Refresher Training on RM Test

Methods
 Raw Material Category & Parameters
Color Moisture
Cocoa Powders
p H Fat
Slip Melting Free Fatty Acid
Point (SMP) (FFA)

Oil & Fat

Peroxide Iodine Value(IV)

Value(PV)

Color Moisture

Milk Products

Acidity Protein
 Melting Point Apparatus
• Melting Point Apparatus is used to analyze
melting point or slip melting point of
samples.
• The Slip melting point (SMP) or "slip point"
is one conventional definition of
the melting point of a waxy solid. It is
determined by casting a 10 mm column of
the solid in a glass tube with an internal
diameter of about 1 mm and a length of
about 80 mm, and then immersing it in a
temperature-controlled water bath.
• The slip point is the temperature at which
the column of the solid begins to rise in the
tube due to buoyancy ,and because the
outside surface of the solid is molten.
• One end open capillaries for solids & both
end open for fat/Oils are preferred.

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 Melting Point Apparatus
• Principle: melting point of a substance.
• This system is a combination of a furnace and
a video camera.
• Melting point is determined by heating the
sample. The furnace is used to control the
temperature during an analysis. Temperature
control and temperature recording is
guaranteed by a digital platinum temperature
sensor.
• The sample sits inside a glass capillary and is
inserted into the furnace. During heating,
when the melting occurs, the light
transmission through the sample changes:
When the material is solid and opaque, less
light is transmitted through the substance.
When a material is liquid and more
transparent, more light is transmitted. This
change in light transmission can easily be
determined with a video camera and is
therefore a reliable way of detecting the

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 Melting Point Apparatus
Wilkote 34 to 36 °C
B.S.
Ultrachoco 34 to 36 °C
Melt the Fat & Fill
the capillary up to
10 mm P’Cote 346 34 to 36 °C

Freeze fat column Wilchox 35 to 38 °C

Record results & Conditioning for
1 Hour
Wilkote 38 to 40 °C
Vanaspati
P’Chox 380 35 to 38 °C

Place Conditioned HVO 29 to 37 °C

Set Initial Temp &
capillaries in Water
Start Machine
filled Vials

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 Melting Point Apparatus
• Precautions:
1.Always fill the column up to 10 mm for correct results.
2.Always set the initial temp of machine as per actual temp of machine with surroundings.
3.Don’t do over conditioning of capillaries.
4.Always set the end temp as per melting point of sample.
5.In case of Fat sample always melt complete sample( Around 60 deg C) and then only fill
capillaries.
6.Always carry out duplicate sample for average readings.

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 Fat estimation by Gerber Method
• Fat estimation can be done by various methods:
• Gerber method
• Soxhlet method
• Mojonier method
• Rose-Gottlieb Method

