Lecture Note On Sample Handling

A.K. BEJ
Asst. Professor of Chemistry

ANALYTICAL CHEMISTRY:
Analytical Chemistry is concerned with the chemical characterisation of matter. It provides the
method and tools needed for insight in to the material world, for answering our basic questions
about a material sample i.e. What? Where? How much?
It has important role in nearly all aspects of Chemistry. Like Pharmaceutical, Manufacturing,
agricultural, forensic, Clinical, metallurgical, environmental. The nitrogen content of a
fertilizer determines its value. The air in the cities must be analysed for CO. Food must be
analysed for contaminates like pesticide residue and for essential nutrients like vitamins. The
quality of manufactured products often depends upon proper chemical compositions, and
measurement of constituents is a necessary part of quality control.
An analytical chemist serves the need of many fields as given bellow

 In medicine, analytical chemistry is the basis of clinical laboratory tests, which help
physicians to diagnose diseases and plan for its recovery.
 Environmental quality often evaluated by testing for suspected contaminants using the
techniques of analytical chemistry.
 The nutritional value of food can be determined by chemical analysis for major
components such as protein, carbohydrates and trace components such as vitamins and
minerals even the calories in food.
 In industry, analytical chemistry provides the mean of testing raw materials and for
assuring the quality of the finished product whose chemical composition is critical.
TYPES OF ANALYSIS:
On the basis of Chemical analysis:
1. Proximate analysis: In this analysis, the amount of each element in a sample is
determined with no concern to the actual compounds present.
2. Partial analysis: It deals with the determination of selected constituents in the sample.
3. Trace Constituent analysis: It is concerned with the determination of specified
components present in very minute quantity.
4. Complete analysis: In this analysis, the proportion of each component of the sample is
determined.
On the basis of sample size:
1. Macro analysis: It is concerned with the quantities of 0.1g or more.
2. Semi micro analysis: Measure the quantities ranging from 10-2g to 10-1g.
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 3. Micro analysis: Measure the quantities ranging from10-3g to 10-2g.
4. Ultra micro analysis: It deals with the quantities below 10-4g.
CLASSIFICATION OF ANALYTICAL METHOD:

Analytical method
↓

Classical Method Instrumental Method Non-destructive Method

1. Gravimetry 1. AAS 1.XRD

2. Titrimetry 2. HPLC 2.XRF

3. TGA 3.EDX

4. DTA

SAMPLING:
Sampling is an important operation to analyse a material or substance. Sampling forms an
integral part of the research design as this method derives the quantitative data and the
qualitative data that can be collected as part of a research study.
Sampling two types
1. Statistical sampling-A sample for analysis may either be selected according to a through
plan, which provides every particle and portion of the substance an equal chance of appearing
in the sample
2. Random sampling-Sample are taken at randomly.
Types of sample
1. Solid
2. Liquid
3. Gas
4. Mixture
SAMPLE PREPARATION:
On first impression, sample preparation may seem the routine aspect of an analytical protocol.
However, it is critical that analysts realize and remember that a measurement is only as good
as the sample preparation that has preceded it. If an aliquant taken for analysis does not
represent the original sample accurately, the results of this analysis are questionable. Generally,
the error in sampling and the sample preparation portion of an analytical procedure is
considerably higher than that in the methodology itself. One goal of laboratory sample
preparation is to provide, without sample loss, representative aliquants that are free of

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laboratory contamination that will be used in the next steps of the protocol. Samples are
prepared in accordance with applicable standard operating procedures (SOPs) and laboratory
SOPs using information provided by field sample preparation.
General Guidance for Sample Preparation
 Sample Losses during Preparation
 Contamination
 Control of contamination
 Control programme

