Chemical Analysis

Proximate analysis:
Estimation of the main components of a food using procedures that allows a reasonably rapid
and acceptable measurement of various food fractions without the need for sophisticated
equipment or chemicals.
Aim: Quality evaluation of raw material

1. Moisture
Material:
 Sample

Apparatus:
 Infrared Moisture Meter

Operational procedure:

 To start drying process with use of program libraries, switch on access to

programs libraries P6-01 Libr
 Moisture analyzer has 20 libraries, which can be freely configured, saved and use
by entering specific library
 To change parameters settings, enter to drying menu with key Test menu
 Group marker way of drying process finish will start flashing. Next using
navigation key Right, start settings of parameters group
 With navigation keys Up & Down, set one of options and accept changes
(pressing twice key Print/Enter) or continue setting the parameters (pressing once
key Print/Enter)
 Start drying procedures by pressing key Start/Stop and next, after unloading the
pan, press key TARA
 Open drying chamber, place tested sample on pan of moisture analyzer and close
drying chamber
 Moisture analyzer will finish drying process with beep sound
 To change during process displayed data press key Display

2. Ash
Material:
 Sample

Apparatus:
 Muffle Furnace
 Silica Crucibles
 Weighing Balance
 Desiccator
  Heating Source

Operational procedure:

 The total ash content of the sample was determined using muffle furnace
 5 g of sample was accurately weighed into a crucible (which was previously
heated to about 600 °C and then cooled)
 The crucible was placed on a tripod and get heated first over a low flame till all
the material was completely charred, followed by heating in a muffle furnace for
about 3-5 hours at 600 °C
 It was then cooled in desiccator and weighed
 The percentage of ash was calculated by using the following equation

Weight of Ash(g)
% Ash = Weight of Sample(g) × 100

3. Fat
Materials:
 Sample
 Petroleum ether

Apparatus:
 Socs Plus Apparatus
 Thimble
 Beaker
 Weighing Balance
 Desiccator
 Hot Air Oven

Operational procedure:

 Rinse all the beakers and place them in oven with temperature about 110 °C and
also the samples
 After removal of moistures from the beakers, place them in desiccator about 5
minutes to bring them to room temperature
 Now weight the empty beaker and let the weight be W 1. This is initial beaker
weight (IBW)
 Now insert the thimble holder and place it on the beaker
 Weigh the samples and transfer them to the thimble. Let the sample weight be
W. Sample weight may be 1 to 2 grams
 Pour the solvent in the beaker. The volume may be 80 ml
 Load all the beakers in the system. Switch ON the system and set the boiling
point of solvent as the boiling temperature. The boiling temperature may be 10
°C to 20 °C higher than that of solvent’s boiling point. Ex: Boiling point of
 Ether is 60 °C than Boiling temperature can be 80 °C
 Leave the process about 45 to 60 minutes. After the process time, increase the
temperature to recovery temperature (boiling point X 2) EX: If the boiling point
of Ether is 60 °C, recovery temperature can be 120 °C
 Now do the rinsing about 2 times in order to collect the remaining fat that may
present in the sample
 Now takeout all the beakers from the system and put in the hot air oven. After 15
to 20 minutes, for removal of traces of solvent. Take out all the beakers and
place them in to desiccator about 5 minutes
 Take out all thimble holders and weigh the beaker. This is the final weight of the
beaker (FBW). Let the weight be W2. By substituting W, W1, W2 in the
following formula, the amount of fat present in the sample can be calculated
using following equation

W 2−W 1
% Fat = ×
W

4. Fibre
Materials:
 Sample
 1.25% H2SO4
 1.25% NaOH
 Distilled water

Apparatus:
 Fibra plus apparatus
 Weighing balance
 Fibra plus crucibles
 Hot air oven
 Muffle furnace
 Desiccator

Operational Procedure:
 Weigh the samples accurately and note down
 Transfer the weighed samples in to oven dried
 Place the crucibles in to the metal adaptors of Fibra Plus hot extraction unit and
ensure proper sealing of crucibles against
Acid wash:

