Chemical properties Claudia Gadizza Perdani STP, M.Si.

References on Analytical Techniques

Official Methods;
- Association of the Official Analytical Chemists (AOAC)

- American Oil Chemists Society (AOCS)

- American Association of Cereal Chemists (AACC)

- Standar Nasional Indonesia (SNI)

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Criteria for Selecting an Analytical Technique

There are many techniques to analyze foods but

each has drawbacks or compromises.
You must select the technique that is required or
fits into your system.
For example, the most accurate techniques
generally take longer to perform and you may not
have the time if the food product you are making
requires real time results such as in the
formulation of processed meats.

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Criteria for Selecting an Analytical Technique
Precision Sensitivity
Accuracy Specificity
Reproducibility Safety
Simplicity Destructive/ Non-destructive
Cost On-line/off-line
Speed Official Approval

SAMPLING AND SAMPLE PREPARATION

What is the Purpose of the Analysis

Official Samples
Raw Materials
Process Control Samples
Finished Products 3
Sampling Plan

A sampling plan is a predetermined procedure for

the selection, withdrawal, preservation,
transportation and preparation of the portion to be
removed from a lot as samples.
The sampling plan should be a clearly written
document containing details such as;
- Number of samples selected
- Sample location (s).
- Method of collecting samples

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Factors Affecting a Sampling Plan
Purpose of inspection
-acceptance/rejection, variability/average

Nature of the product

-homogenous, unit, cost

Nature of the test method

-Critical/minor, destructive, cost, time

Nature of the population

-uniformity, sublot

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Developing a Sampling Plan
Number of samples selected
-Variation in properties, cost, type of analytical techniques
Sample location
-random sampling vs systematic sampling vs judgment sampling
Manner in which the samples are collected
-manual vs mechanical device

The Bottom Line in Sampling

Depending upon the nature of the material to be analyzed, you must
determine a method of taking small subsamples from a large lot (
5,000 lb blender, 20 combos on a truck etc) that accurately reflect the
overall composition of the whole lot.
An inaccurate sample of a large lot may actually be worse than no
sample at all. 6
Preparation of Laboratory Samples

You may have taken as much as 10 lbs of sub-

samples from a lot that now needs to be further
reduced in size;
-Make the sample homogeneous by mixing and grinding
and then more sub-sampling.
-Be aware of any changes that might occur between sampling and
analysis and take proper action ( e.g. enzymatic action, microbial
growth etc).
-Properly label the final sample with name, date/time, location, person
and other pertinent data.

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FOOD COMPONENTS
Food consists primarily of water( moisture), fat (or oil), carbohydrate,
protein and ash (minerals).
Since food consists of these 5 components, it is important that we
understand how these components are measured.

COMPOSITION OF FOODS
COMPONENT % Water %Carbohydrates %Protein % Fat % Min/Vit

Milk 87.3 5.0 3.5 3.5 0.7

Beef 60.0 0 17..5 22.0 0.9
Chicken 66.0 0 20.2 12.6 1.0
Fish 81.8 0 16.4 0.5 1..3
Cheese 37.0 2.0 25.0 31.0 5.0
Cereal grains 10-14 58-72 8-13 2-5 0.5-3.0
Potatoes 78.0 18.9 2.0 0.1 1.0
Carrots 88.6 9.1 1.1 0.2 1.0
Lettuce 94.8 2.8 1.3 0.2 0.9
Apple
84.0 15.0 0.3 0.4 0.3
Melon
92.8 6.0 0.6 0.2 0.4

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 pH DETERMINATION
pH Determination

pH refers to the relative amounts of acid and base

in a product.
It is scientifically defined as the negative log of the
hydrogen ion concentration.
pH ranges from 0 to 14 with pH of 7 being neutral.
pH values below 7 are considered acids and pH
values above 7 are basic or alkaline.
pH is generally determined with a pH meter
although litmus paper can also be used.

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 Moisture Determination
Moisture or water is by far the most common component in foods
ranging in content from 60 95%.
The two most common moisture considerations in foods is that of
total moisture content and water activity.

Moisture Content

The total moisture content of foods is generally determined by some

form of drying method whereby all the moisture is removed by heat
and moisture is determined as the weight lost.