1. Sample Losses during Preparation

i. Losses as Dust or Particulates- When a sample is dry ashed, a fine residue (ash) is often
formed. The small particles in the residue are resuspended readily by any air flow over the
sample. Air flows are generated by changes in temperature (e.g., opening the furnace while it
is hot) or by passing a stream of gas over the sample during heating to assist in combustion.
These losses are minimized by ashing samples at as low a temperature as possible, gradually
increasing and decreasing the temperature during the ashing process, using a slow gas-flow
rate, and never opening the door of a hot furnace . If single samples are heated in a tube furnace
with a flow of gas over the sample, a plug of glass or quartz wool can be used to collect
particulates or an absorption vessel can be used to collect volatile materials. At a minimum, all
ash or finely ground samples should be covered before they are moved.
ii. Losses through Volatilization-The loss of volatile elements during heating is minimised by
heating without exceeding the boiling point of the volatile compound. Ashing aids can reduce
losses by converting the sample into less volatile compounds. These reduce losses but can
contaminate samples. During the wet ashing process, losses of volatile elements can be
minimised by using a reflux condenser. If the solution needs to be evaporated, the reflux
solution can be collected separately. Volatilization losses can be prevented when reactions are
carried out in a properly constructed sealed vessel.
iii. Losses due to Reactions between Sample and Container-Specific elements may be lost from
sample materials from interaction with a container. Such losses may be significant, especially
for trace analyses used in radio analytical work. Losses due to adsorption may be minimised
by using pre-treated glassware with an established hydrated layer. Soaking new glassware
overnight in a dilute nitric or hydrochloric acid solution will provide an adequate hydrated
layer. Glassware that is used on a regular basis will already have established an adequate
hydrated layer. The use of strong acids to maintain a pH less than one also helps minimize
losses from adsorption.

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2. Contamination

i. Air borne- Air borne contamination is most likely to occur when grinding or pulverizing solid
samples. Very small particles (~10 μm) may be produced, Suspended in air, and transported in
the air before settling onto a surface. Other sources of potential airborne contamination include
samples that already consist of very small particles, volatile radionuclides (including tritium),
or radionuclides that decay through a gaseous intermediate (i.e., 226Ra decays to 222Rn gas and
eventually decays to Pb). Therefore, the grinding or pulverizing of solid samples or the
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handling of samples that could produce airborne contamination should be carried out under a
laboratory hood or ventilated enclosure designed to prevent dispersal or deposition in the
laboratory of contaminated air particulates. These particles easily can contaminate other
samples stored in the area. To prevent such cross-contamination, other samples should be
covered or removed from the area while potential sources of airborne contamination are being
processed.
ii. Reagents- Contamination from radiochemical impurities in reagents is especially
troublesome in low-level work. Care must be taken in obtaining reagents with the lowest
contamination possible. Due to the ubiquitous nature of uranium and thorium, they and their
progeny are frequently encountered in analytical reagents.
iii. Glassware/equipment -Other general considerations in sample preparation include the
cleaning of glassware and equipment. Criteria established in the planning documents or
laboratory SOPs should give guidance on proper care of glassware and equipment (i.e.,
scratched glassware increases the likelihood of sample contamination and losses due to larger
surface area). Glassware should be routinely inspected for scratches, cracks, etc., and discarded
if damaged.

3. Control of contamination

i. Labware and Glassware -Some labware is too expensive to be used only once (e.g., crucibles,
Teflon. beakers, separatory funnels). Labware that will be used for more than one sample should
be subjected to thorough cleaning between uses. A typical cleaning protocol includes a
detergent wash, an acid soak (HCl, HNO3, or citric acid), and a rinse with deionized or distilled
water. Scrubbing glassware with a brush aids in removing contaminants.
Practical advice on washing and cleaning laboratory glassware:
 Always clean your apparatus immediately after use. It is much easier to clean the
glassware before the residues become dry and hard. If dirty glassware cannot be washed
immediately, it should be left in water to soak.

