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TURBIDIMETRY AND NEPHELOMETRY

When light passes through an essentially clear liquid, the light beams travel along relatively undisturbed paths.
However, some distortion occurs as light is scattered by molecules present in the pure fluid. As shown in Figure 1, when
light passes through a liquid containing suspended solids, the light beam interacts with the particles, and the particles
absorb the light energy and re-radiate light in all directions.

Figure 1. Scattered light at 900C.

Particle size, configuration, color, and refractive index determine the spatial distribution of the scattered light intensity
around the particle.
Turbidity is an expression of the optical property of a medium, which causes light to be scattered and absorbed
rather than transmitted in straight lines through the sample. The International Organization for Standardization (ISO)
defines turbidity as the reduction of transparency of a liquid caused by the presence of undissolved matter. It is,
therefore, the opposite of clarity.
Turbidity can be measured using the techniques of turbidimetry or nephelometry. Turbidimetry is the
measurement of the degree of attenuation of a radiant beam incident on particles suspended in a medium, the
measurement being made in the directly transmitted beam. It is usually applied to media of fairly high turbidity in which
the scattering particles are relatively large.
Nephelometry is the measurement of the light scattered by suspended particles, the measurement usually being
made perpendicularly to the incident beam. It is more appropriate for media of lower turbidity in which the suspended
particles are small.

Principles
Scattering occurs when a minute particle interacts with incident light by absorbing the light energy and then, as
if a point light source itself, reradiating the light energy in all directions. Absorption takes place when light is converted
to other energy forms within the particle. Scattered light includes that reflected from the surface of the particle and that
refracted within the particle, possibly after many internal reflections. Scattering is often accompanied by absorption.
The direct relationship between turbidity data and suspended solids concentrations is weakened by the complex
interactions of light energy with suspended particles. It is heavily dependent on many factors, including:
concentration of scattering particles suspended in the medium
size distribution of scattering particles
shape, orientation, and surface condition of the scattering particles
refractive index of the scattering particles
refractive index of the suspension medium
wavelength of the light source employed
The Nephelometric Turbidity Unit (NTU) is the most common unit employed for the measurement.

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Instrumentation
Ultraviolet-visible spectrophotometers may be used for turbidimetric measurements by measuring the
absorption of light by particles at a fixed wavelnegth or full spectrum.

Figure 2. Three basic designs of turbidity meter.

(A) Nephelometer, which directly measures light
scattered (usually at 900 to the beam direction) by
suspended particles; (B) Turbidimeter, where
transmitted light is detected, in relation to initial
beam intensity; (C) Ratio Turbidimeter, in which
both transmitted and scattered light are detected.

Modern measurement devices use photosensitive cells to quantify scattered and/or transmitted light. Figure 2
illustrates that most laboratory bench instruments usually have five basic components: (1) a light source of known,
constant intensity, and given wavelength characteristics; (2) a lens to collimate the light beam; (3) a sample cell;
(4) photosensor/s; and (5) a meter or logger to record the output signals from the photosensors/s.
A nephelometer measures directly the intensity of light scattered by the sample, which is proportional to the
amount of matter suspended in the light path, though the influence of size, shape, and refractive index of scattering
particles are also important. Nephelometers usually provide better precision and sensitivity than turbidimeters.
A turbidimeter, sometimes called a transmissometer, absorptiometer, or turbidity meter, measures intensity of
the beam after is has passed through the sample, i.e. it quantifies the amount of transmitted light remaining. These
instruments are more appropriate for relatvely turbid samples in which the scattering particles are large in relation to the
light wavelength used. This is because a significant reduction in the intensity of incident light is needed to yield precise
results.
Ratio turbidimeters, incorporate measurement systems for light which is side-scattered (usually at 900), forward-
scattered, and transmitted. The turbidity value is obtained as the ratio of the 900 signal to the sum of forward-scattered
and transmitted values. Advantages include: increased long-term stability of the sensor (by reducing effects of
instrumental drift); compensates for ageing of, and deposits on, the optics; reduces influence of temperature changes in
the electronics; minimizes need for repeated recalibration; and limits the effect of sample color on readings. This can be
more appropriate for strongly and/or variably colored liquuids, or for samples of high turbidity.

Typical Applications
Turbidity values can act as simple and surrogate measures of the concentration of suspended solids, sulfate ions
and other particulate material. In quantitative chemical and biological analysis, applications include the calculation of the
absolute molecular weights and dimensions of polymers in solution, as well as particle size determination. Chemical
profiles can also be obtained by observing turbidity changes deliberately induced by the addition of specific substances to
the solution. Within microbiology, cell and bacteria growth can be monitored through the media turbidity changes such
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activity causes. In foodstuff manufacturing, turbidimetry us often used to monitor product quality and treatment process
efficiency, especially in the dairy and brewing industries. In addition, nephelometry is used in the (1) determination of
immunoglobulins in biological fluids, (2) determination of the concentrations of individual serum proteins, and
(3) determination of the size and number of particles.

