ADVANCED ANALYTICAL TECHNIQUES IN PHARMACOGNOSY FOR HERBAL PRODUCT QUALITY ASSESSMENT.pptx

ADVANCED ANALYTICAL TECHNIQUES
IN PHARMACOGNOSY FOR HERBAL
PRODUCT QUALITY ASSESSMENT
GROUP MEMBERS
SYBIL T MURENDO R219111F
NYASHA FEIGHLEY MURENJEKWA R212899G
MUTERO RUTENDO R212868D
HAZEL NCUBE R212915M
TAFADZWA T NYANDORO R212898Q
MURINGISI ZVIKOMBORERO R212906Z
NGWAERUDZA E VICTOR R212888K
TRISH MUSHIPE R212876B

INTRODUCTION
 Herbal medication products are medical products obtained from plant materials.
 employ the therapeutic attributes of several plant species, encompassing their leaves, flowers, roots, stems, or
extracts, to enhance health and address or avert ailments.
 comprise a mixture of active constituents, including alkaloids, flavonoids, terpenes, and phenolic compounds,
which enhance their pharmacological effects..
 exhibit therapeutic efficacy for many health illnesses, including digestive disorders, respiratory issues, chronic pain,
and immune system enhancement.
 The increasing demand for herbal pharmaceutical products raises issues regarding quality control, safety, and
efficacy.
 Maintaining the uniform quality and standardisation of herbal products is essential to ensure their safety, efficacy,
and reproducibility to safeguard consumer health and safety.
 minimise potential dangers such as contamination, adulteration, and dangerous compounds, hence ensuring the
safety of herbal pharmaceutical products for ingestion. Inadequate quality control can present significant safety
hazards to consumers.
 Pharmacognosy, the analysis of medicinal substances derived from plants or other natural sources, has advanced
significantly with the advent of advanced analytical techniques.
 These technologies are crucial for assessing the quality, efficacy, and safety of herbal products.The increasing
demand for herbal remedies need reliable testing methods to ensure adherence to regulatory standards.

IR SPECTROSCOPY - PRINCIPLES OF (IR)
 Infrared spectroscopy involves the examination of infrared light's interaction with a molecule.
 The segment of the infrared spectrum most pertinent for the examination of organic molecules
spans a wavelength range of 2,500 to 16,000 nm, correlating to a frequency range of 1.9 x 10^13 to
1.2 x 10^14 Hz.
 The photon energy in this infrared range (1 to 15 kcal/mole) are insufficient to excite electrons but
can elicit vibrational excitation of covalently bound atoms and groups.
 Molecules undergo a diverse range of vibrational motions, distinctive of their constituent atoms, in
addition to the simple rotation of groups around single bonds.
 As a result, nearly all organic compounds will absorb infrared light that matches the energy of these
vibrations.
 Infrared spectrometers, analogous to conventional spectrometers, enable chemists to acquire
absorption spectra of substances that uniquely reflect their chemical structure.
 IR Spectroscopy quantifies atomic vibrations, enabling the identification of functional groups.
Typically, robust bonds and light atoms oscillate at elevated stretching frequencies (wavenumbers).

APPLICATIONS
 yields information regarding chemical structures, functional groups, and concentrations.
 allows for direct analysis of materials without the need for preprocessing
 generate a distinctive fingerprint for each herbal substance, facilitating comparison with reference
standards.
 quantifies moisture content, essential for assessing the stability and shelf-life of herbal products.
 identifies microbiological contamination, including bacteria and fungi, that may jeopardise product
safety.
 detects possible allergens, including proteins and polysaccharides.
 identifies hazardous substances, including aflatoxins and pyrrolizidine alkaloids. Analysis of flavonoids
and phenolic acids that contribute to antioxidant and medicinal effects.