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  Thoroughly rinse all soap or other cleaning agent residue after washing glassware to
prevent possible contamination. If the surface is clean, the water will wet the surface
uniformly; if the glassware is still soiled, the water will stand in droplets.
 Use brushes carefully and be certain that the brush has no exposed sharp metal points
that can scratch the glass. Scratched glassware increases the likelihood of sample
contamination and losses due to larger surface areas. Moreover, scratched glassware is
more easily broken, especially when heated.
ii. Equipment -In order to avoid cross-contamination, grinders, sieves, mixers and other
equipment should be cleaned before using them for a new sample. Additional cleaning of
equipment prior to use is only necessary if the equipment has not been used for some time.
Another practical approach is to brush out the container, and briefly process an expendable
portion of the next sample and discard it. For more thorough cleaning, one may process one or
more batches of pure quartz sand through the piece of solid processing equipment, and then
wash it carefully. The efficacy of the decontamination is determined by monitoring this sand
for radionuclide contamination. An effective cleaning procedure for most grinding containers
is to grind pure quartz sand together with hot water and detergent, then to rinse and dry the
container. This approach incorporates a safety advantage in that it controls respirable airborne
dusts. It is important to note that grinding containers become more difficult to clean with age
because of progressive pitting and scratching of the grinding surface. Hardened steel containers
can also rust, and therefore should be dried thoroughly after cleaning and stored in a plastic
bag containing a desiccating agent. If rust does occur, the iron

4. Laboratory Contamination Control Program

The laboratory should establish a general program to prevent the contamination of samples.
Included in the program should be ways to detect contamination from any source during the
sample preparation steps if contamination of samples occurs. The laboratory contamination
control program should also provide the means to correct procedures to eliminate or reduce
any source of contamination. Some general aspects of a control program include:

 Appropriate engineering controls, such as ventilation, shielding, etc., should be in

place.
 The laboratory should be maintained clean and good laboratory practices should be
followed. Personnel should be well-trained in the safe handling of radioactive materials.
 Counter tops and equipment should be cleaned and decontaminated following spills of
liquids or dispersal of finely powdered solids. Plastic-backed absorbent bench top
coverings or trays help to contain spills.
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  There should be an active health physics program that includes frequent monitoring of
facilities and personnel.
 Wastes should be stored properly and not allowed to accumulate in the laboratory
working area. Satellite accumulation areas should be monitored.
 Personnel should be mindful of the use of proper personnel protection equipment and
practices (e.g., habitual use of lab coats, frequent glove changes, routine hand washing).
 Operations should be segregated according to activity level. Separate equipment and
facilities should be used for elevated and low-level samples whenever possible.
 SOPs describing decontamination and monitoring of labware, glassware, and
equipment should be available.

SOLID SAMPLES:

General procedures such as exclusion of unwanted material in the sample; drying, charring,
and ashing of samples; obtaining a constant weight (if required); and homogenization are
important. Before a solid sample is prepared, the specific procedures given in the planning
documents should be reviewed. This review should result in a decision that indicates whether
materials other than those in the intended matrix should be removed, discarded, or analyzed
separately. Any material removed from the sample should be identified, weighed, and
documented. To ensure that a representative aliquant of a sample is analyzed, the sample should
first be dried or ashed and then blended or ground thoroughly. Homogenization should result
in a uniform distribution of analytes and particles throughout the sample. The size of the
particles that make up the sample will have a bearing on the representativeness of each aliquant.

1. Exclusion of Material
Exclusion of material by size and composition:

During solid preparation, some particles may be identified in the sample that are not a part of
the matrix intended for analysis. Examples of such particles are rocks and pebbles or fragments
of glass and plastic. Depending on the specific procedures given in the planning documents on
the constitution of the sample taken, rocks and pebbles can be removed and analyzed separately
if desired. The sample should be weighed before and after any material is removed. Other
materials that are not a part of the required matrix can also be removed and analyzed separately.
If analysis of the material removed is necessary, applicable SOPs should be used to prepare the
material for analysis.

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Exclusion of organic material:
Leaves, twigs, and grass can easily be collected inadvertently along with samples of soil or
sediment. Because these are not usually intended for analysis, they are often removed and
stored for future analysis, if necessary. The material removed should be identified, if possible,
and weighed.

2. Principles of Heating Techniques for Sample Pre-treatment

Applying elevated temperatures during sample preparation is a widely used technique for the
following reasons:

 To remove moisture or evaporate liquids, raise the temperatures to 60 to 110 ˚C, which
will not significantly alter the physical composition of the sample.
 To prepare a sample containing organic material for subsequent wet ashing or fusion,
char the material by heating to medium temperature of 300 to 350 ˚C.
 To prepare the sample for subsequent determination of non-volatile constituents, dry
ash at high temperature of 450 to 750 ˚C. This may significantly change the physical
and chemical properties of the sample.

