Unit 1

Basic Concepts of Spectroscopy

General Idea of Spectroscopy

Spectroscopy is the scientific study of the interaction between matter and
electromagnetic radiation as a function of wavelength or frequency. It is based
on the principle that every atom or molecule possesses distinct energy levels,
and transitions between these levels occur when they absorb, emit, or scatter
specific amounts of energy (quanta) from electromagnetic radiation.

In simple terms, spectroscopy provides a way to ―read‖ the unique energy

fingerprint of a substance. When radiation interacts with matter, the resulting
spectral pattern (lines or bands) reflects the structural, electronic, and chemical
characteristics of that material. By analyzing these patterns, scientists can
identify substances (qualitative analysis) and determine their amount
(quantitative analysis).

Spectroscopy covers multiple techniques such as absorption, emission,

fluorescence, Raman, and nuclear magnetic resonance each sensitive to different
parts of the electromagnetic spectrum (radio waves, microwaves, infrared,
visible, ultraviolet, X-rays, gamma rays).

In forensic science, spectroscopy plays a vital role in examining evidence like

drugs, explosives, inks, paints, gunshot residues, fibers, and trace metals. Its
non-destructive nature, high sensitivity, and precision make it one of the most
trusted analytical tools in modern investigations.
The title ―Father of Spectroscopy‖ is most often credited to Joseph von
Fraunhofer (1787–1826).
He was a German optician and physicist who made precise measurements of the
solar spectrum and discovered hundreds of dark lines (later called Fraunhofer
lines).
His work transformed spectroscopy from a simple optical curiosity into a
scientific measurement technique by combining experimental accuracy with
optical engineering.
Fraunhofer also developed high-quality diffraction gratings, which became key
tools in spectral analysis.

The term comes from two Latin-derived roots:

―Spectrum‖ (from Latin spectrum, meaning ―appearance‖ or ―image‖) —
referring to the array of colors or wavelengths obtained when light is dispersed.
―-scopy‖ (from Greek skopein, meaning ―to look‖ or ―to examine‖).
Thus, spectroscopy literally means ―the examination of spectra‖. Initially, it
referred to studying the visible light spectrum, but over time it expanded to
include all forms of electromagnetic radiation and even particle-based
spectroscopy (like mass spectroscopy).
Electromagnetic Specturm

Electromagnetic Spectrum
The electromagnetic spectrum is the complete range of all possible frequencies
and wavelengths of electromagnetic radiation from the longest radio waves to
the shortest gamma rays. All electromagnetic waves travel at the speed of light
(c = 3 × 10⁸ m/s) in a vacuum, but they differ in energy, frequency (ν), and
wavelength (λ).
It is divided into distinct regions based on wavelength and frequency, each
interacting differently with matter and having specific applications in
spectroscopy:
1. Radio Waves
Wavelength: > 1 m (up to thousands of km)
Frequency: < 3 × 10⁸ Hz
Energy: Very low
Applications: Nuclear Magnetic Resonance (NMR) spectroscopy,
communication systems.
 2. Microwaves
Wavelength: 1 mm – 1 m
Frequency: 3 × 10⁸ – 3 × 10¹¹ Hz
Applications: Rotational spectroscopy, radar, molecular structure analysis.
3. Infrared (IR)
Wavelength: 700 nm – 1 mm
Frequency: 3 × 10¹¹ – 4 × 10¹⁴ Hz
Applications: Vibrational spectroscopy (IR spectroscopy), identifying molecular
functional groups, forensic analysis of fibers, drugs, explosives.
4. Visible Light
Wavelength: 400 – 700 nm
Frequency: 4 × 10¹⁴ – 7.5 × 10¹⁴ Hz
Applications: UV-Vis absorption and emission spectroscopy, color analysis in
forensic samples.
5. Ultraviolet (UV)
Wavelength: 10 – 400 nm
Frequency: 7.5 × 10¹⁴ – 3 × 10¹⁶ Hz
Applications: Electronic spectroscopy, detecting drugs, inks, and fluorescent
substances in forensics.
6. X-Rays
Wavelength: 0.01 – 10 nm
Frequency: 3 × 10¹⁶ – 3 × 10¹⁹ Hz
Applications: X-ray diffraction (XRD), X-ray fluorescence (XRF) for elemental
analysis, material identification.
7. Gamma Rays
Wavelength: < 0.01 nm
Frequency: > 3 × 10¹⁹ Hz
Applications: Nuclear spectroscopy, radioactive isotope detection.

