Paper No.

: 06 Atomic Spectroscopy

Module :04 Sample preparation in Atomic Absorption spectrometry

Principal Investigator: Dr.NutanKaushik, Senior Fellow

The Energy and Resouurces Institute (TERI), New Delhi

Co-Principal Investigator: Dr. Mohammad Amir, Professor of Pharm. Chemistry,

JamiaHamdard University, New Delhi

Paper Coordinator: Dr. MymoonaAkhtar, Associate professor, Dept. of Pharm.

Chemistry, JamiaHamdard, New Delhi.

Content Writer: Dr. MymoonaAkhtar, Associate professor, Dept. of Pharm.

Chemistry, JamiaHamdard, New Delhi.

Content Reviwer: Dr. Mohammad Amir, Professor of Pharm. Chemistry,

JamiaHamdard University, New Delhi

Analytical Chemistry Atomic Spectroscopy

/ Instrumentation
Sample preparation in Atomic Absorption spectrometry
 Description of Module
Subject Name Analytical Chemistry / Instrumentation

Paper Name Atomic Spectroscopy

Module Name/Title Sample Prepration in Atomic Absorption spectrometry (AAS)

Module Id 04

Pre-requisites

Objectives Why sample preparation is required in atomic absorption spectrometry. What is the
relationship between sample preparation and determination methods. What Types
of Samples are used and Steps for Consideration before carrying out the analysis.
Proper Selection of Sample Preparation Method depending upon the sample
available. You will learn about various techniques like
Dry Decomposition.
Fusion and dry ashing
Wet Decomposition or Acid Decomposition
Thermal,
Ultrasonic
Radiant (infrared, ultraviolet and microwave).
Keywords Atomic Absorption spectrometry (AAS), Dry Decomposition, thermal
decomposition, Sample Preparation

Analytical Chemistry Atomic Spectroscopy

/ Instrumentation
Sample preparation in Atomic Absorption spectrometry
 Introduction

Nowadays the goals to be achieve best results, in the minimum time, with least
contamination, minimum consumption of reagent and generation of minimal waste or
residue.

Table 1: Relationship between methods of determination and preparation of sample

Analytical Chemistry Atomic Spectroscopy

/ Instrumentation
Sample preparation in Atomic Absorption spectrometry
 Types of Samples

For routine analysis by atomic spectroscopic techniques, which are all dedicated to work with
aqueous samples, the analysis of other liquids must be adapted and the solids are generally
converted into a solution by an appropriate dissolution method

Samples analyzed may be divided as

• already in an aqueous solution:(e.g. various water samples, beverages, blood, serum,

and urine.),in other liquid form (e.g. oils, fuels, and organic solvents.)
• in solid form:(e.g. soils, sediments, plants, animal tissues, metals, and plastics.),
Solid samples may contain a high proportion of organic matter (e.g. plants, animal tissues,
and plastics.) or have more inorganic composition (e.g. soils, sediments, and metals.).

Steps for Consideration

Steps, which should be taken, into consideration for sample preparation, to achieve the real
objectives of the analysis are

• Simplification in sample manipulation,

• use of high purity water and reagents in suitable amounts,
• Proper cleaning of recipients
• and blank preparation in parallel to the samples are desirable.
• validation of the methodology is necessary usually with certified reference materials
Selection of Sample Preparation Method

The selection of method dependent upon following things

i. the analyte(s),
ii. the analyte concentration level(s),
iii. the sample matrix,
iv. the instrumental measurement technique,
v. the required sample size.

Analytical Chemistry Atomic Spectroscopy

/ Instrumentation
Sample preparation in Atomic Absorption spectrometry
 The term matrix here refers to the collection of all of the various constituents making up an
analytical sample.In addition to the analyte, the sample matrix includes all of the other
constituents of the sample, which aresometimes referred to as concomitants. The major
problem in preparing samples for trace analysis is the risk of contamination. Contamination
isassociated with several probable causes, i.e. the grade of reagents used, sample storage
container, steps ofdigestion or dilution of the sample and their previous history, and human
intervention.Losses are a particularly significant problem in trace analysis. Container surfaces,
for example, maypresent a significantly large area on which the analyte can be adsorbed. At
higher levels such a smallabsolute loss would have little effect on the concentration but at
trace levels a large proportion of theanalyte may be stripped from the solution.