INDUCTIVELY COUPLED PLASMA MASS
SPECTROMETRY (ICP-MS)
 detecting and quantifying trace elements, heavy metals, and other inorganic pollutants in a
sample.
 use an argon (Ar) plasma, to ionise the sample, which is subsequently analysed by a mass
spectrometer.
 Prevalent Herbal Preparations Analysed by ICP-MS are dietary supplements, Ayurvedic
medicines, herbal teas such as fennel, linden, roots, chamomile, green tea, thyme, sage,
rosemary, rosehip, ginger, balm, echinacea, blue tea, etc and essential oils.
 apparatus comprises an ion source (the ICP), a mass spectrometer (MS)—typically a scanning
quadrupole mass filter—and a detector.

PRINCIPLE
 A sample is delivered into the ICP in a nebulised state using a nebuliser, which transforms the liquid
sample into a fine aerosol.
 The aerosol sample is introduced into the ICP for ionisation.
 ICP is generated by applying a radiofrequency (RF) field to argon gas, resulting in a high-
temperature plasma of up to 10,000 K.
 The sample is atomised and ionised within this plasma. The elevated temperatures in the ICP
induce the atoms in the sample to ionise.
 The predominant ions possess a positive charge resulting from electron loss. Positively charged ions
are removed from the ICP and directed into the mass spectrometer for subsequent examination.
 Within the mass spectrometer, ions are propelled through a magnetic or electric field.
 The magnetic field in a magnetic sector instrument or the electric field in a quadrupole or time-of-
flight instrument induces the separation of ions according to their mass-to-charge ratio (m/z).
 The detector identifies the separated ions and quantifies their amount. The detector output is
utilised to quantify the concentration of components inside the sample.

APPLICATIONS
 ICP-MS is extensively employed to assess herbal items for heavy metal contamination.
 It identifies hazardous metals such as lead, mercury, arsenic, cadmium, and chromium,
which pose risks to human health, including Cu, Cd, Pb, As, Cr, Mn, and Hg, with a focus on
the determination of Cu. Heavy metal elements denote metals possessing a density over 4.5
g/cm3, such as Au, Hg, Pb, and Cr.
 It is employed for quality control to ascertain the authenticity and purity of botanical
products, assuring adherence to regulatory norms.
 Nutrient analysis measures vital minerals such as calcium, iron, magnesium, and potassium.
 Toxicity assessment to ascertain the existence of hazardous materials, guaranteeing the
product's safety for ingestion.

Chemometrics techniques
 It is the extraction of information from chemical systems using data-driven
methods.
 Use of Applied mathematics, multivariate statistics and computer science
principles to solve both descriptive and predictive problems in chemistry
and other related fields.
 Techniques include:
 Artificial Neural Networks (ANNs)
 Principal Component Analysis (PCA)
 Cluster Analysis
 Partial Least Squares Regression (PLS-R)

Artificial Neural Networks (ANNs)
 ANN models involving computations and mathematics to simulate
electrical activity of the brain and nervous system.
 The artificial neural networks emulate a biological neural network and uses
a reduced set of concepts from biological neural systems
 Types:
 Feedback ANN
 Recurrent Neural Networks (RNNs)
 Generative Adversarial Networks (GANs

Principles and applications
 Principles:
 ANNs consists of interconnected units called neurons that are organized in layers as input
layers, hidden layers and output layers.
 ANNs learn through adjusting weights that are based on the error of the output compared
to the expected result.
 For feedforward networks, the data moves in one direction from input to output and
backpropagation is used to update the weights by propagating the error backward
through the network
 Applications:
 ANNs are trained on historical data so as to predict therapeutic efficacy and potential
adverse effects of herbal drugs
 ANNs classify herbal drugs into different quality categories which are based on chemical
profiles and is useful in quality assurance, providing rapid and accuracy in classification

Principal Component Analysis (PCA)
 It is a data transformation technique used to reduce the complexity of high dimensional
data sets (such as mass spectroscopy data) while retaining most of the variation in the data
set
 Principles:
 Data standardization
 Capturing maximum variance
 Application:
 Identification of patterns and correlations in chemical composition of herbal drugs to
ensure consistency and quality
 Visualization and identification of potential outliers, causing toxicity of herbal drugs

Instrumentation
 Covariance matrix
 Eigenvalues and Eigenvectors
 Sorting eigenvalues
 Selection of principal component
 Projecting data

Cluster analysis
 Cluster analysis is a statistical method for processing data and organizing items based on how closely associated they
are and is typically used when there is no assumption made about the likely relationships within the data.
 It is concerned with data collection in which the variables have not been partitioned beforehand into criterion versus
predictor subsets.
 Principles:
1. Evaluating the quality of the clusters using methods like silhouette scores and elbow method
2. It is an iterative process that involves multiple runs with different parameters and algorithms to find meaningful clusters
3. Standardizing and normalizing data is often necessary to ensure that each feature contributes equally to the distance
calculations
 Applications:
 Detection of adulteration herbal products that do not fit into any cluster
 Identification of unique formulations through classification of herbal drugs into chemical properties category

Instrumentation
 Prepare data
 Choosing the right clustering algorithm
 Determining the optimal number of clusters
 Applying the clustering algorithm

Partial least squares regression
 It is a method which reduces the variables which are used to predict to a smaller set of predictors.
 A machine learning technique which combines the advantages of integrating principal component analysis,
typical correlation analysis and linear regression analysis.
 Principles:
 PLS regression identifies a set of latent variables that capture the maximum covariance between the
predictor variables (X) and the response variables (Y)
 Cross-validation is used to help avoid overfitting and ensuring that the model generalizes well to new data
 To ensure that all variables are on the same scale, data is standardized so that each variable has a mean of 0
and a standard deviation of 1.
 Applications:
 Identify key compounds responsible for therapeutic effects in herbal drugs
 Ensures only high-quality herbal products reach the market

Instrumentation
 Calculations of PLS components
 Regression modeling
 Cross-Validation

NUCLEAR MAGNETIC RESOLUTION
 It is based on how an external magnetic force and electromagnetic
radiation causes atomic nuclei to reorient from base energy to high energy
levels
 The principle involves three basic steps;
 There is alignment of the nuclear spin when a external magnetic force is
applied, analysis of the alignment by radio frequency waves and detection
and analysis of the electromagneic waves emitted by the nuclei in the
sample.

Application
 i.Confirming presence of the active phytochemicals.
 ii.Toxicity checks
 iii.Assessing ease of metabolism
 Identifying such functional groups helps to decide on whether the product
should
 Be completely withdrawn or be structurally modified but at the same time
maintaining its initial ability to bring about the desired physiological effect.

UV/VIS SPECTROSCOPY
 The interaction between matter and radiation within the ultraviolet visible
region of the electromagnetic spectrum. Chemical compounds absorb radiation and
reflect UV and visible light at different wavelengths within the same region.
INSTRUMENTATION

Application
 UV/VIS spectrophotomers are mostly used to investigate samples that contain
coloured compounds such as carotenoids and chlorophyll which are highly
pigmented.
 Absorbance UV/VIS spectroscopy uses the wavelength absorption characteristics of
samples of organic compounds to quantify specific chemical compounds ranging
from visible light to the ultraviolet region.
 It makes sure that key chemical compounds are present and fall within the specified
range.
 UV/VIS spectroscopy can also be used to monitor enzyme kinetics by measuring
changes in absorbance over time.
 Absorbance spectroscopy measures the optical density and absorbance of biological
components, which is helpful for real-time fermentation and bioprocess monitoring.
 Absorbance spectroscopy measures the optical density and absorbance of biological
components, which is helpful for real-time fermentation and bioprocess monitoring.

MOLECULAR TECHNIQUES
POLYMERASE CHAIN REACTION
 This approach enables the amplification of specific DNA sequences,
making it easier to detect and identify different components in herbal
preparations.
 PCR is important in protecting the integrity of herbal remedies since it allows
for the fast detection of contaminants, the authentication of species, and
the quantitative analysis of active compounds.

