Basic Environmental Engineering and Pollution Abatement

Instrumental techniques of environmental analysis 1

DR. PRASENJIT MONDAL

CHEMICAL ENGINEERING

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 CONTENTS
 Environmental analysis
 Instrumental analysis
 Electrochemical analysis
 Chromatographic analysis
 Spectroscopic analysis
 Other methods

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 Environmental analysis
 Environmental analysis encompasses matrices from the hydrosphere, atmosphere,
lithosphere, and biosphere, and target analytes include naturally occurring chemicals and
anthropogenically derived contaminants. Different methods used for chemical analysis are:
• Qualitative analysis
Identification/detection of elements or compounds present in in a sample
• Quantitative analysis
Determination of quantities of components or elements contained in a sample
 Volumetric method
o Performed by volumetric calibration / titration
 Gravimetric method
o Performed by using weighing of a material
• Instrumental analysis
 Use of instrument for the detection and quantification

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 Instrumental analysis
 Instrumental analysis can provide fast, sensitive, and reliable analytical
techniques which can be carried out with minimum environmental sample
handling. High resolution in some instrument provide additional structure
 Electrochemical
information analysis
of the sample
• Based on the electrochemical properties of solutions
 Some common example : analysis for pH/ORP, Conductivity, Dissolved Oxygen, Titration
 Chromatographic separation analysis
• Based on the separation of sample components according to chemical and/or physical
characteristics
 Some common example : GC, HLPC, Ion chromatography
 Spectrophotometric analysis
• Based on the interaction of matter with light
 Some common example : UV Visible, FTIR spectroscopy, fluorescence, AAS, MS
 Others Brunauer–Emmett–Teller (BET)
• Furnish information on chemical composition, structure, and decomposition of the sample
 Some common example : TGA, BET analyzer, XRD, SEM, GC-MS, LC-MS
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 GC
 Chromatographic analysis contd.
• Chromatography is a technique that separates components in a mixture by the
difference in partitioning behavior between mobile and stationary phases.
• Gas chromatography (GC) is one of the popular chromatography techniques to
separate volatile compounds or substances.
• The mobile phase is a gas such as helium, and the stationary phase is a high-
boiling liquid that is adsorbed on a solid.
• Because of its simplicity, high sensitivity, and the ability to effectively separate
mixtures, gas chromatography has become one of the most important tools in
chemistry.

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  Chromatographic analysis contd. GC

There are three main GC system Source: Stauffer et al., 2008

components:
Sample injection unit

Heats the liquid sample and vaporizes it

Column
GC components
Used to separate each compound
Detector Components in the mixture are distributed
Detects the compounds and outputs between two phases, one of which is a
their concentrations as electrical stationary phase, and the other is a mobile
signals
phase gas, or carrier gas, that carries the
mixture through the stationary phase.

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  Chromatographic analysis contd. GC

First, the
sample is Sample
When the
introduced into components
Substances that components are
a stream of move through
interact more eluted from the
inert gas, or a the packed
with the column, they
carrier, which column at a
stationary can be
is usually rate affected by
phase are quantified
helium or the degree of
delayed and and/or
argon. For a interaction of
thus separated collected
liquid sample, each
from through the
it needs to be component
substances that detector for
evaporated with the
interact less. further
before being stationary non-
analysis.
injected into volatile phase.
the carrier.

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  Chromatographic analysis contd. GC

Principle of gas chromatography

• Compounds in the mobile phase
interact with the stationary phase as
they pass through.
• Due to the differences in properties
and structures of each component,
the size and affinity of each
interaction with the stationary phase
are different. Therefore, under the
same driving force, the retention
time of different components differs
in the column, thus moving out of
the column in different orders.

Source: shimadzu.com Source: Jam and Waters, 2014 8

  Chromatographic analysis contd. Column GC

• GC columns may be of packed columns or capillary columns

• Short, thick columns made of glass or stainless-steel tubes; packed columns have been
used since the early stages of gas chromatography
• Currently the prevailing column type, capillary columns produce sharp peak shapes,
achieve excellent separation performance, and are suited to high-sensitivity analysis
• An optimized chromatographic separation begins with the column

The selection of the proper capillary column for any application should be
based on four significant factors:

• Stationary phase
• Column length
• Colum ID
• Film thickness

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 Chromatographic analysis contd. Column GC

Tube •Internal Diameter: 2 to 4 mm

•Length: 0.5 to 5 m (most commonly 2 m)
Packing
•Packing: Support material with 0.5 to 25
% liquid phase (partition material) or no
Packed column liquid phase (adsorbent material)
•Liquid Phase: Multiple types
Fused silica or
stainless
•Internal Diameter: 0.1, 0.25, 0.32, 0.53 mm
Liq. phase •Length: 5 to 100 m (most commonly 30 m)
•Material: Fused silica glass
Capillary column •Liquid Phase: Good separation, multiple types

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  Chromatographic analysis contd. Detectors GC

Different types of detectors used in GC are:

Flame ionization detector (FID)

Thermal conductivity detector (TCD or hot wire detector)
Electron capture detector (ECD)
Photoionization detector (PID)
Flame photometric detector (FPD)
Thermionic detector
A new variant of the FPD called the pulsed flame photometric detector (PFPD)
Atomic emission detector (AED)
Ozone- or fluorine-induced chemiluminescence detectors.