Various Sources of Radiation – Region & Utility in Spectroscopy

Radiation sources are selected based on the wavelength range needed, the type
of interaction studied, and the required intensity & stability. Below is a detailed,
point-wise explanation with region, description, and forensic/analytical uses.

1. Tungsten Filament Lamp

Region: Visible (400–700 nm) and Near-Infrared (700–2500 nm)
Description: A heated tungsten filament emits continuous radiation similar to a
blackbody source in the visible and near-IR range.

Utility:
Common in visible absorption spectroscopy (colorimetry, UV-Vis
spectrophotometers in visible mode).

Used for determining concentration of colored compounds and dyes.

Limitation: Cannot produce UV radiation; filament gradually evaporates and

blackens the bulb.
2. Deuterium / Hydrogen Discharge Lamp
Region: Ultraviolet (160–375 nm)
Description: Gas discharge in deuterium or hydrogen generates continuous UV
radiation with stable output.
Utility:
UV-Vis spectrophotometers (for UV range) to detect aromatic compounds,
conjugated double bonds, proteins, nucleic acids.

Widely used in forensic drug and ink analysis.

Limitation: Requires high-voltage starter; intensity drops with prolonged use.

3. Xenon Arc Lamp

Region: UV to Visible (~200–1000 nm)

Description: High-pressure xenon gas arc discharge gives intense, continuous
light across UV and visible spectrum.
Utility:
Fluorescence spectroscopy for detecting trace body fluids, dyes, or explosive
residues.
Photoluminescence studies in material analysis.

Limitation: High heat output; expensive; requires cooling.

4. Mercury Vapor Lamp

Region: Strong UV lines at 254, 313, 365 nm; some visible lines.
Description: Mercury atoms emit characteristic line spectra when excited
electrically.
Utility:
Calibration of wavelength scales in spectrometers.
Excitation source for fluorescence microscopy and certain photochemical
reactions.
In forensics, used for document examination under UV light.

Limitation: Only discrete wavelengths; not suitable for continuous absorption

measurements.

5. Lasers

Region: Depends on laser type —

UV Lasers: e.g., Excimer lasers (~193–351 nm)

Visible Lasers: e.g., He-Ne laser (632.8 nm)

IR Lasers: e.g., CO₂ laser (~10.6 μm)

Description: Highly coherent, monochromatic, and intense light source.

Utility:
Raman spectroscopy for pigments, inks, explosives.
Laser-induced fluorescence for ultra-trace detection.

3D scanning and micro-profiling in evidence analysis.

Limitation: Fixed wavelength; expensive; safety precautions required.

6. Globar (Silicon Carbide Rod)

Region: Mid-Infrared (4000–200 cm⁻¹)
Description: Electrically heated silicon carbide rod emits continuous IR
radiation.
Utility:
FTIR spectroscopy for functional group identification in organic and inorganic
compounds.
Forensic fiber, paint, and polymer analysis.
Limitation: Needs stable electrical supply; strong heat output.

7. Nernst Glower

Region: Mid-IR to Far-IR

Description: A ceramic rod of rare-earth oxides heated electrically; used before
globar became common.
Utility:

IR spectroscopy of solid samples and inorganic materials.

Limitation: Fragile; requires preheating before emission.

8. X-ray Tubes
Region: X-rays (~0.01–10 nm)
Description: High-energy electrons strike a metal target (anode), producing
continuous and characteristic X-rays.
Utility:
X-ray diffraction (XRD) for crystal structure determination.
X-ray fluorescence (XRF) for elemental composition analysis in glass, metals,
paints, soils.
Limitation: Hazardous without shielding; requires high voltage.

Interaction of Radiation with Matter

When electromagnetic radiation encounters matter, it can be reflected, absorbed,

transmitted, scattered, or cause emission. The exact interaction depends on the
energy of the radiation and the electronic, vibrational, and rotational structure of
the material.

Here, we focus on Reflection, Absorption, and Fluorescence — all highly

relevant to spectroscopy.

1. Reflection
Definition: Reflection occurs when incident radiation is redirected away from a
surface without significant absorption.