PRINCIPLES OF PCR
 The PCR technique is based on the enzymatic replication of DNA.
 In PCR, a short segment of DNA is amplified using primer mediated
enzymes.
 DNA Polymerase synthesizes new strands of DNA complementary to the
template DNA.
 The DNA polymerase can add a nucleotide to the pre-existing 3’-OH group
only. Therefore, a primer is required. Thus, more nucleotides are added to
the 3’ prime end of the DNA polymerase

COMPONENTS OF PCR
 DNA Template– The DNA of interest from the sample.
 DNA Polymerase– Taq Polymerase is used. It is thermostable and does not
denature at very high temperatures.
 Oligonucleotide Primers- These are the short stretches of single-stranded DNA
complementary to the 3’ ends of sense and anti-sense strands.
 Deoxyribonucleotide triphosphate– These provide energy for polymerization and
are the building blocks for the synthesis of DNA. These are single units of bases.
 Buffer System– Magnesium and Potassium provide optimum conditions for DNA
denaturation and renaturation. It is also important for fidelity, polymerase
activity, and stability.

PROCESS OF PCR
 The main processes in polymerase chain reaction are
 1. Denaturing
 The double helix structure of DNA is thermally denatured into two single strands.
 The reaction mixture is heated to 94-95 degrees Celsius between 15 to 30 seconds.
 2. Annealing
 The primer anneals the single stranded DNA templates at their complementary sites.
 For annealing to occur the heat should be reduced 55 to 70 degrees Celsius for
about 30 to 60 seconds.
 3. Elongation
 In the final step, temperature is raised to 72 degrees Celsius so that the Taq DNA
polymerase enzyme begins synthesizing new DNA strands in the 5’ to 3’ direction

APPLICATIONS
 1. Detection of contaminants and adulterants.
 PCR is very useful for detecting microbial contamination in herbal products. For example, studies have shown that it
can quickly detect microbiological contamination in herbal medications, which is critical for quality control. PCR is
a speedier alternative to traditional contamination detection methods, ensuring product safety through timely
actions.
 2. Authentication of Herbal Species
 The procedure is also used to authenticate herbal species. Using particular primers that target unique DNA
sequences of the chosen plant species, PCR can authenticate the identity of herbal compounds. This is critical for
preventing adulteration and ensuring that customers obtain authentic herbal goods.
 3. Real-time PCR for quantitative analysis.
 Real-time PCR (qPCR) expands the capabilities of classical PCR by providing DNA measurement in real time. This
approach can be used to determine the quantity of active substances or contaminants in herbal products, which
provides useful information for quality assurance.
 4. DNA Mini-barcoding
 DNA mini-barcoding is a derivative technique that uses PCR to amplify short DNA sequences for rapid identification
of herbal species. This method is especially effective for complicated herbal combinations in which numerous
species may be present, ensuring that the product is both safe and excellent quality.

DNA BARCODING
 DNA barcoding is a molecular technique that has gained popularity in the
analysis of herbal products, primarily to ensure quality and safety. This
method involves sequencing a short, standardized region of DNA from a
specimen, which is then compared to a reference database to determine
the species present in herbal formulations.

PRINCIPLES
 DNA barcoding uses a short, standardized DNA sequence from a specific
region of an organism’s genome to identify species.
 This is used as an identification method. The unknown specimen's DNA is
compared to a reference database of known DNA sequences to
determine species identity.
 It’s used for comparison with reference data. Each species has its own
unique DNA barcode, similar to how a supermarket scanner uses UPC
barcodes to identify items

PROCESSES OF DNA BARCODING
 1. Sample Collection
 The first step is to collect a sample from the plant. This may involve collecting a tissue sample such as a leaf.
 2. DNA extraction
 The next step is to extract the DNA from the sample.
 This can be done using a variety of techniques such as phenol chloroform extraction or automated DNA
extractors.
 3. PCR amplification
 Once the DNA has been extracted, it is typically amplified using the polymerase chain reaction to obtain
enough DNA for analysis. The PCR reaction is performed using specific primers that are designed to amplify the
DNA region of interest.
 4. DNA sequencing
 The amplified DNA is then sequenced to determine the exact order of the DNA base pairs. This can be done
using a variety of techniques.
 5. Data analysis
 Its then compared to a reference database of known DNA sequences to determine the identity of species

