Laser Ablation Inductively Coupled Plasma Mass ...joics.org/gallery/ics-1890.pdf3.3.1. Inductively...

Laser Ablation Inductively Coupled Plasma Mass Spectroscopy-A Review

*Dr. Bhavyasri. Khagga1, Dhana Lakshmi.Chinta2, Swetha sri. R3

1Department of pharmaceutical analysis, Associate Professor, RBVRR women’s college of

pharmacy, Barkatpura, Hyderabad, Telangana, India.

2Department of pharmaceutical analysis, Research Student, RBVRR women’s college of

pharmacy, Barkatpura, Hyderabad, Telangana, India.

3Department of pharmaceutical analysis, Assistant Professor, RBVRR women’s college of

Pharmacy, Barkatpura, Hyderabad, Telangana, India.

Corresponding Author Email:[email protected],

Abstract:

Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) has received

significant attention over the last 10 years and has been widely used for analysis of biological

samples. The technique allows elements and isotopes determination in biological tissues and related

materials with a spatial resolution typically ranging from 10-100 µm. When compared to other

techniques usually employed to obtain bio images, the greater advantage of LA-ICP-MS is its

higher sensitivity. Applications of isotope ratio (IR), including trace experiments, and isotope

dilution (ID) are reviewed for biological samples (briefly for proteins, only in order to show the

utility of LA-ICP-MS). Bio imaging methods, studies and applications to animal and plants tissues

are emphasized, demonstrating the importance bio images of metals and metalloids in biomedical

research, bioaccumulation and bioavailability studies for ecological and toxicological risk

assessment in humans, animals and plants.

Key words: LA-ICP-MS, Plasma, Photons, Energy, Elements, Isotopes, Atomizing, Ionizing.

1. Introduction:

LA-ICP-MS (Laser Ablation Inductively Coupled Plasma Mass Spectrometry) is a powerful

analytical technology that enables highly sensitive elemental and isotopic analysis to be performed

directly on solid samples. LA-ICP-MS begins with a laser beam focused on the sample surface to

generate fine particles – a process known as Laser Ablation. The ablated particles are then

transported to the secondary excitation source of the ICP-MS instrument for digestion and

ionization of the sampled mass. The excited ions in the plasma torch are subsequently introduced to

a mass spectrometer detector for both elemental and isotopic analysis [1].

1.1. Benefits of LA-ICP-MS:

LA-ICP-MS is one of the most exciting analytical technologies available because it can

perform ultra-highly sensitive chemical analysis down to ppb (parts per billion) level —

without any sample preparation.

Samples can be both conducting or non-conducting, and the analysis can be performed

in the air without the need for a complex vacuum system. Results are available within

seconds; therefore, LA-ICP-MS delivers that fastest analysis speed of all analytical

techniques with the limit of detection approaching ppb level.

Journal of Information and Computational Science

Volume 9 Issue 12 - 2019

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The sample mass size required for LA-ICP-MS analysis is sub-microscale —

picograms to femtograms. Traditional liquid nebulization approaches for ICP-MS

require the removal of milligrams of sample mass in order to be effective [2].

When applied with optimized laser ablation conditions and ICP-MS data acquisition

protocols, LA-ICP-MS allows versatile solid sampling schemes that include:

Bulk analysis

Local inclusion and defect analysis

Depth profiling

Elemental/isotope mapping

The two most commonly used laser-based methods are bulk analysis with a typical laser

spot size of 100 ~ 350μm and microanalysis with the laser spot size as small as a few

microns.

2. Principle:

Laser is an acronym for Light Amplification by Stimulated Emission of Radiation. As a

consequence of its light-amplifying property, a laser produces spatially narrow and extremely

intense beams of radiation having identical frequency, phase, direction and polarisation properties.