Selection of method on the basis of Samples

If measured concentrations satisfy the principal criteria of the spectroscopic method used
(sensitivity, dynamic range) and possible interferences are under control, the analysis of
solutions may be performed automatically with all modern atomic spectroscopic systems.

Two types of samples can be available

• Liquid
• Solid
Aqueous samples can be generally introduced for analysis directly and without any prior
treatment. The important concern about liquid sample is their collection and storage.
Concerning atomic spectroscopic analysis itself, no particular precautions have to be taken.
Non-aqueous samples can sometimes be run directly, but this depends significantly on their
viscosity. Organic solvents like ethanol or methyl isobutyl ketone, fulfill this condition
therefore used for dilution of organic liquids. In flame atomic absorption spectroscopy (FAAS)
analysis, the viscosity should be similar to that of water for which most nebulizers are
designed.

Major drawback

• The dilution factor, which reduces the metal content per unit volume.

Analytical Chemistry Atomic Spectroscopy

/ Instrumentation
Sample preparation in Atomic Absorption spectrometry
 • Standards can be prepared in the pure solvent. Elements in organic solvents usually
give an FAAS analysis response similar to that given by the same element in aqueous
solution.

Solid Samples

• Many steps are required, including

sampling,
sub-sampling,
grinding, and
dissolution.
• The risk of contamination is higher than the case of liquid samples.
Sample Preparation Methods

 Preparation of liquid sample can be performed by

• simple dilution,
• With heating for pre-concentration
• partial or total dissolution.
 Sample preparation normally involves two basic procedures
• Dry decomposition.
• Wet decomposition. Or Acid digestion
Dry Decomposition.

• Is usually carried out by destroying the combustible (organic) portion of the sample
with the help of heat in presence of an ashing aid, using a muffle furnace.
• Temperature required for ashing ranges from 450 to 550°C at atmospheric pressure
Loss due to degree of volatilization is a limiting factor and depends ontemperature applied,
the form of analyte in which it is present in the sample, and

the chemical environment in the ashing stage.

Fusion and dry ashing

• Normally used for silicates, refractory materials, some mineral oxides and iron alloys

Analytical Chemistry Atomic Spectroscopy

/ Instrumentation
Sample preparation in Atomic Absorption spectrometry
 • sample and flux is mixed and then fused to form products which are dissolved with
water or dilute acid.
• High temperature nearly (300 to 1000oC) is required which is achieved by flame,
conductive or microwave assisted heating.
• Few examples of fluxes are presented in Table 1
 Disadvantages of this approach
• There may be interference of impurities present in the fluxes,
• There may be high content of electrolytes in the resulting solution,
• And losses by volatilization.

Table 2: Fusion sample preparation

Temperature 300-1000ºC

Fluxes Lithium metaborate, Sodiumcarbonate Potassium nitrate,

Alkaline hydroxides

Crucible Platinum, nickel, silver, iron, graphite, porcelain

Disadvantages High content of dissolved solids

Prior to mineral element determination dry ashing is used for the elimination or minimization
of organic compounds by its ignition in air or in a stream of oxygen. Additives are used to
avoid losses of volatile elements such as As, Cd, Hg and Pb.

Table 3:Additives used for dry ashing of sample during sample

preparation

Temperature 500-800ºC

Volatile elements (As, Cd, Hg,Pb) Magnesium nitrate

Analytical Chemistry Atomic Spectroscopy

/ Instrumentation
Sample preparation in Atomic Absorption spectrometry
 Volatile chlorides (Zn, Sn etc.) Sulfuric acid

and volatile elements

Wet Decomposition or Acid Decomposition: Wet decomposition of metal samples is done by

oxidizing agents or minerals in order to have accuracy in determination of metal content in
sample.

• Concentrated acid is used along with heating for wet decomposition

 Important aspects to consider are
• Acid strength,
• its oxidizing and complexing power,
• Boiling point of the acid used
• Solubility of the resulting salts,
• Safety in manipulation,
• and purity.
Methods of Wet Decomposition: there are three methods known

• Thermal,
• Ultrasonic
• Radiant (infrared, ultraviolet and microwave).
Wet decomposition-thermal energy.

Decomposition in open vessel by using heat from a Bunsen burner, hotplate, digestion block,
oven or muffle

Limitations of Wet decomposition-thermal energy

• Time consuming
• Requires large amounts of reagents,
• Contamination from the environment,
• pre-concentration of reagent impurities,
• Strong oxidizing agents required, such as hydrogen peroxide,
• Constant supervision.

