Kwame Nkrumah University of

Science & Technology, Kumasi, Ghana

Ash and Mineral

Analysis

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Ash and Minerals Content of Foods

• Ash content is a measure of the total amount

of minerals present within a food

• Mineral content is a measure of the amount

of specific inorganic components present
within a food such as Ca, Na, K and Cl

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 Determination of Ash Content
• Ash refers to the inorganic residue remaining after
either ignition or complete oxidation of organic matter
in a foodstuff.
• Principle:
– Minerals are not destroyed by heating, and that they
have a low volatility compared to other food
components.
• Two major types of ashing used:
– Dry ashing: primarily for proximate composition and some
type of specific mineral analysis
– Wet ashing (oxidation): as a preparation for the analysis of
certain minerals
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 Sample Preparation
• Solid food are finely ground and mixed
• Sample with high moisture content are often
dried to prevent spattering during ashing
• Sample with high fat content are usually
defatted
• Mineral from grinders, glassware or crucible
should be prevented to come into contact with
sample.
• Deionized water is recommended to avoid
incorrect results.
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 Dry Ashing
• Dry ashing refers to the incineration at high
temperatures of 500-600°C by the use of muffle
furnace

• Water and volatile materials are vaporized and organic

compounds are burned in the presence of oxygen in
air to CO2, H2O and N2

• Most minerals are converted to oxides, sulfates,

phosphates, chlorides or silicates

• The sample is weighed before and after ashing

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• The ash content can be expressed on either a dry or
wet basis.

% ash (wet basis) = weight after ashing x 100

weight before ashing (wet)
% ash (dry basis) = weight after ashing x 100
weight before ashing (dry)

% ash (dry basis) = wt after ashing – tare wt of crucible x 100

original sample wt x dry matter coefficient

dry matter coefficient = %solids/100

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• There are different types of crucible available for ashing food
samples: quartz, pyrex, porcelain, steel and platinum crucibles.
• Selection of an appropriate crucible depends on the sample
being analyzed and the furnace temperature used.
• Quartz crucibles are resistant to acids and halogens but not
alkali, at high temperatures.
• Vycor brand crucibles are stable to 900 oC
• Pyrex Gooch crucibles are limited to 500oC
• Porcelain crucibles - most widely as relatively inexpensive,
can be used up to high temperatures (< 1200oC) and are easy
to clean. They are similar to quartz crucibles in properties but
will crack with rapid temperature changes.
• Platinum crucibles – very inert but far too expensive
• Quartz fiber crucibles - disposable, unbreakable and can
withstand temperatures up to 1000 oC
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 Dry Ashing
• Advantages:
– Safe, few reagent are required, many samples
can be analyzed simultaneously, not labor
intensive, and ash can be analyzed for specific
mineral content
• Disadvantages:
– Long time required, muffle furnace are quite
costly to run due to electrical costs, lost of
volatile minerals at high temperature e.g., Cu,
Fe, Pb, Hg, Ni and Zn

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 Wet Ashing
• Also called wet oxidation or wet digestion

• Primarily used in the preparation of samples for

specific mineral analysis and metallic poisons

• It breaks down and removes the organic matrix

surrounding the minerals so that they are left in an
aqueous solution.

• A dried ground food sample is usually weighed

into a flask containing strong acids and oxidizing
agents (e.g. nitric, sulfuric acid) and then heated.
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 Wet Ashing
• Heating is continued until the organic matter is
completely digested, leaving only mineral oxides
in solution.

• Advantages: little loss of volatile minerals due to

low temperature used, more rapid than dry ashing

• Disadvantages: labor intensive, requires a special

fume-cupboard, low sample throughput

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 Low Temperature Plasma Ashing
• A sample is placed into a glass chamber which is
evacuated using a vacuum pump
• A small amount of oxygen is pumped into the chamber
and broken down to nascent oxygen by application of
an electromagnetic radio frequency field
• The organic matter in the sample is rapidly oxidized by
the nascent oxygen and the moisture is evaporated
because of the elevated temperatures
• Advantages: Less chance of losing trace elements by
volatilization
• Disadvantages: relatively expensive equipment and
small sample throughput
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 Other Ash Measurements
1. Soluble and Insoluble Ash in Water
– An index of fruit content of preserves and jellies
2. Ash Insoluble in Acid
– Use to measure surface contamination of fruits and
vegetables, wheat and rice coating. Contaminants are
generally silicates and remain insoluble in acid except HBr
3. Alkalinity of Ash
– Used as quality index of fruit and fruit juices
– Ash of fruits and vegetables is alkaline, ash of meats and
some cereals is acid.
4. Sulfated Ash
– Applied to sugars, syrups, and colour additives
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 Determination of Specific Mineral Content
• Methods include: complexometric titations,
precipitation titrations, colorimetry, atomic ion-
selective electrodes or absorption spectroscopy

