 Trace elements

Element is considered essential if a deficiency impairs a biochemical or functional Trace elements

process  Example: Iron, Copper, and Zinc
 replacement of the element corrects this impairment  Found in milligram per liter or parts per million concentration

Deficiencies Ultratrace elements

 typically impair one or more biochemical functions and excess  Example: Selenium, Chromium and Manganese
concentrations are associated with at least some degree of toxicity  Found in microgram per liter or parts per billion concentrations

Conditions that could result in deficiency: Nonessential trace elements

 Decreased intake, Impaired absorption, Increased excretion and Genetic  are of medical interest primarily because many of them are toxic
abnormalities
Essential trace elements
Dietary requirement  are often associated with an enzyme (metalloenzyme) or another
 the smallest amount of the nutrient needed to maintain optimal function protein (metalloprotein) as a cofactor
and health

 Methods and Instrumentation

1. Atomic Emission spectroscopy  Chemical analysis that uses intensity of light Particularly used in: the form of inductively coupled
emiited from a flame, plasam or spark in plasma atomic emission spectroscopy (ICP-AES) for
wavelength to determine the quantity of an atomization and excitation
element in a sample

The three most important components of atomic
emission spectrophotometer are as follows:
1. Source
 the sample is atomized at a sufficient
temperature to produce an excited-state
species
 Those species will emit radiation
upon relaxation back to the ground
state
2. Wavelength selecting device (monochromator)
 for the spectral dispersion of the radiation
and separation of the analytical line from
other radiation
3. Detector permitting measurement of radiation
intensity
 A liquid sample is converted into an aerosol
and delivered into the source where it
receives energy sufficient to emit radiation
The intensity of the emitted radiation is correlated to
the concentration of an analyte and is the basis for
quantitation
The most commonly used sources:
1. Flame - are capable of producing
temperatures up to 3,000 K
2. Inductively coupled plasma (ICP)
Gases
 are combined in a specially designed mixing
chamber
 A sample is also introduced into the mixing
chamber using a nebulizer that converts
liquid into a fine spray
Typical fuel gases:
 include hydrogen and acetylene
Oxidant gases:
 include air, oxygen and nitrous oxide
Both atomic and ionic excited states can be produced
 depending on the element and the source
 leads to the production of complicated
emission spectra
Emission spectrum
 is composed of a series of very narrow peaks
(sometimes known as “lines”) with each line at
a different wavelength and each line matched
to a specific transition
 Each element has its own characteristic
emission spectrum
 Example: sodium can be detected by tuning the
monochromator to a wavelength of 589 nm
Interference-free wavelength
(atomic or ionic line)
 wavelengths producing suitable analytical
performance
 Example: limit of quantitation, freedom from
interferences, and robustness are selected
The first detectors in AES uses photographic film
Contemporary AES instruments feature
photomultiplier tubes or array-based detector
systems

2. Atomic absorption spectrometry (AAS)  an analytical procedure for the quantitative  most commonly used instrumentation for trace
determination of elements through the absorption and toxic metal analysis
of optical radiation by free atoms in the gas phase

Spectra of the atoms Typical radiation sources for AAS: Most common sources for AAS:
 are line spectra that are specific for the
absorbing elements a) Hollow cathode lamps (HCLs) 1) FAAS
 contains a quantity of the target element in  Flame atomic absorption spectroscopy
Absorption is governed by the Beer-Lambert law: the form of a hollow cylinder  Measures: Copper, Iron, and Zinc
 are an ideal source for determining most  Graphite tubes are the most commonly used
elements by AA atomizers
 Tubes are made of high-purity
b) Electrodeless discharge lamps (EDLs) polycrystalline electrographite and
 for volatile elements coated with pyrolytic graphite and
The four most important components of AA can be heated to a high
spectrophotometer are as follows: temperature by an electrical
1. Radiation (light) source current
o which emits the spectrum of  A small aliquot (usually 20 µL) of
the analyte element liquid sample is placed in the tube
2. Atomizer at the ambient temperature
o in which the atoms of the  Heating program: dry the sample -> pyrolyze ->
element of interest in the vaporize -> atomize the sample -> clean
sample are formed
 3. Monochromator 2) GFAAS
o for the spectral dispersion of  Graphite furnace atomic absorption
the radiation and separation of spectroscopy
the analytical line from other  also called flameless or electrothermal AAS)
radiation  Measures: Selenium, Cadmium, and Lead
4. Detector permitting measurement of  Allows for measurements of both liquid and
radiation intensity solid samples
 Common problem: analyte volatility depends
on the molecular form of the analyte and the
Mixing chamber burner: sample matrix
 produces laminar flames of high optical  Treatment: Chemical modifiers (palladium
transparency nitrate, magnesium nitrate, or a mixture of
both)

 Sample Collection and Processing

Specimens Specimen consideration

 Must be collected with details:
 anticoagulant, collection apparatus, and specimen type (urine, serum,
plasma, or blood)
 Extraordinary measures are required to prevent contamination of the
specimen:
 low concentration in biologic specimens and the ubiquitous presence in
the environment
 includes using special sampling and collection devices, specially cleaned
glassware, and water and reagents of high purity
Sample collection
 Venipuncture can be used to obtain serum, plasma and whole blood
 24 – hour urine can also be used
 Rarely, hair, nails or biopsy samples may be obtained
 Care arefully evaluate the following for the use in trace and ultratrace
analyses:
o selection of needles, evacuated blood collection tubes, anticoagulants
and other additives, water and other reagents, pipettes, and sample
cups
 Recommended measures include:
o placing the trace elements laboratory in a separate room incorporating
rigorous contamination control features
o Example: sticky mats at doors, non-shedding ceiling tiles, carefully
controlled air flow to minimize particulate contamination, disposable
booties worn over shoes, and particle monitoring equipment