X-RAY DIFFRACTION

X-ray diffraction (XRD) is a rapid analytical technique primarily used for phase identification of a crystalline
material and can provide information on unit cell dimensions. Atoms of crystalline substances are arranged in a regular
pattern, and there is a smallest volume element (unit cell) that by repetition in three dimensions describes the crystal.
Diffraction is a phenomenon that light undergoes especially in passing by the edges of opaque bodies or through narrow
openings, and in which the rays appear to be deflected. The x-ray diffraction pattern of a pure substance is like a
fingerprint of the substance.

Principles
X-ray diffraction is based on constructive interference of monochromatic x-rays and a crystalline sample. These
x-rays are generated by a cathode ray tube, filtered to produce monochromatic radiation, collimated to concentrate, and
directed toward the sample. The interaction of the incident rays with the sample produces constructive interference (and
a diffracted ray) when conditions satisfy

This law relates the wavelength of electromagnetic radiation to the diffraction angle and the lattice spacing in a crystalline
sample. These diffracted X-rays are then detected, processed and counted. By scanning the sample through a range of 2
angles, all possible diffraction directions of the lattice should be attained due to the random orientation of the powdered
material. Conversion of the diffraction peaks to d-spacings allows identification of the mineral because each mineral has a
set of unique d-spacings. Typically, this is achieved by comparison of d-spacings with standard reference patterns.

Instrumentation
X-ray diffractometers consist of three basic elements: an X-ray tube, a sample holder, and an X-ray detector.
X-rays are generated in a cathode ray tube by heating a filament to produce electrons, accelerating the electrons toward
a target by applying a voltage, and bombarding the target material with electrons. When electrons have sufficient energy
to dislodge inner shell electrons of the target material, characteristic X-ray spectra are produced. The specific
wavelengths are characteristic of the target material (Cu, Fe, Mo, Cr). Filtering, by foils or crystal monochrometers, is
required to produce monochromatic X-rays needed for diffraction. These X-rays are collimated and directed onto the
sample. As the sample and detector are rotated, the intensity of the reflected X-rays is recorded. When the geometry of
the incident X-rays impinging the sample satisfies the Bragg Equation, constructive interference occurs and a peak in
intensity occurs. A detector records and processes this X-ray signal and converts the signal to a count rate which is then
output to a device such as a printer or computer monitor.
The geometry of an X-ray diffractometer is such that the sample rotates in the path of the collimated X-ray beam
at an angle while the X-ray detector is mounted on an arm to collect the diffracted X-rays and rotates at an angle of 2.
The instrument used to maintain the angle and rotate the sample is termed a goniometer. For typical powder patterns,
data is collected at 2 from ~5 to 70, angles that are preset in the X-ray scan.
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Applications
X-ray diffraction is used to identify unknown crystalline materials, to characterize crystalline materials, identify
fine-grained minerals such as clays that are difficult to determine optically, determinen unit cell dimensions and
measurement of sample purity.

Strengths
Powerful and rapid (< 20 min) technique for identification of an unknown mineral
In most cases, it provides an unambiguous mineral determination
Minimal sample preparation is required
XRD units are widely available
Data interpretation is relatively straight forward

Limitations
Homogeneous and single phase material is best for identification of an unknown
Must have access to a standard reference file of inorganic compounds (d-spacings, hkls)
Requires tenths of a gram of material which must be ground into a powder
For mixed materials, detection limit is ~ 2% of sample
For unit cell determinations, indexing of patterns for non-isometric crystal systems is complicated
Peak overlay may occur and worsens for high angle 'reflections'

References:
EPA Guidance Manual: Basic Turbidimeter Design and Concepts. April 1999.
The International Pharmacopeia 4th Ed. Fourth Supplement. 2014.
2005 Turbidimetry Encyclopedia full article 10.1016BO-12-369397-700718-4.pdf (downloaded from
www.researchgate.net on September 01, 2014).
http://www.kau.edu.sa/Files/0013791/Subjects/Turbidimetry&Nephemometry.pdf (downloaded on August 29, 2014).
http://www.merriam-webster.com/dictionary/diffraction (accessed on September 01, 2014)
http://serc.carleton.edu/research_education/geochemsheets/techniques/XRD.html (accessed on August 23, 2014).
http://www.vscht.cz/clab/RTG/dokumenty/thermo/xrd/Introduction%20to%20powder%20diffraction.pdf (downloaded
on September 01, 2014).
http://www-outreach.phy.cam.ac.uk/camphy/xraydiffraction/xraydiffraction7_1.htm (accessed on September 01, 2014)

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