What is UV Spectroscopy?

Spectroscopy is the measurement and interpretation of electromagnetic

radiation absorbed or emitted when the molecules or atoms or ions of a
sample move from one energy state to another energy state. UV spectroscopy
is a type of absorption spectroscopy in which light of the ultra-violet region
(200-400 nm) is absorbed by the molecule which results in the excitation of
the electrons from the ground state to a higher energy state.
Principle of UV Spectroscopy
1. Basically, spectroscopy is related to the interaction of light with matter.
2. As light is absorbed by matter, the result is an increase in the energy
content of the atoms or molecules.
3. When ultraviolet radiations are absorbed, this results in the excitation
of the electrons from the ground state towards a higher energy state.
4. Molecules containing π-electrons or nonbonding electrons (n-
electrons) can absorb energy in the form of ultraviolet light to excite
these electrons to higher anti-bonding molecular orbitals.
5. The more easily excited the electrons, the longer the wavelength of
light they can absorb. There are four possible types of transitions (π–
π*, n–π*, σ–σ*, and n–σ*), and they can be ordered as follows: σ–σ* >
n–σ* > π–π* > n–π*
6. The absorption of ultraviolet light by a chemical compound will
produce a distinct spectrum that aids in the identification of the
compound.
Instrumentation or Parts of UV Spectroscopy
Light Source
 Tungsten filament lamps and Hydrogen-Deuterium lamps are the most
widely used and suitable light sources as they cover the whole UV
region.
 Tungsten filament lamps are rich in red radiations; more specifically they
emit the radiations of 375 nm, while the intensity of Hydrogen-
Deuterium lamps falls below 375 nm.
Monochromator
 Monochromators generally are composed of prisms and slits.
  Most of the spectrophotometers are double beam spectrophotometers.
 The radiation emitted from the primary source is dispersed with the help
of rotating prisms.
 The various wavelengths of the light source which are separated by the
prism are then selected by the slits such the rotation of the prism
results in a series of continuously increasing wavelengths to pass
through the slits for recording purposes.
 The beam selected by the slit is monochromatic and further divided into
two beams with the help of another prism.
Sample and reference cells
 One of the two divided beams is passed through the sample solution
and the second beam is passé through the reference solution.
 Both sample and reference solution is contained in the cells.
 These cells are made of either silica or quartz. Glass can’t be used for the
cells as it also absorbs light in the UV region.
Detector
 Generally, two photocells serve the purpose of the detector in UV
spectroscopy.
 One of the photocells receives the beam from the sample cell and the
second detector receives the beam from the reference.
 The intensity of the radiation from the reference cell is stronger than the
beam of the sample cell. This results in the generation of pulsating or
alternating currents in the photocells.
Amplifier
 The alternating current generated in the photocells is transferred to the
amplifier.
 The amplifier is coupled to a small servometer.
 Generally, the current generated in the photocells is of very low
intensity, the main purpose of the amplifier is to amplify the signals
many times so we can get clear and recordable signals.
Recording devices
 Most of the time amplifier is coupled to a pen recorder which is
connected to the computer.
  The computer stores all the data generated and produces the spectrum
of the desired compound.
Applications of UV Spectroscopy
Detection of Impurities
 It is one of the best methods for the determination of impurities in
organic molecules.
 Additional peaks can be observed due to impurities in the sample and it
can be compared with that of standard raw material.
 By also measuring the absorbance at a specific wavelength, the
impurities can be detected.
Structure elucidation of organic compounds
It is useful in the structure elucidation of organic molecules, such as in
detecting the presence or absence of unsaturation, the presence of
heteroatoms.
1. UV absorption spectroscopy can be used for the quantitative
determination of compounds that absorb UV radiation.
2. UV absorption spectroscopy can characterize those types of
compounds that absorb UV radiation thus used in the qualitative
determination of compounds. Identification is done by comparing the
absorption spectrum with the spectra of known compounds.
3. This technique is used to detect the presence or absence of a
functional group in the compound. The absence of a band at a
particular wavelength is regarded as evidence for the absence of
particular group.
4. Kinetics of reaction can also be studied using UV spectroscopy. The UV
radiation is passed through the reaction cell and the absorbance
changes can be observed.
5. Many drugs are either in the form of raw material or in the form of the
formulation. They can be assayed by making a suitable solution of the
drug in a solvent and measuring the absorbance at a specific
wavelength.
6. Molecular weights of compounds can be measured
spectrophotometrically by preparing the suitable derivatives of these
compounds.
 7. UV spectrophotometer may be used as a detector for HPLC.

