Process Instrumentation

Module-II
Dr. Lipika Parida
Module-II
Composition measuring instruments:
•Chromatography
•Spectroscopic methods
•Thermal conductivity cells
•Carbon dioxide analyzer
•Colorimetry

•Humidity measurement
•pH meter
•Polarograph
•Refractometer
 Composition measurement by chromatography
Chromatography is a physical process to analyze identify separate purify and quantify a
complex mixture.

• Gas chromatography

• Liquid chromatography
 Chromatography

• Chromatography is a physical separation method in which the components of a mixture is separated by

differences in their distribution between two phases, one of which is stationary known as stationary phase

while the other mobile phase moves through it in a definite direction.

• The components in the mixture must interact with the stationary phase to be retained and separated by it.

• Type of interaction may be based on adsorption, solubility, chemical bonding, polarity, and molecular

filtration.

• Mobile phase: is the gas or liquid that carries mixture of components through the stationary phase.

• Stationary phase: is the part of the apparatus that holds the components as they move through it

separating them. A tube packed with stationary phase is called column.

• Retention time: It is the characteristic time it takes for a particular analyte to the pass through the system

(from the column inlet to the detector) under set conditions.

 Chromatography
• Chromatograph: is the instrument employed for a chromatography.

• Chromatogram: is the detector versus time signal.

• Eluent: fluid entering a column

• Eluate: fluid exiting the column

• Elution: the process of passing the mobile phase through the column

• Flow rate: how much mobile phase passed per unit time

• Linear velocity: distance passed by mobile phase per unit time.

 Gas Chromatography
• Gas chromatography is common technique for separating and analyzing components of a mixture.

• A given liquid mixture is first vaporized and then entrained by a carrier gas. the carrier gas commonly used a helium, argon,

hydrogen, and dry nitrogen.

• The gas mixture is passed through a tube which is known as column. In the terminology chromatography containing a stationary

phase: it may be microscopic layer of liquid or polymer on an inert solid support.

• Various components of the mixture travel through the stationary phase at different speeds which cause them to separate.

• Since different components emerge from the column at different times, they can be identified by a detector at the outlet of the

column. A common detector is Thermal Conductivity Detector.

• A gas chromatography system generates a plot of the detector signal as a function of time. This plot is refer to as a chromatogram. As

an analyte appears in the detector, it’s presence is signaled by a peak. Thus, a gas chromatogram consists of series of peaks, one for

each of the components of the sample. The chromatogram is displayed on a chart recorder or computer screen.

• Different peaks on a chromatogram corresponds to different components and the areas under these peaks can be used to quantify

the mole fraction of each component.

 Gas Chromatography

Common
Adsorbents
Alumina, Molecular
sieves, Silica,
Activated carbon.

Carrier Gas
Helium, Hydrogen, Dry nitrogen,
Argon
Gas Chromatography
 Gas Chromatography

Retention time means the time elapsed between the sample injection and the point of
detection by the detector.
tM = retention time of the mobile phase
tR = retention time of the analyte
 Gas Chromatography : Detectors

• After the components of a mixture are separated using gas chromatography, they must be detected as they exit
the gas chromatography column.

Thermal conductivity detector

• The temperature of the sensing

element depends on the thermal
conductivity of the gas flowing around
it. Thermal conductivity changes with
presence of other components in the
mobile phase.
• One resistor is in contact to the mobile
phase leaving the column. As the solute emerge from the column,
Change in thermal conductivity→Change in amount of heat
• another is in contact with the reference
removed from the resistor → Change in register’s temperature
stream of pure mobile phase. and resistance → Change in voltage difference between points
Liquid Chromatography
 Liquid chromatography is identical in principle
to gas chromatography, instead of carrier gas,
liquid solvent is used.
 The mobile phase is passed with a pressure of
1000 psig, which is 7 mega pa almost.
 Detector : UV spectrophotometer or
refractive index-based detector.
 Very good quality pumps are required for
liquid chromatography, because the mobile
phase which is solvent has to be passed
through the column with a very accurate flow
rate at high pressures.
 The typical column length is 1m, diameter is 2
mm, and packed with about 20 micrometre
sized particles, which are adsorbents.
 Liquid Chromatography
• The heart of HPLC system is the column. The column contains the particles that contains the stationary phase.
• The mobile phase is pumped through the column by a pump at a specific flow rate. Solvents must be degassed
to eliminate formation of bubbles.
• The injector serves to introduce the liquid sample into the flow system of the mobile phase, this may be auto
sampler or manual.