• Gerber Method :The Gerber Method is a primary and historic chemical test to determine the fat
content of substances, most commonly milk and cream. Milk fat is separated from proteins by adding
sulfuric acid. The separation is facilitated by using amyl alcohol and centrifugation. The fat content is
read directly via a special calibrated butyrometer.
• Problems encountered during Gerber fat estimation: The two major defects associated with the
Gerber method include:
• Charring - is observed as tiny black specs at the fat/non-fat interface in the butyrometer. This problem
is due to charring of the milk proteins by the concentrated Gerber acid. It could also arise as a result of
pouring the milk sample directly onto the acid in the butyrometer. To avoid charring, first, ensure the
acid concentration is right. Secondly, while adding the milk sample, let it flow on the side of the
butyrometer to avoid violent reaction when it drops on the surface of the acid.
• Light colored fat column (with or without brown specs at the interface) - may affect your ability to
read the fat content correctly. This problem is due to weak acid, which fails to dissolve all the milk non-
fat content. To avoid this problem, use sufficient volumes of the Gerber acid at the correct
concentration.
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 Fat estimation by Gerber Method
• Principle: The free fat globules rise to the surface by subsequent application of centrifugal
force to this mixture and heat produced due to mixing of acid and milk, causing melting of
fat. It facilitates the fat particles to come to the surface freely. The specific gravity of fat is
0.9 and that of acid milk mixtures is 1.43.
• Precautions:
1. Carryout through mixing of milk before testing.
2. Amyl alcohol must be pure.
3. Sulphuric acid is to be added gently by the sides of the butyrometer without wetting
the neck of the butyrometer. Avoid direct pouring of milk on acid.
4. The three fluids viz. Sulphuric acid, milk and amyl alcohol should be added gently, so
that they form three distinct layers.
5. Rubber stopper should be dry, clean and without crack.
6. Before centrifuging the butyrometer, see that there is no curdy white material left
undissolved.
7. The centrifugal machine must be properly balanced.
8. Always carry out the test in duplicate.
9. Butyrometer should be free from Na2Co3 (soda ash) if cleaned by Na2Co3 otherwise
it lowers the specific gravity and strength of Sulphuric acid.
10. Use butyrometer stand for shaking of butyrometer contents to dissolve the SNF
content of milk.
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 Fat Estimation by Soxhlet Apparatus
• Soxhlet Method: A Soxhlet extractor was to solubilize fat and fat is extracted from
originally designed for the extraction of food in combination with the solvent.
a lipid from a solid material. Typically, Later the fat is collected by evaporating
Soxhlet extraction is used when the the solvent .Al most all the solvent is
desired compound has a limited solubility distilled off and can be reused.
in a solvent , and the impurity is insoluble
in that solvent. It allows for unmonitored
and unmanaged operation while
efficiently recycling a small amount of
solvent to dissolve a larger amount of
material.
• Principle: Lipid in food present in various
forms like mono glycerides, di glycerides,
triglycerides and sterol and free fatty acid
and phospholipid and carotenoids and
fat-soluble vitamins. Lipid is soluble in
organic solvent and insoluble in water,
because of this, organic solvents like
hexane, petroleum ether have the ability

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 Fat Estimation by Soxhlet Apparatus
• Required Glass wares & Apparatus: knob.
• 1.Soxhlet Apparatus • 8.Continue extraction of sample for 6 hours.
• 2.Round Bottom Flask • 9.After extraction close the heating knob &
• 3.Extractor water supply .
• 4.Condensing unit • 10.Before removing round bottom flask try to
• 5.Thimble evaporate petroleum ether maximally for reuse.
• 6.Cotton • 11. After maximum collection of petroleum
ether remove the round bottom flask & place it
• 7.Filter Paper
in Hot air oven (at 105 deg c for1 hour) for
• 8.Hot Air Oven complete removal of petroleum ether.
• Procedure: • 12.After drying cool the flask and weigh the
• 1.Weigh 10 gm (W)sample in filter paper. flask(W2)
• 2.Place a filter paper in thimble & add weighed • 13.Note down the readings for calculation.
sample in it. • Calculation: W2-W1/W*100
• 3.Place cotton plug at top of the thimble. • Precautions:
• 4.Initially weigh empty round bottom flask(W1 ) • Due to the continuous running of water and a
& then fill about 150-200 ml of Petroleum ether heat source, it is not advisable to leave
in round bottom flask. the apparatus unattended.
• 5.Attach extractor to round bottom flask &
place thimble in extractor.
• 6.Attach condensing unit with extractor.
• 7.Start water in condensing unit & start heating 10
 Viscosity Meter
• Viscosity: Viscosity is a measure of a fluid's resistance to flow. It describes the internal
friction of a moving fluid. A fluid with large viscosity resists motion because its molecular
makeup gives it a lot of internal friction. A fluid with low viscosity flows easily because its
molecular makeup results in very little friction when it is in motion.
• Principle: Viscosity is the measure of a substance's resistance to motion under an applied
force. These viscometers use the principle of 'rotational viscometery', i.e. their
measurement of product viscosity is based upon immersing a specifically selected spindle
within a sample of the product followed by measurement of the torque required to rotate
the spindle at a set speed whilst immersed within the product sample.
• Factors affecting Viscosity:
• 1. Temperature
• 2.Concentration
• 3. Particle Size

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 Viscosity Meter

• For RM
Fill Beaker with (Sample
solution as per
FG Sample specification's
)

• Correct
Set Spindle No & spindle as per
Speed specifications

Dip Sample up to •
Record results
groove Mark & in Cp.
RUN it.

