SAMPLE DIGESTION

METHODS
Reasons for sample preparation
• To degrade and solubilize the matrix, to release all metals for analysis.
• To extract metals from the sample matrix into a solvent more suited to
• the analytical method to be used.
• To concentrate metals present at very low levels to bring them into a
• concentration range suitable for analysis.
• To separate a single analyte or group of analytes from other species
• that might interfere in the analysis.
• To dilute the matrix sufficiently so that the effect of the matrix on the
• analysis will be constant and measurable.
• To separate different chemical forms of the analytes for individual
• determination of the species present.
Definition of sample digestion
• It refers to the process of decomposing a solid sample into a liquid
state by dissolving it using reagents such as strong acids, alkalis, or
enzymes.
Digestion Methods
• Many metal analyses are carried out using atomic spectroscopic
methods such as flame or graphite furnace atomic absorption or
inductively coupled plasma atomic emission spectroscopy (ICP-AES).
• These methods commonly require the sample to be presented as a
dilute aqueous solution, usually in acid.
WET DIGESTION METHODS
The common methods used for dissolving samples for metals analysis
are digestion in an open flask, digestion in a pressurized, sealed
container, and microwave assisted decomposition.
Samples to be analyzed for elemental metal content are usually
prepared
by digesting the matrix in a strong acid.
In the case of organic matrices, an oxidizing mixture is used to
destroy the entire organic matrix and solubilize the sample.
This yields a clear solution containing the metals for analysis by such
techniques as AA, ICP, or ICP-MS.
WET DIGESTION METHODS
Nitric acid is commonly used because there is no chance of forming
insoluble salts as might happen with HCl or H2SO4. Hydrogen
peroxide may be added to increase the oxidizing power of the
digestion solution.
Inorganic samples, soils, sediments, ores, rocks, and minerals may be
digested in dilute or concentrated acids or mixtures of acids, which may
be sufficient to leach out the analytes.
However, if total digestion is required. hydrofluoric acid can be used
as a final digestion step to dissolve silicates.
Refractory materials such as cements, ceramics, and slags may
require fusion or flux digestion, which involves melting the ground
sample with a salt such as sodium carbonate or sodium peroxide.
WET DIGESTION METHODS
The resulting solid is then more easily dissolved for analysis.
However, the method is not generally suitable for metals, which tend
to be lost by volatilization because of the high temperatures required.
In addition, the material of the container becomes more critical as the
aggressiveness of the digestion process increases.
Reagents that are commonly used in sample
digestion
Acid Digestion—Wet Ashing
The simplest method for wet digestion is carried out in an open container.
Samples are dried, weighed, and placed in a beaker. The digestion reagent
is
Added, and the beaker is covered with a watch glass and placed on a hot
plate.
The sample is allowed to boil very gently to avoid spattering. More
solution may be added from time to time to prevent the sample from
drying out.
Hydrogen peroxide may be added at a point during the digestion to help
oxidize organic materials.
When the sample has been digested completely, it is evaporated to near
dryness and then taken up in a dilute acid solution and diluted to volume
for analysis.
Acid Digestion—Wet Ashing
Samples are generally not allowed to dry completely, as species even
less soluble may form.
Filtration at this point is often necessary, as many matrices will leave
some insoluble matter, such as silica.
The filter must be rinsed carefully to avoid the loss of analyte.

