Republic of the Philippines

BATANGAS STATE UNIVERSITY

The National Engineering University
Alangilan Campus
Golden Country Homes, Alangilan, Batangas City, Batangas, Philippines 4200
Tel Nos.: (+63 43) 425-0139 loc. 2121 / 2221
E-mail Address: [email protected] | Website Address: http://www.batstate-u.edu.ph

College of Engineering
Department of Electronics

SUMMARY OF REPORT
“ INFRARED SPECTROSCOPY”

Presented to:
Engr. Apryll Joy A. Hernandez
Batangas State University
Alangilan, Batangas City

In Partial Fulfillment
of the Requirements for
Che 437 - Analytical Instrumentation
Bachelor of Science in Instrumentation and Control Engineering

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WHAT IS INFRARED SPECTROSCOPY?

Infrared spectroscopy is a scientific technique that involves shining infrared light onto a
material and then observing how the material interacts with that light. Different molecules
absorb infrared light in specific ways, providing valuable clues about their composition and
structure. By analyzing these interactions, scientists can identify substances and learn more
about their properties, helping in fields ranging from chemistry to materials science and beyond.
Infrared spectroscopy has the ability to examine samples in various states (liquid, solid, or gas)
and can be employed independently or in conjunction with other devices using diverse sampling
techniques. In addition to primary vibrational modes, secondary features like overtone and
combination bands, Fermi resonance, coupling, and vibration-rotational bands are also evident
in the spectrum. Given the wealth of information contained in its spectrum, infrared spectroscopy
has emerged as a widely utilized and beneficial method for deducing molecular structure and
identifying substances.

HOW DOES INFRARED SPECTROSCOPY WORK?

Infrared spectroscopy works by exposing a sample to infrared radiation and then observing
how the molecules in the sample interact with this radiation. When infrared light is passed through
or reflected off a sample, the molecules absorb certain wavelengths of the infrared light, causing
them to vibrate. These vibrations correspond to the specific chemical bonds within the molecules.
The energy required for these vibrations is characteristic of the types of bonds present in the
molecule. By measuring which wavelengths of infrared light are absorbed, scientists can identify
the types of bonds and functional groups present in the sample, providing valuable information
about its chemical composition and structure.

PRICIPLE OF INFRARED SPECTROSCOPY

The principle of infrared spectroscopy lies in the interaction between infrared radiation and
matter, particularly the molecules of a sample under analysis. When infrared radiation, which
consists of wavelengths longer than visible light but shorter than microwaves, is passed through
or reflected off a sample, the molecules within that sample absorb specific wavelengths of the
radiation. This absorption occurs because the energy of the infrared radiation matches the
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energy needed to excite the molecular vibrations, specifically the stretching and bending of
chemical bonds within the molecules. Different types of chemical bonds absorb infrared radiation
at characteristic frequencies, allowing for the identification of functional groups and molecular
structures present in the sample. By analyzing the pattern of absorption, scientists can obtain
valuable information about the chemical composition, structure, and properties of the sample.

SAMPLES IN INFRARED SPECTROSCOPY

Infrared spectroscopy can be applied to analyze a wide range of samples across various
states of matter, including liquids, solids, and gases.

 Liquid Samples: Liquids are frequently analyzed using infrared spectroscopy. These
samples may include organic solvents, chemical solutions, oils, pharmaceutical formulations,
beverages, and biological fluids such as blood or urine. Liquid samples are typically placed in
a suitable container, such as a cuvette, and analyzed directly.

 Solid Samples: Solids represent another common category of samples for infrared
spectroscopy. Solid samples may include polymers, plastics, minerals, pharmaceutical tablets,
food products, fibers, and various materials used in manufacturing. Solid samples are often
analyzed as pellets or thin films, which are prepared by mixing the solid with a suitable matrix
material like potassium bromide (KBr) and compressing it into a disc or coating it onto a
surface.

 Gaseous Samples: Infrared spectroscopy is also employed to analyze gases and vapors.
Gaseous samples may include atmospheric gases, industrial emissions, volatile organic
compounds (VOCs), combustion products, and gases evolved from chemical reactions.
Gaseous samples are typically introduced into the spectrometer using specialized gas cells or
sample chambers designed to contain and analyze the gas.

