Key Points on Chromatography Techniques and Their Principles

Chromatography is a powerful analytical technique used to separate and analyze complex mixtures. This post will provide an overview of some common chromatography methods, their principles, steps, uses and examples.

What is Chromatography?

Chromatography is a laboratory technique in which a mixture is separated into its individual components. It relies on the differential distribution of the sample components between a moving fluid mobile phase and a stationary phase to achieve separation.

Chromatography Definition:

Chromatography is a laboratory technique for the separation of a mixture into its constituent parts. 

Stationary Phase: The stationary phase is the immobile phase fixed in place in a chromatography column. It can be a solid or liquid.

Mobile Phase: The mobile phase is the solvent that moves through the chromatography column carrying the sample. 

1. Affinity Chromatography

- Principle: Based on specific biological interactions between antibody and antigen, enzyme and substrate, etc.

- Steps: The sample is applied, target binds to stationary phase, impurities are washed away, target is eluted.

- Uses: Purification of biomolecules like proteins.  

- Example: Purification of IgG antibodies using Protein A affinity column.

2. Anion Exchange Chromatography 

- Principle: Based on interaction between positively charged stationary phase and negatively charged sample ions.

- Steps: The sample is applied, anions bind to positively charged sites, impurities are washed away, anions are eluted by increasing salt concentration or changing pH.

- Uses: Separation of anions and polar molecules.

- Example: Separation of proteins, nucleic acids, carbohydrates.

3. Cation Exchange Chromatography

- Principle: Based on interaction between negatively charged stationary phase and positively charged sample ions. 

- Steps: The sample is applied, cations bind to negatively charged sites, impurities are washed away, cations are eluted by increasing salt concentration or changing pH.

- Uses: Separation of cations and polar molecules.

- Example: Separation of proteins, peptides, amines.

4. Column Chromatography

- Principle: Based on differential partitioning between stationary and mobile phase.

- Steps: The sample is applied, components separate as they travel down the column at different rates, fractions are collected.

- Uses: Analytical and preparative separation and purification of chemicals.

- Examples: Separation of plant pigments, lipids, drugs, etc.

5. Flash Chromatography 

- Principle: A faster version of column chromatography by using pressurized gas to push the mobile phase through a short column.

- Steps: Sample is loaded, pressure pushes mobile phase, components separate quickly, fractions are collected.

- Uses: Quick analytical and preparative separation of organic compounds.

6. Gas Chromatography

- Principle: Based on partitioning between mobile gaseous phase and stationary liquid or solid phase.

- Steps: Sample is vaporized, carried by inert gas through the column, separates based on affinity for stationary phase, detected.

- Uses: Separate and analyze volatile mixtures.  

- Examples: Analyze essential oils, detect air pollutants, etc.

7. Gel Filtration Chromatography

- Principle: Based on size-exclusion separation technique. Larger molecules cannot enter pores and elute first.

- Steps: Sample loaded, molecules separate based on size as they pass through column, smaller molecules elute later.

- Uses: Separate proteins and other biomolecules based on size.

- Example: Fractionate proteins and estimate their molecular weight.

8. High Performance Liquid Chromatography (HPLC)

- Principle: Improved column chromatography with optimized stationary phase, high pressure delivery of mobile phase, sensitive detectors.

- Steps: Sample injected, carried by mobile phase at high pressure through column, separates based on affinity, detected.

- Uses: Qualitative and quantitative analysis of compounds.

- Example: Analyze pharmaceuticals, foods, biomarkers, etc.

9. Hydrophobic Interaction Chromatography

- Principle: Based on interaction between hydrophobic sample and hydrophobic stationary phase.   

- Steps: Sample applied in high salt buffer, hydrophobic molecules bind, changing to low salt buffer elutes sample.

- Uses: Separate proteins and biomolecules based on hydrophobicity.

- Example: Purify monoclonal antibodies, hormones, enzymes etc.

10. Ion Exchange Chromatography

- See Anion and Cation exchange chromatography.

11. Liquid Chromatography 

- Principle: Separation based on differential partitioning between liquid mobile phase and solid or liquid stationary phase.

- Steps: Sample injected, carried through column by mobile phase, separates based on affinity for stationary phase.

- Uses: Separate and analyze non-volatile mixtures.

- Examples: Amino acid analysis, purification of drugs, vitamins, proteins etc. 