% water = wet weight of sample-dry weight of sample

wet weight of sample
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Methods of Moisture Loss Measurement

Convection or forced draft ovens (AOAC)

- Very simple; Most common
Vacuum Oven
-Sample is placed in oven under reduced pressure thereby reducing
the boiling point of water.
Microwave Oven
-Uses microwave as a heat source; Very fast method
Infrared Drying
-Uses infrared lamp as a heat source; Very fast

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Water Activity (aw)

Water Activity (Aw) is the amount of free water in a sample that is

not bond and therefore free for microbial growth, enzyme and
vitamin decomposition and can reduce color, taste and flavor
stability.
Two general types of sensors:
Capacitance sensor: electrical signal
Chilled-mirror dew point method (AquaLab): dew point temperature change
due to ERH change.

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WATER ACTIVITY

Foods
Aw Microorganism Meat, fish, sausage, milk
1.0-0.95 Bacteria
Cheese, cured meat (ham), fruit juice
0.95-0.91 Bacteria conc

Fermented sausages (salami), dry

cheeses, margarine
0.91-0.87 Yeasts

Juice conc, syrups, flour, fruit cakes,

honey, jellies, preserves
0.87-0.80 Molds

Cookies, crackers, bread crusts

0.30-0.20 No microorganism
proliferation

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 PROTEIN ANALYSIS
PROTEINS
Proteins are made up of amino acids.
Amino acids are the building blocks of protein.
Nitrogen the most distinguishing element versus other food
components (carbohydrates, fats etc)
Nitrogen ranges in proteins : 13.4 - 19.1%
Non-protein nitrogen: free amino acids, nucleic acids, amino
sugars, some vitamins, etc.
Total organic nitrogen = protein + non-protein nitrogen

Types of Protein Analysis

Kjeldahl measures the amount of nitrogen in a sample.
Lowry- measures the tyrosine/tryptophan residues of
proteins. 14
Total organic nitrogen - Kjeldahl method
Crude protein content
Johan Kjeldahl (1883) developed the basic process
Principle: total organic N released from sample and absorbed by acid
Digestion: sulfuric acid + catalyst
Neutralization and distillation; Sodium hydroxide
Titration; Hydrochloric acid

Total organic nitrogen - Kjeldahl method

Digestion

Protein Sulfuric acid (NH4)2SO4

Heat, catalyst (ammonium sulfate)

Protein N NH4+ + H2SO4 (NH4)2SO4 15

Total organic nitrogen - Kjeldahl
method
Neutralization and distillation

(NH ) SO
4 2 4 + 2NaOH 2NH3 + Na2SO4 + 2H2O

-
NH3 + H3BO3 NH4+ : H2BO3 + H3BO3
(boric acid) (ammonium-borate complex)
excess

Color change

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Total organic nitrogen - Kjeldahl
method
Titration (direct titration)
-
H2BO3 + H+ H3BO3
(HCl)
Calculation
moles HCl = moles NH3 = moles N in the sample

%N = N*(HCl)
(mL acid sample-mL acid blank) 14g N
%N = N*(HCl) 100
N*=Normality of HCl 1000 g sample mole
(mL acid sample-mL acid blank)
1.4
g sample
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Total organic nitrogen - Kjeldahl
method
Calculation
%Protein = %N conversion factor
Conversion factor: generally 6.25
most protein: 16% N

Conversion factor
egg or meat 6.25
milk 6.38
wheat 5.33
soybean 5.52
rice 5.17
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Kjeldahl Apparatus

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Total organic nitrogen - Kjeldahl
method
Advantages:
applicable to any foods
simple, inexpensive
accurate, official method for crude protein content
Disadvantages:
measuring total N not just protein N
time consuming
corrosive reagents

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Lowry Method

Principle: Color formation between tyrosine and

tryptophan residues in protein and Biuret reagent
and Folin-Ciocalteau phenol reagent (750 nm or 500
nm).
Procedure
protein solution + biuret reagent
(20-100 g) room temp10 min
+ Folin reagent
50C 10 min
650 nm

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Lowry Method

Advantages
most sensitive (20-200g)
more specific, relatively rapid
Disadvantages
color development not proportional to protein
concentration
color varying with different proteins
interference (sugars, lipids, phosphate buffers, etc)

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Infrared Spectroscopy

Principle: absorption of radiation of peptide bond at

mid-infrared (MIR) and near-infrared (NIR) bands
Advantages
NIR applicable to a wide range of foods
rapid, nondestructive
little sample preparation
Disadvantages
expensive instruments
calibration for different samples

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 Crude Fat Analysis
Fats
Fats refers to lipids, fats and oils.
The most distinguishing feature of fats versus other components
( carbohydrates, protein etc) is their solubilty. Fats are soluble in organic
solvents but insoluble in water.

Solvent Extraction Methods

Sample preparation: Best under nitrogen & low temperature
Particle size reduction increases extraction efficiency
Predrying sample to remove water is common.