INFRARED SPECTROSCOPY INSTRUMENTATION

The instrumentation for infrared (IR) spectroscopy consists of several key components that
work together to analyze samples. The instrumentation for infrared spectroscopy is designed to
provide accurate and reliable measurements of the infrared radiation absorbed or emitted by
samples, allowing for the identification and analysis of chemical compounds based on their
unique spectral signatures. The following are the key components:

IR Radiation Source: This component emits infrared radiation, which is directed towards the
sample being analyzed. Common sources include heated filaments, globars, or lasers,
depending on the specific requirements of the instrument.

Sample Holder: The sample holder securely holds the sample in place during analysis.
Samples may be in various forms, such as liquids, solids, or gases, depending on the nature of
the analysis.

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Monochromators: Monochromators are used to isolate specific wavelengths or ranges of
infrared radiation emitted by the source. They ensure that only the desired wavelengths pass
through to the sample, improving the specificity of the analysis.

Detectors: Detectors measure the intensity of infrared radiation that interacts with or passes
through the sample. Common types of detectors include thermal detectors, photoconductors,
and photovoltaic detectors, each with its own sensitivity range and response time.

Recorder: The recorder captures and records the data obtained from the detector. This data is
typically displayed as an infrared spectrum, which shows the intensity of infrared radiation as a
function of wavelength or wavenumber. Modern instruments often use computer software for
data acquisition and analysis, allowing for more advanced processing and interpretation of the
results.

TYPES OF INGRARED SPECTROMETER

Dispersion-Based IR Spectrometers: In dispersion-based spectrometers, infrared radiation

from the source is passed through a monochromator, which disperses the radiation into its
component wavelengths. The selected wavelength is then directed onto the sample, and the
resulting spectrum is measured. These spectrometers offer high spectral resolution but may be
slower compared to other types.

Fourier Transform Infrared (FTIR) Spectrometers: FTIR spectrometers utilize

interferometers to measure the entire infrared spectrum simultaneously. The interferometer
generates an interferogram, which is then Fourier-transformed to obtain the spectrum. FTIR
spectrometers offer rapid data acquisition and high sensitivity, making them well-suited for a
wide range of applications.

Near-Infrared (NIR) Spectrometers: NIR spectrometers operate in the near-infrared region of

the electromagnetic spectrum, typically from around 700 nm to 2500 nm. While not strictly
infrared spectroscopy, NIR spectroscopy shares some similarities and is used for similar
purposes, such as chemical analysis and material identification. NIR spectrometers are often
used in industries such as agriculture, pharmaceuticals, and food processing for rapid analysis
of samples.

Mid-Infrared (MIR) Spectrometers: MIR spectrometers operate in the mid-infrared region of

the spectrum, typically from around 2.5 μm to 25 μm. These spectrometers are particularly
useful for analyzing chemical bonds and functional groups in organic and inorganic compounds.

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APPLICATIONS

Infrared (IR) spectroscopy finds a wide range of applications across numerous scientific
disciplines and industries due to its ability to provide valuable insights into the composition,
structure, and properties of materials.
Chemical Analysis: IR spectroscopy is extensively used for qualitative and quantitative
analysis of chemical compounds. It enables the identification of functional groups, determination
of molecular structures, and quantification of analytes in complex mixtures. Applications include
pharmaceutical analysis, environmental monitoring, forensic science, and quality control in
various industries.

Material Characterization: IR spectroscopy is invaluable for characterizing the composition

and properties of materials. It is used to analyze polymers, plastics, fibers, coatings, ceramics,
metals, and composites. By studying molecular vibrations, IR spectroscopy helps elucidate
material properties such as chemical composition, crystallinity, purity, and thermal stability.

Biomedical Research: In biomedical research and clinical diagnostics, IR spectroscopy is

employed for studying biological tissues, cells, proteins, lipids, and other biomolecules. It
enables the detection of disease markers, assessment of tissue pathology, monitoring of drug
interactions, and characterization of biomolecular structures. Applications include cancer
diagnosis, drug development, and understanding biochemical processes.