 Pour 150 ml of 1.25 % H2SO4 in to the extractors from top

 Switch on the instrument and set the initial temperature to 500 °C
 After boiling starts, reduce the temperature to 400 °C
 Allow the samples to boil for 45 minutes in acid
 After 45 minutes boiling, drain the acid and wash the samples twice or thrice
with distilled water (till free from acid. During draining, ensure that the knob is
in vacuum mode
 If the draining is not effective due to clogging of sample in the crucible, then
keep the knob in pressure mode, press the pressure button twice or thrice and
immediately turn the knob to vacuum mode

Alkali wash:

 Pour 150 ml of 1.25 % NaOH into the extractors from top

 After 45 minutes boiling, drain the alkali, wash the samples twice or thrice
with distilled water (till free from alkali)
 During draining ensure that the knob is in vacuum mode
 If the draining is not effective due to clogging of sample in the crucible, then
keep the knob in pressure mode, press the pressure button twice or thrice and
immediately turn the knob to vacuum mode
 After alkali wash take out crucibles and dry it in hot air oven until the
crucibles are free from moisture
 Cool down the hot crucible to room temperature using a desiccator
 Now weigh the crucibles and record the weight (CWAA = W2)
 Place all the crucibles in the muffle furnace at 500 °C for ashing
 Cool down the hot crucible after ashing

Calculation:

 Sample weight = W
 Crucible weight before ashing (CWBA) = W1
 Crucible weight after ashing (CWAA) = W2
 W3 = W1 - W 2

W3
% Crude Fibre = ×100
W
5. Protein

Materials:
 Sample
 Sodium/Potassium Sulphate
 Copper Sulphate
 Concentrated Sulphuric Acid
 Distilled Water
 Alkali
 Boric Acid
 KMnO4

Apparatus:
 Kelplus/ kelvac/ kelflow system
 Digital weighing balance
 Digestion tubes
 Auto titrator/manual burette
 Digestion system

Operational Procedure:

 Grind the sample

 Weigh the sample in a digital precision balance
 Transfer the sample to the Digestion tube
 Add digestion mixture (Sodium or Potassium Sulphate and Copper
Sulphate) to the sample (digestion tube)
 Add required volume of concentrated sulphuric acid to the digestion tube
 Using SS insert rack, take samples to the digestion system
 The manifolds are permanently connected to Kelflow or Kelvac
 Place the manifold with manifold rack over the digestion tube
 The water inlet to Kelflow is connected to water tap
 Load the insert rack with tube and manifold into the block digestion system
 Open the water tap for switch on the Kelvac system
 Ensure the water drains out through the outlet of the Kelflow system
 Ensure proper sealing of exhaust manifold to avoid any leakage of acidic
fumes
 Switch ON the power switch
 Set the temperature in the temperature controller
 If frothing is observed in the sample, digest the sample, in low temperature
for some time and increase the set temperature
 After the digestion of samples are over, separate the sample from the system
 Remove the insert rack with samples and place it over the tube support rack
 Allow water to flow through the Kelflow for some time, to remove the
 fumes from the tube
 Remove the manifold from the tubes
 Place the manifold in the manifold support rack
 Take out the sample one by one and dilute with distilled water
 Dilution is necessary to avoid solidification of sample on cooling
 The diluted sample is placed in the distillation system for Ammonia
recovery
 Keep the Alkali, Boric acid and KMnO 4 reservoirs to the rear of the
distillation system. Immerse the hose coming out from the system in to the
respective reservoir tanks. Select the program number by operating the
PGM Key
 Wait for the “READY” indication in the Control Panel
 Open the water inlet to the water condenser
 Press “RUN” key to activate the program
 Ensure addition of reagents and steam
 Reagents can also be added manually
 Water refilling to the boiler takes place automatically
 During refilling of water ensure an alarm and an indication
 Ensure the distilled water reservoir always filled with sufficient distilled
water
 Ensure collection of Ammonia in the receiver solution
 Remove the receiver with Ammonia and take to an Auto Titrator / manual
burette for titration. Titrate either by direct or back titrate to estimate the
Nitrogen Percentage

Weight of Nitrogen (g)

% Nitrogen = Weight of Sample (g) ×100

 Once the nitrogen content has been determined it is converted to a

protein content using the appropriate conversion factor

% Crude Protein = % Nitrogen × 6.25