Name of LC Method Type of Stationary phase

Adsorption chromatography Solid, Underivatized support
Partition liquid chromatography Liquid coated or derivatized support
Ion exchange chromatography Support containing fixed charges
Size exclusion chromatography Porous support
Affinity chromatography Support with immobilized ligand
Composition Measurement by UV-Vis spectroscopy
 Absorption of Light
• Certain molecules absorb light in a characteristic way. This property can be used to identify and quantify
molecules.
• Absorption occurs when the energy contained in a photon is absorbed by an electron resulting in a
transition to an excited state.
• Absorption corresponds to the amount of light that is absorbed and it is plotted with various wavelengths.
 UV-Vis Spectroscopy

• Spectroscopy measures the interaction of molecules with electromagnetic

radiation.

• Light in the near ultraviolet and visible range of the electromagnetic

spectrum has an energy of about 150 to 400 kilojoules per mole. The
energy of the light is used to promote electrons from the ground state to an
excited state. A spectrum is obtained when the absorption of light is
measured as a function of it is frequency or wavelength.

• Molecules absorb light in the near UV or in the visible region; that means,
in the wavelength range of 150 to 400 nanometer or 400 to 800
nanometre.
 UV-Vis Spectroscopy
• In UVV spectroscopy, a beam of light with a

wavelength in UV-Vis region passes through a solution

taken in a cuvette.

• The sample in the cuvette absorbs this UV or visible

radiation.

• The amount of light that is absorbed by the solution

will depend on

1. The concentration of the solution

2. The path length of the light through the cuvette

3. How well the analyte absorbs light at a certain

wavelength.
UV-Vis Spectroscopy
 UV-Vis Spectroscopy: Components
 Sources used in the UV-Vis spectrophotometer are continuous sources. Continuous sources emit radiation or of all
wavelengths within the spectral region for which they are to be used. Sources of radiation should be stable and of
high intensity. The commonly used visible light sources tungsten filament lamp, and the commonly used ultraviolet
sources deuterium discharge lamp.

 Ideally speaking the output of a wavelength selector would be a radiation of a single wavelength. Since, no real
wavelength selected is ideal, a band of radiation is always obtained. The performance of the instrument will be
better if this bandwidth is narrow. Filters and monochromators are used as wavelength selector.

 Sample compartment :For visible and UV spectroscopy, a liquid sample is usually contained in a cell called cuvette.
Glass is suitable for visible, but not for UV spectroscopy. Because glass absorbs UV radiation, quartz can be used in
UV spectroscopy, as well as in visible spectroscopy.

 The detectors convert radiant energy into electrical signal. Our detector should have high sensitivity, and fast
response over a range of wavelengths. The electrical signal produced by the detector must be directly proportional
to the transmitted intensity.
 Carbon dioxide analyzer
• Carbon dioxide analyzers are also called CO2 gas analyzers.
• They are devices that can detect and quantify the amount of carbon dioxide in a sample.
• The commonest technologies applied are based on Infrared detectors, owing to the fact that CO 2 is an excellent absorber of infrared
energy.
• Carbon dioxide can be measured in percent (%) and parts per million (ppm) depending on the requirements of the application.
Different Types of CO2 Analyzers
1. Mass Spectrometry
2. Raman Scattering
3. Colorimetric Analyzer
4. Infrared Absorption Spectrophotometry (IRAS)

Applications:
5. Pharmaceutical industry
6. Food and beverage industry
7. Research and development
8. Chemical and Medical laboratories
9. Environmental sciences for monitoring purposes and quality control
 Mass Spectrometry
• Measures concentration of CO2 largely on basis of

molecular weight.

• CO2 passes through an ionizer creating ion

fragments.

• These fragments are accelerated in a vacuum

through a high magnetic field.

• The magnetic field separates the fragments by their

mass and charge.

• The fragments are deflected onto a detector plate,

where CO2 has a specific landing site.

• The processor section calculates the concentration

 Raman Scattering

• Photons generated by a high-intensity argon laser collide with CO2. After impact, CO2 is
momentarily excited to unstable vibrational and rotatory state. When CO 2 returns to
the normal state, photons of a characteristic frequency are emitted. The scattered
photons are measured as peaks in a spectrum that determines the concentration.
 Colorimetric Analysis
• Colorimetric analysis is a method of determining the concentration of a chemical
element or chemical compound in a solution with the aid of a color reagent. It is
applicable to both organic compounds and inorganic compounds and may be used with
or without an enzymatic stage.
 Infrared spectrophotometry

• Infrared spectrophotometry is based on the fact

that molecules absorb specific frequencies that are

characteristic of their structure.

• These absorptions are resonant frequencies, i.e. the

frequency of the absorbed radiation matches the

transition energy of the bond or group that

vibrates.

• The sample absorbs the incident Laser energy at a

given frequency and emits light, which is

characteristic to the composition of sample

Infrared spectrophotometry

• An infrared light source - produces infrared

energy.

• CO2 absorbs light with a characteristic peak

at a wavelength close to 4300nm.

