Application of Spectroscopy in Isolation, Purification and Identification

• Application of Spectroscopy in Isolation, Purification and Identification helps detect, separate, and purify plant compounds precisely.
• Explore the Application of Spectroscopy in Isolation, Purification and Identification to analyze structure and ensure compound purity.
• Spectroscopic methods play a crucial role in identifying the structure, functional groups, and composition of bioactive compounds in crude extracts.
• Common techniques include:

1. UV-Vis Spectroscopy

   • Principle:

     • Measures absorption of ultraviolet or visible light, typically in conjugated systems (e.g., aromatic rings, unsaturated bonds).

   • Application:

     • Rapid screening for characteristic chromophores (e.g., flavonoids, alkaloids).
     • Preliminary quantification of known markers.

2. Infrared (IR) Spectroscopy

   • Principle:

     • Detects molecular vibrations, revealing functional groups (e.g., –OH, –NH, –C=O).

   • Application:

     • Confirmation of functional groups in isolated fractions.
     • Fingerprint region (600–1500 cm⁻¹) aids compound identification.

3. Nuclear Magnetic Resonance (NMR) Spectroscopy

   • Principle:

     • Uses magnetic fields to analyze hydrogen (¹H NMR), carbon (¹³C NMR), and other nuclei environments.

   • Application:

     • Structural elucidation of purified natural products.
     • Determination of stereochemistry and molecular connectivity.

4. Mass Spectrometry (MS)

   • Principle:

     • Ionizes molecules and separates ions based on mass-to-charge ratio (m/z).

   • Application:

     • Molecular weight determination.
     • Identification of characteristic fragmentation patterns.
     • Used with chromatographic techniques (GC-MS, LC-MS) for rapid profiling of complex mixtures.

• These techniques complement each other, providing a comprehensive understanding of compound identity and purity.

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