Solvent Extraction Methods

Solvent selection nonflammable
Ideal solvent nontoxic
high solvent power for lipids good penetration into sample
low solvent for other components single component
easy to evaporate inexpensive
low boiling point non-hygroscopic 24
Solvent Extraction Methods
Common Solvents
Ethyl ether - best solvent for fat extraction, more expensive, explosion, fire hazard,
hygroscopic
Petroleum ether - cheaper, more hydrophobic, less hygroscopic
Hexane - soybean oil extraction

Types of Fat Analysis

Extraction Methods
Continuous Goldfinch
Semi-Continuous- Soxhlet
Discontinuous- Mojonnier

Instrumental Methods
Dielectric
Infrared
Ultrasound 25
 Solvent Extraction Methods
Continuous extraction: Goldfish method
Principle: Solvent continuously flowing over the sample with no build-up
Advantages: fast, efficient.
Disadvantages: channeling not complete extraction.

Semicontinuous extraction: Soxhlet method

Principle: Solvent building up in extraction chamber for 5-
10 min before siphoning back to boiling flask.
Advantages: no channeling
Disadvantages: time consuming
Discontinuous extraction: Mojonnier method (wet method
extraction)
Principle: a mixture of ethyl ether and petroleum ether in a Mojonnier flask
Advantages: no prior removal of moisture
Disadvantages: constant attention

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Instrumental Methods
Dielectric method
Principle: low electric current from fat
Infrared method
Principle: Fat absorbs infrared energy at a wavelength of 5.73 m
Ultrasound method
Principle: sound velocity increases with increasing fat content

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 CARBOHYDRATE ANALYSIS

Introduction
Next to water, carbohydrates are the most abundant food component
%carbohydrate=100% - (H2O + ash + fat + protein)
Types of carbohydrates include;
monosaccharide: glucose, fructose, galactose
disaccharide: sucrose, lactose, maltose
oligosaccharids: raffinose
polysaccharide: starch, cellulose

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 Ash and Mineral Analysis
Definitions
Ash: total mineral content; inorganic residue
remaining after ignition or complete oxidation of
organic matter
Minerals:
Macro minerals (>100 mg/day)
Ca, P, Na ,K, Mg, Cl, S
Trace minerals (mg/day)
Fe, I, Zn, Cu, Cr, Mn, Mo, F, Se, Si
Ultra trace minerals
Va, Tn, Ni, Sn, B
Toxic mineral
lead, mercury, cadmium, aluminum
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Ash Contents in Foods

Wheat flour, whole grain 1.6%

Macaroni, dry, enriched 0.7%
Milk, whole, fluid 0.7%
Butter, with salt 2.1%
Apple, raw with skin 0.3%
Banana, raw 0.8%
Egg, whole, raw 0.9%
Hamburger, regular, plain 1.7%

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Methods for Determining Ash
Dry ashing
high temperature
Wet ashing
oxidizing agent and/or acid
Low-temperature plasma ashing
dry ashing in partial vacuum at low temperature

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Dry Ashing
Principles
High temperature (>525C) overnight (12-18 hr)
total mineral content
Instrumentation
Muffle furnace
Crucible
quartz
porcelain
steel
nickel
platinum

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General Procedure for Dry Ashing

1. 5-10g pretreated sample into a crucible

2. Ignite crucible to constant weight at ~550C for 12-18 hr
3. Cool in desiccator
4. Weigh cooled crucible

% ash (db) = 100

wt after ashing - crucible wt

Sample wt solid%/100

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Dry Ashing

Advantages
safe and easy
no chemical
many samples handled at one time
resultant ash for further mineral analysis
Disadvantages
loss of volatiles
interaction
long time and expensive equipment

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Ion-Selective Electrodes
Direct measurement via chemical potential of cations (Ca, Na, K),
anions (Br, Cl, F), or even dissolved gases (O2, CO2)
Components:
sensing electrode
reference electrode
readout device
Types: glass membrane, polymer-body, solid-state

Activity (A) vs. Concentration (C)

A=C =activity coefficient
A: chemical activity
C: a measure of ions in solution
is a function of ionic strength; ionic strength is a function of concentration
and charge on all ions
AC

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Ion-Selective Electrodes
Advantages Applications:
more precise, rapid, processed meats: salt,
practical nitrate
direct measurement of a butter and cheese: salt
wide range of ions milk: Ca
inexpensive and simple low-sodium products:
Disadvantages sodium
inability to measure below soft drink: CO2
2-3 ppm wine: Na, K
unreliable at low can vegetable: nitrate
concentration (10-4M)

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