APPLICATIONS
 1. Species Identification
 One of the primary applications of DNA barcoding in herbal products is the precise identification of plant species. This is critical
for determining whether the herbal product contains the correct species, particularly in cases where adulteration or mislabeling
may occur. Manufacturers and regulators can verify the authenticity of herbal ingredients by comparing the DNA sequences
extracted from a sample to known sequences in a database.
 2. Detection of contaminants
 DNA barcoding is also useful for identifying contaminants in herbal products, such as unwanted plant species or microbial
contamination. This capability is critical for ensuring
 consumer safety is paramount, as contaminated herbal products can pose serious health risks. The method enables quick
identification of contaminants, making quality control measures more effective.
 3. Quality Control
 The use of DNA barcoding in quality control processes contributes to the maintenance of high manufacturing standards for
herbal products. Manufacturers can improve their offerings' overall quality and safety by ensuring that they contain the correct
species and are free of harmful contaminants. This is especially important in the herbal medicine industry, where product quality
is critical.
 4. Next-Generation Sequencing (NGS).
 Advances in sequencing technologies, such as next-generation sequencing (NGS), have improved the utility of DNA barcoding.
NGS enables the simultaneous analysis of multiple samples and provides a more complete picture of the genetic diversity found
in herbal products. This technology can improve the detection of adulterants and the accuracy of species identification.

NEXT GENERATION SEQUENCING
Principles of NGS
 DNA polymerase catalyzes the deoxyribonucleotide triphosphates (dNTPs)
into a DNA template strand during successive cycles of DNA synthesis.
Fluorophore excitation is used to identify the nucleotides during each step
of the process. The main distinction is that, rather than sequencing a single
DNA fragment, NGS extends the process across millions of fragments. NGS
provides high accuracy, a higher yield of error-free readings, and a high
percentage of base calls that exceed Q30.

PROCESSES
 1.DNA extraction
 It involves isolating DNA from the biological sample using chemical extraction.
 2.Library Preparation
 The genetic material is fragmented into smaller pieces after DNA extraction.
 These are then tagged with adapters and barcodes allowing the sequencer to
differentiate individual DNA molecules.
 3.Sequencing
 The library is then loaded into a sequencer which reads the sequence of fragment.
 4.Data Analysis
 This involves aligning the short sequencing reads to a reference genome or
assembling them de novo to reconstruct the original genetic sequence.

APPLICATION
 1. Thorough Species Identification
 NGS can identify multiple species within a single herbal product at the same time. This capability is especially useful in
complex mixtures, where traditional methods may struggle to accurately identify all components. NGS ensures the
presence of the correct species by analyzing the genetic material, which is critical for product authenticity and
consumer safety.
 2.. Detection of contaminants
 One of the key benefits of NGS is its ability to detect contaminants such as unwanted plant species and microbial
pathogens. This is critical for quality control in herbal products, as contamination can pose serious health risks. NGS makes
it easier to identify these contaminants, allowing manufacturers to act quickly to correct the situation.
 3. Quality control and assurance.
 NGS improves quality control processes by providing detailed information about the genetic composition of herbal
products. This information assists manufacturers in ensuring that their products meet safety requirements and contain the
intended active ingredients. The ability to analyze fragmented DNA also enables the evaluation of degraded samples,
which is frequently a challenge in herbal product testing.
 4.Meta-barcoding
 NGS can be combined with met barcoding techniques which sequence specific DNA regions to analyze species
diversity in a sample. This approach is especially useful for herbal products because it provides a comprehensive
overview of the plant species present, ensuring that the products are both safe and effective.

Real-Time Polymerase Chain Reaction
(qPCR)
Principle
 It states that the amplification and quantification of the target nucleic acid
occurs in a single reaction. Either a labeled probe or fluorescent dye is used
to monitor the reaction. The more the DNA is amplified the more
fluorescence is released and detected using the fluorescence detector
present in the machine.