• An optical path: A detection system -

calibrated to determine the concentration
of CO2 by creating electric currents.
 Humidity measurement
• Humidity is the amount of water vapor present in the air.

• Humidity is an important factor for operating certain equipment (e.g., high-resistance electronic

circuits, electrostatic-sensitive components, high-voltage devices, fine mechanisms, etc.).

• Humidity is used to determine the likelihood of precipitation , fog or dew.

• Determining the humidity of a certain place provides a way to gauge the weather .

• Relative humidity is the percentage of water vapour in the air at a given temperature.

• Relative humidity is used for climate control and how it affects the health, comfort and safety of

humans .

• Relative humidity is also used to ensure of machines, vehicle and buildings.

 Humidity measurement
1. Psychrometer (Wet- and Dry-Bulb Hygrometer)
2. Mechanical Hygrometers
3. Electrical Hygrometers
4. Dew-Point Sensors
 1. Psychrometer (Wet- and Dry-Bulb Hygrometer)
• This constitutes a pair of temperature sensors. These are placed in a wet air
stream. One of the thermometers (the wet bulb) has a wet wick around it;
evaporation causes this thermometer to read a lower temperature than the
other. The difference in temperatures, and the absolute temperature of the
wet thermometer can be used to calculate the relative humidity of the air.

• pw is the experimental partial vapor pressure of water

• ps is the saturation vapor pressure at temperature Tw

• A is the psychrometer constant (typically, values of A, for Tw above 0°C, are

around 5×10-4 to 10-3)

• P is the experimental pressure

• TD is the dry bulb temperature
• TW is the wet bulb temperature
 2. Mechanical Hygrometers
• Mechanical hygrometers
utilize the change in
dimensions of various porous
materials (such as wet paper
and hair) as they absorb water
vapor.

• This change in dimensions can

be used to move a needle or
dial; which, when calibrated,
will give the relative humidity.
 3. Electrical Hygrometers

• Electrical impedance sensors measure the changes in electrical capacitance or resistance of a

hygroscopic material.

• The material will absorb or desorb water depending on the partial vapor pressure in the
atmosphere around it, thus changing its electrical properties.

• Types:
1. Resistive hygrometers
2. Capacitive hygrometers.
 3.1. Electrical Hygrometers : Resistive Type
• The resistance-type hygrometer utilizes the principle that
electrical resistance varies in a material that absorbs
moisture.

• Special resistive sensors measure the electrical resistance

between the electrodes.

• The conducting film of the resistive hygrometer is made by

the lithium chloride and the carbon.
• The resistance of the conducting film varies with the
change in the value of humidity present in the surrounding
air.
• If the relative humidity is high, the lithium chloride will
absorb more moisture and their resistance decreases.
• The change in the value of resistance is measured by
applying the AC to the bridge
 3.2. Electrical Hygrometers : capacitive Type
• A capacitance-type hygrometer measures
humidity by detecting the change in
capacitance of a thin polymer film.

• This type of sensor is sufficiently accurate in

most cases and is used frequently in industry.

• It is made by placing the hygroscopic material

between the metal electrodes, quickly
absorbing water and decreasing the
capacitance value.
 4. Dew-Point Sensors
• Dew-point sensors measure the temperature of the surrounding air, and the dew-point of a small
sample there of.
• The humidity of the air is then measured from the dew-point temperature and the initial air
temperature.
• Optical sensors cool a surface (usually a mirror) until condensation starts to form. This indicates that the
surface is at the air's dew-point.
• Other methods of measuring dew-point include measuring the oscillation rate of a quartz crystal. When
condensation forms on the crystal, this rate will change.
pH measurement
pH measurement
 pH measurement: Chemical Indicators

• pH is normally measured by chemical indicators or by pH meters.

• The final color of chemical indicators depends on the hydrogen

ion concentrations.

• For indication of acid alkali or neutral water litmus paper is used,

it turns pink when acidic; blue when alkaline; and stays white if
neutral.
 pH measurement: pH meter
• The most common method of measuring pH is
by glass cell and calomel cell electrodes used
with a potentiometer device.

• An electrode is immersed in the solution. An

electric potential is produced at the electrode
which forms an electrolytic half-cell. This is the
measuring cell.

• A second electrode is required to provide a

standard potential and to complete the cell.
This is the reference cell.

• The algebraic sum of the potentials of the 2 half

cells is proportional to the pH of the solution.
pH measurement: pH meter, Nernst equation
 Polarograph

 When a solution is subjected to electrolysis, the electrical properties like emf,

current, resistance etc. depend on nature and concentration of various species
present in that solution.

 Principle: Study of solutions or of electrode processes by means of electrolysis

with two mercury electrode, one polarizable and one non polarizable, the former
formed by mercury regularly dropping from capillary tube.