Other Methods and Instrumentation:

Coupled Plasma Mass Spectrometry ICP-MS : a state-of-the-art analytical technique for elemental analysis

Plasma Argon plasma

 refers to an ionized gas (typically argon) in which a certain proportion of  induced by commercial ICP instruments (both ICP-AES and ICP-MS)
electrons are free  generates temperatures ranging from 6,000 to 10,000 K
 measures the mass-to-charge ratio (molecular mass divided by ionic  Steps:
charge [m/z]) of selected analyte ions) and includes the following 1. dries and then vaporizes the droplets produced by the nebulizer
components: 2. atomization of any molecular species
(1) an ion source 3. atoms are thermally ionized at which point they are ready for
(2) a mass analyzer introduction into the mass spectrometer
(3) an ion detector
Nearly, all ICP torches consist of three concentric quartz tubes surrounded by a coil
Quantitative analysis for clinical samples carrying radiofrequency (RF) power The middle tube of the torch carries the argon
 is best performed with the use of an internal standard (Ar) that forms the plasma

All patient samples, calibrators, and controls

 are diluted with an internal standard, usually a solution of an uncommon
element such as yttrium

Quadrupole Mass Spectrometers  the typical mass spectrometer used for ICP-MS

 Analyzer consists of four parallel conducting rods arranged in a square
array
 Applying RF and constant (DC) voltages to the rods, the instrument can
be tuned so that only ions of a specific m/z ratio can pass through the
device to reach the detector
 Able to well resolve peaks separated by one m/z unit but not able to
resolve peaks separated by a small fraction of an m/z unit
 Tends to be relatively simple to use and maintain, but the resolution (the
ability to discriminate between closely spaced m/z values) is limited

High-Resolution Mass Spectrometers  an ICP-MS instruments

 are usually “double focusing sector field” instruments
 separate ions of different m/z values via deflection in a magnetic field,
with ions of greater m/z being deflected to a lesser degree than those of
lower m/z
 The magnetic field is adjusted to allow only ions of a selected m/z to
reach the detection system at any given point in time
Commercially available high resolution ICP-MS instruments
 are capable of a resolution of 10,000 (10% valley)
 This is enough to resolve
 Example: 75As+ from 75ArCl+, both nominally 75 m/z units,
but which differ by 1 × 103 units when viewed at high
resolution
Electrostatic analyzer Magnetic sector instruments
 corrects for certain nonideal effects, allowing the instrument to achieve  are not able to resolve elemental isobaric interferences
high resolution  Example: 115Sn/115In and 40Ca/40Ar would require resolution much
higher than 10,000

 Interferences
 are classified as spectroscopic or nonspectroscopic

1. Spectroscopic

Spectral interferences:
 generally result from a spectral overlap with the spectrum of the target
analyte
 Example: in AA certain molecular species may have broad absorption
spectra that may overlap the line spectra of the elements of interest,
leading to false elevations of the target element concentrations
Various strategies are used to deal with spectral interferences:
 A continuum source background corrector may be included in the
instrument design
 Alternative is Zeeman background correction, which relies on shifting
the atomic spectral lines by the application of a magnetic field

A much less common occurrence would be for the absorption spectrum of one
element to overlap with that of another

2. ICP-MS

 spectral interferences include:

a) polyatomic species whose m/z may overlap m/z of the target
analyte
b) arises from nearby elements in the periodic table
c) comes from doubly charged ions

3. Nonspectroscopic

Matrix interferences Properties of significance

 involve the bulk physical properties of the sample to be analyzed  viscosity, presence of easily ionized elements, and presence of carbon
 aqueous samples may behave differently than organic and biological
specimens depending upon the technology used and the analyte of
interest
 Overcome matrix interference by: Matrix matching of the calibrators,
controls, and specimens
 Minimize matrix effect by: Dilution of the specimens but it is only
applicable to certain analytical techniques and to the determination of
analytes with higher concentrations

4. Nonspectral

 anything that could interfere with atomization of the sample  Examples:

 In AA: a flame may not be hot enough for efficient atomization
 Difference in sample viscosity between standards and unknown samples,
resulting in differing rates of sample introduction
 In AES: anything that would prevent the efficient excitation or emission
of spectral lines used for the analysis

 Alternative Analytical Techniques

Voltammetric methods Ion chromatography Gas chromatography–mass Laser ablation ICP-MS (LA-ICPMS)
spectrometry (GC-MS)

 Example: anodic stripping
voltammetry (ASV) and adsorptive
stripping voltammetry
 can be used in determination of
selected metals and are the basis
for some point-of-care devices
 can be used for the determination
of copper, iron, and zinc in blood,
serum, and plasma
 for the determination of zinc in
urine
 is capable of determination of  a method accommodating direct
cadmium, chromium, cobalt, analysis of solid samples
copper, lead, and selenium in
urine, copper in serum, and lead
in blood

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