MCQs on UV-Visible Spectrophotometry

1. What is the principle of UV-Visible spectrophotometry?

A. Absorption of light by a molecule depends on its atomic weight.

B. Absorption of light by a molecule occurs when its energy matches the energy of a photon.
C. Absorption of light by a molecule is proportional to its polarity.
D. Absorption of light by a molecule occurs when its energy is lower than the photon's
energy.

Answer: B
Explanation: UV-Visible spectrophotometry is based on the principle that molecules absorb
light at specific wavelengths corresponding to electronic transitions. The absorbed energy
must match the energy of a photon for this to occur.

2. Which electronic transitions are most commonly observed in UV-Visible

spectroscopy?

A. σ→π∗
B. π→π∗
C. n→σ∗
D. n→n

Answer: B
Explanation: The π→π∗ transitions occur in compounds with conjugated systems, and they
fall within the UV-Visible range, making them the most common transitions observed in this
technique.

3. What is the role of the monochromator in a UV-Visible spectrophotometer?

A. Detects the absorbance of light.

B. Splits light into individual wavelengths.
C. Amplifies the signal from the detector.
D. Transmits all wavelengths of light simultaneously.

Answer: B
Explanation: The monochromator splits the light from the source into its component
wavelengths and selects the desired wavelength for measurement.

4. Which detector is commonly used in modern UV-Visible spectrophotometers?

A. Thermocouple
B. Photomultiplier tube (PMT)
C. Bolometer
D. Flame ionization detector

Answer: B
Explanation: Photomultiplier tubes are highly sensitive and capable of detecting low light
intensities, making them suitable for UV-Visible spectrophotometry.

5. According to Beer-Lambert's Law, absorbance (A) is directly proportional to:

A. Wavelength and concentration.

B. Molar absorptivity, path length, and concentration.
C. Path length and wavelength.
D. Molar absorptivity and wavelength.

Answer: B
Explanation: Beer-Lambert's Law states that A=ϵ⋅b⋅cA = \epsilon \cdot b \cdot c, where ϵ\
epsilon is molar absorptivity, bb is the path length, and cc is the concentration.

6. Which of the following compounds will show absorbance in the UV region?

A. Benzene
B. Water
C. Sodium chloride
D. Ethanol

Answer: A
Explanation: Benzene has a conjugated π\pi-electron system, which undergoes π→π∗\pi \
rightarrow \pi^* transitions, absorbing light in the UV region.

7. What is a limitation of UV-Visible spectrophotometry?

A. It cannot measure colored compounds.

B. It requires sophisticated sample preparation.
C. It provides limited structural information about compounds.
D. It is unable to measure high absorbance values.

Answer: C
Explanation: UV-Visible spectrophotometry provides information about electronic
transitions but does not give detailed structural data like IR or NMR spectroscopy.

8. Which of the following is an application of UV-Visible spectrophotometry?

A. Determining molecular weight.
B. Quantifying DNA and proteins.
C. Measuring ionization energy.
D. Determining bond angles.

Answer: B
Explanation: UV-Visible spectrophotometry is widely used to quantify biomolecules like
DNA and proteins by measuring their absorbance at characteristic wavelengths (e.g., 260 nm
for DNA).

9. What happens when a sample exhibits turbidity during a UV-Visible

spectrophotometric analysis?

A. Absorbance increases accurately.

B. Scattering of light occurs, leading to errors.
C. Light intensity remains unchanged.
D. Absorbance decreases, and accuracy improves.

Answer: B
Explanation: Turbidity scatters light, causing errors in absorbance measurements, as the
spectrophotometer assumes no scattering occurs.

10. Which factor does NOT influence UV-Visible absorbance?

A. Concentration of the sample

B. Temperature
C. Detector material
D. Wavelength of light

Answer: C
Explanation: The detector material does not influence absorbance directly; absorbance
depends on sample concentration, temperature, and wavelength.

Certainly! Here are more MCQs on UV-Visible Spectrophotometry, keeping in mind the
style and level of questions that might appear in Graduate Pharmacy Aptitude Test (GPAT)
exams:

11. In UV-Visible spectrophotometry, the absorption maximum (λmax) of a

compound typically corresponds to:

A. The lowest energy transition

B. The highest energy transition
C. The highest wavelength transition
D. The lowest wavelength transition
Answer: A
Explanation: The absorption maximum (λmax) is where a compound absorbs light most
strongly, usually corresponding to the lowest energy transition.