Spectroscopy is the branch of chemistry concerned with the investigative

measurements of the spectrum. UV-VIS (ultraviolet-visible) spectroscopy or
spectrophotometry is the study of the interaction of light with matter at electronic
levels. It ranges from the vacuum level ultraviolet region i.e. 180nm to the visible
region i.e. 780nm. UV spectrum extends from 180nm to 400nm whereas the visible
region ranges from 400nm to 780nm.

UV-VIS spectroscopy is an instrumental technique used for chemical analysis. It is

used for qualitative as well as quantitative analysis. This technique has several
applications for a large number of compounds. It is used to observe the optical
behavior of chemical compounds, identification of various species, and
quantification of specific analytes.

UV-Visible spectroscopy
In UV-VIS spectroscopy, the transition of electrons at various levels by absorption
of radiation from ultraviolet to visible region is plotted in a graph. This line graph of
various absorptivities on specific levels of radiations is because of the absorption
capacities of compounds at certain levels. These levels are called regions of
absorption and the compounds are termed as chromophores.

The chromophores are present in almost every compound. This can be deduced by
the fact that almost all compounds and especially organic compounds can be
identified and quantified by the uv-vis spectroscopy.

Working principle of UV-visible spectroscopy

When a chemical compound absorbs light, some excitation and de-excitation
processes of electrons occur in atoms which result in the production of the distinct
spectrum.

Electromagnetic spectrum
The electromagnetic spectrum is the division of electromagnetic radiation based on
the energy, frequency, or wavelength of a photon.
Interaction of radiation and matter
In spectroscopic analysis, when radiations interact with a chemical species, they can
cause transition at different energy levels. The type of transition depends upon the
energy of the radiation and the detection mode.

The transition of electrons always occurs from the ground state of low energy
HOMO (highest occupied molecular orbital) to a higher energy excited state LUMO
(lowest unoccupied molecular orbital). The overlap of atomic orbitals forms three
types of molecular orbitals. This whole concept arises from molecular orbital theory
(MOT).

 Bonding molecular orbitals of lower energy.

 Non-bonding molecular orbitals of intermediate energy.
 Anti bonding molecular orbitals are of higher energy.

The electronic transitions that require the highest energy are σ-σ* electronic
transitions, while the transitions requiring the least energy are n-π* electronic
transitions.

The energy order of different types of electronic transitions is as under.

σ-σ* > n-σ* > π-π* > n-π*

This phenomenon is studied under Beer-Lambert law.

Lambert law
Lambert’s law implies that the absorbance ‘A’ of incident monochromatic light is
directly proportional to the path length (cell length) ‘ℓ’. This means that equal
portions of absorbing material absorb equal fractions of incident light.

A∝ℓ

Beer law
Beer law implies that the absorbance ‘A’ of incident light or electromagnetic
radiation is directly proportional to the concentration ‘c’ of solution. This law gives
the quantitative relationship between the intensity of radiation and the
concentrations of chemical species.

A∝c

Beer-lambert law
Collectively Beer and Lambert’s laws state that the absorbance ‘A’ of an incident
monochromatic beam is directly proportional to concentration ‘c’ of the solution
and path length ‘ℓ’.

The rate of decrease in intensity of monochromatic light is proportional to the

thickness of medium ‘ℓ’ and concentration ‘c’ of absorbing substance in dilution.

