pH Meter
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pH is defined as the “negative logarithm of hydrogen ion concentration”.
pH= -log [ H+ ]
p = power
H = hydrogen
[H+ ] = hydrogen ion concentration
The greater the concentration of hydrogen ions, the stronger the acid.
First commercial pH meter was built in 1936 by Arnold Beckman in United
States.
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Measurement of pH
pH can be measured by:
1. pH meter
2. pH scale
3. pH strips/ indicators
The most important difference between a pH scale, pH strip and a pH meter is
the accuracy of the measurement.
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pH Meter
A pH meter is an electronic instrument used for measuring the pH (acidity or
alkalinity) of a liquid.
pH meter consists of a measuring probes (glass electrode and reference electrode)
connected to an electronic meter that measures and displays the pH reading.
It measure hydrogen-ion activity in water based solution.
The greater the concentration of hydrogen ions, the stronger the acid.
First commercial pH meter was built in 1936 by Arnold O. Beckman in United States.
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Types of probes / electrodes
1. Glass electrode
2. Reference electrode or calomel electrode
The glass electrode is an half cell & the
reference electrode is another half cell.
This glass electrode sense the pH as
concentration of H+.
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pH meters measures the potential difference between the two electrodes.
The principle of a pH meter is based on measurement of the hydrogen ion
concentration in a solution.
When the pair of electrodes (glass electrode and calomel electrode) dipped in
an aqueous solution, a potential is developed across the glass electrode.
Potential difference recorded on the potentiometer scale.
Then the potentiometer converts this potential difference into a pH reading.
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Types of pH meter
1. Manual pH meter
2. Digital pH meter
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Glass electrode
The main components of Glass electrode are:
1. Silver-silver chloride molecules
2. Hydrochloric acid (HCl) solution
3. Platinum wire
The silver-silver chloride molecules form the electrode's half-cell. It
maintains a constant potential necessary for pH measurement.
HCl in a glass electrode helps to maintain the concentration of the
solution inside the bulb and prevents the glass membrane from
drying out.
The platinum wire allows for electrical contact to the external
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circuit.
Reference electrode or calomel electrode
The main components of reference electrode are:
1. Mercury and mercury-chloride molecules
2. Saturated potassium chloride (KCl) solution
3. Platinum wire
The mercury and mercury-chloride molecules form the
electrode's half-cell. It provide a stable voltage.
The KCl solution provides a stable ionic environment for
the electrode.
The platinum wire allows for electrical contact to the
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external circuit.
Precautions
• Instrument should be well calibrated.
• Probe properly dipped in solution.
• pH electrodes are sensitive and fragile so must be handle with care.
• Always keep pH electrode moist, usually kept dipped in KCL.
• Temperature must be set before measurement.
Why is KCl used?
KCI is a salt in nature which shows that it has a cation of strong
base and anion of strong acid and they both have no effect on the
pH paper and it is has a standard value which shows that its pH is
7.0 which is a neutral figure. 11
pH Meter used for:
A pH meter is used to measure hydrogen ion activity in solutions.
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pH Scale
• Less accurate.
• It measures the acidity or alkalinity of a substance.
• The scale ranges from 0 (the most acidic) to 14 (the most basic).
• A pH of 7 is neutral.
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pH scale
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pH Strip
Litmus dye is a natural pH indicator.
Litmus paper is a small strip of paper that has
litmus dyes in it.
There are two types of litmus paper: red and blue
1. Red litmus paper: Turns blue when exposed to basic or alkaline materials.
2. Blue litmus paper: Turns red when exposed to acidic materials.
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How to use pH test strips
1. Dip the pH strip for two seconds in the fluid that you want to measure.
2. Wait for ten seconds.
Take out the paper and then,
the strip color is compared with a indicator scale.
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Photometry
• Introduction
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Introduction
• Greek word: Photo: light
Meter: measure
Photometry: Means Measurement of intensity of light or amount of light.
Photometry is of two types:
a) Colorimetry: Device that measures the absorbance of light in visible range.
b) Spectrophotometry: Device that measures the absorbance of light in
ultraviolet, visible and infrared range. 18
Difference between Colorimeter and Spectrophotometer
COLORIMETER SPECTROPHOTOMETER
Measures the absorbance of light in Measures the absorbance of light in
visible range. ultraviolet, visible and infrared range.
