol-
90% volume by volume.

Procedure – 1) Weigh 5 g of the flour sample. Put in conical flask.

2) Add 50 ml of neutral ethyl alcohol.

3) Stopper the flask, shake and allow to stand for 24 hours.

4) filter the alcoholic extract (through an ordinary dry filter paper).

5) Titrate 10 ml of alcoholic extract against standard NaOH solution using phenolphthalein as

indicator.

6) Calculate the percentage alcoholic acidity as sulphuric acid.

7) Calculate the alcoholic acidity of the sample using formula.

Gluten Content –
The strength of the flour is associated with the amount of gluten present in the flour. The
dough obtained by mixing wheat flour with water possesses the characteristics of viscosity.
Plasticity and elasticity which are significant during handling the dough as well as in its end
performance. These properties are due to the interaction of water soluble proteins of wheat
with water. The albumins, globulins and other smaller proteins as well as starch are washed
along with water leaving behind a cohesive, elastic and rubbery mass called as "Crude wet
gluten". Colour of the gluten varies from pale yellow to greenish grey. Weaker gluten
generally have yellow colour, while strong gluten will have deeper greenish colour. Gluten
from strong wheat are generally extensive and elastic. Hydrated glutenins and gliadin differs
in their mechanical property. Gliadin is viscous, sticky mass which is extensible; while
glutenins is tough, rubbery elastic mass. Gliadin is more extensible, but less elastic. Gluten
exhibits the properties of both the components. Gluten in a sample of flour can be estimated
by hydrating the flour with water and later washing the dough free of starch, sugars, water
soluble proteins and other minor components. The wet cohesive, viscos elastic mass obtained
is referred to as wet gluten while the dried product obtained from it is referred to as dry
gluten.

Apparatus/ Chemicals - Hot air oven, Aluminium dishes/petri plates (small), Analytical
balance (least count 0.001 g), Desiccator, Aluminium foil, Plastic bowl ,Measuring cylinder,
Glass rod, Fine sieve.

Procedure – Weigh out 25g flour into a plastic bowl.

1) Add l5 mL water, mix to get a dough ball.

2) Immerse the dough ball in water for one hour to ensure proper hydration.
3) Wash out the starch by kneading gently in a gentle stream of water over a fine sieve.
4) Continue washing till the washed out liquid is clear.
5) Squeeze the washed water into clean water.
6) If water turns turbid, continue washing.
7) The cohesive mass obtained is called wet gluten
8) After pressing as dry as possible place in a petri dish containing a small piece of
aluminium foil.
9) The wet gluten so obtained is dried in a hot air oven at 100DCfor 24 hr.
10) Weigh the gluten to constant weight.

7. Starch Damage –

The milling of wheat causes physical damage to a proportion of the starch granules of

the flour. The level of starch damage directly affects water absorption and dough mixing
properties of the flour and is thus of technological significance. In the current procedure,
damaged starch granules are hydrated and hydrolysed to maltosaccharides plus α-limit
dextrins by carefully controlled treatment with purified fungal α-amylase. The fungal α-
amylase treatment is designed to give near complete solubilisation of damaged granules with
minimum breakdown of undamaged granules. This reaction is terminated on addition of
dilute sulphuric acid, and aliquots are treated with excess levels of purified amyloglucosidase
to give complete degradation of starch-derived dextrins to glucose. The glucose is
specifically measured with a high purity glucose oxidase/peroxidase reagent mixture.
Determined values are presented as starch (damaged) as a percentage of flour weight on an
“as is” basis.
Apparatus - Glass test tubes (round bottomed, 16 x 100 mm, 12 mL capacity). , Micro-
pipettes, 100 μL, Positive displacement pipette, e.g. Eppendorf Multipette, Bench centrifuge
(required speed 3,000 rpm, i.e. approx.1000 g), Analytical balance, Spectrophotometer set at
510 nm, Vortex mixer, Thermostat water bath set at 40°C, Stop clock.

Procedure –
1. Accurately weigh 100 ± 10 mg of wheat flour or milled gelatinised starch sample
into a thick walled glass centrifuge tube (16 x 120 mm; 12 mL capacity).
2. Pre-equilibrate the tubes plus contents at 40°C for approx. 5 min.
3. Pre-equilibrate fungal α-amylase solution (50 U/mL) at 40°C for approx. 5 min in
a small glass beaker.
4. Add 1.0 mL of pre-equilibrated fungal α-amylase solution (50 U/mL) to each
tube, stir the tube on a vortex mixer for approx. 5 sec and incubate at 40°C for
exactly 10 min (from time of addition of the enzyme).
5. Add 8.0 mL of dilute sulphuric acid solution (0.2% v/v) to each tube after exactly
10 min from the time of addition of the fungal α-amylase and stir the tube
vigorously for approx. 5 sec. This inactivates the enzyme and thus terminates the
reaction.
6. Centrifuge the tubes at 3,000 rpm (1,000 g) for 5 min or filter the slurry through
what man No. 1 (9 cm) filter paper.
7. Carefully and accurately transfer 0.1 mL aliquots of the supernatant solution (or
filtrate) to the bottom of two test tubes.
8. Add 0.1 mL of amyloglucosidase solution (2 U) to each tube, stir the tubes on a
vortex mixer and incubate them at 40°C for 10 min.
9. Add 4.0 mL of GOPOD reagent solution to each tube (including glucose standards
and reagent blank tubes) and incubate the tubes at 40°C for 20 min.
10. Measure the absorbance of all solutions at 510 nm against a reagent blank.

1. Colorimeter
Lab hunter colorimeter is used for wheat flour colour analysis. The
lightness value, L*, also referred to as "Lstar," defines black at 0 and white at 100.a*
axis is relative to the green–red opponent colors, with negative values toward green
and positive values toward red, and b* is the yellow/blue coordinate with positive
value towards yellow and negative towards blue. Standard value of a = -2.4 and b =
+6.9.
2. Farinograph
In baking, a farinograph measures specific properties of flour. The farinograph is a
recording dough mixer that measures and records the torque developed by the action
of the mixer blades on the dough during mixing. It is used to estimate the water
absorption of flours, the relative mixing time, the stability to overmixing, and
rheological properties of the dough during mixing.

3. Extensograph
An extensograph is a tool used for measuring the flour quality, gluten strength and
stretching behaviour of dough. Extensional properties, which determine the course of
dough expansion during proofing and baking, have a direct effect on: Loaf volume.

4. Alveograph
Alveograph is a tool for flour quality measurement. It measures the flexibility of the
dough produced from the flour, by inflating a bubble in a thin sheet of the dough until
it bursts. The resulting values show the strength of the flour, product consistency
during production and thus its suitability for different uses.

5. Mixograph
The mixograph is a dough testing equipment used to assess the baking quality of
flours from soft, hard and durum wheat. It provides information on the mixing and
absorption characteristics of flour. And indicate Dough Gluten Strength.

6. Glutomatic
The Glutomatic System is the global standard for determination of gluten quantity and
quality. The effect of gluten quantity and quality in the wheat flour used for breads,
cookies, crackers and pasta is noticeable.

7. Amilograph
Amilograph is instrument that measures and records the gelatinization temperature,
Enzyme Activity and viscosity of pastes of starch and wheat flour.

8. Rapid Visco Analyser(RVA)

The main use of the RVA has been in the routine analysis of cereals (e.g. wheat,
barley) to determine the effects of sprouting on grain quality at the delivery point, as
well as to simulate the commercial processing conditions in the Brewhouse, including
the pH, malt to water ratio, time to temperature profiles.