Analytical Chemistry Atomic Spectroscopy

/ Instrumentation
Sample preparation in Atomic Absorption spectrometry
 To overcome the problem

wet decomposition in a pressure decomposition container was first time performed by

Carius,
The sample and reagents were mixed and the tube was sealed
Tölg and coworkers and Bernas proposed

• The use of pressurized PTFE bombs

• Small sample masses in addition with acid or a mixture of acids,
• And pointed out increase in temperature increases the decomposition.
• They are especially used for the analysis of trace elements with organic, biological and
inorganic samples, alloys, metals, and minerals.

• An adapted pressure decompositionTölg’s bomb is shown in figure 1 as employed

nowadays containsCap, Pressure spring,Metal lid,PTFE lid,PTFE insert and a pressure
vessel

Figure 1: Scheme of Tölg’s PTFE bomb for sample preparation (Source: J. Braz. Chem. Soc . 14,
2, 2003)

Tölg’s bomb: Adapted pressure decomposition method:

It is placed on an aluminum heating block with a thermal probe, a cold water connection and
a heating current connection with an over-heating fuse. In the PTFE vessel during the process
of decomposition

Analytical Chemistry Atomic Spectroscopy

/ Instrumentation
Sample preparation in Atomic Absorption spectrometry
 Temperature limit is about 170oC
The sample must contain carbon up to 100 mg for the 35ml vessel.
A high-pressure asher was more recently presented by Knap which performs decompositions
at 180 to 300oC, practically generating carbon-free solutions

Figure 2:Scheme of Knapp’s High Pressure Asher for sample preparation.

(Source: J. Braz. Chem. Soc . 14, 2, 2003)

Table 4: Type of metal analyzed in a particular analyte and the reagents used for digestion.

Samples Analytes Reagents

Silicates Traces HF +H2SO4

Chromium ore Traces H2SO4+ HCl + NaCl a

Magnetite Ge H3PO4+ KMnO4

Pyrite Traces HCl+HNO3

Magnesium ores Traces HCl

(digestion of the residue)

Fluorite Traces H2SO4 + HCl

(digestion of the residue)

Analytical Chemistry Atomic Spectroscopy

/ Instrumentation
Sample preparation in Atomic Absorption spectrometry
 Mercury ores Traces HCl + HNO3 +H2SO4 c

Bauxite Traces HCl+ HNO3 + H2SO4

Sulfide ores Cu H3PO4+ HCl

a NaCl is used to form and remove volatile CrOCl, c Evaporation of the HgCl.

Table:4 Examples of wet dissolution-thermal energy for several ores.

Wet decomposition/extraction-ultrasonic energy.

• Is a simple and in expensive alternative method for sample preparation for biological
and inorganic matrices
• The extraction effect is considered as being caused by acoustic cavitation, that is,
bubble formation and subsequent disruptive action.
• To avoid this, Sonication of solutions is used to collapse the bubbles which results in
extremely high temperature (ca.5000K) and pressure (ca.10 GPa) gradients which
helps sample preparation.
• To reducing both reagent and time consumption a diluted acid medium is normally
used in comparison with traditional wet digestion
• For decomposition of organic compounds in environmental samples ultrasound
probes are generally employed

Wet decomposition-radiant energy-infrared radiation

This technique has been employed for organic and biological samples. Infrared radiation
(1.2x10-14 to 6.0x10-12 Hz) are known to increases molecular vibration and variation in
molecular rotation which results in generation of heat and also its propagation. A small
amount of biological sample,200 mg, plus 2 mL of nitric acid in a small glass flask (4 mL) is
placed in the focus of three IR lamps for 120 to180s, and 10 V are applied, then 1ml of
hydrogen peroxide is added followed by application of 10V for60s. To avoid contamination of
the solution, analytes are directly analyzed in the same tube.

Other applications of the technique are –

Analytical Chemistry Atomic Spectroscopy

/ Instrumentation
Sample preparation in Atomic Absorption spectrometry
 To determine volatile analytes by the heating of liquid and solid samples and preparation of
sub-boiling acids.