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 Ash Content
• Ash content is the fraction in biomass that is composed of incombustible mineral
material. Ash is present in the biomass in the form of certain inorganic compounds like
sodium, potassium, calcium, silicon, phosphorous, and chlorine.

• Principle: Ash is the residue remaining after the organic matter of a food has been
destroyed by incineration ,and it is the most convenient assessment of the total mineral
matter in the material.

• Appratus:
• 1.Muffale Furnace
• 2.Silica Crucible
• 3.Tong
• 4.Dessicator
• 5.Analytical Balance

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 Ash Content
• Procedure:
• 1. About 5 - 10 gm of the material is weighed (W1) accurately in a tared weighed (W) silica
crucible.
• 2. Placed silica crucible inside a muffle furnace at 5500C + 200C till the ash is free from any blank
carbon particle.
• 3. After completion of ashing the silica crucible is kept inside the desiccator to attain the room
temperature.
• 4. After cooling, the final weighment (W2) is noted.
• 5. The ash inside the silica crucible is kept for the determination of acid insoluble ash.(If
Required)
• 5.Calculation:
•
• Total ash (on dry basis), percent by weight = (W2 – W) x 100 x 100
• (W1 – W) x (100 – M)
•
• W1=Weight in g of the dish with the material taken for the test.
• W2 =Weight in g of the silica crucible with the ash,
• W=Weight in g of the empty silica crucible, and
• M=Moisture % of the sample.
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 Refractometer(Brix & RI)
• Refractometer: Refractometer is an optical device that can be used to measure the
refractive index of a substance (usually fluid) and, by converting the refractive index,
one can get the specific gravity, sugar content (Brix).
• Refractive Index n is the speed of light in vacuum relative to the speed of light in the
considered medium.
• Principle: A refractometer is an instrument that measures the concentration of liquid by
using the principle of light refraction.

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 Peroxide Value
• The peroxide value is defined as the amount of peroxide oxygen per 1 kilogram of fat or
oil. Traditionally this was expressed in units of milliequivalents.(Meq).
• Peroxide value is the most widely used method to measure the rancidity in fat /Oils.
(Primary Oxidation).
• Autoxidation: Autoxidation is a free radical reaction involving oxygen that leads to
deterioration of fats and oils which form off-flavors and off-odors .The double
bonds found in fats and oils play a role in autoxidation. Oils with a high degree
of unsaturation are most susceptible to autoxidation. The best test for autoxidation
(oxidative rancidity) is determination of the peroxide value. Peroxides are intermediates
in the autoxidation reaction.
• Principle :The peroxide value (PV) determines the concentration of hydroperoxide, the
primary oxidation products. The principle involves peroxides liberating iodine from
potassium iodide, i.e. The amount of peroxides is calculated back by the amount of
sodium thiosulfate (Na2S4O6) consumed.

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 Peroxide Value

Add 30 ml Solvent(G2:C3)
Place flask in Dark for 1
Weigh 3-5 gm Sample & Saturated Potassium
Min
Iodide

Add 30 ml D/W ,Mix and Titrate against 0.01N Thio

Add 1 ml Starch indicator sulphate(T.V.*N*1000/W)

Wilkote B.S. 0.5 Max.

All other Fat Variants 1 Max.(Meq./Kg)

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 Free Fatty Acid
• The acid value (AV) is a common
parameter in the specification of fats and Weigh 20 gm
oils. melted oil
• It is defined as the weight of KOH in mg sample
needed to neutralize the
organic acids present in 1g of fat. it is a
measure of the free fatty acids (FFA)
present in the fat and oil. Add 50 ml Ethyl
 FFA is a measurement of hydrolytic Alcohol & warm
oxidation of Fat. it

 End Point will be light pink color persist for

at least 30 sec. Mix it & Titrate
 Calculation: T.V.*N*28.2/Wt of sample against 0.1 N
 FFA for All Fat Variants 0.1 % Max. NaOH

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Rancidity

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 Iodine Value
• The iodine value (or iodine adsorption value or iodine number or iodine index) in
chemistry is the mass of iodine in grams that is consumed by 100 grams of a chemical
substance. Iodine numbers are often used to determine the amount of unsaturation in
fatty acids.
• Required Chemicals for Iodine Value :
 Potassium Iodide (15 % solution Freshly prepared)
 Distill water
 Starch as indicator (1% solution freshly prepared)
 0.1 N sodium Thio sulphate
 Wijs Solution
 Chloroform
• Required Glassware for PV:
 Iodine Flask
 Pipettes 1 ml.
 Measuring Cylinder
 Burette

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 Iodine Value
Wilkote B.S. 4 to 6

Wilkote 380 4 Max.