• The most effective means of determining the type

and concentration of specific minerals in foods is to
use atomic absorption or emission spectroscopy

• A near-infrared spectroscopy allows for of mineral

estimation without destruction of carbohydrates, fats,
protein and vitamins that make up foods
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 Sample Preparation
• Factors such as pH, sample matrix, temperature, and
other analytical conditions and reagents can interfere
with the analytical method to quantify a mineral
• The sample need to be ashed using one of the method
previously described
– Reason: to isolate the minerals from the organic
matrix surrounding them
• Presence of contaminants in the water, reagents or
glassware
– Recommended to use ultrapure water and/or
reagents
– Use the same glassware and reagents for blank
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 Gravimetric Analysis
• The element to be analyzed is precipitated from
solution by adding a reagent that reacts with it to form
an insoluble complex with a known chemical formula

• The precipitate is separated from the solution by

filtration, rinsed, dried and weighed

• The amount of the element present in the sample is

determined from the chemical formula of the
precipitate

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 Gravimetric Analysis
• Example: the amount of chloride in a solution can be
determined by adding excess silver ions to form an
insoluble silver chloride precipitate.
– It is known that Cl is 24.74% of AgCl.

• Not suitable to determine trace elements in food

because the balances are not sensitive enough to
accurately weigh small amount of precipitate formed.

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 Colorimetric Methods
• Chromogens are chemicals that, upon reaction with a
compound of interest, form a colored compound
• Chromogens - selectively reacts with a wide variety of
minerals
• Each chromogen reacts with a specific mineral to
produce a soluble coloured product that can be
quantified by measuring the absorbance of the solution
at a specific wavelength using a spectrophotometer
• Example: yellow-orange color forms when
molybdovanadate is added to phosphorus containing
sample. The phosphorus then can be quantified by
measuring the absorbance of the solution at 400nm and
comparing with a calibration curve.
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 EDTA complexometric titration
• Ethylenediaminetetraacetate (EDTA) forms strong complexes with
multivalent metallic ions.
• The stability of mineral-EDTA complexes generally increases with
valence of the ion, although there is variation among ions with similar
valence due to their coordination chemistry
• The complexation equilibrium is strongly pH dependent. With
decreasing pH, the chelating sites of EDTA become protonated,
thereby decreasing its effective concentration
• Endpoints are detected using mineral chelators that have
coordination constant lower than EDTA (i.e. less affinity for metal
ions)

• Calmargite and Erichrome Black T (EBT) – indicators that change

from blue to pink when complexed with Ca or Mg
• Endpoint of titration complexometric EDTA titration is detected as
colour changes from pink to blue
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• Procedure
1. Sample is diluted with water
2. Made alkaline (pH 12.5 to 13)
3. Indicator is added
4. Titrate with EDTA

• EDTA forms complexes with minerals first then with

the indicator – determine the end point of the reaction
• The calcium content is determined by comparing the
volume of EDTA required to titrate it to the end point
with a calibration curve prepared for a series of
solutions of known calcium concentration
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 Redox Reactions
• Based on coupled reduction-oxidation reactions
• One of the half-reactions leads to a measurable change in the
system that can be conveniently used as an end-point e.g. a
color change
• E.g. permanganate ion is a deep purple color (oxidize form)
while the mangenous ion is a pale pink color (reduced form)
• Procedure
1. Potassium permanganate is titrated into the aqueous
solution of ashed food
2. MnO4- is converted to Mn2+ as Fe2+ still remaining in the food
3. All Fe2+ converted to Fe3+ and remaining MnO4- leads
formation of a purple color (end-point)
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 Precipitation Titrations
• When at least one of a titration reaction is insoluble
in precipitate

• Common method: Mohr method for chloride analysis

• Silver nitrate is titrated into an aqueous solution

containing sample and a chromate indicator
AgNO3 + NaCl → AgCl (s) + NaNO3

• End point: the first hint of orange color

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 Ion-Selective Electrodes
• The principle is similar with pH meters, but the
composition of the glass electrode is sensitive to
specific types of ion

• Principle: ISE only sensitive to one type of ion

(free ions)
– Free ions can be obtained by ashing the food.

• Problem: interference from other types of ions

– Can be reduced by adjusting pH, complexing or
precipitating the interfering ions

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 Ion-Selective Electrodes
• Two electrodes are dipped
into an aqueous solution
containing dissolved
minerals

• Concentration of the
specific minerals are
determined from a
calibration curve of voltage
versus log of concentration

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 Atomic Spectroscopy
• Principle:
• Each element has a unique electronic structure
• Consequently, it absorbs or emits radiation at specific
wavelength
• Absorption: electrons in the ground state are promoted
to various excited states. Atomic absorption spectra
are produced when ground state atoms (or ions)
absorb energy from a radiation source.
• Emission: electrons in an excited state fall back to a
lower energy level. Atomic emission spectra are
produced.
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 Atomic Absorption Spectroscopy
• Atomic absorption spectroscopy (AAS) is an analytical method
that is based on the absorption of UV-visible radiation by free
atoms in the gaseous state.