Open sample digestion using a hot plate

The choice of acid to be used depends on the sample. Relatively
soluble inorganic samples, salts, active metals, or alloys may be
dissolved in water or dilute acid.
Electropositive metals will also dissolve in dilute acid, although
aluminum may need a trace of mercuric chloride added to prevent the
formation of an impervious oxide layer.
The mildest solution that will digest the sample is preferred, as
stronger acids are more likely to add to the blank, attack digestion
vessels, and generally require more care in the laboratory.
Concentrated acids may be used individually, in mixtures, or in
sequence.
Hot concentrated acids will dissolve many metals and alloys. Nitric
acid oxidizes the sample and should be used before a stronger oxidizer
such as perchloric acid, to remove the more readily oxidized material.
Several progressively more aggressive digestion schemes can be
considered.
If the sample is not water soluble, a nitric acid digestion may be
suitable.
In preparation, the glassware should be acid washed and rinsed with
distilled water.
A general procedure is to place the weighed sample in a conical flask
or beaker and add 5 to 10% nitric acid. This is covered with a watch
glass and brought to a slow boil on a hot plate.
If the sample is prone to bumping, a few boiling chips can be added.
The solution is evaporated down to a few milliliters without allowing
it to dry.
Heating is continued and small quantities of concentrated nitric acid
are added until the digested solution is clear and light colored. The
beaker walls are washed down. If filtration is necessary, it is carried
out at this point.
The solution is transferred to a volumetric flask and diluted to volume
before analysis. Sometimes, 1 : 1 hydrochloric acid is added when the
nitric acid digestion is complete, and a further digestion is carried out,
before filtration and dilution.
If the sample is not digested satisfactorily by nitric acid alone, or by
nitric acid followed by HCl, further treatment with sulfuric acid can be
done.
A 2 : 1 mixture of sulfuric and nitric acids is added. The sample is
evaporated to dense white fumes of SO3.
More nitric acid may be added if the solution does not clear. The
solution is again heated to SO3 fumes.
The solution is then cooled, diluted with water, and heated to
dissolve any salts. Then it is filtered, if necessary, diluted to volume,
and is ready for analysis.
An even more aggressive digestion begins with the nitric acid
digestion as described above.
After the sample has been digested and boiled down to a few
milliliters, the sample is cooled and equal volumes of nitric and
perchloric acids are added, cooling the beaker between additions.
It is evaporated gently until dense white fumes of perchloric acid are
seen. If further digestion is needed, nitric acid can again be added.
The cooled sample is diluted with water, filtered if needed, and then
diluted to volume.
For samples that show significant losses of analyte due to the
retention of metals in the silica residues, the sample is first digested
thoroughly with nitric acid in a PTFE (polytetrafluoroethylene) beaker.
Then concentrated perchloric acid and a small amount of hydrofluoric
acid are added.
The sample is boiled until clear and white fumes have appeared. The
sample is cooled and diluted to volume. For even more dissolution
power, mixtures of concentrated acids with oxidizing agents or with
hydrofluoric acid are used.
Aqua regia, a 3 : 1 mixture of concentrated hydrochloric and nitric
acids will dissolve noble metals.
Sulfuric acid with hydrogen peroxide is a powerful oxidizer. A mixture
of an oxidizing acid with hydrofluoric acid provides acidity, oxidizing
power, and complexation to dissolve the sample.
These mixtures will dissolve all metals and alloys and most refractory
minerals, soils, rocks, and sediments.
Microwave Digestion
Digesting a sample in a closed container in a microwave oven has several
advantages over open container dissolution methods.
i. The containers are fabricated of high-temperature polymers, which are less likely
to contain metal contaminants than are glass or ceramic beakers or crucibles.
ii. The sealed container eliminates the chance of airborne dust contamination.
iii. The sealed, pressurized containers reduce evaporation, so that less acid digestion
solution is required, reducing blanks.
iv. The sealed container also eliminates losses of more volatile metal species, which
can be a problem in open container sample decomposition, especially in dry
ashing.
v. The electronic controls on modern microwave digesters allow very reproducible
digestion conditions.
vi. Automated systems reduce the need for operator attention.
vii. The controlled exhaust contains the acid fumes, which can then be scrubbed
in a neutralizing solution.
Microwave digestion consists of a microwave oven, a rotating carousel
holding several sample digestion bombs, and a system for venting these in a
controlled fashion.
It may also provide monitoring and recording of both temperature and
pressure in the containers.
The sample containers are relatively high pressure containers, usually made
of strong, high-temperature-resistant polymers, often polycarbonate for
strength or PTFE for chemical resistance.
Systems designed for the strong acid digestion required for metals analysis
often include a separate liner, which is more resistant to chemical attack and
can be changed as it begins to break down under the very corrosive
conditions of high acidity and high temperature in the digestion bomb.
Each bomb has a pressure relief valve, which vents into a manifold. This
exhausts the acid fumes into a tube, which should be connected to an acid
scrubbing trap.
The relief valves are set so that the sample is heated under pressure, allowing
higher temperatures and more rapid digestion than is possible in an open
container.
Modern microwave digestion systems monitor both pressure and temperature
in the containers. As the temperature or pressure reaches the set point, power
to the oven is cut.
The oven power as well as the maximum pressure and temperatures can be
set. Both digestion time and oven power can be programmed so that each
sample is treated in a reproducible manner.
The initial digestion is done slowly at low temperature, and the temperature is
increased after the majority of the readily digested matrix is dissolved.
Microwave containers for sample digestion are commercially available which
can be used for ashing samples at temperatures up to 300°C or pressures to
800 psi, under controlled pressure and temperature.
Under these conditions, even refractory samples can be digested successfully
in a reason-able time. A method for dissolution of alumina samples uses a
high-boiling mixture of H2SO4 and H3PO4 and digests the sample at 280°C,
with the pressure reaching only 40 psi.
Similar samples can also be digested at 240°C with HCl, but the pressure
reaches as high as 660 psi.
Ovens that do not have the facility to monitor and control the sample
temperature may need to be calibrated, so that different ovens can be used
with similar results. It is not sufficient simply to set the power fraction and
make a correction for the difference in wattage of the ovens.
The easiest way to calibrate a microwave oven is to measure the
temperature rise of 1 L of water at various power settings and times and
compare these between ovens.
Dry ashing
 This refers to the use of a muffle furnace to maintain temperatures
of 500–600°C.
 Water and volatiles are vaporized, and organic substances are
burned in the presence of oxygen in air to CO2 and oxides of N2.
Most minerals are converted to oxides, sulfates, phosphates,
chlorides, and silicates. Elements, such as Fe, Se, Pb, and Hg, may
partially volatilize with this procedure, so other methods must be
used because ashing is a preliminary step for specific 
elemental analysis.
Procedure for determining ash content in food sample
Apparatus:
1.Porcelain dish
2.Analytical balance
3.Bunsen burner
4.Oven
5.Muffle furnace
6.Tongs

Procedures:
1. A shallow porcelain dish is dried in the oven at 105˚C for 3 hours.
2. The dish is then cooled in the desiccator and weighed after attaining the room temperature.
3. 5g of the homogenized food sample is weighed into the porcelain dish.
4. The sample is dried in an oven at a temperature of 130˚C for one day.
5. The dried sample is charred gently over a Bunsen burner until the smoking ceased.
6. The dish will be put in muffle furnace and was heated at 550˚C for 3 hours.
7. The sample will be ashed until a whitish or greyish ash is obtained.
8. The dish Will be removed and cooled in a desiccator and weighed soon after attaining room temperature.
9. The muffle furnace will be replaced and the heating will continue until a constant weight is obtained.
10. The total ash content of food sample will be calculated