Page 110 - CatalogNEP-PS

 Understand the underlying assumptions of the most common chromatographic separation
techniques and approaches to method validation.

THEORY (45 Hours)

UNIT – I: (6 Hours)
Qualitative and quantitative aspects of analysis: Sampling, evaluation of analytical data, errors,
accuracy and precision, methods of their expression, normal law of distribution if indeterminate
errors, statistical test of data; F, Q and t test, rejection of data, and confidence intervals.
UNIT-II: (6 Hours)
Optical methods of analysis: Origin of spectra, interaction of radiation with matter, fundamental
laws of spectroscopy and selection rules, validity of Beer-Lambert's law.
UV-Visible Spectrometry: Basic principles of instrumentation (choice of source, monochromator
and detector) for single and double beam instrument.
UNIT-III:. (6 Hours)

Basic principles of quantitative analysis: Estimation of metal ions from aqueous solution,
geometrical isomers, keto-enol tautomers. Determination of composition of metal complexes
using Job's method of continuous variation and mole ratio method.

UNIT – IV: Chromatography (10 Hours)
Classification of chromatographic methods: Principles of differential migration, description of
chromatographic process, distribution coefficients, modes of chromatography, Column
chromatography (elution time and volume) capacity factor, column efficiency and resolution,
sample preparation.

UNIT – V: Paper Chromatography (05 Hours)
Experimental modifications, various modes of developments, nature of paper, detections of spots,
retardation factors, factors that affect reproducibility of Rf values (due to paper, solvent system,
sample, development procedures), selection of solvent, quantitative analysis, applications.
UNIT – VI: Thin Layer Chromatography (06 Hours)

Stationary phase, adsorbents, liquid phase support, plate preparation, mobile phase, sample
application, development, saturation of chamber, detection of spot, Rf values (effect of adsorbent,
solvent, solute, development process), quantitative analysis, applications.

UNIT – VII: Solvent Extraction (07 Hours)
Distribution law, determination of distribution ratio, batch extraction, continuous extraction,
discontinuous extraction, counter-current extraction.

PRACTICAL (30 Hours)

1. Separation and identification of amino acids present in the given mixture by ascendingpaper
Chromatography.
2. Separation of ortho-nitrophenol & para-nitrophenol and o- and p-amino phenol by thin layer
chromatography (TLC) and calculation of their Rf values.
3. Separation of constituents of leaf pigments by thin layer chromatography and paper
chromatography.
4. Separation of a mixture of compounds by solvent extraction.
5. Analysis of soil samples (at least three soil samples to be collected for analysis).
i. Determination of pH of soil samples.
ii. Determination of total soluble salts.

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