• Requires that atoms of the element of interest be in the atomic

state (not combined with other elements in a compound)
• In foods, all elements are present as compounds or complexes
and therefore must be converted to neutral atoms (atomized)
before absorption or emission measurement can be made

• Atomization – separating particles into individual molecules

(vaporization) and breaking molecules into atoms
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 Atomic Absorption Spectroscopy
• Procedure
1. Ashed sample is dissolved in an aqueous solution
2. Placed in the instrument where it is heated to
vaporize and atomize the minerals
3. A beam of radiation is passed through the
atomized sample
4. Absorption of radiation at specific wavelength

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Schematic Diagram of AAS
Atomised
sample in
flame

Monochromator
Lens Lens

Flame
Solution Amplifier

Display

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 AAS - Instrumentation
• Radiation Source
– The lamp emits radiation characteristic of the metal in the cathode
– i.e. if the cathode is made of sodium, a sodium emission spectrum
is produced.
– When this radiation passes through a sample containing sodium
atoms it will be absorbed because it contains radiation of exactly
the right wavelength to promote transition from one energy level to
another
– Thus, a different lamp is needed for each type of element analyzed
• Chopper
– Radiation arriving at the detector comes from two different
sources
1. Radiation emitted by the filament of the lamp
2. Radiation that is emitted by the atoms in the sample
– Purpose: To isolate the signal generated by the light from that
emitted by the atoms in the sample
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• Atomizer
– To convert sample to be analyzed into individual atoms
– Atomization process: exposing the sample to high
temperatures
– involves 3 stages: water removal, conversion molecules
into a gas and atomization
– Ionization of atom is undesirable
• Two types of atomizers:
• Flame-atomizers consist of a nebulizer and a burner
– Nebulizer converts the solution into a fine mist or aerosol
• Electrothermal AAS
– The sample is placed in a small graphite cup which is
electrically heated to a temperature (2000 – 3000°C) to
produce volatilization and atomization
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 AAS - Instrumentation
• Wavelength selector
– To select the radiation of the desired wavelength
– Typical wavelength selectors: monochromatic or
filters

• Detector
– A photomultiplier tube that converts electromagnetic
energy reaching it into an electrical signal.

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 Atomic Absorption Spectrometry
• Measures small concentrations of metal ions in
solution
– Al, As, Au, B, Ca, Cd, Co, Cr, Cs, Cu, Fe, Ge, K, Li, Mg, Mn, Mo, Na, Ni,
Pb, Si, Sr, Ti, V, W and Zn
• Applications:
– analysis of ores for metal content
– quality control of metals in steel
– testing water for metals ions
– analysing food and pharmaceuticals for metal ions

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 AAS
• Advantages
– very sensitive:
can detect concentrations as small as g / L (ppb)
– generally very specific:
set wavelength is strongly absorbed by the particular
metal ion being analysed (and not by other components)
• A source of error
– Another species may be absorbing at the same
wavelength.

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 Atomic Emission Spectroscopy
• Atomic Emission Spectroscopy (AES) utilizes the
emission of radiation by a sample
• Several ways that energy can be supplied: heat, light,
electricity and radio waves

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 Instrumentation
• The sample itself acts as the source of the detected
radiation
• Radiation source or chopper are not needed
• Atomic emissions are produced when the electrons in
an excited state fall back to lower energy levels
• Each atom have characteristic emission spectrum as
they have different allowed energy levels
• The emitted radiation passed through a wavelength
selector and the intensity of the peak isolated is
measured using detector.

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• Atomization-Excitation Source
– Two most common sources
1. Flame - AES: similar to flame AAS
2. Inductively Coupled Plasma (ICP) – AES: heat the sample
in the presence of argon ions
• Argon ions lead to the release of electrons that push the
equilibrium towards the non-ionized form of the mineral
• Wavelength selectors
– To isolate particular spectral lines
– Types of wavelength selectors:
– Filters – can only be used to measure the intensity at a
particular fixed wavelength (one type of mineral)
– Gratings – can be used to measure the intensity at many
different wavelengths (many different types of minerals)
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 Practical Consideration
• Sample must be ashed, dissolved in a suitable
solvent (water or diluted HCl) before injecting it into
the instrument
• Vegetable oils can be analyzed by dissolving them
in acetone or ethanol and injecting them directly
into the instrument
• Use very pure reagents when preparing samples
for analysis
• Use glassware that is in very clean and dry so that
it contains no contaminating elements
• No interfering substances present (absorb at the
same wavelength) in the samples
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