Proximate Analysis

UDAA1124
AGRICULTURAL CHEMISTRY
 Accuracy vs Precision

Accuracy Precision
How closely a How closely
measured value repeated
Proximate analysis

agrees with the measurements of

correct value the same quantity
agree with another

Measurements are frequently repeated to

improve accuracy and precision
Average values obtained from several
measurements are usually more reliable than
individual measurements
 Accuracy vs Precision

Accuracy Precision
How closely a How closely
measured value repeated
Proximate analysis

agrees with the measurements of

correct value the same quantity
agree with another

Measurements are frequently repeated to

improve accuracy and precision
Average values obtained from several
measurements are usually more reliable than
individual measurements
 Moisture determination

Air oven
Purpose:
The method is used for the
quantitative determination of
Proximate analysis

moisture in all foods except

food samples high in sugar and
fat (> 10%)

Definition:
Moisture in this method refers to the amount of free water and volatile
substances that are lost by drying the food under controlled
temperature in an air oven

Principle:
The method is based on the drying of food sample under controlled
temperature until constant weight is obtained. Moisture content is
required to express the nutrient content per dry weight basis.
In some foods, moisture is used to indicate their quality.
 Moisture determination

Vacuum oven
Purpose:
The method is used for the
quantitative determination of moisture
in food products high in sugar and
Proximate analysis

fat (> 10%)

Definition:
Moisture in this method refers to the amount of free water and volatile
substances that are lost by drying the food under vacuum and
controlled temperature

Principle:
Weighed samples are placed under reduced pressure (typically 25-100
mm Hg) in a vacuum oven for a specified time and temperature and
their dried mass is determined. The boiling point of water is reduced
when it is placed under vacuum. Lower temperatures can be used to
remove the moisture.
 Crude protein determination

Kjeldahl method
Purpose:
The method is used for the
quantitative determination of crude
protein in all types of foods.
Proximate analysis

Definition:
Crude protein is total nitrogen multiplied by protein factor. Total
nitrogen content includes nitrogen primarily from proteins and to a
lesser extent from all organic nitrogen containing non-protein
substances

Principle:
Digestion
The method is based on the digestion of proteins and other organic
food components in the sample with sulfuric acid in the presence of
catalyst e.g. sodium or potassium sulfate to release nitrogen from
protein and retain it as ammonium salt.
 Crude protein determination

Kjeldahl method

Principle:
Distillation
Ammonia gas is liberated upon addition of
Proximate analysis

excess alkali (concentrated sodium hydroxide)

and is distilled into a boric acid solution to form
ammonium-borate complex.

Titration
The ammonia liberated from the complex is
titrated with standardised hydrochloric acid. The
amount of nitrogen in the sample is determined
from the milligram equivalent of the acid used.

Crude protein is determined by multiplying the nitrogen content

with a conversion factor specific to the food matrix.
 Ash determination

Ash drying
Purpose:
The method is used for the quantitative
determination of ash in raw, cooked and
Proximate analysis

processed foods

Definition:
Ash content refers to the total mineral
residue left after incineration of organic
matter. The value for ash is a useful check
in summing up the proximate composition
of food and a measure of its mineral
content
 Ash determination

Ash drying

Principle:
The method involves oxidation of all organic matter by incineration in a
furnace at a specified temperature (<550°C). Ashing above 650°C will
Proximate analysis

volatilise inorganic salts like alkali chloride and a portion of ash will fuse
and enclose some carbon, preventing them from being ignited. The
residue left after incineration is the ash content of the sample

If ash samples are to be used for mineral analysis, temperature

of the furnace should not exceed 450°C. Too high a temperature may
cause the volatilisation of certain elements particularly Fe, K, Na, S, Cl
& P. It may also cause the mineral matter to fuse and melt.