Take 1 gm
Sample in Place
RBD Palm Oil 45-50 Iodine Flask & iodine flask 15 ml Chloroform & 25
add in Dark for ml Wijs Solution
Chloroform 1 hour.
All other Variants 2 Max. Wijs Solution

Take out it Add 150 ml

from Dark D/W &
Use Starch as an
& add 20 titrate
indicator
ml KI against 0.1 N
Solution Thiosulphate

End Same
point is Procedure
repeat for .(VB-VS)*12.69*N/w
colorless Blank as
solution well

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 Protein Estimation By Pynes Method

Principle: When formaldehyde is added to milk which has previously titrated against
standard alkali to the end point of an indicator like phenolphthalein, it binds the amino groups
of the milk proteins and release an equivalent amount of proteins (H-ions), which could be
titrated against the alkali to the same end point. The amount of alkali used in the second
titration is a measure of the amino groups originally present in the proteins.
• Reagents :
• 1.Neutral formalin(Formaldehyde solution neutralized with NaOH)
• 2.Saturated Potassium Oxalate solution(Excess Potassium oxalate shall be present after
dissolution )
• 3. NaOH (0.1N )
• 4.Phenolphthalein indicator

• Apparatus :
• 1.Pipette 10ml, 2ml and 1ml
• 2.Graduated Burette
• 3.Conical flask 250ml

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 Protein Estimation By Pynes Method
• Procedure :
• 1.Pipette out 20ml sample of ice cream mix/10 % Solution of powdered sample
(e.g.SMP, WPC)
• 2. Add 5 drops of Phenolphthalein indicator .
• 3.Add 0.8ml /0.4 ml of Saturated Potassium Oxalate and keep it aside for 5 mins without
disturbing. 4.Titrate the sample against 0.1N NaOH soln to its end point pink color.
• 5. Record the burette reading V1 ml.
• 6. Add 4ml / 2 ml of neutral formalin, after addition of formalin, pink color will disappear
• 7.Again titrate with alkali till pink color persists for 10-15 secs. Record reading V2ml.
• Calculation :
• % of Protein in Ice-cream Mix =[(V2 - V1) x N x 9.04] – 0.14
• % of Protein in Milk= Vx1.7 where V is the corresponding Titre value after
neutralization where 1.7 is Pyne’s Constant

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 Bursting Strength
• Bursting Strength of materials tells about the property of the material to bear the stress
when an external pressure is applied to it.

• Procedure: Prepare a circular sample of dimensions larger than the annular clamp. The
dimensions of the sample are very important so that it does not slip during the testing.
• Clamp the sample appropriate on the annular clamp
• Using the ‘push to burst’ switch to apply the hydraulic pressure on the sample.
• The pressure is applied via a rubber diaphragm which inflates when hydraulic pressure is
increased.
• The point at which sample burst, the machine notes down the pressure applied.
• Using this bursting strength, bursting factor and bursting index can be determined.
• Bursting Factor=Bursting strength /Grammage (gf/cm2).

• Difference between BF & BS: Bursting factor is calculated to determine the amount of
pressure that a paper or cardboard can tolerate easily before it collapses. ... On the
other hand, bursting strength is measured by providing maximum hydrostatic pressure
to rupture the paper used to produce corrugated boxes.

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 GSM
• GSM (Gram Square Meter)
• The grammage of substance is defined as the weight of one square meter of
the paper or paperboard under standard test conditions and expressed as
grams per square meter.
• First Place template/Meter scale on test sample .
• Measure accurately 10 X 10 cm area with template/scale.
• Cut accurately five samples of the corrugated fiber board /Pure packs .
• Weigh the five samples correctly to 3 significant figures.

• Grammage (g/m2) = 10000 x W

AxB

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