Mechanism:
Governed by the law of reflection: the angle of incidence equals the angle of
reflection.
Occurs because the electromagnetic wave interacts with the electron cloud at the
surface, inducing oscillations that re-emit light in a specific direction.
Types:
Specular Reflection: Smooth surfaces (like mirrors or polished metals) reflect
light in a single direction.
Diffuse Reflection: Rough surfaces scatter reflected light in multiple directions.
Spectroscopic Relevance:
Reflectance spectroscopy measures the intensity of reflected light to analyze
opaque or solid materials.
Used in forensic paint analysis, fiber identification, and surface coating studies.
Forensic Example: Comparing paint layers from a crime scene with a suspect
vehicle using visible/UV reflectance spectra.
2. Absorption
Definition: Absorption occurs when photons transfer their energy to atoms or
molecules, promoting them to higher energy states.
Mechanism:
Photon energy () must match the difference between two quantized energy
levels in the atom or molecule.
Depending on radiation energy, this may cause electronic, vibrational, or
rotational transitions.
Spectroscopic Relevance:
Absorption spectroscopy measures the decrease in radiation intensity as it
passes through a sample.
The resulting absorption spectrum reveals information about molecular structure
and concentration (Beer–Lambert Law).
Forensic Example:
UV-Vis absorption to identify and quantify drugs, dyes, inks.

IR absorption to detect functional groups in fibers, explosives, or polymers.

3. Fluorescence
Definition: Fluorescence is the emission of light by a substance that has
absorbed electromagnetic radiation, usually at a longer wavelength (lower
energy) than the absorbed radiation.
Mechanism:

1. Absorption: Molecule absorbs high-energy photon (UV/visible), exciting

an electron to a higher singlet state.
2. Internal Conversion: Electron loses some energy as heat
3. Emission: Electron returns to the ground state, releasing a photon of
lower energy (Stokes shift).
Time Scale: Very short-lived — occurs within nanoseconds (10⁻⁹ s).

Spectroscopic Relevance:
Fluorescence spectroscopy is highly sensitive, detecting even trace levels of
substances.
Works best for aromatic and conjugated compounds.
Forensic Example:
Locating biological fluids (semen, saliva, urine) on fabrics under UV light.
Detecting latent fingerprints using fluorescent powders or dyes.
Identifying altered or forged documents with fluorescent ink analysis.

💡 Key Difference Between Absorption & Fluorescence:

Absorption is the process of taking in energy from incident radiation.
Fluorescence is a form of emission that follows absorption, but only in certain
molecules with specific electronic structures.
Detection of Radiation in Spectroscopy
Detection of radiation is the process of converting incoming electromagnetic
radiation into a measurable signal (visual record, electrical output, etc.).
The choice of detector depends on wavelength range, sensitivity required, and
type of spectroscopy.
1. Photographic Detection
Principle:
Radiation interacts with a photosensitive film or plate coated with silver halide
crystals (e.g., AgBr).
Photons cause the reduction of some silver ions to metallic silver atoms in the
crystal, forming a latent image.
After chemical development, exposed areas appear dark, creating a permanent
record of the radiation pattern.
Features:
Works over a wide spectral range (UV, visible, X-rays) with proper coating.
Produces a permanent record of the spectrum (good for archival evidence).
Forensic Applications:
Capturing UV fluorescence of biological stains (semen, saliva) for later
comparison.
Recording emission spectra from trace elements in forensic samples.
X-ray photography for bullet fragment localization in forensic pathology.
Limitations:
Not real-time; requires chemical processing.
Lower sensitivity compared to modern electronic detectors.

2. Photoelectric Detection
Principle:
Based on the photoelectric effect: when photons strike a photosensitive surface,
electrons are ejected.

These electrons are collected, amplified, and measured as an electric current

proportional to the light intensity.
Types of Photoelectric Detectors:
Photomultiplier Tube (PMT): Extremely sensitive; amplifies photoelectrons via
multiple dynodes. Ideal for very weak signals (fluorescence, Raman
spectroscopy).
Photodiodes: Semiconductor devices where light generates electron–hole pairs;
used in UV-Vis spectrophotometers.
Charge-Coupled Devices (CCD): Arrays of photodiodes; convert light into
digital images; widely used in modern spectrometers.
Features:
Real-time measurement.
High sensitivity and dynamic range.
Can be integrated with computers for data processing.
Forensic Applications:
Real-time analysis of drug or explosive residues using portable Raman or
fluorescence detectors.
UV-Vis spectrophotometry of inks and dyes in questioned documents.
Trace detection of GSR (Gunshot Residue) via emission spectroscopy.
Limitations:
Wavelength-dependent response (detector must be suited to the spectral region).
Some types require cooling to reduce noise.