PROCESSES
 The two steps involved are
 1. Amplification
 i. Denaturation
 ii. Annealing
 iii. Extension
 2. Detection
 It is based on fluorescence technology
 The specimen is first kept in proper well and subjected to thermal cycle like
in the normal PCR.

APPLICATION
 1. Detection of contaminants
 qPCR is especially effective at detecting microbial contaminants in herbal products like fungi and bacteria. For
example, multiplex qPCR has been used to identify fungi that produce toxic mycotoxins like aflatoxins and
ochratoxin A. This rapid screening capability is critical for ensuring the safety of herbal medicines because
contamination can pose serious health risks to consumers. This is essential for preventing adulteration and ensuring
that products contain the correct ingredients, which is critical for both efficacy and safety.
 2. Quality Control.
 qPCR improves quality control processes by accurately quantifying active ingredients and contaminants in herbal
products. This quantitative aspect enables manufacturers to ensure that their products meet safety standards and
have the appropriate levels of active compounds. For example, studies have shown that qPCR can be used to
assess the quality of herbal medicines by analyzing specific genetic markers.
 3. Rapid Results
 One of the most significant advantages of qPCR is its ability to produce results faster than traditional methods. This
speed is critical for timely decision-making in quality assurance processes, allowing manufacturers to address any
related issues related to contamination or ingredient authenticity quickly.

CHROMATOGRAPHIC TECHNIQUES
 Chromatography – a technique of isolation and identification of
components or compounds or mixture into individual components
by using the stationery phase and mobile phase.
 Chromatographic techniques - a set of analytical methods used to
separate, identify, and quantify components in complex mixtures,
such as herbal products.
 Chromatographic methods enable the detection of active
pharmacological compounds, assessment of purity, and
identification of potential contaminants (like pesticides, heavy
metals, and microorganisms). By providing detailed profiles of the
constituents within herbal products, these techniques play a crucial
role in ensuring their quality, efficacy, safety, and compliance with
regulatory standards.

HIGH-PERFOMANCE LIQUID
CHROMATOGRAPHY

Principles of High Performance Liquid
Chromatography
 Separation: separates complex mixtures into individual components
based on differences in affinity for stationary and mobile phases.
 Adsorption: interactions between analytes and stationary phase
influence separation.
 Partitioning: Distribution of analytes between analytes between
mobile and stationery phases affects separation.
 Detection- HPHC detectors measure analyte concentrations.

HPHC Applications in herbal analysis
 Identification and qualification of bioactive compounds.
 Detection of adulterants and contaminants.
 Standardization of herbal extracts.
 Fingerprinting for quality control
 Stability testing.

DETERMINATION OF THE SAFETY OF
HERBAL PRODUCTS
 Detection of adulterants and Contaminants.
 Quantification of active ingredients
 Detection of toxic compounds
 Stability testing
 Comparison with reference standards
 Detection of illegal additives

OTHER CHROMATOGRAPHIC METHODS
 Thin Layer Chromatography
 Gas Chromatography
 Gas Chromatography-Mass Spectrometry
 Liquid Chromatography-Mass Spectrometry

Physical Analytical Techniques
Scanning electron microscopy
Transmission electron microscopy
Thermal analysis
Scanning Electron Microscopy
It is a powerful imaging technique that uses a focused beam of electrons to produce
high-resolution images of sample’s surface
Principle
Electron beam scans sample surfaces
Interactions between electrons and sample produce signals that can be used to obtain
information about the surfaces topography and composition.
Signals detected and processed to form image and the images are viewed in real-time
on an external monitor.