12. Paper Chromatography

- Principle: Based on partition between water held in cellulose paper (stationary phase) and mobile solvent phase.

- Steps: Spot sample on paper, place in solvent, components separate as solvent moves up paper.

- Uses: Separation and identification of amino acids, carbohydrates, etc. 

- Example: Identify amino acids in protein hydrolysate.

13. Reverse Phase Chromatography

- Principle: Based on hydrophobic interactions with a non-polar stationary phase and polar mobile phase.

- Steps: Polar sample injected, interacts weakly with non-polar stationary phase, elutes quickly. Less polar compounds elute more slowly.

- Uses: Commonly used HPLC method for separation of organic compounds. 

- Example: Separate lipids, steroids, vitamins, etc.

14. Thin Layer Chromatography (TLC) 

- Principle: Based on partition between a thin stationary phase immobilized on a plate and a mobile phase.

- Steps: Spot sample on plate, place in solvent tank, components separate as solvent moves up plate.

- Uses: Analytical separation and identification of organic and biomolecules.

References:

- Skoog, Holler and Crouch. Principles of Instrumental Analysis.

- Wilson and Walker. Principles and Techniques of Biochemistry and Molecular Biology.  

- Mohrig et al. Techniques in Organic Chemistry.

An Introduction to Spectroscopy Techniques and Their Applications in Analysis

 Spectroscopy is the study of the interaction between matter and electromagnetic radiation. It is a technique used to analyze the composition and structure of matter by examining how light or other electromagnetic radiation is absorbed, emitted, or scattered by that matter.

A spectrometer is an instrument used to measure spectra. It can split light into its constituent wavelengths and measure the intensity at each wavelength. 

A spectrophotometer is a specific type of spectrometer that measures the intensity of light as a function of wavelength. It can be used to measure the absorption, transmission, or reflection of light.

A spectroscope is a simple spectrometer used to observe spectral lines and bands. It usually consists of a prism or diffraction grating to disperse light and view a spectrum.

A spectrograph is a spectroscope that can record the spectrum onto a photographic plate or detector. It produces a spectral graph or spectrogram.

Spectra refers to the characteristic patterns of frequencies/wavelengths in electromagnetic radiation emitted or absorbed by atoms and molecules. Each atom or molecule has a unique spectral signature.

Some key types of spectroscopy and their basic principles, steps, and uses:

1. Absorption Spectroscopy

- Principle: Based on the absorption of electromagnetic radiation by the analyte at characteristic wavelengths. The absorption follows Beer-Lambert law.

- Steps: 1) Shine light on sample, 2) Sample absorbs light at specific wavelengths, 3) Measure transmitted light intensity at different wavelengths, 4) Relate absorption to analyte concentration.

- Uses: Determine concentration of analytes, identify analytes, quantitative and qualitative analysis.

2. Astronomical Spectroscopy 

- Principle: Analysis of electromagnetic radiation from stars and other celestial objects. Allows determination of chemical composition, motion, temperature, etc.

- Steps: 1) Light from celestial source dispersed into spectrum, 2) Sensitive detectors used to measure intensity at different wavelengths.

- Uses: Determine composition, temperature, radial velocity of astronomical objects. 

3. Atomic Absorption Spectroscopy

- Principle: Sample is vaporized and light of specific wavelength is passed through vaporized atoms. Amount of light absorbed is proportional to analyte concentration.

- Steps: 1) Atomize sample, 2) Irradiate sample with light source specific to analyte, 3) Measure absorption.

- Uses: Quantitative determination of specific metal elements in samples.

4. Circular Dichroism Spectroscopy

- Principle: Differential absorption of left and right circularly polarized light due to structural asymmetry of molecules.

- Steps: 1) Irradiate sample with circularly polarized light, 2) Measure difference in absorption between left and right circularly polarized components. 

- Uses: Study chiral molecules, determine protein secondary structure.

5. Electrochemical Impedance Spectroscopy

- Principle: Apply small amplitude AC signal to electrochemical cell and measure current response across a range of frequencies. 

- Steps: 1) Apply AC potential to cell, 2) Measure current response as a function of frequency, 3) Analyze impedance spectrum.

- Uses: Study electrode processes and complex interfaces.

6. Electron Spin Resonance Spectroscopy

- