M.Pharm: Spectral Analysis & Analytical Techniques

• The material covers advanced spectral analysis topics, problem-solving using spectral data, and various analytical techniques.
• It also provides instructions for a M.Pharm Semester-2 examination.
• The exam requires answering any five questions, making assumptions where needed, and noting that figures indicate full marks.

IR Spectroscopy and Compound Differentiation

• Differentiation of compounds like hex-2-yne/hex-1-yne, cis-but-2-ene/trans-but-2-ene, and acetaldehyde/acetone can be achieved using IR spectra.

Flash Chromatography

• Flash Chromatography to be covered, short notes to be written.

ATR-IR

• ATR-IR's principle and working should be explained.
• Advantages and disadvantages of ATR-IR compared to transmittance and diffuse reflectance modes should be outlined.

2-D NMR Spectrum

• A 2-D NMR spectrum definition is required.
• Understanding of COSY, NOESY, HETCOR, and INADEQUATE (full forms) is being tested.

Bioassay

• The term Bioassay should be explained.
• Principles and methodology for bioassay of a drug are required.

Structure Elucidation

• Structure elucidation of an organic compound is tested based on provided spectral data, including:
  • Molecular Formula: C8H19N
  • IR (cm⁻¹): 3410, 2928, 1461, 1132
  • ¹H NMR (ppm): 2.57 (t), 1.45 (pent), 1.31 (sextet), 1.10 (exchangeable), 0.90 (t)
  • ¹³C NMR (ppm): 49.8, 32.3, 20.4, 13.9
  • MS (m/z): 129, 86, 44

Index of Hydrogen Deficiency

• Determining the index of hydrogen deficiency for given compounds is needed.

McLafferty Rearrangement

• The McLafferty rearrangement needs to be explained with examples.
• Usefulness of McLafferty rearrangements in mass spectrum interpretation should be explained.

EI Mass Spectra

• General fragmentation rules for EI mass spectra should be described.

Structure Elucidation Rules

• Nitrogen rule, Rule of thirteen, and Isotope peaks help in structure elucidation, along with examples

Supercritical Fluid Chromatography

• Supercritical fluid definition and instrumentation in chromatography are to be described.

HPTLC

• Principles, instrumentation, and applications of HPTLC should be described.

IR Spectra and Isomers

• Prediction of main absorption bands in IR spectra of isomers is required.

NMR Techniques Compared

• Comparison and contrast is needed between ¹H and ¹³C NMR techniques.

Raman Spectroscopy

• The principle and applications of Raman Spectroscopy should be explained.

UV Absorption Maxima

• Calculation of UV absorption maxima for compounds from their structures is tested.

ELISA

• Principle, methodology, and applications of ELISA should be explained.

DSC

• Instrumentation and applications of DSC should be described.

LC-MS, LC-FTIR and LC-NMR

• Applications of LC-MS, LC-FTIR, and LC-NMR techniques should be described.

Spectral Definitions

• Definitions are needed for: Base peak, Fermi resonance, Chemical shift, Chromophore, and Glass transition temperature.

NMR Spectroscopy Q&A

• Questions related to NMR spectroscopy are asked:
  • Ethyl acetate mixed with TMS: expected spectrum depiction with multiplicity.
  • PMR chemical shift values for methane, chloromethane, dichloromethane, and chloroform explanation (0.2 ppm, 3.0 ppm, 5.3 ppm, 7.2 ppm respectively).

Structure Elucidation with Spectral Data

• Structure elucidation of an organic compound based on spectral data is being tested:
  • Molecular Formula: C8H9OCl
  • IR (cm⁻¹): broad absorption from 3600-3400
  • ¹H NMR (ppm): δ 7.50 (d, J= 9.0, 2H), 7.10 (d, J= 9.0, 2H), 3.70 (t, J= 5.0, 2H), 3.00 (s, 1H), 2.70 (t, J= 5.0, 2H)
  • ¹³C NMR (ppm): δ 138, 131, 129, 127, 65, 39
  • MS (m/z): 154, 152, 137, 135, 109, 105, 73

RIA and ELISA Techniques

• Explanation of RIA and ELISA techniques.

Flash Chromatography

• Principle, instrumentation, and applications of flash chromatography.

Hyphenated Techniques

• Definition of hyphenated techniques.
• Brief note on various hyphenated techniques of gas chromatography.

Ionization Techniques in Mass Spectrometry

• Different ionization techniques used in mass spectrometry should be explained.