3. Obtaining a Constant Weight

If required, constant weight is obtained by subjecting a sample to repetitive cycles of drying
and weighing until a series of weights meets specified requirements. An accurately weighed
sample is heated for 2 hours, cooled in a desiccator, and weighed. Drying is repeated at hourly
intervals to attain a constant weight within the same accuracy. The consistent drying of
materials from a large sample set may require a qualitative evaluation of change in the sample
composition. If a qualitative change occurs the drying method may need to be checked for
completeness. One way to do this would be to perform routine dry-to-constant-weight
evaluations on separate samples. Laboratory conditions and handling of the samples by the
analyst during sample weight determinations can increase the uncertainty of the final sample
mass.

4. Subsampling
Laboratories routinely receive larger samples than required for analysis. The challenge then
becomes to prepare a sample that is representative and large enough for analysis, but not so
large as to cause needless work in its final preparation. Generally, a raw sample first is crushed
to a reasonable particle size and a portion of the crushed material is taken for analysis. This
step may be repeated with intermittent sieving of the material until an appropriate sample size
is obtained. Some of the important points to remember include the following:
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  For most practical purposes, a subsample is guaranteed to be unbiased only if every
particle in the sample has the same probability of being selected for the subsample.
 The weight of the subsample should be many times greater than the weight of the largest
particle in the sample.
 The variance associated with subsampling may be reduced either by increasing the size
of the subsample or by reducing the particle sizes before subsampling.
 Grouping and segregation of particles tends to increase the subsampling variance.
 Grouping and segregation can be reduced by increment sampling, splitting, or mixing.

LIQUID SAMPLE:
1. Aqueous Liquid sample:
Aqueous liquids are a common matrix analyzed by laboratories, and are often referred to as
water samples. For certain samples that are not filtered, inversion is a form of homogenization.
Typically, the sample is homogenized by inverting the container several times to mix the
sample thoroughly. If there is some air in the container, the passage of air bubbles through the
sample will create sufficient turbulence to mix the sample thoroughly with three or four
inversions of the sample container. Simply shaking the container will not mix the contents as
thoroughly as inverting the sample container. Mechanical shakers, mixers, or rotators may be
used to homogenize aqueous samples thoroughly.

2. Non- Aqueous Liquid sample:

Nonaqueous liquids can be substances other than water such as organic solvents, oil, or grease.
The appropriate plan document should be reviewed to determine if filtration is necessary.
Homogenization of nonaqueous samples is accomplished in a manner similar to that for
aqueous samples. Visual inspection is typically used as a qualitative measure of homogeneity
in non-aqueous samples. If a quantitative measure of mixing is desired, turbidity measurements
can be performed after a predetermined amount of mixing until a steady level of turbidity is
achieved. Many nonaqueous liquids present a health hazard (e.g., carcinogenicity) or require
special safety considerations (e.g., flammability). Any special handling requirements based on
health and safety considerations should be documented in the planning documents.

GASES SAMPLE:
The samples of gasses are collected by expansion into an evacuated container. Sampling
devices are usually made from glass and are fitted with stop cocks at both the ends. If some
contamination occurs, it can be removed by extensive flushing with the gas to be sampled.
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UNCERTAINTIES:

Uncertainty of measurement, or measurement uncertainty (MU), is defined as

Parameter, associated with the result of a measurement that characterises the dispersion of
the values that could reasonably be attributed to the measurand.

Uncertainty is ‘associated with’ each measurement result. A complete measurement result

