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Electronics and Communication Engineering (ECE) Exam  >  Electronics and Communication Engineering (ECE) Notes  >  Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering

Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering - Electronics and Communication Engineering (ECE) PDF Download

Basic Principles of HPLC
Martin R. Hackman
NJ- DEP
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Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering - Electronics and Communication Engineering (ECE)

 

Basic Principles of HPLC----------------------------------------- Next Slide------------------------------------Martin R. Hackman

H igh
P erformance
L iquid
C hromatography

 

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H igh
P ressure
L iquid
C hromatography

 

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H igh
P riced
L iquid
C hromatography

 

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HPLC Methods

Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering - Electronics and Communication Engineering (ECE)

 

 

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Compounds

Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering - Electronics and Communication Engineering (ECE)

 

 

Basic Principles of HPLC----------------------------------------- Next Slide------------------------------------Martin R. Hackman

 

Compounds

 

Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering - Electronics and Communication Engineering (ECE)

Basic Principles of HPLC----------------------------------------- Next Slide------------------------------------Martin R. Hackman

 

 

Basic Principles of HPLC----------------------------------------- Next Slide------------------------------------Martin R. Hackman

Partitioning

•Separation is based on the analyte’s relative solubility between two liquid phases

 

Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering - Electronics and Communication Engineering (ECE)

Basic Principles of HPLC----------------------------------------- Next Slide------------------------------------Martin R. Hackman

HPLC - Modes

•Normal Phase. - Polar stationary phase and non-polar solvent.
• Reverse Phase. - Non-polar stationary phase and a polar solvent.

 

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Common Reverse Phase Solvents

Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering - Electronics and Communication Engineering (ECE)

 

Basic Principles of HPLC----------------------------------------- Next Slide------------------------------------Martin R. Hackman

Columns

• Analytical - Performs the separation.
•Solid Support - Backbone for bonded phases.
–Usually 10μ, 5μ or 3μ silica or polymeric particles.
•Bonded Phases - Functional groups firmly linked (chemically bound) to the solid support.
–Extremely stable
–Reproducible
•Guard - Protects the analytical column:
–Particles
–Interferences
–Prolongs the life of the analytical column

 

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Bonded Phases

 

Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering - Electronics and Communication Engineering (ECE)

 

Basic Principles of HPLC----------------------------------------- Next Slide------------------------------------Martin R. Hackman

 

Instrumentation

 Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering - Electronics and Communication Engineering (ECE)

 

Basic Principles of HPLC----------------------------------------- Next Slide------------------------------------Martin R. Hackman

Detectors

•UV
–Single wavelength (filter) [610, 8330]
–Variable wavelength (monochromator) [8316, 8325]
–Multiple wavelengths (PDA) [555]
•Fluorescence [610]
•Electrochemical [605]
•Mass Spectrometric [8325]

 

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Chromatograms

 

Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering - Electronics and Communication Engineering (ECE) Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering - Electronics and Communication Engineering (ECE)

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FAQs on Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering - Electronics and Communication Engineering (ECE)

1. What are the basic principles of HPLC?
Ans. High Performance Liquid Chromatography (HPLC) is based on the principles of liquid chromatography, which involve the separation of components in a mixture based on their interactions with a stationary phase and a mobile phase. In HPLC, a sample is injected into a column filled with a stationary phase, and a solvent (mobile phase) is pumped through the column. The components in the sample interact differently with the stationary phase, leading to their separation and subsequent detection.
2. What is the significance of the stationary phase in HPLC?
Ans. The stationary phase in HPLC plays a crucial role in the separation process. It is a solid or liquid material that is packed into the column and provides sites for interaction with the components of the sample. The choice of stationary phase depends on the specific separation requirements, such as the nature of the sample and the desired selectivity. Different stationary phases offer different interactions, such as reversed-phase, normal-phase, ion-exchange, and size-exclusion, allowing for a wide range of separations.
3. How does the mobile phase affect the separation in HPLC?
Ans. The mobile phase in HPLC is a solvent or mixture of solvents that carries the sample through the column. It plays a critical role in the separation process by transporting the components of the sample and facilitating their interactions with the stationary phase. The composition of the mobile phase, including the type of solvent and its concentration, can be adjusted to optimize the separation. Factors such as polarity, pH, and viscosity of the mobile phase influence the separation efficiency and selectivity.
4. What are the different detection techniques used in HPLC?
Ans. HPLC utilizes various detection techniques to quantify and identify the separated components. Some commonly used detection techniques include UV-Visible spectroscopy, fluorescence spectroscopy, electrochemical detection, and mass spectrometry. UV-Visible spectroscopy is widely used due to its simplicity, sensitivity, and compatibility with many compounds. Fluorescence spectroscopy offers higher sensitivity and selectivity for compounds that exhibit fluorescence. Electrochemical detection is suitable for electroactive compounds, while mass spectrometry provides high accuracy and structural information.
5. How is HPLC applied in the field of electronics and communication engineering?
Ans. HPLC finds applications in the field of electronics and communication engineering, particularly in the analysis of electronic components and materials. It can be used to analyze the purity and composition of electronic materials, such as semiconductors, polymers, and conductive coatings. HPLC is instrumental in quality control and failure analysis of electronic components, ensuring their reliability and performance. It can also be employed for the analysis of contaminants or impurities in electronic devices, contributing to the overall assessment of their safety and functionality.
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Dec 14, 2024 Last updated
Document Description: Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering for Electronics and Communication Engineering (ECE) 2024 is part of Electronics and Communication Engineering (ECE) preparation. The notes and questions for Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering have been prepared according to the Electronics and Communication Engineering (ECE) exam syllabus. Information about Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering covers topics like and Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering Example, for Electronics and Communication Engineering (ECE) 2024 Exam. Find important definitions, questions, notes, meanings, examples, exercises and tests below for Chapter : Basic Principles of HPLC, PPT, Chemistry, Semester, Engineering.
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