A∝c.ℓ

A=ε.c.ℓ

where,

ε is the molar absorptivity coefficient constant.

Or

ε is the absorbance for a solution of concentration 1mole/dm-3 and a path length of

1cm.

Absorbance
It is the ratio of the intensity of incident electromagnetic radiation from the source
to that of refracted electromagnetic radiation detected by the detector.

Mathematical representation:

A = Log ℓo / ℓ

A=ε.c.ℓ

So,

Log ℓo / ℓ = ε . c . ℓ
Transmittance
It is the ratio of power of electromagnetic radiation leaving the sample Pt to that of
the incident radiation on the sample from the source.

Mathematical representation in terms of transmittance

T= Pt / Po

% T = T × 100

% T = Pt / Po × 100

Conversion of absorbance to transmittance

A = – log T

A = – log Pt / Po (Since T= Pt / Po)

A = log Po / Pt

A = 2- log T %

% T = antilog (2 – A)

Limitations of lambert beer law

1. The light source used must be monochromatic.

2. This is not suitable for concentrated solutions i.e. It can only be applicable to
dilute solutions.
3. With an increase in dilution, the dissociation of weak acids occurs. The weak
acids reach equilibrium with their conjugate base. The acid (HA) and
conjugate base (A–) cannot have the same absorbance. Hence this law is not
completely applicable to weak acidic solutions.

Instrumentation of UV-visible spectroscopy

The basic instrumentation of the UV-vis spectrometer comprises of

1. Light source
2. Diffraction grating
3. Wavelength selector
4. Sample container or cuvette
 5. Detector

1. Light Source
Light sources that lie in the ultraviolet and visible region are used as UV-visible
spectrometer sources.

1. Hydrogen & deuterium lamps range 160-380nm

2. Xenon arc lamps range 250-600nm
3. Tungsten halogen lamps range 240-2500nm

2. Wavelength selector
UV-Vis spectroscopy requires a single wavelength for proper functioning whereas
the ideal output of a single wavelength is not possible. This is so because no real
wavelength selector is ideal. Although a single wavelength is not possible, a band of
radiation could be used. So an instrument with narrow bandwidth would be better.

Types of the wavelength selectors

1. Filters
2. Monochromators

Filters
Filters are used to permit a certain band of wavelength. The simplest type of filter is
the absorption filter. Most commonly colored glass filters are used. They absorb a
broad portion of the spectrum (complementary colors) and transmit other portions
(its own color).

Advantages

 The filters are inexpensive.

 They possess technical simplicity.

Disadvantages

 The filters are restricted to the visible region only.

 They are not very good wavelength selectors.
 Not useful for research purposes as they allow broad bandwidth. So there are
more chances of deviation from Lambert beer’s law.
Monochromators
A monochromator is an optical device that is used to select a narrow band of a
wavelength of light. It may be a quartz prism or grating.

Uses of monochromators

 Monochromators are used for spectral scanning i.e. varying wavelength of

radiation over a range.
 They can be used for the UV-visible region.

Components of a monochromator

All monochromators are similar in mechanical construction. The essential

components of a monochromator are:

1. Slit
2. Mirror
3. Lense
4. Grating/prism

3. Sample container/cells or cuvettes

Sample containers or cuvettes may be made up of

1. Quartz
2. Borosilicate
3. Plastic

 Only quartz is transparent in both UV & visible regions (200-700nm range).

 Glass & plastic are suitable for the visible region only.
 Glass is not suitable for the UV region because it absorbs UV radiation i.e. it
is not transparent in the UV region.
 Plastic cells are not used for organic solvents.

Cuvette size

The most common cuvette size is 1 cm, although it can vary from 0.1-10 cm.

4. Detectors
Detectors are devices that indicate the existence of some physical phenomenon.
Some examples of simple detectors are
  Transducers
 Photodetectors
 Photographic films
 Mercury level in thermometers (temperature detector)
 Human eye

Transducers
A transducer is a special type of detector that converts signals such as light
intensity, pH, mass, and temperature, etc into electrical signals. This electrical signal
is amplified and manipulated. The signal is represented in numbers (digital form).

Properties of transducers

 Transducers produce a fast response to low levels of radiant energy.

 Suitable for a wide range of wavelengths.
 Electrical signals produced by transducers should have low noise.
 The signal produced by the transducer is directly proportional to the beam of
power.

Photodetectors
Photodetectors are used as:

1. Photo tubes
2. Photomultiplier tubes
3. Silicon diodes
4. Photovoltaic cells

Phototubes

Photo tubes or photoemissive cells are a type of photodetector. They are concave
surfaces of cathode and anode inside a glass bulb. The cathode is coated with
photoemissive material. e.g. cerium, potassium oxide, silver oxide, etc. The anode is
a metallic sheet or ring at high voltage by the battery. The interior of a bulb is filled
with an inert gas at low pressure.

Photomultiplier tubes

Photomultiplier tubes are structured as repeated dynodes at particular angles. The

emitted electrons strike on different dynodes. Every dynode has a high voltage than
the previous ones. This voltage difference accelerates the electrons. These fast
electrons knock out more electrons upon striking the next dynodes.
The repetition of the above process causes up to 106 electrons collected for each
photon, striking the first cathode.

Photomultiplier tube is sensitive and costly with respect to phototubes.

Spectral analysis in UV-VIS spectroscopy

UV-VIS spectrum is a graph that shows absorption at different wavelengths. The relative
maxima are known as lambda max (λmax). Spectra obtained by such techniques can be
useful for extracting information such as purity and composition.

UV-Vis spectroscopic spectra are frequently utilized to check the presence of different
organic and conjugated chemical species that have a wavelength in that range.

For example

Chlorophyll-a absorbs the regions of violet and orange wavelength regions. This is
the fact that makes plants unable to absorb green light making them appear green.
The below graph shows the absorbance activity of chlorophyll-a in visible light and
it proves the fact that it is most active in violet and orange regions.

Amoxicillin is a famous antibiotic that gives an identification peak at approximately

265nm on the wavelength scale.

The spectral analysis of uv-vis spectroscopy is also used for the quantitative analysis of
analytes. The absorbance at a certain wavelength of light is directly proportional to the
concentration by Beer-Lambert law.

Shifting of absorption band and change in intensity

Chromophores
The chromophore is an atom or group of atoms that are responsible for the
absorption of UV-visible radiation.

Types of chromophores
There are two types of chromophores:

1. Chromophores that can only contain π electrons. They undergo π-

π* transitions only e.g. ethylenic group (C＝C) and acetylenic group (C ≡ C)
etc.
2. Chromophores that contain π as well as n (nonbonding) electrons. This type
of chromophore contains lone pair(s) of electrons. So they are responsible for
two types of transitions i.e. n-π* and π-π* e.g. nitro group (-NO2 ), azo group (-
N＝N-), nitro group (-NO3 ), carbonyl group ( >C＝O), nitrite group (-ONO).

Auxochromes
An auxochrome is an atom or group of atoms that do not give rise to an absorption
band of its own but change the absorption characteristics of a chromophore.
Auxochromes may change both intensity and wavelength of chromophore when
added to it. It is also called a color-enhancing group. The replacement of hydrogen
on a basic chromophore changes its absorption characteristics. i.e. Methyl (-CH3,),
chloride (Cl–), hydroxyl (OH–), amino (-NH2), alkoxy (CH3O–), etc.

There exist four types of shifts corresponding to auxochromes:

1. Bathochromic shift (redshift)

2. Hypsochromic shift (blueshift)
3. Hyperchromic shift
4. Hypochromic shift

Bathochromic shift or redshift

The bathochromic shift is the change of position of the absorption band towards the
longer wavelength. This change occurs in the presence of auxochrome or change in
the solvent.

Hypsochromic shift or blue shift

The hypsochromic shift is the change in the position of the spectral band towards a
shorter wavelength. It occurs due to the removal of conjugation or change in
polarity of the solvent.

Hyperchromic shift
Hyperchromic shift is the increase in the intensity of the absorption band i.e. εmax. A
hyperchromic shift occurs due to the presence of an auxochrome.

Hypochromic shift
A hypochromic shift is a decrease in the intensity of the absorption i.e. εmax band.
Factors affecting change in the absorption and wavelength
These are the factors that result in changes in absorption and wavelength:

Effect of conjugation
According to MOT, as the number of pi electrons increases, delocalization increases.
Due to this increase in delocalization, the molecules of the sample get stabilized and
hence reach a state of lower energy. This lowering of energy causes a change in
wavelength towards a higher wavelength known as redshift.

The maximum absorption of 1,2-butadiene is at 210nm but for 1,3-butadiene is at

217nm due to conjugation of double bonds. This wavelength increases further to
260nm for 1,3,5-hexatriene because of conjugation at two points.

Effect of additive characteristics

When a molecule contains two or more chromophores separated by more than one
single bond the total absorption is equal to the sum of absorption characteristics of
each chromophore.

For example, ethylene and 1,5 hexadiene absorb the same wavelength. But the
amount of radiation absorbed for similar concentrations is almost doubled for 1,5
hexadiene than ethylene.

Effect of the aromatic ring

The aromatic ring especially when two or more rings in conjugation (polycyclic
compounds) absorbs a higher wavelength in the visible region, it alters the spectrum
of absorption.

For example

Naphthalene (C6H6 ) absorbs at 268nm

Anthracene absorbs at 311nm

Tetracene absorbs at 476nm

Effect of substitution of auxochrome

Benzene is a less effective chromophore, the substitution of a polar group to it
causes an increase in λmax in the visible region and hence εmax value increase.
 For example

Auxochrome Compound λ(max) ε(max)

--- Benzene 256 250

-NH2 Aniline 280 200

-Cl Chlorobenzene 265 360

Effect of the polarity of the solvent

Polarity causes a pronounced effect on the position and intensity of absorption
bands. This increase is due to the n-π* and π-π* transitions. In the presence of polar
hydrolytic solvent (i.e. water ) hydrogen bonds form with the lone pair of electrons
of auxochrome. As a result, the auxochrome’s energy lowers to an equal amount of
the bond formation energy, and hence the energy gap between HOMO and LUMO
increases, so, a hypsochromic shift is observed for n-π* transition.

Types of UV-Vis spectrometers

There are two types of UV-vis spectrophotometers

1. Single beam UV-Vis spectrometer

2. Double beam UV-Vis spectrometer

Single beam UV-Visible spectrophotometer

Single beam uv-vis spectrophotometer has a single beam as the name indicates. The
incident light coming from the source is passed through a monochromator then that
incident monochromatic light moves through a slit. Then it passes through the
sample solution. Where some of the incident light is absorbed by the sample while
other is transmitted. That transmitted light is detected by the detector. The detected
light is then amplified, recorded, and then displayed on a suitable readout device.
Spectrum is plotted and the λmax is located.

Single beam uv-vis spectrophotometer comprises of:

 Light source
 Lens
 Gratings
  Wavelength selector
 Sample container/cuvette
 Detector
 Digital meter/Recorder

Double beam UV-Visible spectrophotometer

The instrumentation of single and double beam spectrophotometers is almost the
same. The basic difference from a single beam UV-Vis spectrophotometer is that the
beam of incident light is passing simultaneously from the sample and the reference
cells.

The incident light splits and is directed towards both the reference and sample
cuvette. The refracted or transmitted beam is detected by the detectors. A double
beam UV-vis spectrophotometer needs two detectors that detect electron ratio to
measure or calculate absorbance in a test sample. It also requires a stabilized
voltage supply.

