chromatography in analysis of natural products

1. Subject presentation: (BT 418)
submitted in partial fulfillment of the requirements for the award of the Degree
of
DOCTOR OF PHILOSOPHY
in
SCIENCES
by
MADONA MATHEW
(PHD/BE/10054/2023)
Department of Bioengineering and Biotechnology
Birla Institute of Technology, Mesra
Ranchi - 835215, Jharkhand
The Use of Chromatography in the Analysis of Natural Products

2. Chromatography
• Chromatography is a powerful analytical technique
used to separate, identify, and quantify the
components of complex mixtures.
• It plays a crucial role in the analysis of natural
products, providing insights into their chemical
composition and purity.

3. Principles of Chromatography
 Separation of Compounds: Chromatography utilizes differences in the physical and chemical
properties of compounds to separate them into their individual components.
 Mobile and Stationary Phases: The sample is carried through a stationary phase by a mobile
phase, which can be a liquid, gas, or supercritical fluid.
 Analyte Interactions: As the sample passes through the stationary phase, different compounds
interact differently, causing them to separate and elute at different times.

4. Types of Chromatography
• Chromatography encompasses a diverse range of analytical
techniques used to separate, identify, and quantify the
components of complex mixtures.
• Some of the major types include thin-layer
chromatography (TLC), high-performance liquid
chromatography (HPLC), gas chromatography (GC), and
ion-exchange chromatography.
• Each technique offers unique advantages and is suited for
the analysis of different types of natural products, such as
plant extracts, essential oils, and pharmaceutical
compounds.
• The choice of chromatographic method depends on factors
like sample complexity, analyte properties, and the
desired level of separation and sensitivity.

5. Thin Layer Chromatography (TLC) Analysis
 Thin Layer Chromatography (TLC) is a powerful
analytical technique used to separate and identify the
components in natural product extracts.
 It provides a simple, rapid, and cost-effective
method for analyzing the chemical composition of
complex mixtures.
 TLC allows for the visual detection of compounds
based on their distinct migration patterns on a silica
gel-coated plate.
 This technique helps identify the presence and
relative abundance of target compounds in
natural product samples.

6. High-Performance Liquid
Chromatography (HPLC)
Separation Columns
HPLC utilizes specialized
columns packed with absorbent
materials to separate complex
mixtures of compounds.
Mobile Phase
A liquid solvent system is used
to carry the sample through the
column, enabling effective
separation.
Detection
Sophisticated detectors identify
and quantify the separated
compounds based on their
physical and chemical
properties.

7. Gas Chromatography (GC) Analysis
Sample Preparation
Careful sample preparation is crucial
for GC analysis, often involving
extraction, derivatization, and
purification steps to ensure analytes
are compatible with the GC system.
Separation Principles
GC separates analytes based on
their volatility and affinity for the
stationary phase, allowing for high-
resolution separation of complex
natural product mixtures.
Detection Methods
Various GC detectors can be used,
such as flame ionization (FID) or
mass spectrometry (GC-MS), to
identify and quantify the separated
natural product components.

8. Identification and Quantification of
Natural Compounds
Chromatographic techniques play a crucial role in the identification and quantification of natural
compounds. These methods allow for the separation, isolation, and analysis of complex mixtures
found in natural products. The compounds are identified by comparing their retention times and spectral
data with reference standards.
Quantification of natural compounds is achieved through calibration curves and internal standards.
This enables researchers to determine the concentrations of specific compounds in the sample, providing
valuable insights into the chemical composition and potency of natural products.

9. Sample Preparation for Natural Products
1
Extraction
Selecting the appropriate solvent to extract the
desired compounds from the natural source,
such as plant materials or microbial cultures. 2 Purification
Removing unwanted impurities and
contaminants through techniques like liquid-
liquid extraction, solid-phase extraction, or
column chromatography.
3
Concentration
Concentrating the extracted compounds to
increase their relative abundance for more
accurate analysis.

10. Applications of Chromatography in
Natural Product Analysis
1
Quality Control
Ensuring consistent purity of natural products
2
Identification
Detecting the presence and quantity of specific
compounds
3
Purification
Isolating target compounds from complex

11. Applications of Chromatography in Natural
Product Analysis
Herbal Medicine
Analysis
Chromatography is used
to identify and quantify
active compounds in
herbal medicines,
ensuring product quality
and safety.
Essential Oil
Profiling
Chromatography
techniques like GC-MS
allow detailed analysis of
the chemical composition
of essential oils for quality
control.
Nutraceutical
Screening
HPLC is commonly used
to screen plant extracts for
beneficial phytochemicals
for use in dietary
supplements and
functional foods.
Cannabis Potency
Testing
Chromatography is the
gold standard for accurate
quantification of
cannabinoids in cannabis
products for quality
assurance.

12. Advantages and Limitations of
Chromatographic Techniques
1 Advantages
Chromatography is a powerful analytical tool that
can separate, identify, and quantify a wide range
of natural compounds with high precision and
sensitivity.
2 Versatile Applications
The various chromatographic techniques can be
applied to analyze diverse natural products, from
plant extracts to essential oils and secondary
metabolites.
3 Limitations
Proper sample preparation and method
optimization are critical to ensure reliable and
reproducible results, which can be time-consuming
and require specialized expertise.

13. Conclusion and Future Perspectives
 In conclusion, chromatographic techniques have proven invaluable in the analysis and
identification of natural products.
 These powerful tools provide insights into the complex chemical composition of plant,
animal, and microbial sources, paving the way for further research and development.
 Looking ahead, advancements in chromatographic instrumentation and data analysis
software will likely expand the reach and capabilities of these techniques.
 Integrating chromatography with other analytical methods, such as mass spectrometry, will
further enhance our understanding of natural products and their potential applications in fields
like medicine, agriculture, and cosmetics.

14. References
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