According to the resonance condition (∆E = hv) and to the rules of quantum mechanics, an atom

can change its energy level, which leads to the absorption or emission of a photon. Stimulated

emission is the basis of laser behaviour. Stimulated emission leads to the emission of a coherent

radiation with the incoming radiation. In order to have light amplification in a laser it is necessary

that the number of photon produced by stimulated emission exceed the number of photons lost by

absorption. This light amplification is only achieved when a population inversion from the normal

distribution of energy state exists. This population inversion (activation of a laser material) is

created by an external pumping source, so that a few photons of proper energy will trigger the

formation of a cascade of photons of the same energy. This cascade of photons is focussed on a

sample. Interaction between the laser beam allows the conversion of photon energy into thermal

energy, which is responsible for the vaporisation of most of the exposed solid surface. The material

ablated is swept away with an argon stream to an Inductively Coupled Plasma Mass Spectrometer

(ICP-MS) and analysed [3].

3. Instrumentation:

Figure 1. Block diagram of LA-ICP-MS.



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The LA-ICP-MS instrument consists of

Laser ablation chamber

Inductively coupled plasma(ICP)

Mass spectroscopy(MS)

These parts are discussed in detail in the following:

Figure 2. Instrumentation of LA-ICP-MS.

3.1. Concept of LA-ICP-MS:

Laser Energy

↓

Solid sample

↓

Solid vaporization

↓

Dissociation

↓

Atomization

↓ in ICP-MS

Ionization/Excitation of elements



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3.2. What Is Laser Ablation?

Laser ablation or photoablation is the process of removing material from a solid (or occasionally

liquid) surface by irradiating it with a laser beam. At low laser flux, the material is heated by the

absorbed laser energy and evaporates or sublimates. At high laser flux, the material is typically

converted to a plasma. Usually, laser ablation refers to removing material with a pulsed laser, but it

is possible to ablate material with a continuous wave laser beam if the laser intensity is high

enough. Excimer lasers of deep ultra-violet light are mainly used in photoablation; the wavelength

of laser used in photoablation is approximately 200 nm. Lasers are such as the frequency-

quadrupled Nd: YAG (Neodymium-doped yttrium aluminium garnet) in which the atoms that emit

light are fixed within a crystal or a glassy material. Lasers are characterised mainly by their

wavelength (1064 nm for a Nd: YAG), output power and pulse length fixed by the design of the

laser (nanosecond or picosecond) [4-5].

Figure 3. Laser ablation chamber.

3.3. What Is ICP-MS?

It is an inorganic (elemental) analysis technique.

ICP-Inductively Coupled Plasma.

High temperature ion source.

Decomposes, atomizes and ionizes the sample.

MS - Mass Spectrometer.

Featuring quadrupole mass analyser.

Mass range - 7 to 260 amu (Li to U...).

Separates all elements in rapid sequential scan.

Ions measured using dual mode detector.

Ppt to ppm levels.

Isotopic information available.

3.3.1. Inductively coupled plasma:

An inductively coupled plasma is a plasma that is energized (ionized) by inductively heating the gas

with an electromagnetic coil and contains a sufficient concentration of ions and electrons to make



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the gas electrically conductive. The plasmas used in spectrochemical analysis are essentially

electrically neutral, with each positive charge on an ion balanced by a free electron. The plasma

used in an ICP-MS is made by partially ionizing argon gas (Ar → Ar+ + e−). The energy required

for this reaction is obtained by pulsing an alternating electric current in wires that surround the

argon gas. The Sample first atomizes then ionize and detected. Icp for spectrometry is sustained in a

torch [6-8].

3.3.2. Basic functional components-ICPMS:

3.3.3. Instrumentation:

Sample introduction system.

Icp torch.

Interface.

Vacuum system.

Collision/ reaction cell.

Ion optics.

Mass spectrometer.

Detector.

Figure 4. ICP-MS instrument.

a) Sample introduction system:

Composed of a nebulizer and spray chamber. Liquid sample introduced to nebulizer by

peristaltic pump. Aerosol formation. Fine droplets are passed through spray chamber.

Argon gas is used.