All of these except the AED produce an electrical signal that varies with the amount of
analyte exiting the chromatographic column

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  Chromatographic analysis contd. GC
chromatogram :
The X-axis – Retention time of peak
(Rt)
This is calculated from the time the
sample was injected into the column
(t0) till it reaches the detector. Every
analyte peak has a retention time that
is measured from the apex of the peak,
just like tR.
The Y-axis – Detector response
This shows the measured response of the analyte peak within the detector.
The baseline here represents the signal received from the detector where no analyte is eluting from the
column or is beneath the detection limit. It is considered as an indication of a problem or indication to
check the maintenance, in situations where the baseline is found higher than usual.

Measurements such as width at the baseline, width at half height, area, and total height can be withdrawn
from the peak. For better sensitivity and better resolution, narrower, sharper peaks are desired.

Source: technologynetworks.com 12
 Chromatographic analysis contd. Advantages GC
High separation efficiency and analysis speed: for example, gasoline samples can
obtain more than 200 chromatographic peaks in 2 hrs. A general sample analysis can be
completed in 20 minutes.
Small sample consumption and high detection sensitivity: 1 ml of gas sample
consumption, 0.1 µl of liquid sample consumption, a few mg of solid sample
consumption. Proper detectors can detect impurities in the tens to a few parts per
million.
Gas chromatography has good selectivity and can be used to analyze azeotropic
mixtures and samples with close boiling points. For example, some isotopes, cis-trans
isomers, adjacent or intertrans isomers, optical isomers, etc.
Wide range of applications, although mainly used to analyze gases and volatile organic
substances; under certain conditions, it can also be used to analyze high boiling point
substances and solid samples.

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  Chromatographic analysis contd. GC
Limitations
It can only be used to analyze volatile substances. Some highly polar substances can be
derived to increase their volatility for GC analysis, but the process can be complex and may
introduce errors in quantitative analysis.
Issues
One of the major problems with gas chromatography is leakage. As the mobile phase is a gas
that flows through the system, leakage may occur. Therefore, it is crucial to ensure that the
parts and consumables are correctly installed, and the system is regularly checked for leakage
Another problem is the activity for more polar analytes, especially the ones at trace levels.
Issues like irreversible adsorption or reactant breakdown can also occur due to dirt build-up in
the system and silanol groups on the glass liners and columns.
Most problems are seen on the inlet area where the sample is injected, transferred and
vaporized into the GC column. Hence, ensuring proper maintenance of the inlet and use of
correct consumable is essential.
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 Chromatographic analysis contd. GC
Suitability of GC for analysis of different Compounds
• High volatility
• Thermal stability
• Low molecular weights
Compounds that cannot be analyzed
• Compounds that do not vaporize (inorganic metals, ions, and salts)
• Highly reactive compounds and chemically unstable compounds
(hydrofluoric acid and other strong acids, ozone, NOx and other highly reactive compounds)

Compounds that are difficult to analyze

• Highly adsorptive compounds (compounds containing a carboxyl group, hydroxyl group,
amino group, or sulfur)
• Compounds for which standard samples are difficult to obtain (Qualitative and quantitative
analyses are difficult.)
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  Chromatographic analysis contd. GC-MS
• The separated substances emerge from
the GC column opening flow into the MS
• The MS breaks each separated
compound coming from the GC into
ionized fragments
• To do this, a high energy beam of
electrons is normally passed through the
sample molecule to produce electrically • Mass spectrometry identifies compounds by
charged particles or ions the mass of the analyte molecule
• Once the sample is fragmented it is then • A library of known mass spectra, covering
detected, usually by an electron several thousand compounds, is stored on a
multiplier , which essentially turns the computer
ionized mass fragment into an electrical • Mass spectrometry is considered the only
signal that is then detected definitive analytical detector (mass detector)
https://en.wikipedia.org/wiki/Gas_chromatography%E2%80%93mass_spectrometry
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  Chromatographic analysis contd. GC-MS

• 2,4-dichlorophenol (DCP) The concentrations of

• 2,4,5-trichlorophenol (TCP) phenolic compounds in
• pentachlorophenol (PCP) water sample obtained
• bisphenol A (BPA) Compound Concentrati
on (µg/mL)

Phenol 8.15
DCP ND

TCP ND
PCP 11.67
The chromatogram of spiked tap water sample. (a: phenol; b: BPA 7.94
DCP; c: TCP; d: PCP; e: BPA) Source: Meng et al., 2011
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 Chromatographic analysis contd. HPLC
High performance liquid chromatography or commonly known as HPLC is an analytical
technique used to separate, identify or quantify each component in a mixture.
The mixture is separated using the basic principle of column chromatography and then
identified and quantified by spectroscopy.
HPLC is thus basically a highly improved form of column liquid chromatography. Instead of a
solvent being allowed to drip through a column under gravity, it is forced through under high
pressures of up to 400 atmospheres.
It is used for the separation of components of an organic mixture of compounds of type
• Nonvolatile
• Thermally unstable
• Have relatively high molecular weights.
In the column, the stronger the affinity (e.g.; van der waals force) between the component and the mobile
phase, the faster the component moves through the column along with the mobile phase. On the other hand,
the stronger the affinity with the stationary phase, the slower it moves through the column.
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 Chromatographic analysis contd. HPLC