12. The Beer-Lambert Law relates which of the following parameters?

A. Wavelength, refractive index, and concentration

B. Absorbance, path length, and concentration
C. Absorbance, pH, and temperature
D. Wavelength, absorbance, and temperature

Answer: B
Explanation: The Beer-Lambert Law states that absorbance is directly proportional to the
concentration of the solute and the path length of the light through the solution.

13. Which of the following is a key advantage of UV-Visible

spectrophotometry in pharmaceutical analysis?

A. It provides detailed molecular structures.

B. It allows for the simultaneous measurement of multiple components.
C. It requires large sample volumes.
D. It can only be used for liquid samples.

Answer: B
Explanation: UV-Visible spectrophotometry is highly sensitive and allows for the
simultaneous measurement of multiple components in a sample, particularly when they
absorb at different wavelengths.

14. The molar absorptivity (ε) in Beer-Lambert's Law is a constant that

depends on:

A. The wavelength of light used

B. The temperature of the solution
C. The pH of the solution
D. The solvent used

Answer: A
Explanation: Molar absorptivity (ε) is a constant that depends on the wavelength of light, as
different wavelengths are absorbed differently by molecules.

15. In a UV-Visible spectrophotometer, the purpose of a reference blank is to:

A. Correct for the absorbance of the sample
B. Measure the absorbance of the solvent or sample without the analyte
C. Provide a standard to compare absorbance data
D. Calibrate the detector

Answer: B
Explanation: A reference blank is used to zero the spectrophotometer by measuring the
absorbance of the solvent (or the sample without the analyte), ensuring that any absorbance
detected is solely due to the analyte.

16. Which type of light source is typically used in UV-Visible

spectrophotometry?

A. Incandescent bulb
B. Deuterium lamp
C. LED
D. Halogen lamp

Answer: B
Explanation: A deuterium lamp is typically used in the UV range, while tungsten lamps are
often used for the visible range. Both are essential for covering the UV-Visible spectrum.

17. Which of the following is a disadvantage of UV-Visible

spectrophotometry?

A. It is a non-destructive technique.
B. It cannot be used for solid samples.
C. It requires expensive instrumentation.
D. It provides detailed molecular structures.

Answer: B
Explanation: UV-Visible spectrophotometry is generally used for liquid samples and may
require additional sample preparation for solids (e.g., dissolving in a solvent).

18. Which of the following is the primary application of UV-Visible

spectrophotometry in pharmaceuticals?

A. Quantification of active pharmaceutical ingredients (APIs)

B. Determination of molecular weight of APIs
C. Elucidation of 3D molecular structures
D. Monitoring of chemical reactions in real-time
Answer: A
Explanation: UV-Visible spectrophotometry is widely used in pharmaceutical analysis for
quantifying active pharmaceutical ingredients (APIs) by measuring their absorbance at
specific wavelengths.

19. A sample absorbs at 280 nm. The sample most likely contains:

A. DNA
B. Proteins
C. Both A and B
D. None of the above

Answer: C
Explanation: Both DNA and proteins absorb strongly at 280 nm, as aromatic amino acids
(like tryptophan) and nucleic acids have absorption maxima in this region.

20. The primary disadvantage of using a photodiode array (PDA) detector in

UV-Visible spectroscopy is:

A. Limited spectral range

B. High cost and complexity
C. Slow response time
D. Low sensitivity

Answer: B
Explanation: Photodiode array detectors (PDA) are more expensive than traditional detectors
and require more complex electronics, but they provide rapid, simultaneous multi-wavelength
measurements.

21. In a UV-Visible spectrophotometer, the monochromator’s primary role is

to:

A. Increase the intensity of the light beam.

B. Filter out stray light.
C. Separate the light into its constituent wavelengths.
D. Detect the absorbance.

Answer: C
Explanation: The monochromator separates the light into individual wavelengths and selects
the appropriate wavelength to pass through the sample.
22. The relationship between absorbance and concentration in UV-Visible
spectrophotometry is:

A. Exponential
B. Linear, according to Beer-Lambert's law
C. Inversely proportional
D. Logarithmic

Answer: B
Explanation: Beer-Lambert’s Law states that absorbance is directly proportional to
concentration, allowing for the determination of concentration from absorbance data.

23. What is the wavelength range of UV light typically used in UV-Visible

spectrophotometry?

A. 100 - 400 nm
B. 400 - 700 nm
C. 700 - 1000 nm
D. 10 - 200 nm

Answer: A
Explanation: The UV range in UV-Visible spectrophotometry typically covers wavelengths
from 100 nm to 400 nm, while the visible range is from 400 nm to 700 nm.

