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Instrumentation_Answers

The document provides an overview of various analytical techniques including UV-Visible Absorption Spectrophotometry, Spectrofluorometry, Flame Photometry, and Nephelometry. Each technique is described in terms of its principle, instrumentation, working mechanism, applications, advantages, and limitations. These methods are essential tools in fields such as chemistry, biology, and environmental monitoring.

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Sonali Sri
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8 views4 pages

Instrumentation_Answers

The document provides an overview of various analytical techniques including UV-Visible Absorption Spectrophotometry, Spectrofluorometry, Flame Photometry, and Nephelometry. Each technique is described in terms of its principle, instrumentation, working mechanism, applications, advantages, and limitations. These methods are essential tools in fields such as chemistry, biology, and environmental monitoring.

Uploaded by

Sonali Sri
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Instrumentation Answers for Mid-Semester Exam

UV-Visible Absorption Spectrophotometry

Introduction:
UV-Visible spectrophotometry measures the absorption of UV and visible light by a substance based on
Beer-Lambert?s law (A = ?cl).

Principle:
- Molecules absorb light at specific wavelengths.
- Absorbance follows Beer-Lambert?s law.

Instrumentation:
- Light source: Deuterium & Tungsten lamps
- Monochromator: Selects specific wavelengths
- Detector: Photodiode or photomultiplier tube

Working Mechanism:
1. Light passes through a monochromator.
2. Selected wavelength reaches the sample.
3. Detector measures absorbance.

Applications:
- Pharmaceutical analysis
- Water quality testing
- Food industry

Advantages & Limitations:


Advantages: Simple, rapid, and accurate; Non-destructive.
Limitations: Interference from other absorbing species; Requires clear solutions.

Conclusion:
UV-Visible spectrophotometry is a fundamental analytical tool in chemistry and biology.

Spectrofluorometry

Introduction:
Spectrofluorometry measures fluorescence emitted by a substance after excitation by light.

Principle:
- Excited molecules emit fluorescence.
- Intensity is proportional to concentration.

Instrumentation:
- Excitation source: Xenon/mercury lamp
- Detector: Photomultiplier tube

Working Mechanism:
1. Sample absorbs UV light.
2. Fluorescence is emitted.
3. Detector measures emitted light.

Applications:
- Biomedical research
- Forensic science
- Environmental monitoring

Advantages & Limitations:


Advantages: Highly sensitive; Selective for fluorescent compounds.
Limitations: Quenching effects reduce accuracy; Background fluorescence interference.

Conclusion:
Spectrofluorometry is a powerful tool for detecting fluorescent compounds.

Flame Photometry

Introduction:
Flame photometry analyzes alkali and alkaline earth metals.

Principle:
- Sample in flame emits light.
- Emission intensity is measured.

Instrumentation:
- Flame source: Air-acetylene or LPG
- Detector: Measures light intensity

Working Mechanism:
1. Sample introduced into flame.
2. Atoms emit characteristic light.
3. Emission intensity is recorded.

Applications:
- Clinical analysis
- Agriculture
- Food industry

Advantages & Limitations:


Advantages: Simple & cost-effective; Rapid & selective.
Limitations: Detects only specific metals; Ion interference possible.

Conclusion:
Flame photometry is essential for metal ion analysis.

Nephelometry

Introduction:
Nephelometry measures light scattering by particles.

Principle:
- Scattered light is measured at an angle.
- Intensity is proportional to particle concentration.

Instrumentation:
- Light source: LED or laser
- Detector: Measures scattered light

Working Mechanism:
1. Light passes through suspension.
2. Particles scatter light.
3. Detector measures intensity.

Applications:
- Clinical testing
- Environmental monitoring
- Industrial quality control

Advantages & Limitations:


Advantages: High sensitivity; Rapid analysis.
Limitations: Cannot analyze clear solutions; Requires calibration.

Conclusion:
Nephelometry is useful in medical and environmental applications.

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