typically includes an indication of the uncertainty in the form x±U, where x is the measurement
result and U an indication of the uncertainty. This form of expressing a result is an indication
to the end user of the result that, with reasonable confidence, the result implies that the value
of the measurand is within this interval.
The measurand is simply a quantity, such as a length, mass, or concentration of a material,
which is being measured. The term ‘value of the measurand’ is closely related to the traditional
concept of ‘true value’ in classical statistical terminology. From this alternative viewpoint
‘uncertainty’ has also been defined as
“An estimate attached to a test result which characterises the range of values within which the
true value is asserted to lie”
The metrological definition asserts that uncertainty expresses ‘the dispersion of the values that
could reasonably be attributed to the measurand’. This is a particularly important phrase. It
indicates that although the uncertainty is associated with a measurement result, the range
quoted must relate to the possible range of values for the measurand. For example, the
measurand could be the total mass of gold in a geological deposit. This is quite different from
a statement of precision, which would describe the range of results that might be observed if
the measurement were repeated. In requesting information about ‘where the measurand value
might be’, this definition of uncertainty implicitly requires the measurement scientist to
consider all the effects that might influence the measurement result. These effects obviously
include the causes of random variation from one measurement to the next over the
measurement timescale. But it is also essential to consider sources of bias during the
experiment, and very often, these generate larger effects than can be observed by repeated
measurement alone. That is, measurement uncertainty automatically asks for a range that
includes an allowance for both random and systematic effects.
Sampling and physical preparation as sources of uncertainty of measurement:
The act of taking a sample introduces uncertainty into the reported measurement result
wherever the objective of the measurement is defined in terms of the analyte concentration in
the sampling target and not simply in the laboratory sample. Sampling protocols are never
perfect in that they can never describe the action required by the sampler for every possible

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eventuality that may arise in the real world in which sampling occurs. The location in space (or
time) for the taking of a sample is rarely specified exactly (e.g. to the nearest millimetre or
second). The sampler has to make such decisions (ideally on objective criteria), but as
heterogeneity is inevitable (in space or time) such decisions will affect the estimated
concentration. An appreciation of these sources of uncertainty is important in the design and
implementation of methods for the estimation of uncertainty. When duplicate samples are
taken, for example, taking them at exactly the same place and time may not reflect the
uncertainty of the measurement that really exists.
Heterogeneity always gives rise to uncertainty. If the sampling target were perfectly
homogeneous then this contribution would be zero, but nearly all materials are heterogeneous
to some extent at some scale. If the test portion is a few micrograms, then nearly all material
will be heterogeneous and the sampling step will contribute to the uncertainty in the
measurement of an analyte concentration. Heterogeneity can be quantified in a separate
experiment, but if the aim is to estimate the analyte concentration in the larger sampling target,
then this heterogeneity is just one cause of measurement uncertainty.
Similar arguments can be made for the uncertainty that arises in the processes of
physical preparation (e.g. transportation, preservation, comminution, splitting, drying, sieving,
homogenisation) that happen after the act of sampling and before the chemical preparation of
the test sample. Each step can introduce errors from a range of mechanisms, such as loss of
analyte, loss of fine particles, or contamination from equipment or previous samples. The
methods employed, and training given, should aim to reduce these errors to a minimum. In
addition, however, procedures are required to estimate the uncertainty that all of these steps,
when applied in practice, generate in the final measurement value.
Heterogeneity as a source of uncertainty:
IUPAC currently define both homogeneity and heterogeneity as ‘The degree to which a
property or constituent is uniformly distributed throughout a quantity of material.’ So defined,
heterogeneity is among the most important factors contributing to uncertainty associated with
sampling. Increments from different locations in the sampling target will have different
concentrations of analyte in a heterogeneous material and there will be a sample-to-sample
variation in analyte concentration – usually visible as a contribution to the observed variation
of results. In general, the exact dependence of concentration on location is unknown, so no
correction can be made. This results in uncertainty in any given result or, in general, any
average of such results.

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 Source Description
Fundamental sampling error (FSE) A result of the constitutional heterogeneity
(the particles being chemically or
physically different)
Grouping and segregation error (GSE) A result of the distributional heterogeneity
Long-range point selection error (PSE1) Trends across space or over time
Periodic point selection error (PSE2) Periodic levels across space or over time
Increment delimitation error (IDE) Identifying the correct sample to take.
Considers the volume boundaries of a
correct sampling device
Increment extraction error (IXE) Removing the intended sample. Considers
the shape of the sampling device cutting
edges
Increment and sample preparation error Contamination (extraneous material in
(IPE) sample)
Losses (adsorption, condensation,
precipitation etc.)
Alteration of chemical composition
(preservation)
Alteration of physical composition
(agglomeration, breaking of particles,
moisture etc.)