24. A higher molar absorptivity (ε) value indicates:

A. A molecule absorbs less light at a given concentration.

B. A molecule absorbs more light at a given concentration.
C. The molecule is colorless in the UV-Visible range.
D. The molecule absorbs at a longer wavelength.

Answer: B
Explanation: A higher molar absorptivity indicates a greater ability to absorb light at a given
wavelength, meaning the molecule is more efficient at absorbing light.

Here are more problem-oriented MCQs based on UV/Visible Spectrophotometry to help

with understanding practical applications, data interpretation, and problem-solving. These
questions are designed to test knowledge on common issues and troubleshooting scenarios
that may arise in the lab.
25. A sample of an unknown drug solution shows absorbance at 280 nm when
measured using UV-Visible spectrophotometry. The concentration of the drug
solution is calculated to be 0.5 mg/mL. What will be the concentration of the
solution if the absorbance is doubled, assuming Beer-Lambert’s Law is
followed?

A. 1 mg/mL
B. 0.25 mg/mL
C. 0.5 mg/mL
D. 2 mg/mL

Answer: A
Explanation: According to Beer-Lambert’s Law, absorbance is directly proportional to
concentration. If the absorbance is doubled, the concentration will also double. Therefore, the
new concentration will be 1 mg/mL.

26. In a UV-Visible spectrophotometry experiment, the blank solution absorbs

at 300 nm, but the sample does not. Which of the following is the most likely
cause of this?

A. The sample has a chromophore that absorbs at 300 nm.

B. The solvent used in the blank absorbs at 300 nm.
C. The detector is faulty.
D. The lamp is malfunctioning.

Answer: B
Explanation: The blank should contain only the solvent, which is used to account for any
absorbance by the solvent or container. If the blank absorbs at 300 nm, it suggests that the
solvent itself absorbs at that wavelength, causing an error in the readings.

27. A UV-Visible spectrophotometer is calibrated using a standard solution of

a compound with known absorbance at a specific wavelength. After
calibration, the same solution is measured, but the absorbance is lower than
expected. Which of the following could explain this discrepancy?

A. The standard solution was diluted.

B. The wavelength of the spectrophotometer was changed.
C. The lamp is not operating at full power.
D. The cuvette was not cleaned properly.

Answer: C
Explanation: A decrease in absorbance could occur if the light source is not operating at full
power, leading to a weaker signal. This can cause lower readings than expected.
28. A drug is dissolved in ethanol, and the absorption spectrum is recorded.
The UV-Visible spectrum shows a peak at 230 nm. However, when the same
solution is measured in a different solvent (water), the peak shifts to 250 nm.
What is the likely reason for this shift?

A. The ethanol solvent absorbs at 230 nm, masking the peak.

B. The drug undergoes a chemical change in water, shifting its absorbance.
C. The polarity of the solvents affects the electronic transitions of the drug.
D. The path length in water is longer than in ethanol.

Answer: C
Explanation: The shift in absorbance is likely due to the change in polarity between the
solvents, which can affect the electronic structure of the drug, thereby altering its absorption
wavelength.

29. A solution of a compound has an absorbance of 0.75 at 350 nm. The

concentration of the compound is 1 mg/mL. If the path length is halved, what
will be the new absorbance assuming Beer-Lambert's Law holds?

A. 0.75
B. 0.375
C. 1.5
D. 0.50

Answer: B
Explanation: According to Beer-Lambert's Law, absorbance is directly proportional to the
path length. If the path length is halved, the absorbance will also be halved, giving a new
absorbance of 0.375.

30. In an experiment, the absorbance of a sample solution was measured at

280 nm, and the following data were obtained: Absorbance = 0.5, Molar
absorptivity (ε) = 1.2 L·mg⁻¹·cm⁻¹, and the path length (b) = 1 cm. What is
the concentration (in mg/mL) of the sample?

A. 0.42 mg/mL
B. 0.55 mg/mL
C. 0.38 mg/mL
D. 0.67 mg/mL

Answer: A
Explanation: Using Beer-Lambert's Law, A=ϵ⋅b⋅cA = \epsilon \cdot b \cdot c, where:

 A=0.5A = 0.5
  ϵ=1.2 L\cdotpmg−1⋅cm−1\epsilon = 1.2 \, \text{L·mg}⁻¹·\text{cm}⁻¹
 b=1 cmb = 1 \, \text{cm}

Solving for cc (concentration):

c=Aϵ⋅b=0.51.2⋅1=0.42 mg/mLc = \frac{A}{\epsilon \cdot b} = \frac{0.5}{1.2 \cdot 1} = 0.42

\, \text{mg/mL}

31. During a UV-Visible spectrophotometric measurement, the sample

absorbance is very high (>2.0), resulting in saturation. What should be done
to correct this issue?