Application
🠶 Identifying contaminants
🠶 Analysing particle size and shape thereby ensuring uniformity an identifying potential
allergens
🠶 Detecting heavy metals, pesticides or other harmful substances
🠶 Examining plant morphology thereby verifying authenticity and identifying adulterant.
🠶 Detecting microbial contamination e.g. bacteria, yeast
🠶 Monitors manufacturing processes, ensuring consistency and quality

Transmission Electron Microscopy
This is a powerful technique that uses abeam of electrons to focus on a specimen producing a
highly magnified and detailed image of the specimen producing a highly magnified and
detailed image of the specimen.
Principle
The working principle of the Transmission Electron Microscope (TEM) is similar to the light
microscope. The major difference is that light microscopes use light rays to focus and produce
an image while the TEM uses a beam of electrons to focus on the specimen, to produce an
image.
Applications
Transmission Electron Microscopy (TEM) plays a crucial role in analyzing herbal products,
ensuring safety and quality.

 1. Morphological analysis: Studies shape, size, and structure of herbal particles.
 2. Contamination detection: Identifies microorganisms, insect fragments, and foreign
particles.
 3. Authentication: Verifies herbal material identity through ultrastructural
characteristics.
 4. Purity assessment: Detects adulterants and contaminants.
 5. Nanoparticle analysis: Studies size, shape, and distribution of nanoparticles in herbal
products
 6. Chemical composition analysis: Uses Energy-Dispersive Spectroscopy (EDS) for
elemental analysis

Thermal analysis
 Thermal analysis (TA) is a group of physical techniques in which the chemical or physical
properties of a substance, a mixture of substances or a reaction mixture are measured as a
function of temperature or time, while the substances are subjected to a controlled
temperature programmed heating or cooling rate.
 The program may involve heating or cooling at a fixed rate of temperature change, or
holding the temperature constant at different time spam. The graphical results obtained are
called the thermogram. These methods are usually applied to solids, liquids and gels to
characterize the materials for quality control.
 Principle of thermal analysis
 Thermal Analysis includes all the methods of measuring the sample properties while the
sample temperature is program controlled.

Application
 Qualitative and quantitative analysis
 Characterisation of physical, mechanical and chemical properties of herbal medicines
 Providing structure and properties of herbal products
 Provide information about thermal stability of herbal products
 Solvent detection and quantification of additives
 Investigate the drug excipient compatibility in herbal products

Near-Infrared Spectroscopy (NIRS)
 Principles:
 Measures absorbance of near-infrared light by molecular vibrations.
 Utilizes the overtone and combination bands of molecular vibrations.
 Applications:
 Identification and quantification of active compounds in herbal products.
 Rapid quality control in production settings.
 Advantages:
 Non-destructive analysis.
 Fast results with minimal sample preparation.
 Limitations:
 Limited sensitivity for trace components.
 Requires calibration with known standards.

Raman Spectroscopy
 Principles:
 Based on inelastic scattering of monochromatic light (Raman effect).
 Provides molecular fingerprinting by measuring vibrational modes.
 Applications:
 Characterization of complex mixtures in herbal products.
 Detection of adulteration and authenticity verification.
 Advantages:
 Minimal sample preparation; can analyze solids, liquids, and gases.
 High specificity for molecular structures.
 Limitations:
 Fluorescence interference can complicate spectra.
 Higher cost and complexity of equipment.

Fourier Transform Infrared
Spectroscopy (FTIR)
 Principles:
 Measures the absorption of infrared light, providing a spectrum that represents molecular vibrations.
 Utilizes Fourier Transform for rapid data analysis.
 Applications:
 Identification of functional groups and molecular structures in herbal extracts.
 Quality assessment for consistency and standardization.
 Advantages:
 High resolution and sensitivity.
 Broad applicability across different sample types.
 Limitations:
 Requires careful sample preparation (e.g., solid samples may need dilution).
 Interpretation of spectra can be complex.

Handheld Spectrometers
 Principles:
 Portable devices utilizing various spectroscopic techniques (often NIRS or FTIR).
 Designed for on-site analysis.
 Applications:
 Field testing for rapid quality assessment of herbal products.
 Useful for practitioners to verify product authenticity.
 Advantages:
 Portability and ease of use.
 Immediate results aiding in quick decision-making.
 Limitations:
 Limited performance compared to laboratory-grade instruments.
 May have restrictions in terms of resolution and sensitivity.

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