A USE CASE DOCUMENTATION

“Inductive Coupled Plasma – Mass Spectroscopy”

At

“KONKAN SPECIALITY POLYPRODUCTS PVT LTD”

Industrial area, Baikampady, Mangalore
Submitted by

“SHRUTHI D”
103CH20054
Department of Chemical Engineering

DEPARTMENT OF CHEMICAL ENGINEERING

KARNATAKA (GOVT.) POLYTECHNIC
KADRI HILLS MANGALORE-575004
 A USE CASE DOCUMENTATION
“Inductive Coupled Plasma – Mass Spectroscopy”
At

“KONKAN SPECIALITY POLYPRODUCTS PVT LTD”

Industrial area, Baikampady, Mangalore
Submitted by

“ROOPASHREE”
103CH21704
Department of Chemical ngineering

DEPARTMENT OF CHEMICAL ENGINEERING

KARNATAKA (GOVT.) POLYTECHNIC
KADRI HILLS MANGALORE-575004
Serial Table of Content Page
No. No.

1 Synopsis

2 Introduction

3 Sample Preparation

4 Instrumentation and Working

5 Applications

6 About AP-89

7 Results
Serial Table of Content Page
No. No.

1 Synopsis

2 Introduction

3 Sample Preparation

4 Instrumentation and Working

5 Applications

6 About IS9833

7 Results
Inductively Coupled Plasma – Mass
Spectroscopy:
Synopsis:

Inductively coupled plasma mass spectrometry (ICP-MS) is an

analytical technique that can be used to measure elements at trace
levels in biological fluids. Although older techniques such as atomic
absorption and atomic emission are still in use by some laboratories,
there has been a slow shift toward ICP-MS, particularly in the last
decade. As this shift is likely to continue, clinical scientists should be
aware of the analytical aspects of ICP-MS, as well as the potential for
both spectroscopic and non-spectroscopic interference, and strategies
that can be employed to eliminate or mitigate these issues.

Introduction:

The measurement of trace elements in biological samples is useful in

a large number of clinical settings. Lists a number of elements of
clinical interest, along with an approximate guide to the concentration
ranges which may be encountered in biological samples. Elements
monitored for nutritional purposes include essential elements such as
iodine, manganese, copper, selenium and zinc. These elements play
important roles in a wide range of biological processes including
electron transport, oxygen transport, hormone synthesis and catalysis
of biological reactions. Disturbances in normal homeostasis of these
elements may cause (or be a symptom of) one or more
pathophysiological conditions. Other elements such as arsenic,
cadmium, mercury and lead are known to exert toxic effects (often
through a variety of different mechanisms), and are therefore
measured to assess exposure. A wide range of analytical techniques
have historically been used for trace element analysis. This review
will focus on the analytical aspects underlying ICP-MS.
There are six fundamental compartments of a single quadrupole ICP-
MS: the sample introduction system, inductively coupled plasma
(ICP), interface, ion optics, mass analyser and detector. shows a
simple diagram of the instrument. Liquid samples are first nebulised
in the sample introduction system, creating a fine aerosol that is
subsequently transferred to the argon plasma. The high-temperature
plasma atomises and ionises the sample, generating ions which are
then extracted through the interface region and into a set of
electrostatic lenses called the ion optics. The ion optics focuses and
guides the ion beam into the quadrupole mass analyser. The mass
analyser separates ions according to their mass-charge ratio (m/z), and
these ions are measured at the detector.