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b) ICP torch:

Argon plasma @10000K. Copper coil connected to RF generator. Collisions of electron

with argon atoms. Ionization of Ar. Oscillation of ions and electrons. Ion transfer from

atmospheric pressure to vacuum.

c) Interface:

Allow the plasma and ion lens to coexist. Allows ions generated by plasma to pass through

ion lens. It consists of 2-3 cones. Downstream focusing of ion is done by charged ion

lenses. If 3 cones are used, then there is no need for ion lens.

Interface serves to direct the ions toward the mass spectroscopy.

Interface area consists of two conical metal (usually nickel) called sampler cone and

skimmer cone, each with small holes to allow ions to pass through to the ion optics.

Figure 5. Interface.

d) Collision\reaction cell:

Interferences caused when ions carry a mass-to-charge ratio identical to analyte ion. Passing

both through a cloud of inert gas molecules. Kinetic energy discrimination. Collision cell

can be a reaction cell. Inert gas is used. No-gas mode - no gas in the cell - the instrument

performs like a standard ICP-MS. High sensitivity is achieved for all elements. This mode

is typically used for uninterfered elements such as Be, Hg, Pb Helium (Collision) mode -

used for all analytes that suffer matrix-based Interferences are removed based on their

physical size (Reaction) mode - used only for the very few situations where He collision

mode is not efficient enough.

Figure 6. Reaction mode.

e) Ion optics:

Delivers ions toward the quadrupole. Stop or hinder the electrons, protons and neutrons to

the detector analyser[9-14].



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Fig-7: Ion Optics.

3.4. Mass Spectroscopy:

Principle: In this technique the compounds under investigation are bombarded with a beam of

electrons which produce an ionic molecule. The resulting assortment of charged particles is then

separated according to their masses.

1/2mv²=eV. (1)

m=Mass of particle.

v=velocity of particles.

e=charge on electron.

V=accelerating voltage.

3.4.1. Mass Analysers:

After ions are formed in the source region they are accelerated into mass analysers by electric field.

They deflect ions down a curved tube in a magnetic fields based on their kinetic energy determined

by the mass, charge and velocity [15-17].

The magnetic field is scanned to measure different ions.

Types of mass analyser:

(1) Quadrapole mass filter.

(2) Magnetic sectors.

(3) Time of flight.

(4) Ion trap.

1) Quadrapole mass analyser:

In 1955 introduced by W. Paul. A Quadrupole mass filter consists of four parallel metal

rods with different charges. Two opposite rods have an applied + potential and the other two

rods have a – potential. The applied voltages affect the trajectory of ions traveling down the

flight path. For given DC and AC voltages, only ions of a certain mass-to-charge ratio pass

through the quadrupole filter and all other ions are thrown out of their original path.

Potential of 5-10 V applied. High scan rates [18].



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Fig-8: Quadrapole mass analyser.

2) Magnetic sectors:

Designed by mattauch and herzog. Electrostatic field applied. Selection of ions of certain

velocity or K.E. High resolution. Used for determination of precise molecular weight.

Figure 9. Magnetic sector.

3) Time of flight(TOF):

TOF Analysers separate ions by time without the use of an electric or magnetic field. In a

crude sense, TOF is similar to chromatography, except there is no stationary/ mobile phase,

instead the separation is based on the kinetic energy and velocity of the ions.

4) Ion trap:

TOF Analysers separate ions by time without the use of an electric or magnetic field. In a

crude sense, TOF is similar to chromatography, except there is no stationary/ mobile phase,

instead the separation is based on the kinetic energy and velocity of the ions [19-20].

3.4.2. Detector system:

As ions hit the surface they are converted to electrons. They ensure high sensitivity and low

background noise. They are expensive. Light sensitive. Depending on usage last for 6-18

months. Once the ions are separated by the mass analyser, they reach the ion detector,

which generates a current signal from the incident ions [21-23].

The most commonly used detectors in MS are as follows:

Faraday cup.

Electron multiplier.

Photomultiplier dynode.



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1. Faraday cup:

TOF Analysers separate ions by time without the use of an electric or magnetic field. In

a crude sense, TOF is similar to chromatography, except there is no stationary/ mobile

phase, instead the separation is based on the kinetic energy and velocity of the ions [24].