Applications of UV-Vis Spectroscopy

 UV-visible spectroscopy is used to identify organic and inorganic species
present in a solution.
 It is used to find the concentration of the unknown solution.
 For the determination of structure along with other data such as bands and
intensities of functional groups uv-visible spectroscopy is used.
 It is also used in the study of chemical kinetics i.e. disappearance of one
functional group and appearance of another functional group.
 UV-visible spectroscopy is used to study isomers, i.e. in geometric isomerism,
the trans-species absorb a high wavelength with a large molar absorptivity ‘ε’
value than the cis-species.
 It helps in the detection or presence of conjugation.
 UV-vis spectroscopy is used in clinics and hospitals for drug analysis.
 It is used in petrochemical industries.
 UV-visible spectroscopy is used in water quality control labs.
 It is used in forensic labs.
 Another important application of UV-visible spectroscopy is in the field of
chemical and biological plants.
 For qualitative analysis maximum absorption is used and for quantitative
analysis, Lambert beer’s law is used.
Concepts berg
What is a UV-Vis spectrophotometer?

Uv-vis spectrophotometer is an instrument in which the radiation from ultraviolet

and visible regions interacts with the sample solution. The electrons get excited
when absorbing some of the incident radiation and transmitting the other. This
transmitted radiation is detected, amplified, recorded, and displayed on a readout
device in the form of percent transmittance.

What information can be obtained from UV-Vis spectra?

UV-visible spectra explain the radiation absorbance region of a compound. When

UV-visible light strikes with the matter, some of the light is absorbed within the
molecules of the compound while the rest will be transmitted through it. This
transmitted light shows how much light is being absorbed by the sample (absent
from the refracted radiation).

What is the principle of UV visible spectroscopy?

The basic principle of UV-visible spectroscopy is based on the absorption of light

(ranges from -nm) by different chemical compounds. It is the interaction of
ultraviolet and visible light with matter. Every chemical compound has a particular
or distinct spectrum as it only absorbs a specific wavelength of light (radiation).

How is beer Lambert law used in spectroscopy?

Lambert beer’s law is used to determine the transmittance ‘T’ and absorbance ‘A’
of a solution in a transparent cell with path length ‘ℓ’. According to this law, the
concentration of the absorbing analyte is directly proportional to the absorbance
and path length of the cell.

A = – log T = log Po / P = ε . c . ℓ

What is UV visible spectroscopy used for?

UV-visible spectroscopy is used to identify functional groups, water analysis, and

measure an analyte’s concentration using Lambert Beer’s law. It is also used for
quantitative measurements of DNA, RNA, and proteins in the sample solution. UV
visible is widely used in forensic, chemicals, environmental, and clinical
laboratories. UV-visible spectrometer is used as a detector for various analytical
techniques i.e. chromatography.

What are the applications of UV-visible spectroscopy?

UV-visible spectroscopy has various applications in different fields, such as clinical

analysis, food industries, petrochemical industries, forensic laboratories, research,
educational laboratories, pharmaceutical industries, and quality control purposes. It
is also used to detect and identify different organic and inorganic species, functional
groups, isomeric studies, and the presence of conjugation, etc.

Why is quartz cuvette used in UV-visible spectroscopy?

Fused silica and quartz cuvettes are most commonly used in ultraviolet spectroscopy
as they are transparent in the ultraviolet region i.e. quartz can not absorb
ultraviolet light so are used in ultraviolet spectrophotometers. Plastic and glass
materials absorb ultraviolet light which interferes with the results.

Which detectors are used in UV visible spectroscopy?

The most commonly used detector in UV visible spectroscopy is a photomultiplier

tube. Repetition of the dynode is structured with a slight potential difference at a
particular angle. The incoming photon strikes the cathode, after knocking out
several electrons from the dynodes every time.

Which lamp is used in UV-vis spectroscopy?

Hydrogen & deuterium lamps (ranging 160nm-380nm) with a tungsten halogen

lamp (ranging 240nm-2500nm) are commonly used for UV-vis spectroscopy.

What is the range of UV Visible Spectroscopy?

The range of UV-vis is from 180nm to 780nm. The ultraviolet region comprises
180nm-400nm while the visible region extends from 400nm-780nm.