A. Increase the sample concentration.

B. Decrease the path length of the cuvette.
C. Increase the wavelength of measurement.
D. Use a more sensitive detector.

Answer: B
Explanation: When the absorbance is too high, it can cause saturation. To correct this, the
path length of the cuvette should be decreased, as absorbance is directly proportional to path
length. This will reduce the signal and avoid saturation.

32. In a UV-Visible spectrophotometry experiment, the absorption of a

compound is measured at 350 nm, but the measured absorbance is higher
than expected. What could be the most likely cause?

A. The spectrophotometer is not calibrated.

B. The solvent has absorbance at 350 nm.
C. The compound does not absorb at 350 nm.
D. The cuvette is scratched.

Answer: B
Explanation: If the solvent has absorbance at 350 nm, it can contribute to the total
absorbance and give a higher reading than expected. The sample should be measured against
a blank solvent to account for this.

33. If a sample shows a distinct peak at 400 nm in a UV-Visible spectrum, but

a second peak is also observed at 420 nm, what does this most likely indicate?

A. The compound has multiple chromophores.

B. The sample contains impurities absorbing at 420 nm.
C. The solution concentration is too high.
D. The spectrophotometer is faulty.
Answer: A
Explanation: A distinct peak at 400 nm and another at 420 nm suggests that the compound
contains multiple chromophores that absorb at different wavelengths, resulting in multiple
peaks.

34. During a UV-Visible spectrophotometric experiment, you measure the

absorbance of a sample and get a value of 0.35 at 310 nm. If you want to
increase the sensitivity of the measurement, what should you do?

A. Increase the concentration of the sample.

B. Decrease the wavelength of measurement.
C. Increase the path length of the cuvette.
D. Use a different solvent.

Answer: C
Explanation: To increase the sensitivity of the measurement, you can increase the path
length, as absorbance is directly proportional to path length. This will increase the signal for
the same concentration of the sample.

Here are more problem-oriented MCQs on UV/Visible Spectrophotometry with detailed

explanations:

35. A UV-Visible spectrophotometer measures an absorbance of 0.80 at 310

nm for a solution of unknown concentration. The molar absorptivity (ε) of the
compound at 310 nm is 1.5 × 10⁴ L·mol⁻¹·cm⁻¹, and the path length of the
cuvette is 1 cm. What is the concentration of the compound in the solution?

A. 5.33 × 10⁻⁵ M
B. 5.33 × 10⁻⁴ M
C. 8.33 × 10⁻⁵ M
D. 1.33 × 10⁻⁵ M

Answer: A
Explanation: Using Beer-Lambert's Law, A=ϵ⋅b⋅cA = \epsilon \cdot b \cdot c, where:

 A=0.80A = 0.80
 ϵ=1.5×104 L\cdotpmol−1⋅cm−1\epsilon = 1.5 \times 10^4 \, \text{L·mol}⁻¹·\
text{cm}⁻¹
 b=1 cmb = 1 \, \text{cm}

Solving for concentration cc:

c=Aϵ⋅b=0.80(1.5×104)⋅1=5.33×10−5 Mc = \frac{A}{\epsilon \cdot b} = \frac{0.80}{(1.5 \
times 10^4) \cdot 1} = 5.33 \times 10^{-5} \, \text{M}

36. The absorbance of a solution was measured at 280 nm, and the value
obtained was 1.5. If the sample has a concentration of 0.2 mg/mL and the path
length is 1 cm, what is the molar absorptivity (ε) of the compound at 280 nm?

A. 7500 L·mol⁻¹·cm⁻¹
B. 750 L·mg⁻¹·cm⁻¹
C. 0.75 L·mg⁻¹·cm⁻¹
D. 0.75 L·mol⁻¹·cm⁻¹

Answer: B
Explanation: Using Beer-Lambert’s Law A=ϵ⋅b⋅cA = \epsilon \cdot b \cdot c, we need to
rearrange it to solve for ϵ\epsilon:

ϵ=Ab⋅c=1.51⋅0.2=7.5 L\cdotpmg−1⋅cm−1\epsilon = \frac{A}{b \cdot c} = \frac{1.5}{1 \cdot

0.2} = 7.5 \, \text{L·mg}⁻¹·\text{cm}⁻¹

37. You are performing UV-Visible spectrophotometry of a drug solution, and

the reading shows a very low absorbance at the specified wavelength. Which
of the following could be the most likely cause of this?