Forensic Applications of Spectroscopy

Spectroscopy plays a vital role in forensic science because it enables precise,
non-destructive, and rapid analysis of a wide variety of evidence. Different
spectroscopic techniques help in identifying the chemical composition,
structural properties, and origin of materials found at a crime scene.
1. Identification of Unknown Substances
Application: Helps determine the chemical composition of seized drugs,
poisons, and other unknown powders or liquids.

Example Technique: Infrared (IR) Spectroscopy detects characteristic

absorption bands of functional groups in drugs like heroin or cocaine.
2. Analysis of Paint and Polymer Evidence

Application: Determines the chemical structure and pigment composition in

paint chips from hit-and-run accidents or burglary scenes
Example Technique: UV-Visible Spectroscopy and FTIR are used to match
paint layers with a suspect vehicle.

3. Gunshot Residue (GSR) Detection

Application: Detects and identifies metallic residues (lead, barium, antimony)
and organic components from gun discharge.
Example Technique: Atomic Absorption Spectroscopy (AAS) and X-ray
Fluorescence (XRF).
4. Examination of Inks and Documents
Application: Identifies the chemical makeup of inks to detect forgeries or
alterations in cheques, wills, or contracts.
Example Technique: Raman Spectroscopy and UV-Vis Spectroscopy can reveal
different ink formulations.
5. Blood and Biological Stains
Application: Identifies hemoglobin derivatives, semen, saliva, or sweat stains,
even if degraded or washed.
Example Technique: UV-Vis Spectroscopy for hemoglobin, fluorescence
spectroscopy for biological fluids.
6. Explosives and Fire Debris Analysis
Application: Determines the presence and type of explosive residues or
accelerants used in arson.
Example Technique: Fourier Transform Infrared (FTIR) spectroscopy for solid
residues, Raman for molecular identification.

7. Trace Element Analysis

Application: Determines trace metals in hair, nails, soil, or glass for criminal
linkage.
Example Technique: Atomic Emission Spectroscopy (AES) or Inductively
Coupled Plasma–Optical Emission Spectroscopy (ICP-OES).

8. Glass Fragment Analysis

Application: Determines the refractive index and elemental composition of
glass fragments from burglary or accident scenes.
Example Technique: Laser-Induced Breakdown Spectroscopy (LIBS) or ICP-
MS.

9. Counterfeit Currency and Art Authentication

Application: Identifies pigments, dyes, or special printing inks in banknotes or
paintings.
Example Technique: Fluorescence spectroscopy under UV light reveals security
features.

10. Toxicological Examination

Application: Detects toxic metals, drugs, alcohol, or poison in biological
samples.
Example Technique: UV-Vis, Atomic Absorption, or Mass Spectrometry (when
combined with chromatography).

Sample Preparation for Spectroscopic and Chromatographic Evidence

Importance of Sample Preparation
In forensic science, the accuracy of spectroscopic or chromatographic analysis
largely depends on how well the sample is prepared.
Proper preparation ensures representative, uncontaminated, and analyte-rich
samples that give reliable results.
Since forensic samples are often limited or degraded, minimal loss during
preparation is critical.

General Principles of Sample Preparation

1. Prevent Contamination – Use clean glassware, gloves, and avoid contact

with foreign substances.
2. Maintain Integrity – Avoid excessive heating, strong light exposure, or
pH changes that may degrade the analyte.
3. Concentration Adjustment – Adjust analyte concentration to fall within
the detection range of the instrument.
4. Remove Interferences – Remove substances that may obscure or alter
results (dust, pigments, solvents, etc.).

1. Sample Preparation for Spectroscopic Analysis

Spectroscopy involves interaction of radiation with matter; the sample must be
prepared to be optically suitable.
a) Solid Samples
Grinding and homogenization to ensure uniform particle size (reduces scattering
effects).
Pellet Formation – In IR spectroscopy, solids are often mixed with KBr and
pressed into pellets.
Thin Film Preparation – Organic or polymer samples may be cast into thin films
for transmission measurements.
b) Liquid Samples
Filtration – Removes suspended solids that may scatter light.
Dilution – Ensures absorbance is within the instrument’s linear range.
Solvent Selection – Solvent must not interfere with the absorption/emission
spectrum.
c) Gaseous Samples
Gas Purification – Removes dust and moisture.
Gas Cell Filling – Stored in specialized optical cells with suitable path length.

