These techniques are closely related to colorimetry Both the techniques are based on the scattering of light by non

transparent particles in a suspended in a solution. The two techniques differs only in the manner of measuring the scattered radiation When the light is allowed to pass through a suspension, the part of the incident radiant energy is dissipated by absorption,reflection,and refraction while the remainder is transmitted.

Tyndall effect
Light scattering by particles in a colloid or particles in a fine suspension. the longer-wavelength light is more transmitted while the shorter-wavelength light is more reflected via scattering.

PRINCIPLE
NEPHELOMETRY measurement of the light scattered by suspended particles at right angles(900) (perpendicular) to the incident beam. TURBIDIMETRY measurement of the light transmitted by suspended particles to the incident beam.

TURBIDIMETRY & COLORIMETRY

Measurement of the intensity of light transmitted through a medium Light intensity is decreased

NEPHELOMETRY & FLUORIMETRY

Measurement of scattered light at 900 both incident & scattered light are same wavelength -NEPHELOMETRY scattered light wavelength is longer than the incident light - FLUORIMETRY

CHOICE OF THE METHOD

depends upon the amount of light scattered by suspended particles present in solution.

TURBIDIMETRY - high concn. Suspensions

NEPHELOMETRY low concn. Suspensions more accurate results

THEORY
REFLECTION VS SCATTERING
If the dimensions of the suspended particles larger than the wave length of the incident light - REFLECTION If the dimensions of the suspended particles smaller(same order) than the wave length of the incident light - SCATTERING NEPHELOMETRY suspended particles < incident light wave length : smaller particles undergo scattering secondary rays - maximum intensity at 900
- most of the instruments measured at this angle

suspended particles > incident light wave length : larger particles undergo reflection small fraction of light get deviated maximum intensity at < 900 5-200 / 450

NEPHELOMETRY
suspended particles should neither be too large otherwise the scattering efficiency falls off. optimum particle size should be 0.1- 1 micro meters. nor too small

TURBIDIMETRY
suspended particles > incident light wave length : larger particles undergo reflection- measuring transmitted radiation larger particles- absorbance vs concn not linear measurements can not be accurate

Factors affecting measurements

The amount of radiation removed or deviated from the primary radiation beam depends on the following factors

A.Concentration:
TURBIDIMETRY I T=Transmittance = Io Beers law Io S = log I S= turbidence due to scattering K = proportionality constant / Turbidity constant b = path length C = concentration of suspended material = kbc

NEPHELOMETRY

Is = Ks Io c Is = scattered intensity Ks = empirical constant Io = incident intensity c = concentration of the scattered material

Working curve C Vs Is / Io Vs C

log Io / Is

B.Particle geometry
Control of particle size & shape - most critical factor Same distribution

Conditions

concn. Of reactants, temp, agitation, pH, order of mixing, time allowed for particle growth

C.Incident light wave length

TURBIDIMETRY It is an imp factor Select a wave length- sample solution does not absorb strongly If the sample solution is colorless use the incident light of the same color If clear solutions having dark particles light in red / IR

NEPHELOMETRY Absorption is much less- white light is generally used

D.Refractve index difference

Appreciable RI differences between particles & surrounding medium best results Change solvents in order to increase the RI differences

INSTRUMENTATION
SOURCES FILTERS/MONOCHROMATORS CELLS DETECTORS SOURCES White light nephelometers Mercury arc Tungsten lamp

FILTERS / MONOCHROMATORS
mono chromatic radiation

CELLS
cylindrical cells - flat faces to minimize reflections & multiple scatterings cell with a rectangular cross section is preferred semi octagonal faces octagonal faces- 00,450,900,1350

NEPHELOMETER
metal test tube cover to exclude extraneous light.

test tube

reflector to collect the scattered light annular photocell

light source

Filter wheel with a series of colour filters

sensitive micro-ammeter

The test solution (sample) is placed in a test tube (F) that has been duly rested on a light source (A) The scattered light caused by the particles in a turbid or cloudy solution is immediately directed by the reflector (E) on to an annular photocell (D). A series of standard colour filters are usually provided in the form of a filter-wheel (C) so as to facilitate analysis of coloured solutions ; Taking care that the filter chosen must be similar to colour to that of the solution. The current generated after passing through the photocell (i.e., light energy is being converted to electrical energy) is recorded by a sensitive micro-ammeter (B). The test tube is provided with a metallic cover (G) to get rid of any extraneous light.

Usually a nephelometer is provided with zero-setting controls, sensitivity adjusting device and a set of previously matched test tubes.

TURBIDIMETER
Either visual or photoelectric colorimeters may be satisfactorily employed as turbidimeters.

However, the use of the blue filter normally enhances the sensitivity appreciably.

It has been observed that the light transmitted by a turbid solution does not normally obey the Beer-Lambert Law accurately and precisely.

Therefore, as an usual practice it is advisable to construct a calibration curve by employing several standard solutions.

The concentration of the unknown solution may be read off directly from the above calibration curve as is done in the case of colorimetric assays

APPLICATIONS