Pharmaceutical Analysis: IR spectroscopy plays a crucial role in pharmaceutical analysis,

including drug development, formulation, and quality control. It is used to identify and quantify
active pharmaceutical ingredients (APIs), detect impurities, assess drug stability, and analyze
drug delivery systems such as tablets, capsules, and topical formulations.

Environmental Monitoring: IR spectroscopy is applied in environmental science for monitoring

air and water quality, assessing pollution levels, and identifying environmental contaminants. It
enables the analysis of pollutants, such as volatile organic compounds (VOCs), pesticides, and
industrial by-products, in environmental samples. IR spectroscopy also aids in studying soil
composition and characterizing atmospheric gases.

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 SUMMARY OF REPORT “VISIBLE AND ULTRAVIOLET SPECTROSCOPY”

WHAT IS VISIBLE AND ULTRAVIOLET SPECTROSCOPY?

Visible and ultraviolet (UV) spectroscopy are analytical techniques used to study the
interaction of light with matter in the visible and ultraviolet regions of the electromagnetic
spectrum. These techniques provide valuable insights into the electronic structure and
properties of molecules, enabling a wide range of applications in fields such as chemistry,
biology, materials science, and environmental science.

HOW DOES VISIBLE AND ULTRAVIOLET SPECTROSCOPY WORK?

In visible and ultraviolet (UV) spectroscopy, light from a suitable source, such as a tungsten
lamp for visible light or a deuterium lamp for UV light, is passed through a sample. When the
light interacts with the molecules in the sample, some wavelengths are absorbed by the
molecules. The remaining light is transmitted through the sample and detected by a detector.
The absorption of light occurs due to electronic transitions within the molecules. Specifically,
electrons in the molecules can absorb energy from the incident light and move from lower
energy levels to higher energy levels. The energy of the absorbed light corresponds to the
energy difference between these electronic energy levels. The absorption of light at specific
wavelengths corresponds to electronic transitions that are characteristic of the molecular
structure of the sample. Different types of molecules and functional groups absorb light at
different wavelengths, leading to characteristic absorption spectra.

PRICIPLE OF VISIBLE AND ULTRAVIOLET SPECTROSCOPY

The principle of visible and ultraviolet (UV) spectroscopy lies in the interaction of light with
matter, specifically the absorption of light by molecules in a sample. When light passes through
the sample, certain wavelengths are absorbed by the molecules, causing electronic transitions
from lower to higher energy states. By measuring the intensity of absorbed light at different
wavelengths, scientists can obtain an absorption spectrum that reveals information about the
electronic structure and composition of the sample.

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VISIBLE AND ULTRAVIOLET SPECTROSCOPY INSTRUMENTSTION

The instrumentation for visible and ultraviolet (UV) spectroscopy typically includes the following
components:

Light Source: Provides the light that interacts with the sample. For UV spectroscopy, common
light sources include deuterium lamps, which emit light in the UV range (200-400 nm). For
visible spectroscopy, tungsten filament lamps are commonly used, emitting light in the visible
range (400-800 nm).

Wavelength Selector: Selects specific wavelengths of light to pass through the sample. This
component could be a monochromator, which disperses light into its component wavelengths
and selects a narrow band of wavelengths, or filters that selectively transmit certain
wavelengths of light.

Sample Holder: Holds the sample being analyzed. Samples can be liquids, solids, or gases,
depending on the analysis requirements. Liquid samples are often placed in cuvettes, while
solid samples may be prepared as thin films or dissolved in a suitable solvent. Gaseous
samples are typically introduced into the spectrometer using specialized gas cells.

Detectors: Measure the intensity of light transmitted through the sample at different
wavelengths. Photomultiplier tubes (PMTs) or photodiodes are commonly used detectors in UV-
visible spectroscopy. These detectors convert the light signal into an electrical signal, which can
be further processed and analyzed.

Amplifier: The electrical signals produced by the detectors are often weak and need to be
amplified for accurate measurement and analysis. The amplifier boosts the amplitude of these
signals, making them easier to measure and analyze. Amplifiers help improve the signal-to-
noise ratio and enhance the sensitivity of the instrument.

Recorder: Captures and records the data obtained from the detectors. This data is typically
displayed as an absorption spectrum, which shows the intensity of light absorbed by the sample
as a function of wavelength or frequency. Modern UV-visible spectrometers often use computer
software for data acquisition, storage, and analysis, allowing for more efficient and advanced
processing of spectral data.