Sample Preparation:
Sample preparation for ICP-MS is relatively simple; biological
samples are usually diluted or thermally digested before analysis.
Common diluents include dilute acids (e.g. nitric acid, hydrochloric
acid) or alkali (e.g. ammonium hydroxide, tetramethylammonium
hydroxide). Deionised water has been used as a diluent, however
some elements are unstable in pure water, therefore acidic or alkaline
diluents are preferred in most cases. The isoelectric point of many
proteins is approximately 5–6, therefore the addition of an acidic
diluent to a highly proteinaceous sample such as blood may cause
protein precipitation, which may co-precipitate certain analytes or
cause an obstruction in the nebuliser. Proteins tend to be more tolerant
of dilute alkali, however not all elements are soluble at alkaline pH,
therefore a chelating agent such as EDTA is often incorporated into
alkaline diluents. Surfactants such as Triton-X100 are also commonly
added to help solubilise and disperse lipid and membrane proteins in
the sample.
Instrumentation and working of ICP MS

Inductively Coupled Plasma:

A plasma is essentially an ionised gas, consisting of positively-
charged ions and free (unbound) electrons. The role of the plasma
(ICP) in ICP-MS is to ionise the sample. In contrast to so-called ‘soft’
ionisation sources used in other forms of mass spectrometry (such as
electrospray) which impart relatively little energy to the analyte, the
ICP is considered a ‘hard’ ionisation technique because it completely
atomises most molecules in the sample. ICP-MS instruments use an
argon plasma, although helium plasmas have also been
described. Although there are several advantages to using helium,
argon is preferred as the cost of helium is prohibitive.

The plasma is formed in the end of a set of three concentric quartz

tubes, collectively referred to as the torch. Argon gas flows through
all three tubes. The inner tube is called the injector, and contains the
sample aerosol in a stream of argon which delivers the sample to the
plasma. Concentric to this tube is a tangential flow of argon called the
auxiliary gas, which forms the plasma. The outer tube contains a flow
of argon which serves as a cooling layer to prevent the torch from
melting. The far end of the torch is surrounded by a copper induction
coil (or ‘load coil’), which is connected to a radio frequency (RF)
generator. The RF generator supplies power to the load coil, creating
a high-frequency alternating current which in turn induces a time-
varying electromagnetic field in the torch. With argon gas flowing
through the torch, a high-voltage discharge (called a tesla spark) is
applied, which ionises a fraction of the argon atoms generating ions
and electrons. Ions and electrons in the torch are influenced by the
electromagnetic field, and are accelerated and collide with other argon
atoms. If these collisions impart sufficient energy, additional atoms
are ionised creating electrons and ions which propagate the cascade.
The movement of electrons and ions in the torch generate a
tremendous amount of heat. The result is called an ICP, which can
reach a temperature of up to 10,000 Kelvin (hotter than the surface of
the sun).

Nebuliser:
A number of different nebulisers are commercially available,
including pneumatic, ultrasonic and desolvating types. Pneumatic
nebulisers which use gas flow to generate the aerosol are the most
common type for routine clinical applications. There are several
different types of pneumatic nebulisers, including concentric, cross-
flow, Babington and v-groove (a variant of the Babington). Each has
its own advantages and disadvantages; the choice is application
dependent. Concentric nebulisers are ideal for low matrix (low TDS)
samples, whereas cross-flow and v-groove nebulisers are more
rugged, tolerating high matrix samples. In contrast to the pneumatic
design, an ultrasonic nebuliser utilises sound energy (from a
piezoelectric transducer) to generate the aerosol. These nebulisers
improve analytical sensitivity by an order of magnitude compared to
pneumatic nebulisers, however they are significantly more
expensive. Desolvating nebuliser systems use a heated spray chamber
to desolvate the sample before it reaches the plasma. In addition to
increasing sensitivity, desolvating nebulisers also decrease the
formation of oxide species in the plasma which can interfere with the
measurement of certain analytes (discussed in Interference section).

Spray Chamber:
After being aerosolised by the nebuliser, the sample enters the spray
chamber. The spray chamber has a simple design but serves a few
important purposes: it selectively filters out the larger aerosol droplets
that are generated by the nebuliser and acts to smooth out nebulisation
‘pulses’ produced by the peristaltic pump. This is important because
the plasma is inefficient at dissociating large droplets (>10 μm
diameter). In a double-pass spray chamber, aerosol droplets emerge
from the nebuliser and travel down a central tube in the spray
chamber. At the end of the tube, larger aerosol droplets exit the spray
chamber under the influence of gravity and are drained to waste while
smaller droplets, roughly <10 μm diameter, are transferred to the
plasma. Cyclonic spray chambers have a different design but operate
in a similar manner. In contrast to techniques such as graphite furnace
atomic absorption, the sample introduction process in ICP-MS is quite
inefficient. Normally only 1–2% of the sample reaches the plasma;
the remainder is drained to waste. Therefore factors which influence
the efficiency of sample introduction even slightly can have marked
effects on instrument response. One such factor is the spray chamber
temperature, which is normally maintained at around 2°C using a
thermoelectric cooling device or water jacket. Operating the spray
chamber at this temperature minimises the formation of oxides, and
prevents overloading the plasma with solvent.

Detector:
The most common detector used for ICP-MS is an electron multiplier
(EM). Positively-charged analyte ions strike the first dynode of the
detector which is held at a high negative voltage. The impact of the
ion on the detector causes the emission of several electrons from the
surface, which, in turn, strike the next dynode releasing more
electrons. This process (called secondary emission) continues,
generating an amplification cascade that culminates in a signal large
enough to be measured reliably as an ion ‘count’. In this way, an EM
can generate a measurable signal pulse from the impact of a single ion
on the detector, conferring very high analytical sensitivity. In fact,
detection limits in ICP-MS are far superior to flame atomic
absorption, and are comparable (or superior) to graphite furnace
atomic absorption. Typical limits of detection in ICP-MS are in the
nmol/L range for most elements; the exact value being dependent on
the element, the type of biological matrix, the dilution factor
employed during sample preparation, the design of the sample
introduction system, instrument operating conditions (including
plasma temperature) and background signals (reagent purity etc.).
Most detectors are able to operate in both a pulse (digital) and
analogue mode. These so-called dual detectors automatically switch
from pulse to analogue mode when the signal intensity exceeds a
certain threshold, allowing the linear dynamic range of the detector to
be extended to approximately 8–12 orders of magnitude. These two
detector modes require cross-calibration to ensure optimum linear
response across this range.

Application of ICP MS:

 For Quantification of toxic elements

 Study of Metal transport
 Adsorption and metabolic studies of metals and metallo-drugs
 Affinity products having metal tag are used for ICP-MS
immunoassays.
 Means of detection by fluorochromes and radioisotopes suffer
from limited dynamic range and limited ability to detect
multiple protiens simultaneously So, this provides a good
alternative.

It would be advisable to establish that the colourants being used meets

the applicable purity specifications.
In one or more EU member states and in the council of Europe
resolution AP(89) 1 on colourants.

Having regard to the fact the colourants are used to impart a colour to
plastic material coming into contact with food.

Considering that plastic materials coming into contact with food may
by reason of their colouration, pose a risk to human health if not used
under normal conditions or if the colourants used to do not meet
purity criteria based on good manufacturing practice.

Taking the view that each member state faced with the need to
introduce regulations governing this matter would find it beneficial to
harmonise such regulations at European level.

The present standards for pigments (AP89) to ensure that the

pigments and colourants used were not harmful to consumers due to
any leaching of the pigments into the food products

AP(89)1:The use of colorants in plastic materials and articles coming

into contact with food. The Resolution defines purity requirements for
pigments including concentration limits for extractable heavy metals.

Test Procedure
 Approximately 10 gm of sample taken in glass beaker and added
150ml of 0.1M HCl
 Kept the beaker with sample in mechanical agitator for 15
minutes.
 Allowed it to stand for 15mins.
 After 15 minutes filtered the solution.
 Then analysed the same in ICP-MS

Impurities Limit(ppm)

Antimony(Sb) 500ppm

Arsenic(As) 100ppm
 Barium(Ba) 100ppm

Cadmium(Cd) 100ppm

Chromium(Cr) 1000ppm

Lead(Pb) 100 ppm

Mercury (Hg) 50ppm

Selenium(Se) 100ppm