PC for Data
acquisition

Injection

HPLC column Detector Waste

Solvent

Pump

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 Chromatographic analysis contd. HPLC
• The purification takes place in a separation column between a stationary and a mobile phase.
• The stationary phase is a granular material with very small porous particles in a separation
column.
• The mobile phase, on the other hand, is a solvent or solvent mixture which is forced at high
pressure through the separation column via a valve with a connected sample loop, i.e., a
small tube or a capillary made of stainless steel, the sample is injected into the mobile phase
flow from the pump to the separation column using a syringe.
• Subsequently, the individual components of the sample migrate through the column at
different rates because they are retained to a varying degree by interactions with the
stationary phase.
• After leaving the column, the individual substances are detected by a suitable detector and
passed on as a signal to the HPLC software on the computer.
• At the end of this operation/run, a chromatogram in the HPLC software on the computer is
obtained.

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 Chromatographic analysis contd. Chromatogram HPLC
When no compounds are eluted from the
column, a line parallel to the horizontal axis is
plotted. This is called the baseline. The
detector responds based on the concentration
of the target compound in the elution band.
The obtained plot is more like the shape of a
bell rather than a triangle. This shape is called
a “peak”.

• Retention time (tR) is the time interval between sample injection point and the apex of the
peak.
• The required time for non-retained compounds (compounds with no interaction for the
stationary phase) to go from the injector to the detector is called the dead time (t 0).
• The peak height (h) is the vertical distance between a peak's apex and the baseline, and the
peak area (A) is the area enclosed by the peak and baseline.
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 Chromatographic analysis contd. Types of HPLC HPLC
Normal phase:
Column packing is polar (e.g., silica) and the mobile phase is non-polar. It is used for water-
sensitive compounds, geometric isomers, cis-trans isomers, and chiral compounds.

Reverse phase:
The column packing is non-polar (e.g., C18), the mobile phase is water + miscible solvent
(e.g., methanol). It can be used for polar, non-polar, ionizable and ionic samples.

Ion exchange:
Column packing contains ionic groups, and the mobile phase is buffer. It is used to separate
anions and cations.
Size exclusion:
Molecules diffuse into pores of a porous medium and are separated according to their relative
size to the pore size. Large molecules elute first, and smaller molecules elute later.

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 Chromatographic analysis contd. HPLC

The HPLC has developed into a universally applicable method so that it finds its use in
almost all areas of chemistry, biochemistry, and pharmacy.
• Analysis of drugs
• Analysis of synthetic polymers
• Analysis of pollutants in environmental analytics
• Determination of drugs in biological matrices
• Isolation of valuable products
• Product purity and quality control of industrial products and fine chemicals
• Separation and purification of biopolymers such as enzymes or nucleic acids
• Water purification
• Pre-concentration of trace components
• Ligand-exchange chromatography
• Ion-exchange chromatography of proteins
• High-pH anion-exchange chromatography of carbohydrates and oligosaccharides
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 Chromatographic analysis contd. HPLC
Advantages
• Speed
• Efficiency
• Accuracy
• Versatile and extremely precise when it comes to identifying and quantifying chemical
components.

Disadvantages
• Cost: Despite its advantages, HPLC can be costly, requiring large quantities of expensive
organics.
• Complexity
• HPLC does have low sensitivity for certain compounds, and some cannot be detected as
they are irreversibly adsorbed.
• Volatile substances are better separated by gas chromatography.

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 Chromatographic analysis contd. HPLC
Following polyaromatic Technique utilized for both synthetically prepared PAHs
hydrocarbons (PAHs) were containing water samples as well as real surface river water
detected: samples

• Fluoranthene (FlA)
• Pyrene (Pyr)
• Chrysene (Chr)
• Benzo[a]anthracene (BaA)

• Benzo[b]fluoranthene (BbF)
• Benzo[a]pyrene (BaP)

Utilization of solid-phase extraction (SPE) coupled with HPLC

for PAHs detection in water Source: Pan et al., 2011
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 Chromatographic analysis contd. HPLC
Analytical results for PAHs determination in river water samples
Compounds Detected RSD (%) Spiked Recovery RSD (%)
concentrations concentration (%)
(no spiking)/mg s/ mg L-1
L-1
FlA 0.55 1.87 12.00 114.11 10.77
Pyr 0.24 2.77 7.20 115.83 10.39
Chr NDa - 30.00 95.22 4.85
BaA NDa - 3.00 102.22 8.82
BbF NDa - 2.00 79.00 9.95
BbP NDa - 1.00 75.33 8.83
a
ND: not detected
Source: Pan et al., 2011 26
Thanks

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