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TYPES OF UV-VISIBLE SPECTROMETER

Single Beam Spectrophotometer: In a single beam configuration, the sample and detector
are placed in series. Initially, the light intensity (Io) from the monochromator is transmitted
through the sample. The sample absorbs specific wavelengths of light, leading to electronic
transitions from lower to higher energy states. The intensity of the transmitted light (I) is
measured by the detector. This type of spectrophotometer is straightforward but may be prone
to drift and variations in light intensity.

Double Beam Spectrophotometer: In a double beam spectrophotometer, the monochromatic

light is split into two beams by a splitter or chopper. One beam passes through the sample,
while the other beam serves as a reference and passes through a standard. The intensity of the
light transmitted through the sample and the standard is measured simultaneously by two
detectors. By comparing the intensities of the two beams, this configuration compensates for
fluctuations in light intensity, providing more accurate and reliable measurements.

APPLICATIONS

Food and Beverage Industry: UV-visible spectroscopy is used in the food and beverage
industry for quality control, authentication, and safety assessment of food products. It enables
the analysis of vitamins, pigments, antioxidants, and other nutritional components in food
samples. UV-visible spectroscopy is also employed for analyzing flavors, aromas, and additives
in beverages, as well as detecting contaminants and adulterants in food products.

Colorimetric Assays: UV-visible spectroscopy is widely used in colorimetric assays for

quantitative analysis of biochemical substances. It allows for the measurement of absorbance
or transmission of light by colored or chromogenic compounds, enabling the determination of
concentrations of analytes in biological samples. Colorimetric assays are used in enzyme
kinetics, immunoassays, DNA quantification, and various biochemical assays in research,
clinical, and industrial settings.

Biomedical Research: In biomedical research and clinical diagnostics, UV-visible

spectroscopy is employed for studying biological samples, including proteins, nucleic acids,
enzymes, and metabolites. It enables the detection of biomarkers, assessment of sample purity,
monitoring of biochemical reactions, and characterization of molecular structures. Applications
include protein quantification, DNA analysis, enzyme kinetics, and drug interaction studies.

Pharmaceutical Analysis: UV-visible spectroscopy plays a crucial role in pharmaceutical

analysis, including drug development, formulation, and quality control. It is used to identify and
quantify active pharmaceutical ingredients (APIs), detect impurities, assess drug stability, and
analyze drug delivery systems such as tablets, capsules, and topical formulations. UV-visible
spectroscopy is also employed in pharmacokinetic studies and bioavailability assessments.

Environmental Monitoring: UV-visible spectroscopy is applied in environmental science for

monitoring air and water quality, assessing pollution levels, and identifying environmental
contaminants. It enables the analysis of pollutants, such as heavy metals, organic pollutants,
and dissolved gases, in environmental samples. UV-visible spectroscopy is also used for
studying soil composition, analyzing wastewater, and monitoring the effects of pollution on
ecosystems.

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 SUMMARY OF REPORT “FLUORESCENCE SPECTROSCOPY”

WHAT IS FLUORESCENCE SPECTROSCOPY?

Fluorescence spectroscopy is a scientific technique used to study the fluorescence emitted

by substances when they are excited by light of a specific wavelength. In this technique, a
sample is exposed to light of a particular wavelength, known as the excitation wavelength,
which causes the sample to absorb energy and become excited. Subsequently, the excited
molecules return to their ground state by emitting light at longer wavelengths, a process known
as fluorescence emission. The emitted fluorescence light is characteristic of the molecules
present in the sample, providing valuable information about their structure, composition, and
environment. By analyzing the intensity and wavelength of the emitted fluorescence, scientists
can gain insights into various properties of the sample, such as the presence of specific
molecules, their concentration, and their interactions with other substances.

HOW DOES FLUORESCENCE SPECTROSCOPY WORKS?

Fluorescence spectroscopy works by shining light of a specific wavelength onto a sample

containing fluorescent molecules. These molecules absorb the light energy and become excited,
causing their electrons to jump to higher energy levels. As the excited molecules return to their
original state, they release excess energy in the form of light, known as fluorescence emission.
This emitted light has a longer wavelength than the excitation light and is characteristic of the
molecules present in the sample. By measuring the intensity and wavelength of this emitted
fluorescence, scientists can identify the fluorescent molecules, determine their concentration,
and study their interactions with other substances.

