Physico-Chemical Evaluation of Feeds
Physico-Chemical Evaluation of Feeds
Physico-Chemical Evaluation of Feeds
PHYSICO-CHEMICAL
ANALYSIS
by
Margarita T. Arnaiz
OUTLINE
1. Introduction
2. Physical evaluation
3. Chemical evaluation
a. proximate analysis
moisture, crude protein, crude fat, crude fiber, ash, NFE
c. anti-nutrient contents
d. others
4. Exercise (problem solving)
Evaluation methods
Physical rely on the physical characteristics of
feedstuffs but little information on the nutritional value
Chemical characterize or define the chemical nutrients
present
Biological utilized live organisms, i.e., actual feeding
experiment; ascertains the true value of the feedstuff to
the animal; time-consuming, expensive, and requires
facilities for holding the animals
Microbiological use of microorganisms to evaluate
presence of nutrients;
Physical Evaluation
Use of sense
smell rancidity of oil
taste off flavors
visual extraneous matters; homogeneity of ingredients
touch wetness, dryness
Chemical Evaluation
Proximate analysis
Developed by Wendee Experimental Station in
Germany over a 100 years ago
most generally used chemical scheme for describing
feedstuffs
general overview of the chemical composition feeds,
partitioning compounds with common chemical
properties
Proximate analysis
Moisture
Crude protein
Crude fat or ether extract
Crude fiber
Ash
Nitrogen free extract
Moisture
Moisture free sample
Digestion
Distillation
Titration
Ether extraction
Crude protein, CP
Ash
Boil in acid, boil in alkali
Oven dry@105C, incinerate
@ 550C
100-(CP+Cft+Cfb+Ash)
NFE
Sample preparation
formulated feed
as received
Ground feed
grinder
% moisture determination
Moisture
Moisture
analyzer
Spl, 0.1g
Digested sample
Catalyst,
CuSO4+K2SO4
To Kjeltec apparatus
H2SO4,conc
6 mL
Digestion at 420oC
3.1.NaOH added,
3.2.NH3(g) directed towards vessel
with boric acid + indicators
3.3.Titration with std acid (HCl) with
end point electrode sensor
3.4. automatic calculation of %Cp
Conversion Factor
Oil seed
proteins
Cereal
proteins
Plant leaf
Animal or fish
18.5
5.4
17
5.9
15
16
6.6
6.25
Steps;
1. boiling in solvent (petroleum ether)
2. Draining of solvent
3. Recovery of solvent
4. Drying of cups with fat residue at 105oC
Calculation :
Soxtec 2055 (automated)
%Fat = [(Wt cup with fat residue-Wt cup) /Wt sample]x100
Crude fiber
Fibertec
Ash
Inorganic residue
obtained by burning off
organic matter
Residue after burning
allows for mineral analysis
Reference Test Method:
AOAC 942.05
Sample is burned in
furnace for 2 hours at
600oC. The loss in weight
is the amount of ash.
Furnace
Determination
Moisture
Crude fat/ether
extract
Crude fiber
Crude protein
% N x 6.25)
Ash
NFE
By difference
100-(CP+Cfb+Cft+Ash)
Energy content
www.techknow.org.uk
Bomb calorimeter
Amino acids
Fatty acids
Vitamins
Minerals
Anti-nutrient contents
Endogenous (urease,trypsin inhibitors, etc)
Exogenous ( aflatoxins, pesticides, heavy metals, etc)
Feed additives
Antibiotics, antioxidants ( ex. Ethoxyquin)
Amino acids
Amino acids (AA) can be
analyzed using a dedicated
amino acid analyzer or high
performance liquid
chromatograph (HPLC)
Purified proteins are hydrolyzed
and injected in HPLC with
fluorescence detector and cation exchange resin column.
Mobile phase with varying ionic
strength and pH carries the
sample across the column
where separation of different AA
takes place. Identification and
quantification is effected by
calibration of an AA standard
solution
HPLC
Fatty acid
Fatty acids serve as source of
energy for many physiological
functions in organisms
Can be analyzed using gas
chromatograph (GC) with flame
ionization detector
Lipid/fat are extracted from the
sample using organic solvents
(chloroform-methanol),
saponified and formed into
esters prior to injection in the
GC. The sample upon injection
is carried by an inert gas (e.g.
He) across a capillary column
where separation takes place.
Identification and profiling is
aided by a mixed fatty acid
standard.
GC
Vitamins
Variety of nutrients which
do not include proteins,
lipids, and carbohydrates
Required in small
amounts for normal
growth
Can be water soluble
( thiamin, riboflavin,
ascorbic acid, etc)
Some vitamins can be
analyzed using HPLC
Minerals
Inorganic components
which can be analyzed
from the ash (Ca, P, Fe,
etc)
Can be analyzed using
atomic absorption
spectrophotometer
(AAS), inductively
coupled plasma (ICP),
and also by colorimetric
methods
Anti-nutrients determination
Feed additives
Feed components that can affect food safety of
aquaculture products
Antibiotics
- added to feeds to control diseases (treatment and
prophylaxis) or as growth enhancer
Anti-oxidants
- added to feeds to prevent their deterioration, especially
oxidation of fats (prevention of rancidity), ex. Ethoxyquin
Mostly analyzed and widely accepted method of analysis
is by the use of HPLC
Solvent removal
Filter
( 0.22m membrane filter)
Inject in HPLC
Wt of dish
Wt initial
Wt Final
Trial 1
22.0879
24.3992
24.3435
Trial 2
24.8102
26.9212
26.7729
Ash
Wt crucible
Wt Spl
Wt crucible+ash
Trial 1
19.3038
2.0211
19.9151
Trial 2
20.6886
2.0121
21.2946
CP
Wt sample
%N
Trial 1
0.1019
8.857
Trial 2
0.1063
8.8684
CFt
Wt cup
Wt sample
Wt cup +fat
Trial 1
22.397
1.0009
22.4229*
Trial 2
22.3373
1.0422
25.3678*
Cfb
Wt crucible @
105
Wt crucible @500
Net wt of Cfb
Trial 1
29.517
29.49
0.0188*
Trial 2
27.0086
27.0086
0.0139*
%Moisture
%Ash
%CP
%Cft
% Cfb
Average
THANK YOU
Wt of dish
Wt initial
Wt Final
%Moisture
Average(%)
Trial 1
22.0879
24.3992
24.3435
11.31
11.32
Trial 2
24.8102
26.9212
26.7729
11.33
Ash
Wt crucible
Wt Spl
Wt crucible+ash
%Ash
Trial 1
19.3038
2.0211
19.9151
30.25
Trial 2
20.6886
2.0121
21.2946
30.12
CP
Wt sample
%N
%CP
Trial 1
0.1019
8.857
55.36
Trial 2
0.1063
8.8684
55.43
CFt
Wt cup
Wt sample
Wt cup +fat
%Cft
Trial 1
22.397
1.0009
22.4229*
2.59
Trial 2
22.3373
1.0422
25.3678*
2.93
Cfb
Wt crucible @
105
Wt crucible @500
Net wt of Cfb
% Cfb
Trial 1
29.517
29.49
0.0188*
1.88
Trial 2
27.0086
27.0086
0.0139*
1.33
30.18
55.39
2.76
1.61
Problem: Estimate the nitrogen free extract (NFE) of the sample (10.O6%)