MOISTURE IN SAMPLE:
Moisture can be removed from the sample by different method as discussed bellow
 By using desiccator
 By using hot plate
 By using hot air oven
 By using refractometer
% of Moisture = [(W1-W2)/ W1]×100
W1- weight of sample taken for test
W2- weight of sample after heating (final weight)

COMBUSTION METHODS FOR DECOMPOSINGORGANIC SAMPLES:

(a) Combustion over an open Flame (Dry Ashing):

To determine the cations of an organic sample, it has to be heated till red hot in an open dish
or crucible over a flame so that all the carbonaceous matter gets oxidized and converted into
carbon dioxide. Analysis of the non-volatile components follows dissolution of the residual
solid. In addition, volatile metallic compounds may be lost during the ignition process.

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Although dry ashing is the simplest method of decomposing organic compounds, it is often the
least reliable.

(b) Combustion-Tube method:

Several common and important elemental components of organic compounds are converted
into gaseous products as a sample is pyrolyzed in the presence of oxygen. The heating is
commonly performed in a glass or quartz combustion tube through which a stream of carrier
gas is passed. The stream transports the volatile products to the parts of the apparatus where
they are separated and retained for measurement. Elements susceptible to this type of treatment
are carbon, hydrogen, nitrogen, sulphur, halogens and oxygen.

Automated combustion-tube analyzers are now available for the determination of either carbon,
hydrogen, and nitrogen or carbon, hydrogen and oxygen in a sample. This analysis is completed
in less than 15 minutes. In this analyzer, the sample is ignited in a stream of helium and oxygen
and passed over an oxidation catalyst consisting of a mixture of silver vanadate and silver
tungstate. In this process the halogens and sulphur are removed with a packing of silver salts.
A packing of hot copper is located at the end of combustion train to remove oxygen and convert
nitrogen oxides to nitrogen. The exit gas, consisting of a mixture of H2O, CO2, nitrogen and
helium is collected in a glass bulb.

(c) Combustion with oxygen in a sealed container:

The decomposition of many organic materials involves sealed container. Before the reaction
vessel is opened, the reaction products are absorbed in a proper solvent. Schoniger suggested
an apparatus. It contains a heavy-walled flask of 300-1000 mL capacity fitted with a ground
glass stopper. Attached to the stopper is a platinum gauze basket which holds 2 to 200 mg of
sample. If the substance is a solid, it is wrapped in a piece of low-ash filter paper. The liquid
samples are weighed in a gelatin capsule which is then wrapped in a similar manner. The paper
tail helps as the ignition point. A small volume of an absorbing solution is taken in the flask.
The tail of the paper is ignited, the stopper is fitted rapidly into the flask, and the flask is
inverted to prevent the escape of the volatile
oxidation products. The entire reaction is
calalyzed by the platinum gauge surrounding
the sample. This procedure is helpful to
determine the halogens, sulphur, phosphorus,
fluorine, boron, carbon, arsenic, and various
other metals present organic compounds.