A. The solvent absorbs at the wavelength.

B. The concentration of the drug is too high.
C. The sample is not properly mixed.
D. The cuvette path length is too short.

Answer: C
Explanation: If the solution is not properly mixed, it could result in an uneven distribution of
the sample, leading to a low or inaccurate absorbance measurement.

38. During a UV-Visible spectrophotometric measurement of a solution, you

observe a slight deviation from Beer-Lambert’s Law at higher concentrations.
What is the likely cause of this deviation?

A. The instrument is not properly calibrated.

B. The sample absorbs too strongly at the measured wavelength.
C. The path length is too short.
D. The cuvette is scratched.

Answer: B
Explanation: At high concentrations, the solution may exhibit non-linear behavior, causing
deviations from Beer-Lambert’s Law. This is because at high absorbance, scattering and
interactions between molecules can occur, leading to inaccuracies.
39. In UV-Visible spectrophotometry, why is it necessary to use a blank
sample during measurements?

A. To adjust for any light scattering from the solution.

B. To calibrate the spectrophotometer and eliminate the absorbance of the solvent and
cuvette.
C. To ensure that the sample does not undergo any chemical reactions during measurement.
D. To ensure that the path length of the cuvette is consistent.

Answer: B
Explanation: A blank sample, typically consisting of the solvent, is used to zero the
spectrophotometer. This ensures that any absorbance due to the solvent or cuvette is
accounted for, providing accurate readings for the sample.

40. A researcher uses a UV-Visible spectrophotometer to measure a solution

of a compound, but the absorption peak is shifted to a longer wavelength than
expected. What could be the cause of this shift?

A. The solvent used for the measurement is non-ideal.

B. The sample concentration is too low.
C. The path length of the cuvette is too long.
D. The sample is undergoing a chemical transformation.

Answer: A
Explanation: A solvent with a different polarity than the one used in the original
measurement can cause a shift in the absorption peak, as the electronic structure of the
compound may be affected by the solvent.

41. A solution of a drug is analyzed using UV-Visible spectrophotometry, and

the absorbance at 250 nm is measured to be 0.85. The molar absorptivity (ε) of
the compound is 5 × 10³ L·mol⁻¹·cm⁻¹, and the cuvette path length is 1 cm.
What is the concentration of the drug in the solution?

A. 1.7 × 10⁻⁴ M
B. 1.7 × 10⁻³ M
C. 1.7 × 10⁻² M
D. 1.7 M

Answer: A
Explanation: Using Beer-Lambert’s Law A=ϵ⋅b⋅cA = \epsilon \cdot b \cdot c, we can solve
for cc (concentration):
c=Aϵ⋅b=0.85(5×103)⋅1=1.7×10−4 Mc = \frac{A}{\epsilon \cdot b} = \frac{0.85}{(5 \times
10^3) \cdot 1} = 1.7 \times 10^{-4} \, \text{M}

42. During a UV-Visible spectrophotometric analysis, the absorbance of a

drug solution at 350 nm is measured to be 1.3. If the compound’s molar
absorptivity is 2.5 × 10⁴ L·mol⁻¹·cm⁻¹ and the cuvette has a path length of 1
cm, what is the concentration of the solution in mol/L?

A. 5.2 × 10⁻⁵ M
B. 5.2 × 10⁻⁴ M
C. 1.3 × 10⁻⁵ M
D. 1.3 × 10⁻⁴ M

Answer: A
Explanation: Using Beer-Lambert’s Law, solve for concentration cc:

c=Aϵ⋅b=1.3(2.5×104)⋅1=5.2×10−5 Mc = \frac{A}{\epsilon \cdot b} = \frac{1.3}{(2.5 \times

10^4) \cdot 1} = 5.2 \times 10^{-5} \, \text{M}

43. A compound exhibits an absorption peak at 330 nm, but when the same
solution is measured at 400 nm, the absorbance is found to be zero. Which of
the following statements is most likely true?

A. The compound absorbs strongly at 400 nm.

B. The compound does not absorb at 400 nm.
C. The spectrophotometer is malfunctioning at 400 nm.
D. The solvent absorbs at 400 nm.

Answer: B
Explanation: If the absorbance is zero at 400 nm, it indicates that the compound does not
absorb at this wavelength, which is why no signal is detected at that point.