Figure 10. Faraday cup.

2. Electron multiplier:

It is the most common means of detecting ions. It is made up of a series (1 to 24) of

aluminium oxide (Al2O3) dynodes maintained at ever increasing potentials. Ions strike

the first dynode surface causing an emission of electrons. These electrons are then

attracted to the next dynode held at a higher potential and therefore more secondary

electrons are generated. Ultimately, as numerous dynodes are involved, a cascade of

electrons is formed that results in an overall current gain on the order of one million or

higher. The high energy dynode (HED) uses an accelerating electrostatic field to increase

the velocity of the ions and serves to increase signal intensity and therefore sensitivity [25].

Figure 11. Electron multiplier.

3. Photomultiplier dynode:

The photomultiplier conversion dynode detector is not commonly used. It is similar to

electron multiplier in design where the secondary electrons strike a phosphorus screen

instead of a dynode. The phosphorus screen releases photons which are detected by the

photomultiplier and are then amplified using the cascading principle. One advantage of

the conversion dynode is that the photomultiplier tube is sealed in a vacuum, unexposed

to the environment of the mass spectrometer and thus the possibility of contamination is

removed [26].



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Figure 12. Photomultiplier dynode

4. Sample Preparation of LA-ICP-MS:

In general sample preparation is straightforward. Polishing is usually necessary if the electron

microprobe is to be used before loading the sample into the sample cell.

Because the laser sample cell is not under vacuum, outgazing of the support material is not a

problem [27].

Powdered samples should be treated in a similar way to XRF (X-Ray Fluorescence) analysis and

stabilised either by

(a) Compacting the sample into a pellet with or without a binding agent, [Fig:(a)].

(or)

(b) Fusing the sample to a borate glass or using a strip heater. [Fig :(b)]

Figure (a). Pellet Preparation. Figure (b). Fusing of sample.

5. Applications:

Laser ablation has been applied to the determinations of the trace element concentration in a wide

range of materials in many different studies, including volcanology, petrology, mineralogy, ore

deposits, materials and forensic studies, cultural heritage and environmental sciences. In the

environmental science, LA-ICPMS analyses is applied to a variety of biological structures, such as

tree rings, mollusc shells, otoliths, fin ray and fish scales. This is a consequence of two important

factors:

Analyses of solids by laser requires little (or) no sample preparation;

The determination of trace elements content (or) isotopic composition in almost all types of

materials is achieved at high spatial resolution [28]



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1) Analysis of trace elements in lipsticks:

Each lipstick was analysed by LA-ICP-MS to determine the trace element composition.

Four to five smears of lipstick, around 2.4 cm x 5 mm, were applied to clean Teflon squares

(2.5 x 2.5 cm) for laser ablation. Teflon contains minimal trace metal background and thus

was chosen as the substrate for the lipstick smears. The Teflon squares were soaked in 12%

nitric acid with 18 MΩ·cm Millipore water for a minimum of 48 hours before use. The

nitric acid was prepared by in-house sub-boiling distillation. The lipstick smears were

ablated with a femtosecond laser, the laser in a single line scan pattern. Argon was used as

the carrier gas through the ablation cell. The nominal spot size was ~70 µm. e.g.: 7Li+, 25Mg+,

26Mg+, 27Al+. Etc.

2) Imaging of brain tissues:

Images of C, Cu, and Zn measured on a thin section of rat brain tissue by LA-ICP-MS

obtained in the routine mode are summarized. The shape and structure of these LA-ICP-MS

images are in good agreement with the photograph of the tissue from a similar rat brain

stained with Cresyl violet. The LA-ICP-MS images demonstrate that the distribution of the

two metals and one abundant non-metal is quite different. The 13C+ and 64Zn+ images also

clearly demonstrate the localization of the hippocampus in the rat brain. An interesting

multi-layered structure of the cortex is especially visible in the 63Cu+ image.

Figure 13. Imaging of brain tissues.

3) Analysis of soil:

Reference materials: Five Certified Reference Materials: two soils, two lake sediments, and

one stream sediment, were used to determine the best sample preparation method for LA-

ICP-MS of soil samples.