DEPT NMR Spectroscopy

• Brief note on DEPT NMR spectroscopy technique.

UV Spectroscopy for Compound Differentiation

• UV spectroscopy can distinguish between pairs of compounds.

DSC Technique

• Principle and instrumentation of DSC technique should be explained.

Mass Fragmentation Patterns

• Prediction of mass fragmentation patterns of pentan-2-one, isobutanol, and benzyl chloride is being tested.

PMR Spectrum

• Detailed explanation of the expected PMR spectrum (including splitting pattern and approximate δ value) is required for:
  • ethyl acetate
  • 2-chloroacetic acid
  • p-nitrotoluene

LC-MS and CF-MS

• Short note on LC-MS and CF-MS.

Mac Lafferty Rearrangement

• Brief note on Mac Lafferty rearrangement.

Key Terms

• Definitions of key terms: Nitrogen rule, rule of thirteen, index of hydrogen deficiency, relative abundance, molecular ion peak, base peak.

Supercritical Fluid Chromatography

• Note on supercritical fluid chromatography.

Transitions in Organic Compounds

• Explanation of different types of transitions that occur in organic compounds giving suitable examples.

Correlation Spectroscopy

• What is correlation spectroscopy?
• Limitations of 1D NMR spectroscopy.
• Two types of correlation spectroscopy with suitable examples.

Thermal Analysis

• Differentiate between Thermo Gravimetric Analysis (TGA) and Differential Thermal Analysis (DTA).

ATR-IR Spectroscopy

• Brief note on ATR-IR spectroscopy technique.

IR Spectroscopy and Compound Differentiation

• How IR spectroscopy differentiates between the pairs of compounds:
  • benzaldehyde and acetophenone
  • pent-1-yne and pent-2-yne
  • ethylamine and diethylamine
  • nitroethane and ethanenitrile

Chromatography Techniques

• Briefly explain ion and ion exclusion chromatography.

Raman Spectroscopy

• Principle and applications of Raman spectroscopy.

McLafferty Rearrangement

• McLafferty Rearrangement with two examples.

HPTLC Discussion

• HPTLC must be discussed in detail.

IR Spectrum Interpretation

• Interpretation of an IR spectrum in the region: 3200-3650 cm-1 and 2000-2700 cm-1.

LC-NMR

• Write a note on LC-NMR.

ATR-IR Spectrophotometry

• Describe ATR-IR Spectrophotometry.

NOESY NMR

• Explain NOESY NMR in brief.

Mass Analyzers

• Enumerate and explain various types of Mass analyzers.

Raman Spectroscopy

• Write principle, instrument, and application of Raman Spectroscopy.

Fragmentation of Amines

• Write a note on the Fragmentation of amines.

Fragmentation Rules

• Explain Fragmentation Rules in Mass spectroscopy.

HETCOR NMR

• Write a note on HETCOR NMR.

IR Spectra Pair Discrimination

• How to distinguish by IR spectra: (1) CH3COCH3 and CH3CH=CHCH2OH (2) RNH2 and RCONH2.

ELISA Note

• Write a note on ELISA.

GC-MS

• Write principle, instrument, and application of GC-MS.

Woodward-Fieser Rules

• Why solvent corrections are required in Woodward-Fieser rules to calculate λmax of carbonyl compounds?

Acetone Justification

• Justify "acetone shows λmax 279 nm in hexane, whereas in water it shows λmax 264.5 nm."

DTA

• Explain Differential thermal analysis (DTA).

SFC

• Write a note on supercritical fluid chromatography.

MS

• How will you distinguish 2-pentanone and 3-pentanone by using mass spectrometry?

2-D NMR

• Describe 2-dimensional NMR spectroscopy in detail.

DSC Note

• Write a note on DSC.

Difference via IRS

• How can you differentiate Primary, Secondary, and Tertiary amine by IR Spectroscopy?

Woodward-Fieser Rules

• Use the Woodward-Fieser rules to calculate λmax of following compounds

ATR-IR

• Explain principle and instrumentation of ATR-IR.

NMR

• Differentiate between 1D and 2D NMR
• Write a note on NOESY.

COSY and HETCOR

• Explain COSY and HETCOR with suitable examples.

Chemical Shift

• Give probable chemical shift (δ ppm) and splitting pattern in 'H NMR

Molecular Rearrangment

• Explain the McLafferty Rearrangement with example.