Wet decomposition-radiant energy-ultraviolet radiation: This technique is used in complete

removal of organic materialsespecially in polarography or voltametry for the subsequent
elemental determination. Examples include determination of heavy metals in waste-waters
or liquid foodstuffs such as beer and wine. Small amount of acid with hydrogen peroxide is
added to the sample, without a significant increase in the temperature.This results in less
contamination levels of reagents and avoids loss of volatile elements. UV radiation produces
highly reactive chemical radicals and ozone, which can be drawn away using a laboratory
hood.In secondary reactions, organic substances, which bind to heavy metals are degraded.
Sample and reagents containing several quartz tubes can be arranged around a UV emitter
placed in the center.

Wet decomposition-radiantenergy-microwave radiation

For a wide range of applications, Microwave-assisted sample preparation is now used, which
includes decomposition of inorganic and organic materials.

• both ionic migration and dipole rotation is caused by the interaction of microwave
radiation (2450 MHz, 12.2 cm) with sample and reagents resulting in rapid heating of
the mixture with consequent decomposition.
Advantages of this approach

• short time required (minutes) to perform decomposition of the sample,

• direct heating of samples and reagents (the vessels are only indirectly heated by the
hot solution),
• minimal contamination
• and no loss of volatile elements.
• Focused microwave oven can be used at atmospheric pressure for digestion of upto
10kg of sample with high concentration of organic materials
• The tubes used are made of glass, quartz or PTFE, and transparent to microwave
radiation and resist pressure upto 12MPa
• It regulates temperature vs. time or power vs.time (with simultaneous temperature
measurement).

Analytical Chemistry Atomic Spectroscopy

/ Instrumentation
Sample preparation in Atomic Absorption spectrometry
 • Temperature control is programmed in accordance with the boiling points of the acid
or acid mixture used ^
• Different security devices are used in Vessels and microwave ovens depends on the
manufacturer

Figure 3: Microwave Digestion System: a) waveguide type and b) cavity type (Source: J. Braz.
Chem. Soc . 14, 2, 2003)

Table 5: Boiling points of concentrated acid solutions and aqua regia atmospheric pressure

Acid Concentration%(m/v) b.p.°C

HCl 37 110

HF 49 108

HNO3 70 120

Aqua-regia (HCl:HNO3 3:1v/v) 112

Analytical Chemistry Atomic Spectroscopy

/ Instrumentation
Sample preparation in Atomic Absorption spectrometry
 H2SO4 98.3 338

HClO4 72.4 203

The pressure vs. time or power vs. timeor temperature vs. time can be controlled by heating
program. It emphasis should be given to that the selection of temperature be based on the
boiling point of the acid or mixture used (table 6). The relationship between sample amount
andpressure is presented intable 7. Thus, applications include practically all inorganic and
organic samples to be submitted to complete or partial decomposition, especially for
refractory materials and for foods with high contents of carbohydrates, proteins and lipids.
Examples are presented in the literature.

Table 6: Boiling point of concentrated acid solutions and aqua-regia at 7x105 Pa of pressure
in a sealed vessel.

Acid Concentration%(m/v) B.P °C

HCl 37 140

HF 49 175

HNO3 70 190

Aqua-regia (HCl:HNO3 3:1v/v) 146

Table 7: Relationship between pressure vessel and sample amount

%Fat content Sample mass Closed vessel

<75 <0.5g Low pressure (2MPa)

<75 >0.5g High pressure (10MPa)

>75 <0.25g Low pressure (2MPa)

>75 >0.25g High pressure (10MPa)

Analytical Chemistry Atomic Spectroscopy

/ Instrumentation
Sample preparation in Atomic Absorption spectrometry
 Present and Future Trends

Present and future trends are dictated by the techniques that emphasize on minimum
manipulation of the samples. This can be achieved by employing following approaches for
sample decomposition.

• analysis of slurries,
• determinations in situ and on line,
• the principle of single flask
• and automation/ robotics
Summary

• Why sample preparation is required

• Relationship between sample preparation and determination methods
• What Types of Samples are used and Steps for Consideration before carrying out the
analysis
• Proper Selection of Sample Preparation Method depending upon the sample available
• You will learn about various techniques like
Dry Decomposition.
Fusion and dry ashing
Wet Decomposition or Acid Decomposition
Thermal,
Ultrasonic
Radiant (infrared, ultraviolet and microwave).

Analytical Chemistry Atomic Spectroscopy

/ Instrumentation
Sample preparation in Atomic Absorption spectrometry
Analytical Chemistry Atomic Spectroscopy
/ Instrumentation
Sample preparation in Atomic Absorption spectrometry