 Polarized electrode – Dropping Mercury Electrode (DME)

 Depolarized electrode - Saturated Calomel Electrode (SCE)
 Polarograph: Dropping Mercury Electrode (DME)

Dropping mercury electrode (DME) is a working electrode arrangement

for polarography in which mercury continuously drops from a reservoir
through a capillary tube (internal diameter 0.03 - 0.05 mm) into the
solution. The optimum interval between drops for most analyses is
between 1 and 5 s.

A small polarizable electrode is Produced by Passing a steam of mercury

Through a resistance glass capillary.

The assembly consists of –

1. A mercury reservoir.
2. A connecting tube between the mercury reservoir and Capillary
tube.
3. A capillary tube.
4. A glass electrolyte cell.
 Polarograph: Dropping Mercury Electrode (DME)
• The basic component of the experimental set up are the polarograpic cell and the DME. The polarograpic
cell is a H – shaped vessel. The two tubes are connected through an agar agar salt bridge and a sintered glass
tube. One half of the cell act as anode which is usually a reference electrode such as Saturated Calomel
Electrode (SCE). The other half cell contains the solution of the sample analyte and the DME as the cathode.

• The DME consists of a long capillary tube through which mercury is forced under pressure exerted by
approximately 50 cm column or mercury connected to a mercury reservoir.

• The electrical Circuit consists of a DC source, potentiometer, a voltmeter and microammeter usually a
galvanometer.

• The DME is connected to the negative terminal while SCE is connected to the positive terminal of DC source.

• Using the potentiometer an emf upto 3 volt may be gradually applied to the cell.
 Polarogram
• It is a graph of current versus potential
in a polarograpic analysis.

3 categories :
• Collectively referred as residual Current
• Referred to as diffusion current
resulting from reduction of sample
• Called as limiting current
 Ilovic Equation

Ilkovic equation is a relation used in polarography relating the diffusion current (id) and the
concentration of the non-polarisable electrode, i.e., the substance reduced or oxidised at the dropping
mercury electrode (polarisable electrode).
 Applications of Polarography:
1. Qualitative analysis: It helps in characterization of organic matter and various metal interactions from half wave
potential of the current v/s voltage graph.

2. Qualitative analysis: Polarography is used in the determination of concentration of drugs, metal ions etc. in the
given sample.

3. Determination of inorganic compounds: Polarography is used in determination of cations and anions in the
presence of interfering ions.

4. Determination of organic compounds: Polarography is used in determination of structure, quantitative analysis of

mixture of organic compounds.

5. Estimation of dissolved oxygen: Amount of oxygen dissolved in aqueous solution or organic solvent can be
calculated with the help of Polarography.

6. Pharmaceutical applications: Tetracycline antibiotics, sulphonamides can be analysed by Polarography.

 Refractometer
• Refractometry is the method of measuring
the refractive index of substances.

• Refractive index is defined as the ratio of the

speed of light in a vacuum to the speed of
light in another substance.

• The refractive index is a useful physical

property of liqiuid

• Often a liquid can be identified from a

measurements of its refractive index.

Principle: Measurement of RI of unknown

substance by measuring angle of refraction made
when the substance is brought into contact with
the medium prism of a unknown refractive index
 REFRACTIVE INDEX
• Measurement of the refractive index can be used to characterize liquid and solid
sample.

• Measure the concentration of solutions.

• the refractive index allows you to control the quality of multi- component
mixtures and check samples for purity

• The refractive index of a substance is dependent on the wavelength λ of the light

and the temperature T of the substance.
 REFRACTIVE INDEX

• When a ray of light passes from one medium to

another It suffer refraction. Snell’s Law

• If it case from a less dense to more dense medium

like from air to water it is refracted towards
normal so that angle of refraction is less than
angle of incidence .

• The Refractive index of the second medium with

respect to first is given by The R.I and is defined as
ratio of velocity of light in vacuum & air to that in
medium
 INSTRUMENTATION : Abbe’s Refractometer
 It uses for measurement of an Index of
refraction of liquid & Solid sample.
 It consist of stationary telescope & two prism
held in together in contact with each other in
metal case.
 The prism at lower position is called as
illuminating prism where as the prism at
upper position is refracting prism.
 The sample liquid to be analyzed is
sandwiched into a thin layer between. the
two prism.
 The prism system is attached with an arm
which is watched c which moves over the
scale to read RI.
 The Detector is placed on backside of
refractometer to measure position of border
between bright & dark area.
 APPLICATION

 The RI measurement can be used for qualitative & quantitative analysis as well
as in structural analysis.
 It is determining the concentration of Aqueous & Alcohol solution

 Determination of sugar content, alcohol content in fermentation is Routinely

carried out by Refractometer.