2. Sample Preparation for Chromatographic Analysis

Chromatography separates components based on chemical and physical
properties, so sample must be compatible with mobile and stationary phases
a) Solids
Extraction – Solid-phase extraction (SPE) or liquid-solid extraction to isolate
analyte.
Grinding and Sieving – Ensures uniform sample for consistent elution.
b) Liquids
Filtration or Centrifugation – Removes particulate matter.
Dilution – Prevents overloading of column.
Derivatization – Chemically modifying analyte to make it more detectable or
better separated (e.g., silylation for GC).
c) Biological Samples (Blood, Urine, Tissue)
Deproteinization – Precipitates proteins using organic solvents.
Extraction – Liquid-liquid extraction (LLE) or SPE for isolating drugs, toxins,
or metabolites.
pH Adjustment – Enhances analyte solubility or stability before injection.

Storage and Transportation of Samples:

Use airtight containers to avoid volatilization.
Store in dark, cool conditions to minimize degradation.
Maintain chain of custody documentation for forensic validity.

Concepts of Chromatography

Chromatography, derived from the Greek words chroma meaning ―colour‖ and
graphein meaning ―to write,‖ is a separation technique first introduced by the
Russian botanist Mikhail Tsvet in 1903 while separating plant pigments such
as chlorophyll and carotenoids. It is based on the principle that the components
of a mixture distribute themselves differently between a stationary phase and a
mobile phase, causing them to move at different speeds and thus separate. The
stationary phase can be a solid or a liquid supported on a solid, while the mobile
phase can be a liquid or a gas. In forensic science, chromatography is an
essential tool used for the qualitative and quantitative analysis of a wide range
of evidence. Techniques such as Thin Layer Chromatography (TLC), Gas
Chromatography (GC), and High-Performance Liquid Chromatography
(HPLC) are applied to detect and identify drugs, poisons, inks in questioned
documents, explosive residues, and accelerants in arson investigations. The high
sensitivity, accuracy, and ability to analyze complex mixtures make
chromatography an indispensable method in modern forensic investigations.

Chromatographic Methods

Chromatography encompasses a group of analytical techniques designed to

separate, identify, and sometimes quantify the components in a mixture.
Although the principle remains the same—distribution of analytes between a
stationary phase and a mobile phase—different types of chromatography use
varying physical states, equipment, and mechanisms of separation.
1. Thin Layer Chromatography (TLC)

Principle:
TLC is based on the separation of components according to their differing
affinities for the stationary phase (a thin layer of adsorbent like silica gel or
alumina coated on a glass, plastic, or aluminum plate) and the mobile phase (a
liquid solvent or mixture of solvents).

Procedure:

1. Sample spotting – A small amount of sample is applied as a spot near the

base of the plate.
2. Development – The plate is placed upright in a developing chamber
containing the mobile phase. The solvent moves upward by capillary
action.
3. Separation – Components with higher affinity for the mobile phase
travel further; those with higher affinity for the stationary phase remain
closer to the baseline.
4. Visualization – Spots are visualized using UV light or chemical reagents.
5. Retention Factor (Rf) – .

Utility in Forensics:

 Detection of illicit drugs in biological fluids.

 Examination of inks and dyes in questioned documents.
 Analysis of explosives and accelerants.

2. Gas Chromatography (GC)

Principle:
GC separates volatile compounds based on their distribution between a mobile
phase (an inert carrier gas like helium or nitrogen) and a stationary phase (a
liquid or polymer coated on a solid support inside a column).

Procedure:

1. Sample Injection – Sample is vaporized in the injection port.

2. Separation – Compounds are carried by the gas through the column,
separating due to differences in boiling point and interactions with the
stationary phase.
3. Detection – Detectors such as Flame Ionization Detector (FID) or Mass
Spectrometer (GC–MS) identify and quantify compounds.

Utility in Forensics:
  Analysis of toxic substances and drugs.
 Detection of arson accelerants.
 Examination of volatile organic compounds in environmental samples.

3. High-Performance Liquid Chromatography (HPLC)

Principle:
HPLC separates components in a liquid sample using a high-pressure pump to
force the mobile phase through a densely packed stationary phase column.

Procedure:

1. Sample Introduction – Liquid sample is injected into the system.

2. High-Pressure Separation – Components interact differently with the
stationary phase, causing separation.
3. Detection – Detectors such as UV–Vis, fluorescence, or mass
spectrometry analyze eluted compounds.

Utility in Forensics:

 Detection of drugs, poisons, and metabolites in biological fluids.

