Analytical techniques

Pharmaceutical analysis is defined as a branch of chemistry, which involves the series of process for the identification, determination, quantitation, and purification.
Based upon the determination type, there are mainly two types of analytical methods. They are as follows:
a. Qualitative analysis: This method is used for the identification or recognition of the chemical compounds by means of:
• Colour, taste, viscosity and solubility
• Reaction producing a colour
• Reaction producing a precipitate
• Reaction involving a change of a physical parameter.
b. Quantitative analysis: This method is used for the determination of the amount of the sample. Also the quantitative chemical analysis carried out by determining the volume of a solution of accurately known concentration which is required to react quantitatively with a measured volume of the substance to be determined.

1. Analytical Techniques By Dr. Sachin M. Hiradeve Associate Professor Nagpur College of Pharmacy, Nagpur Dr. Sachin M. Hiradeve

2. Introduction to Analysis Pharmaceutical analysis is defined as a branch of chemistry, which involves the series of process for the identification, determination, quantitation, and purification.  Based upon the determination type, there are mainly two types of analytical methods. They are as follows: a. Qualitative analysis : This method is used for the identification or recognition of the chemical compounds by means of: • Colour , taste, viscosity and solubility • Reaction producing a colour • Reaction producing a precipitate • Reaction involving a change of a physical parameter. b. Quantitative analysis : This method is used for the determination of the amount of the sample. Also the quantitative chemical analysis carried out by determining the volume of a solution of accurately known concentration which is required to react quantitatively with a measured volume of the substance to be determined. Dr. Sachin M. Hiradeve

3. Different Techniques of Analysis Dr. Sachin M. Hiradeve

4. Titration Techniques It is based on the complete chemical reaction between the analyte and the reagent (titrant) of known concentration. Analyte + Titrant → Product Terms used in titration Analyte :- Unknown concentration of solution T itran t :- The solution of known concentration. Standard solution:- A solution of known concentration is called the standard solution. Dr. Sachin M. Hiradeve

5. Types of standard solution Primary st a nd a r d Extremely pure. Highly stable. Can be weighed easily. For e.g. Na 2 CO 3 , KHP Se c o n d a ry st a n d a r d: - Less pure than primary standard. Less stable than primary standard. Can not be weighed easily. For e.g. NaOH , HCl Dr. Sachin M. Hiradeve

6. Euivalence Point :- Point where the amount of two reactants are just equivalent . End point :- Point at which the reaction is observed to be complete, this point is usually observe with the help of indicator. Indicator:- An auxiliary substance which helps in the usual detection of the completion of the titration process at the end point. For ex a mple s : - M et h yl or a n g e, Phenolphthalein, Cresol red, Thymol blue. Dr. Sachin M. Hiradeve

7. Concentration Terms :- The concentration of standard solutions (titrants) are generally expressed in units of either molarity (C M , or M) or normality (C N , or N). Molarity (M):-It is the number of moles of a solute per liter of solution. Normality :- It is the gram equivalent weight of solute dissolved per litre of solution. Molality :- It is the number of moles of solute present in per kilogram of solvent. Dr. Sachin M. Hiradeve

8. T itrimetric c a lcul a tion :- It is based on the following law of equivalence:- NaVa = or NsVs MaVa = M sVs Whe r e, Na is the norm a lity o f a n a lyte. Va is the volume of the analyte. Ns is the normality of standard solution. Vs is the volume of standard solution used. Ma is the molarity of analyte. Ms is the molarity of standard solution. Dr. Sachin M. Hiradeve

9. Titrimetric apparatus :- Dr. Sachin M. Hiradeve

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11. Instrumental methods Spectroscopy Spectroscopy measures the interaction of the molecules with electromagnetic radiation. Spectroscopy consists of many different applications such as atomic absorption spectroscopy, atomic emission spectroscopy, ultraviolet-visible spectroscopy, x-ray fluorescence spectroscopy, infrared spectroscopy, Raman spectroscopy, dual polarisation interferometry, nuclear magnetic resonance spectroscopy, photoemission spectroscopy, Mössbauer spectroscopy and so on. Block diagram of an analytical instrument showing the stimulus and measurement of response Dr. Sachin M. Hiradeve

12. Summary of the spectroscopic Techniques Radiation Absorbed Effect on the molecule of the substance and information obtained Ultra-violet (190-400 nm) and Visible (400-800 nm) Changes in electronic energy levels within the molecule, conjugated unsaturation, conjugation with non-bonding electrons, extent of π electron system. Infra-red 667-4000 cm -1 Changes in the vibrational and rotational movements of the molecule. Detection of almost all functional groups which has specific vibrational frequencies such as C=O, O-H, NH 2 etc. Radio-frequency Frequency 60-300 MHz Nuclear magnetic resonance, induces changes in the magnetic properties of certain atomic nuclei, notably that of hydrogen. Hydrogen atom that of different environments can be detected, counted and analysed for structure determination Electron Beam Impact 70 eV, 6000 kJ/mol Ionisation and fragmentation of the molecule into a spectrum of fragment ions (determination of molecular weight and deduction of molecular structure from the fragments obtained) Dr. Sachin M. Hiradeve

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14. UV Spectra of Isoprene Dr. Sachin M. Hiradeve

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16. Mass Spectrum Dr. Sachin M. Hiradeve

17. NMR Spectrum Dr. Sachin M. Hiradeve

18. Dr. Sachin M. Hiradeve INTRODUCTION TO CHROMATOGRAPHY AND ITS APPLICATIONS 1 Dr. Sachin M. Hiradeve

19. I n t r o d u c t i o n  Chromatography, literally "color writing", was first employed by Russian-Italian scientist Mikhail Tsvet in 1900, primarily for the separation of plant pigments such as chlorophyll, carotenes, and xanthophylls.  The word chromatography is derived from two Greek words  Chroma ….……..color  Graphos ………..writing 2 Dr. Sachin M. Hiradeve

20. D e f i n i t i o n  ‘A technique by which a mixture is separated into its components on the basis of relative ability of each component to be moved along/through a stationary phase by mobile phase’ The technique of chromatography is based on the differences in the rate at which the components of a mixture move through a porous medium (called stationary phase) under the influence of some solvent or gas (called moving/mobile phase). Chromatography is a nondestructive procedure Applied both for both qualitative and quantitative studies 3 Dr. Sachin M. Hiradeve

21. P r i n c i p l e  Like-dissolve-like or like-prefer-like.  The basis-partition or distribution coefficient ‘K’ which describes the way in which a compound distributes itself between two immiscible phases.  Defined as the molar concentration of analyte in the stationary phase divided by the molar concentration of the analyte in the mobile phase  𝐾 = 𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑜𝑓 𝑐𝑜𝑚𝑝𝑜𝑛𝑒𝑛𝑡 𝑖𝑛 𝑠𝑡𝑎𝑡𝑖𝑜𝑛𝑎𝑟𝑦 𝑝ℎ𝑎𝑠𝑒 𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑜𝑓 𝑐𝑜𝑚𝑝𝑜𝑛𝑒𝑛𝑡 𝑖𝑛 𝑚𝑜𝑏𝑖𝑙𝑒 𝑝ℎ𝑎𝑠𝑒 4 Dr. Sachin M. Hiradeve

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23. Commonly Used Terms 7 Analyte The substance to be separated during chromatography Analytical chromatography Determines the existence and the concentration of analyte(s) in a sample Chromatogram Visual output of the chromatograph Dr. Sachin M. Hiradeve

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