Mass Interpretation

• Give the Mass interpretation pattern for the following organic compounds

Chromatographies

• Write principle and applications of ion exchange chromatography, and supercritical fluid chromatography

GC-MS

• Give principle and applications of GC-MS.

DSC

• Describe the principle and instrumentation of DSC.

RIA

• Explain RIA with suitable example.

Raman Spectroscopy

• Write principle and applications of Raman spectroscopy.

Write a not on Spectrometry

• Write a note on LC-MS, LC-FTIR and LC-NMR.

DTA

• Write principle and methodology of DTA.

Compounds

• How will you differentiate following compounds using IR spectra?

Hydrogen Deficiency Index

• Explain Hydrogen deficiency index, ring rule and nitrogen rule.

ELISA

• Explain ELISA with suitable example.

Flash Chromatography

• Write a note on ion chromatography and flash chromatography.

Supercritical Fluid Chromatography

• Explain the principle, instrumentation, and applications of Supercritical Fluid Chromatography.

Thermal Techniques

• What is the basic difference in DTA and DSC technique?
• How is DSC used to study the stability of a drug?

Spectrophotometry

• Explain principle and instrumentation of ATR-IR spectrophotometry.

ELIZA/ RIA

• Explain RIA and ELISA and Describe radioimmunoassay of digitalis.

Calorimetry

• Explain principle, instrumentation and applications of differential scanning calorimetry.

Chromatography

• Explain principle and applications of ion exchange chromatography.

Chromatography

• Explain various hyphenation techniques of liquid chromatography and Describe GC-MS.

Chromatography and Distinguish them

• Differentiate: HPLC and HPTLC.

Isotopic Abundance

• What is isotopic abundance?
• Explain importance of isotopic peak in mass spectra giving example.

Raman Spectrometry

• Explain principle, instrumentation and applications of Raman spectroscopy.

COSY and HETCOR.

• Explain COSY and HETCOR.

Commentary of IR spectrum

• An IR spectrum is considered as a finger print of molecule, comment.

Conversion

• Convert 5.8 µm into wave number in cm-¹.

Compounds Absorption and Why

• Acetone absorbs at 1715 cm¯¹ whereas acetaldehyde absorbs at 1730 cm-¹. Why?

Comment on rearrangement and C13

• Explain different types of rearrangements observed in MS with example and Explain NOE and what are the problems in integration in C13 spectra?

NMR

• Write a brief note on 2 dimensional NMR spectroscopy and Describe the methods used in simplification of complex NMR spectra.

Differentiation

• Differentiate the following pair of compounds on the basis of IR Spectra, Acetophenone and p- methyl benzaldehyde ,CH3CH=CH-COOH and C6H5CHОНСООН

Identy spectral data

• Identify the following compound on the basis of the spectral data

Analysis

• UV,IR, NMR,C13NMR, MS

Predict fragmentaion pattern

• Predict fragmentation pattern of butyrophenone and benzyl alcohol

How masses a produced

• Predict the masses of the ions produced in the mass spectrum of CH3C6H4CH2CH2CH2CH3, by (a) benzylic fission and (b) the McLafferty rearrangement

Explain Woodward – Fieser rule

• Explain Woodward – Fieser rule for a, ẞ-carbonyl compounds.

Explain transitions

• Explain and give examples of the types of transition occur in organic compounds.

Explain Spectroscopy

• Explain principle and application of UV-Vis Spectroscopy.

Discuss Notes

• Discuss these notes
  • ATR –IR
  • Super critical fluid Chromatography
  • Write principal, instrument and application of CEMS

Factors affecting the chemical shift

• Describe the factors affecting the chemical shift. ###Principle spectroscopy
• Write the principle of NOESY spectroscopy and HECTOR NMR.

Types of Mass analyzer

• Enumerate various types of Mass analyzer.
• Discuss quadrupole mass analyzer with appropriate diagram.

Principal instrument

• Write principal, instrument and application of LC-FTIR and Mc Lafferty rearrangement. ###What is method analysis
• What is thermal method of analysis? Discuss principle and applications of differential thermal analysis (DTA) ###Define analysis
• Define and describe TGA.
• Discuss the thermogravimetric curve of Calcium oxalate monohydrate and describe double sandwich ELISA technique for antigen measurement

Discuss Notes

• Write principal, instrument and application of Flash Chromatography,Raman Spectroscopy and Ring rule.

RIA of Digitalis

• Write note on RIA of Digitalis.

Write notes on Spectroscopy

• Write principal, instrument and application of GCMS and HPTLC.