How do you read the UV-VIS spectroscopy spectrum?

To read the UV-vis spectrum the graph is plotted between the wavelength and the
absorption. The wavelength at which maximum absorption occurs is called the λmax.

What is maximum absorbance?

The characteristic wavelength at which a chemical species shows the maximum
absorption is called maximum absorption. It is represented by λmax.

What are chromophores?

Chromophores are atoms or groups of atoms that are responsible for the absorption
of incident radiation (UV-visible radiation mainly).

What is the application of UV visible spectroscopy in natural products?

Uv-vis spectroscopy is used in the analysis of many natural products. It is used to

identify organic and inorganic species present in a solution. It is also used for
quantitative measurements of different components in natural compounds, for
example, to find the concentration of vitamin C in orange juice, etc.

What are the advantages and disadvantages of UV visible spectroscopy?

UV-Vis spectroscopy gives accurate results. The instrument is easy to use and
handle. Besides this, it is a time-consuming technique, its preparation is difficult and
effort is required because external light and small vibration can cause interference
with results.

Why is methanol a good solvent for IR and UV spectroscopy?

Methanol is transparent in both ultraviolet and visible regions. It has a cutoff point
below these two regions hence it cannot cause any interference in results. So, it is a
good solvent for UV and IR spectroscopies.

Why are most absorption bands in the visible UV spectra very broad?

Many electronic transitions occur by various molecules, each of which slightly

differs from one another. A large number of closely spaced lines are difficult for the
instrument to detect or resolve individually hence it gives a broad spectrum.

What is the use of a UV-Vis spectrophotometer?

UV-Vis spectrophotometer is used for the quantitative determination and

measurement of different analytes i.e. organic, inorganic, biological molecules, etc.

Why the UV spectrum is broader than the IR spectrum?

In the ultraviolet spectrum, changes in all the energy levels (i.e. rotational,
vibrational, and electronic energy levels) are observed while in IR only vibrational
energy levels are observed. So, a UV spectrum is broader than IR one.

What is the difference between an absorbance, emission, and excitation spectrum

for UV visible spectroscopy?

The main difference between absorbance, excitation, and the emission spectrum is
the type of radiation understudy to analyze the respective chemical compounds. In
absorption spectra, the light being absorbed is studied with transmitted light. That
transmitted light shows the absorbed or missing light as the difference in the blank
and sample transmittance. While in the emission spectrum, such as fluorescence
spectrum the fluorescence intensity is taken as a function of excitation wavelength.
The energy that is first absorbed by the electron of an atom is emitted back. As the
energy is quantized so electrons that excite, gain a certain amount of energy and
release the same when going back to their original state.

Why is ethanol a good solvent for UV measurement but not for IR?

Ethanol is good for UV measurement because its cutoff point is 210 nm which is
near the lower limits of UV (190 nm- 400nm) so it does not interfere with the results
of ultraviolet regions. Whereas, ethanol has a dipole moment, which means it is IR
active, so it interferes with the results.

What is the difference between colorimetry and spectrophotometry?

Colorimetry is only used for colored compounds while spectrophotometry is used

for various compounds either colored or colorless. Spectrophotometry comprises a
wide range of wavelengths i.e ultraviolet, visible, and IR.

Why UV absorption value goes negative?

The negative value of absorption indicates that the sample is having an impurity in
it, which causes interference with the result. The fluorescence caused by the
impurity can enhance the value of transmitted radiation as compared to incident
radiation. That is the reason it gives a negative absorption value.

Why are solid forms of the sample not suitable for UV vis spectroscopy?
For UV-vis spectroscopy, the analyte must be in solution form because the
interaction of radiation is effective this way. The light interacts with all the
molecules of the analyte in solution form and there are very low chances for losses
as in solid form.

What is the difference between zero and baseline in UV Vis spectroscopy?

The zero in UV spectroscopy indicates the total transmittance while baseline is the
amount of radiation absorbed by the cuvette and the sample solution.