By SANDEEP KUMAR THATIKONDA Assistant professor Care college of pharmacy Oglapur, Warangal.

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.

COLORIMETER Similarity:  transmitted light

measured
 measured at 1800

TURBIDIMETER Similarity:
transmitted light measured  measured at 1800

Difference:
 Absorption of radiation

Difference:
 Scattering of radiation

FLOURIMETRY Similarity:  Emergent radiation

measured at 900

NEPHELOMETRY Similarity: Emergent radiation measured at 900 Difference:

Scattered radiation measured emitted radiation same WL as that of incident light

Difference:
emitted radiation measured emitted radiation longer WL than incident light

CHOICE OF THE METHOD depends upon the amount of light scattered by suspended particles present in solution.

TURBIDIMETRY - high conc. Suspensions

NEPHELOMETRY low conc. 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 nor too small otherwise the scattering efficiency falls off. optimum particle size should be 0.1- 1 micro meters.

TURBIDIMETRY suspended particles > incident light wave length : larger particles undergo reflection- measuring transmitted radiation larger particles- absorbance vs. con. not linear relationship 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 B. Particle geometry C. Wave length of incident light D. RI difference

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

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 C

log Io / Is Vs

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

NEPHELOMETRY
Absorption is much less- white light is generally used If clear solutions having dark particles light in red / IR

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

DETECTORS

Nephelometers - Photomultiplier tubes

Turbidimeters - Phototubes

NEPHELOMETER

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

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
1. Determination of inorganic substances Sulphate barium chloride Ammonia Nesslers reagent Phosphorus Strychine molybedate 2.Analysis of water 3.Determination of CO2 clarity, conc. of ions

4.Biochemical Analysis 5.Quantitative Analysis (ppm level)

6. Miscellaneous Water treatment plants, sewage work, refineries, paper industry

7. Atmospheric pollution 8.Turbidimetric titrations

smokes & fogs

9. Determination of mole. Wt of high polymers

10. Phase titration