Pellet preparation:

• Pellets without binder: Well homogenized samples were pressed into a pellet of 1.2 cm

diameter and 2 mm height.

• Pellets with boric acid: Pellets with boric acid were prepared as described. This approach

has been developed for ED-XRF analysis of soils. The samples (0.1 g) were mixed with

H3BO3 ratio 1:5, well homogenised and pressed into pellets. Blank pellets, containing only

boric acid were prepared in the same way.

• Pellets with organic solvents: To a soil sample (0.6 g) placed in an agate mortar, 1 ml of

hexane is added. The mixture is stirred for 10 min and dried in a heater block at 50oC.

Afterwards, 1 ml of dichloromethane is added, stirred for 10 min and dried in a heater block

at 50oC. Soil pellets are prepared as described above.

Determined isotopes:7Li, 23Na (0.013), 24Mg (0.013), 27Al (0.015), 28Si (0.017), 31P (0.012), 39K (0.016), 42Ca (0.013), 47,49,50Ti (0.012), 51V,52,53Cr.



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4) Ornithology:

The elemental (Cd, Cu, Pb, Pd, Pt, Rh and Zn) profiles along feather shafts of various bird

species in Sweden, namely, peregrine falcon (Falco peregrinus), sparrowhawk (Accipiter

nisus), willow grouse (Lagopus lagopus) and house sparrow (Passer domesticus) were

investigated by LA-ICP-MS. For analysis, the feathers were cut to lengths of 3–4 cm and

fixed on sticky tape without additional preparation. A line was ablated along shaft sections

using a spot size of 50–100 mm using a laser energy of 0.7–0.8 mJ, a repetition rate of 20

Hz, and a translation rate of 20 mms. Internal standardisation using the Ca signal intensity

was employed. Ablation of feathers revealed that Pb, Cu and Cd levels were highest in

urban habitats. The profile of Pt, Pd and Rh showed that contamination of feathers consists

almost exclusively of externally attached particles. The highest total metal concentrations

were found to be in the feathers of sparrowhawk and house sparrow because of their urban

habitats.

5) Marine studies:

Elemental analysis of marine samples is still a major area of research utilising LA-ICP-MS,

primarily for providing a chemical archive of environmental changes by the analysis of

shells, coral and fish otoliths.

Coral samples were cut into 45×25 mm sections and cleaned with pure water. The outer

tissue surface was then cleaned with ultrasonic agitation in a 30% hydrogen peroxide

solution and again in water, and dried at 41oC.

The samples were preablated at a repetition rate of 50 Hz and a laser energy of 100 mJ

using pulses from a 193-nm ArF excimer laser. A VG Elemental PG2+ICP-MS was used

for the analyses.

To obtain representative sampling over the structural elements of the coral a mask of 100

mm parallel to the growth axis by 500 mm (perpendicular to the growth axis) was found to

be essential.

For analysis, the laser was pulsed at 10 Hz at 100 mJ. Rare earth elements (La, Ce, Pr, Nd,

Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb and Lu) were determined using the same laser system

at a repetition frequency of 20 Hz but with an Agilent 7500s mass spectrometer. [29-30]

6. Conclusion:

LA-ICPMS is a fast and effective characterization technique for trace element analysis of solid

samples. Although the laser-material interaction is complex, it was observed that an increase in the

fluence, repetition rate and spot size can increase the signal intensity for an element at the ICP-MS.

Impurity concentrations were measured at the grain boundary and grains for high-performance

multi-Si wafers and it was found that ablated grain boundaries contain higher concentrations of

metals, which could have resulted from preferential segregation of impurities at grain boundaries

during crystal growth. Lifetime samples fabricated from standard p-type multi-Si show spatially

non-uniform degradation characteristics during LID. Spatial trace element analysis was performed

on two regions that differed in their response to light induced degradation.

7. Acknowledgement:

I would like to thankful to our college “RBVRR Women’s college of pharmacy” for giving me this

great opportunity.



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