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DECOMPOSING SAMPLES WITH INORGANIC ACIDS:
The mineral acids are generally used to decompose the inorganic samples in open-vessel.
Sometimes ammonia and aqueous solutions of the alkali metal hydroxides are used for this
purpose. Usually, a suspension Of the inorganic sample is prepared in the mineral acid and then
it is heated by flame or a hot plate until the dissolution is conformed to be complete by the total
disappearance of a solid phase.
In this process the temperature of decomposition is adjusted at the boiling point of the used
acid reagent.
The following acids can be used for the purpose:
(a) Hydrochloric Acid
(b) Nitric Acid
(e) Sulphurie Acid
(d) Perchloric Acid
(e) Oxidising Mixture of acids
(f) Hydrofluoric acid.
(a) Hydrochloric Acid: Concentrated HCl is an excellent solvent for inorganic samples. It is
widely used to dissolve the metals which are more easily oxidized than hydrogen as well as
many metal oxides. Hydrochloric acid has limited applications in decomposition of organic
substances. The Strength of concentrated HCI is about 12 M.
(b) Nitric Acid: To dissolve all common metals except aluminium and chromium (owing to
sufrace oxide formation), hot concentrated nitric acid is a potent oxidant. For the determination
of the trace metal contents in samples, hot nitric acid is used alone or in combination with other
acids and oxidizing agents. For decomposition of organic samples, hot nitric acid is used with
hydrogen peroxide and bromine. In this process organic sample converts into CO2 and water.
This decomposition process is known as wet ashing process.
(c) Sulphuric Acid: Many materials are decomposed and dissolved by hot concentrated H2S04.
As the boiling point of sulphuric acid is very high (about 340˚C), most of the organic
compounds are dehydrated and oxidized at this temperature and thus eliminated from the
sample as CO2 and H2O by this wet ashing process.
(d) Perchloric Acid: Perchloric acid is a very strong acid which can attack even on iron alloys
and stainless steel (which are not affected by other mineral acids). Due to its high explosive
nature, perchloric acid is used with great care. Although the dilute heated acid and cold
concentrated acid is not explosive in nature, violent explosion occurs when hot concentrated
perchloric acid comes in contact with organic materials or easily oxidised inorganic substances.
Because of this property, this acid is heated in a special hood, lined with glass or stainless steel

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and its own fan system. The hood for perchloric acid should be independent of all other
systems. Commercially perchloric acid is marketed as 60% to 72% acid.
(e) Hydrofluoric Acid: The main use of hydrofluoric acid is for the decomposition of silicate
rocks and minerals in the analysis of elements other than silica. Hydrofluoric acid finds
occasional use with other acids in dissolving steel which is difficult to dissolve in other
solvents. As hydrofluoric acid is extremely toxic, therefore dissolution of samples and
evaporation to remove excess reagent should always be carried out in a well-ventilated hood.
Hydrofluoric acid causes serious damage and painful injury when brought in contact with skin.
If the acid comes in contact with skin, the affected area should be immediately washed with
large quantity of water.
(f) Oxidising Mixture of acids: To decompose the samples, some oxidising agents are used as
the mixture of acids, such as:
(i) Aquaregia, a mixture containing 3 volumes of concentrated HCI and I volume of nitric acid.
(ii) The addition of bromine or hydrogen peroxide to mineral acids generally increases their
solvent action.
(iii) Mixtures of nitric acid and perchloric acid are also useful for this purpose and are less
dangerous than perchloric acid alone.
DECOMPOSITION OF INORGANIC MATERIAL BY FLUXES:

Some substances can not decompose easily such as some mineral oxides, silicates and some
iron alloys. In such cases fused-salt medium is mixed with the sample and then this
combination forms a water soluble product called melt.

Flux is an alkali-metal salt, used to decompose the substances which are attacked by the reagent
very slowly.

For a sample containing a small fraction of substance which dissolves with difficulty, is mixed
with a liquid reagent, the undecomposed residue is then separated by filtration and is fused
with a small quantity of proper flux. After cooling the above combination, the melt is dissolved
and combined with the major fraction of the sample.

Procedure of Fusion:

Firstly, the sample is prepared by grinding and fine powder is obtained. It is mixed with a
suitable flux. The quantity of flux should be 10 times of the sample. Mixing of flux and sample
is carried out in a crucible. The fusion is done in the crucible for a few minutes to a few hours.
The formation of a clear melt indicates the completion of decomposition. After completing the
fusion, the mass is allowed to cool, before solidification.

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The properties of some common fluxes are given below:

(i) Sodium Carbonate: With sodium carbonate the sample containing silicates and some other
refractory materials can be decomposed by heating at 1000˚C to 1200˚C. By this fusion, the
cationic constituents of the substance (sample) generally convert into acid-soluble carbonates
or oxides and non-metallic constituents are converted into soluble sodium salts. Carbonate
fusion is usually performed in platinum crucible.

(ii) Potassium Pyrosulphate: Potassium pyrosulphate is a strong acidic flux which is used for
attacking the more intractable metal oxides. With it flux fusion is carried out at 400˚C.