Solution need to be colored. Solution may or may not be colored.
Filter is used to select wavelength Prism is used to select wavelength
Less sensitive Highly sensitive instrument
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When a monochromatic light passes through a solution, it is either-
Reflected
Absorbed
Transmitted
Photometry measures absorbance of transmitted light by:
Colorimeter
Spectrophotometer
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Photometry is based on two laws
The measurement of color intensity of a colored solution by photometry is
governed by two laws:
1. Beer’s law
2. Lambert’s law
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Photometry is based on two laws
Beer’s law: When a parallel beam of monochromatic light passes through a
solution, the absorbance of the solution is directly proportional to
concentration of the compound in solution.
A∝C
Where,
A: absorbance (A) of the solution
C: concentration (c) of the compound
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Lambert’s law: When a parallel beam of monochromatic light passes through a
solution, the absorbance of the solution is directly proportional to length of the
light path through the solution.
A∝L
Where,
A: absorbance (A) of the solution
L: length of the light path
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Beer’s law = A ∝ c
Lambert’s law = A ∝ l
By combining above these two equation, we get:
A=ꜫcl
Where,
ꜫ is proportional constant called molar absorption coefficient.
c is concentration of absorbing compound,
l is length of light path.
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Colorimeter
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Colorimeter
Colorimeter: Instrument used for the measurement of optical density of colored
substance in solution.
Simplest type of instrument used for OD measurement.
Less sensitive
Measures the absorbance of light in visible range (400-750nm).
A substance to be estimated colorimetrically, must be colored or it should be capable
of forming chromogens (colored complexes) through the addition of reagents.
Colorimeter
• Principle of colorimeter is based: Beer & Lambert’s law.
• According to which: When a monochromatic light passes through a colored
solution, the absorbance of the solution is directly proportional to
concentration of the compound in solution and the length of a light path
through the solution.
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Components of the colorimeter
Light source
Slit
Condensing lenses
Filter
Cuvette(sample holder)
Detector (photocell)
Output (galvanometer)
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Light source: tungsten filament lamp.
Slit: allows only a beam of light to pass through it. It prevents unwanted or
stray light.
Condensing lenses: light after passing through slit, falls on condenser lenses
which gives a parallel beam of light.
Filter :
Filters are used for selecting light of specific wavelength.
It will absorb light of unwanted wavelength and allow only monochromatic light
to pass through. 29
Cuvette(sample holder): the monochromatic light from the filter passes through the colored
solution placed in a cuvette.
Cuvette is made up of glass/plastic/quartz material.
It has fixed diameter (usually 1 cm) & having uniform surface.
Detector (photocell):
Detector are photosensitive elements which converts light energy into electrical
energy.
The electrical signal is directly proportional to intensity of light falling on the detector.
Output: the electrical signal generated in photocell is measured by galvanometer, which
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displays as optical density.
Application of colorimeter
• To determine the optical density or absorbance of a colored compound.
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Preparation of solution for OD estimation
In colorimetric estimation it is necessary to prepare 3 solutions
Blank (B)
Standard (S)
Test (T)
Blank test tube: To eliminate the OD or the absorbance contributed by the
reagent and solvent. 32
How to calculate concentration of test
1. The instrument is set to zero (0) with the blank.
2. Take OD of blank, test and standard.
3. Calculate the concentration of test.
Concentration of test =
ODT : OD of test
ODB : OD of blank
ODS : OD of standard
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Precautions
1. Instrument should be well calibrated.
2. Make sure that the cuvettes are clean .
3. Dry the surfaces of cuvettes with cotton before measuring OD.
4. Set the colorimeter to zero with blank before taking OD.
5. Fill the cuvettes with equal volume for OD measurement of blank, test and
standard.
6. Remove the cuvettes from the instrument when not in use.
7. Switch off the colorimeter after use.
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spectrphotometer
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SPECTROPHOTOMETER
Principle: Same principle as a colorimeter.
1. Beer’s law
2. Lambert’s law
Measures the absorbance of light in ultraviolet (360-450nm), visible and infrared
range(750-1000nm).
Prism is used to select wavelength
Highly sensitive instrument 36
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