PRINCIPLE

The principle behind fluorescence spectroscopy lies in the ability of certain molecules,
called fluorophores, to absorb light energy at a specific wavelength and then emit light at a
longer wavelength. When a sample containing fluorophores is exposed to light of a particular
wavelength, known as the excitation wavelength, the fluorophores absorb photons of this light
and become excited. This excitation causes the electrons within the fluorophores to move to
higher energy levels. Subsequently, the excited fluorophores return to their ground state by
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releasing excess energy in the form of light, a process known as fluorescence emission. The
emitted light has a longer wavelength than the excitation light and is characteristic of the
specific fluorophores present in the sample. By measuring the intensity and wavelength of the
emitted fluorescence, scientists can gain insights into various properties of the sample,
including the presence of specific molecules, their concentration, and their interactions with
other substances. Fluorescence spectroscopy is widely used in research, industry, and clinical
applications for its sensitivity, selectivity, and versatility in analyzing fluorescent materials.

INSTRUMENTATION

Fluorometers are essential tools in fluorescence spectroscopy, consisting of an excitation

and emission monochromator that enable users to acquire excitation and emission spectra. The
measurements obtained from a fluorometer are unique to each instrument due to variations in
the excitation and emission monochromators.

Excitation: When a sample containing fluorophores is exposed to light of a specific wavelength,

typically in the ultraviolet (UV) or visible range, the fluorophores absorb photons of this
excitation light. This absorption causes the electrons within the fluorophores to move to higher
energy levels, entering an excited state.

Relaxation: Once in the excited state, the fluorophores are unstable and seek to return to their
ground state. This transition involves the release of excess energy, which can occur through
various relaxation processes. These processes may include the emission of photons
(fluorescence), heat generation, or non-radiative decay pathways.

Fluorescence Emission:In the case of fluorescence, the excited fluorophores emit photons of
light as they return to their ground state. This emitted light has a longer wavelength than the
excitation light and is characteristic of the specific fluorophores present in the sample. The
emitted light can be detected and measured to generate a fluorescence spectrum.

Detection and Analysis: The emitted fluorescence light is collected using a detector, such as a
photomultiplier tube or photodiode, and converted into an electrical signal. By analyzing the
intensity and wavelength of the emitted fluorescence, scientists can gain insights into the
properties of the sample, such as the presence of specific molecules, their concentration, and
their interactions with other substances.

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APPLICATIONS

Fluorescent Probes: Fluorescence spectroscopy is widely used to detect and quantify

biological compounds present in low concentrations. Fluorescent probes are employed to
label specific molecules or cellular structures, enabling their visualization and
measurement in biological samples.

Protein Structure Analysis: Proteins contain intrinsic fluorophores such as tryptophan,

tyrosine, and phenylalanine. By analyzing the fluorescence emitted by these
fluorophores, along with solvent properties and the presence of co-factors, fluorescence
spectroscopy can provide valuable insights into protein structure and conformational
changes.

Fluorescence-Activated Cell Sorting (FACS): FACS is a powerful technique used to

sort a mixture of cells based on their fluorescent properties. Cells labeled with
fluorescent markers or dyes can be selectively isolated and sorted using fluorescence
spectroscopy, enabling researchers to analyze and characterize heterogeneous cell
populations.

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 SUMMARY OF REPORT “NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY”

WHAT IS NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY?

Nuclear magnetic resonance (NMR) spectroscopy utilizes magnetic fields and

electromagnetic radiation to examine the local magnetic fields surrounding atomic nuclei.
This resonance phenomenon occurs when atomic nuclei are subjected to both an external
magnetic field and specific-frequency electromagnetic radiation, resulting in the detection of
absorption signals and the generation of an NMR spectrum.
This technique is employed to analyze the content and purity of nanoparticles (NPs) in a
sample, as well as to determine the chemical structure of molecules present. Over time, the
development of NMR spectroscopy has witnessed significant advances in instrumentation,
along with improvements in instrument performance, experimental techniques, spectral
analysis, and theoretical understanding.