Potassium pyrosulphate can be prepared by heating potassium hydrogen sulphate, in this way:

2KHSO4→ K2S2O7 + H2O

(iii) Lithium Metaborate: LiBO2 [lithium metaborate], is mixed with lithium tetraborate, and
fused with refractory silicate and alumina minerals. This fusion is carried out in platinum or
graphite crucible at 900˚C.

MICROWAVE DECOMPOSITION:

The use of microwave ovens for the decomposition of organic and inorganic compounds was
first proposed in 1970. Microwave digestions can be carried out in both closed as well as open
vessels, but closed vessels are often used because higher pressure and temperature can be
achieved in them. The decomposition of compounds in microwave oven is very fast and
speedy, even in case of typical and difficult samples it takes only 5 to 10 minutes. In contrast,
the same results require several hours when sample is heated over a flame or a hot plate.

In Microwave Oven:

The energy is transferred directly to all the molecules of the solution almost simultaneously
without heating up the Vessel. Thus, boiling temperatures are reached throughout the entire
solution very quickly. An advantage of using closed vessels for microwave decomposition is
the higher temperature that develops as a consequence of increased pressure. A further
advantage of micro-wave decomposition is that loss of volatile components of samples is
virtually eliminated. Closed vessel microwave decomposition is often easy to automate, thus
reducing operation time required to prepare samples for analysis.

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Moderate Pressure Vessels:

Microwave digestion vessels are constructed from low loss materials. These vessels are
transparent, thermally stable, and resistant to chemical attack by various acids used for
decomposition. The best material for microwaves is Teflon. It is transparent, inert against
sulphuric acid and phosphoric acid, having a melting point about 300˚C. Quartz or borosilicate
glass vessels are sometimes used in place of Teflon containers. A closed digestion vessel of
teflon body consists of a cap and a safety relief valve to operate at 120 ± 10 Psi pressure is used
pressure commercially in a microwave oven. At this pressure, the safety valve can open and
then resealed.

High Pressure Microwave vessels:

For dissolving highly refractory materials which are incompletely decomposed in the moderate
pressure vessels, the microwave bomb is particularly useful, In this bomb, the heavy-wall body
is composed of a polymeric material which is transparent to microwaves. The decomposition
of the material is carried out in a Teflon cup supported in the bomb body. It has a teflon O—
ring in the liner cap that seats against a narrow rim on the exterior of the liner. The O—ring
distorts, and the excess pressure then compresses the sealer disk, which allows the gases to
escape into the surroundings. The internal pressure in the bomb can be judged by the distance
to which pressure screw protrudes from the cap. A commercial Microwave bomb is designed.to
operate at 80 atm or about 10 times the pressure that can be tolerated by the moderate pressure
vessels as described above.

Atmospheric Pressure Vessels:

Atmospheric pressure Vessel are open-vessel systems, these systems do not have an oven but
use a focussed microwave cavity. They have tubing for the insertion and removal of reagents.
There is no safety because of formation of gas during the reaction of digestion process since
the systems operate at atmospheric pressure.

Microwave Furnaces:
In microwave furnace an organic material can be fused before acid dissolution. These furnaces
consist of a small volume Chamber which accommodates only an ordinary-sized crucible. This
small cavity-like chamber is made up of silicon carbide, which is surrounded by quartz
insulation. The advantage of this type of furnace over a conventional muffle furnace is the
speed, at which the high temperature (1000˚C) is reached in 2 minutes.

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Applications of Microwave Decomposition:

The applications of closed vessel decomposition in microwave oven fall into two categories:
(i) Oxidative decomposition of organic and biological samples.
(ii) Decomposition of refractory inorganic materials in industry.
In both the above cases, the new microwave technique is replacing older conventional methods
because of the large economic gains that result from significant saving in time.

Source:
1. Laboratory sample preparation manual, volume II by MARLAP, 2004.
2. Measurement uncertainty arising from sampling: A guide to methods and approaches
by Eurachem, EUROLAB, CITAC, Nordtest and the RSC Analytical Methods.
3. Analytical Chemistry by G.D.Christian, Purnendu K. Dasgupta, Kevin A. Schug,
seventh edition, Wiley.
4. Analytical Chemistry by W.T.Hall, John Wiley & Sons.
5. Analytical Chemistry by Alka Gupta, Pragati Prakashan.

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