 Analysis of inks, dyes, and synthetic fibers.
 Identification of anabolic steroids and performance-enhancing drugs.

4. Paper Chromatography(Classical Method)

Principle:
Paper acts as the stationary phase, and a liquid solvent moves through it by
capillary action, carrying analytes at different rates.

Utility in Forensics:

 Used for educational demonstrations and simple pigment separations.

 Rarely used in advanced forensic labs due to lower sensitivity compared
to TLC and HPLC.

5. Column Chromatography(Preparative Method)

Principle:
A column is packed with a stationary phase (silica/alumina) and the mobile
phase (liquid) passes through, separating components based on polarity and
adsorption.

Utility in Forensics:
  Isolation of pure compounds from mixtures.
 Purification before further spectroscopic analysis.

Key Advantages of Chromatographic Methods:

 High sensitivity and selectivity.

 Ability to analyze mixtures without prior separation.
 Compatibility with various detectors for qualitative and quantitative
analysis.

Fundamental principles and Forensic Applications of TLC,GC and LC

1. Thin Layer Chromatography (TLC)

Fundamental Principle:
TLC is based on the principle of adsorption chromatography, where
separation occurs due to differences in the adsorption of components on the
surface of a thin stationary layer. A thin film of adsorbent material (silica
gel, alumina, cellulose) is coated onto a glass, plastic, or aluminium plate,
which acts as the stationary phase. The mobile phase is a suitable liquid
solvent or mixture of solvents that moves up the plate by capillary action.
The rate at which each component travels depends on its affinity toward
the stationary phase versus its solubility in the mobile phase. The distance
travelled by each component is expressed as Rf (Retention factor),
calculated as:
Rf = {Distance travelled by the solute}/{Distance travelled by the solvent
front}}
Forensic Applications:
Drug Analysis: Identification of narcotics, amphetamines, LSD,
barbiturates from seized samples.
Ink and Dye Examination: Differentiation of inks in questioned documents
or forgery cases.
Poison Detection: Screening plant toxins, pesticides, and chemical poisons
in biological samples.
Explosive Residue Analysis: Preliminary identification of organic explosive
components.
Counterfeit Detection: Separation of pigments and dyes in counterfeit
currency and artworks.
 2. Gas Chromatography

Fundamental Principle:
GC is based on partition chromatography where separation occurs due to
differences in the distribution of components between a stationary phase (a
high-boiling liquid coated on an inert solid support) and a mobile phase (an
inert carrier gas like helium, nitrogen, or hydrogen). The sample is
vaporized and carried by the mobile phase through a column, where each
component interacts differently with the stationary phase, leading to
separation based on boiling point and polarity. Detection is done using
detectors like Flame Ionization Detector (FID), Thermal Conductivity
Detector (TCD), or Mass Spectrometric Detector (GC-MS) for precise
identification.

Forensic Applications:
Arson Investigations: Detection of accelerants (petrol, kerosene, diesel)
from fire debris.
Toxicology: Identification and quantification of volatile poisons such as
cyanide, methanol, ethanol.
Drug Profiling: Analysis of seized narcotics to determine purity and origin.
Explosives Analysis: Identification of volatile components in explosive
residues.
Environmental Forensics: Detection of pollutants in air samples related to
industrial crimes.
 Liquid Chromatography (LC) / High-Performance Liquid
Chromatography (HPLC)

Fundamental Principle:
LC is based on partition and adsorption chromatography where the
separation of components occurs between a liquid mobile phase and a solid
or liquid stationary phase. In HPLC, high pressure is used to push the
liquid mobile phase through a packed column containing the stationary
phase, allowing rapid, high-resolution separation of complex mixtures. The
choice of stationary phase (normal phase, reverse phase, ion exchange, size
exclusion) depends on the nature of the analytes. Detection is done using
UV-Visible Detectors, Fluorescence Detectors, or Mass Spectrometers (LC-
MS).

Forensic Applications:
Pharmaceutical and Drug Analysis: Identification of illicit drugs and
quality control of medicines.
Poison Detection: Detection of non-volatile and thermally unstable poisons
like alkaloids, glycosides, pesticides.
Blood and Urine Analysis: Quantification of drugs, alcohol metabolites,
and toxic chemicals in biological samples.
Explosives and Ink Analysis: Characterization of dyes, pigments, and
complex explosive mixtures.
Food Forensics: Detection of adulterants and contaminants in suspected
food crime cases.