HOW DOES NUCLEAR MAGNETIC RESONANCE SPECTROSCOPYWORKS?

Nuclear magnetic resonance (NMR) spectroscopy operates by exploiting the magnetic

properties of atomic nuclei. When a sample containing nuclei with an odd number of protons or
neutrons is placed within a strong external magnetic field, these nuclei align themselves either
parallel or antiparallel to the field. Upon exposure to radiofrequency electromagnetic radiation at
a specific frequency, the nuclei absorb energy and temporarily transition to a higher energy
state. This process, known as resonance, occurs when the frequency of the electromagnetic
radiation matches the energy gap between nuclear energy levels. As the nuclei return to their
original state, they release the absorbed energy in the form of radiofrequency signals. These
signals are detected by a receiver coil and converted into a spectrum, known as an NMR
spectrum.

PRINCIPLE

The principle of nuclear magnetic resonance (NMR) spectroscopy is founded on the

property of spin exhibited by atomic nuclei with an odd number of protons or neutrons. When
placed in an external magnetic field, these nuclei generate a magnetic moment, which can
align either parallel or antiparallel to the applied magnetic field. When subjected to
electromagnetic radiation at a specific frequency, a resonance transition occurs between
magnetic energy levels of the nuclei. This resonance transition leads to the absorption of
electromagnetic radiation and the subsequent emission of signals, enabling the acquisition of
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an NMR spectrum. By analyzing these signals, scientists can determine the chemical
structure and composition of molecules in a sample.

INSTUMENTATION
The instrumentation used in nuclear magnetic resonance (NMR) spectroscopy typically
includes the following components:
1. Magnet: NMR spectrometers feature powerful magnets that generate a strong and
uniform magnetic field. This magnetic field is essential for aligning the nuclear spins within
the sample.
2. Radiofrequency (RF) Transmitter: An RF transmitter is used to generate
radiofrequency pulses at specific frequencies. These pulses are applied to the sample to
induce resonance in the nuclear spins.
3. RF Receiver: The RF receiver detects the signals emitted by the sample after
excitation with RF pulses. These signals contain information about the nuclear spins and are
used to construct the NMR spectrum.
4. Gradient Coils: Gradient coils are additional magnetic coils used to apply spatial
gradients to the main magnetic field. These gradients are necessary for spatial localization of
signals within the sample, enabling techniques such as magnetic resonance imaging (MRI).
5. Probe: The probe is the component of the NMR spectrometer that holds the sample. It
typically contains the RF transmitter and receiver coils, as well as mechanisms for controlling
the sample temperature and environment.
6. Data Processing Unit: The data processing unit includes computers and software that
control the spectrometer, acquire NMR data, and process the raw signals to generate the
final NMR spectrum.
7. Console: The console provides a user interface for controlling the NMR spectrometer
and monitoring the experimental parameters. It allows researchers to input experimental
parameters, initiate scans, and visualize the acquired data in real-time.

APPLICATION

Nuclear Magnetic Resonance (NMR) Spectroscopy is a powerful analytical technique used to

determine the structure and composition of molecules. Here are some common applications of
NMR spectroscopy:

Chemical Analysis: NMR spectroscopy is used extensively in chemical analysis to

identify the chemical structure of organic and inorganic compounds. It can determine the

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number of hydrogen atoms in a molecule, their chemical environment, and their
connectivity.
Drug Discovery: NMR spectroscopy is used in drug discovery to identify and
characterize small molecules that interact with target proteins. It can determine the
binding affinity, stoichiometry, and structure of the drug-protein complex.

Materials Science: NMR spectroscopy is used in materials science to study the physical
and chemical properties of materials. It can determine the structure and dynamics of
polymers, glasses, ceramics, and other materials.

Environmental Science: NMR spectroscopy is used in environmental science to study

the composition and structure of natural organic matter in soils, sediments, and water. It
can determine the sources and fate of pollutants in the environment.

Medical Diagnosis: NMR spectroscopy is used in medical diagnosis to detect and

quantify metabolites in biological fluids such as blood, urine, and cerebrospinal fluid. It
can diagnose diseases such as cancer, diabetes, and neurological disorders.

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