The most widely used crucibles porcelain because

• it is relatively inexpensive to purchase,
• can be used up to high temperatures (< 1200oC)
• easy to clean
• resistant to acids but can be corroded by alkaline samples

Prone to cracking if they experience rapid temperature changes

 Ash determination
Wet Ashing
Principle:
Wet Ashing is treatment of the sample with a moderate amount of
strong acids and oxidizing agents (e.g., nitric, perchloric and/or sulfuric
acids) before charring. Charring is performed using an open flame.
Proximate analysis

Liquid samples tend to foam. The resulting solution can then be

analyzed for specific minerals
Breaks down and
removes the organic
matrix surrounding the
minerals (organic matter is
completely digested,
leaving only the mineral
oxide in solution)

Typically, a digestion
takes from 10 minutes
to a few hours at
temperatures of about
350oC
 Crude lipid determination

Semi-continuous solvent extraction

Purpose:
The method is commonly used to increase the
efficiency of lipid extraction from foods.
Proximate analysis

Definition:
Oil and fat from solid material are extracted by
repeated washing (percolation) with an organic
solvent, usually hexane or petroleum ether,
under reflux in a special glassware

Principle:
The basic procedure calls for a solid sample
to be placed in a porous container and
allowing condensed solvent to extract
continuously.
 Crude lipid determination

Semi-continuous solvent extraction

Several solvent extraction systems based on the Soxhlet device are

on the market to allow fast and safe determination of total lipids in
food, soil
Proximate analysis

Soxtherm extractors from Gerhardt GmbH was developed to reduce

extraction times.
 Crude lipid determination

Batch Solvent Extraction

Purpose:
The ideal solvent for lipid extraction would completely
extract all the lipid components from a food, while
leaving all the other components behind
Proximate analysis

Definition:
The total lipid content determined by solvent extraction
depends on the nature of the organic solvent used to
carry out the extraction: the total lipid content
determined using one solvent may be different from
that determined using another solvent
Principle:
The efficiency of solvent extraction depends on the polarity of the lipids
present compared to the polarity of the solvent. Polar lipids (eg.
glycolipids or phospholipids) are more soluble in polar solvents (eg.
alcohols), than in nonpolar solvents (eg. hexane). On the other hand,
nonpolar lipids (eg. triacylglycerols) are more soluble in nonpolar
solvents than in polar ones
 Crude lipid determination

Batch Solvent Extraction

Principle:
Mixing the sample and the solvent in separatory funnel. The separatory
funnel is shaken vigorously and the organic solvent and aqueous phase
are allowed to separate (either by gravity or centrifugation). The
Proximate analysis

aqueous phase is then decanted off, and the concentration of lipid in the
solvent is determined by evaporating the solvent and measuring the
mass of lipid remaining

A solvent should be inexpensive, relatively low boiling point, nontoxic

and nonflammable.
It is difficult to find a single solvent which meets all of these requirements
Proximate analysis
 Dry matter basic

Why dry matter determination?

Dry matter content of a feedstuff is important because it reveals the
actual amounts of various nutrients available
to the animal consumig the feed.
Feed analysis
 Dry matter basic

Moisture content = 11.1200%

Dry matter = ?
88.8800%
Feed analysis

Ash % = 4.3500%
Ash % (dry matter) = ?
43.500/88.8800%
=4.8942
 Exercise
1.
Below are the various sets of results expressed in
mean ± s.e., choose the result(s) with high precision.
I. 1.0089 ± 0.0020
s.e standard error
II. 2.0030 ± 0.0002
Feed analysis

III. 3.3333 ± 0.0030

IV. 1.0000 ± 0.9999

A. I only
B. IV only
C. I, II
D. I, II, III
 Exercise
2.
Choose the correct statement(s).
I. Average values obtained from several
measurements are usually more reliable than
individual measurements.
II. Measurements are frequently repeated to improve
Feed analysis

accuracy and precision.

III. Accuracy is a determination of the reproducibility of
a measurement.
IV.Precision is a measure of how closely the value
determined agrees with a known or accepted value

A. I, II
B. III, IV
C. I, II, IV
D. All of the above
 Exercise
3.

Vacuum oven is commonly used for the quantitative

determination of moisture in _____________.

A. all foods
Feed analysis

B. food samples high in sugar and fat (> 10%)

C. food samples with lower sugar and fat (<10%)
D. food samples high in protein content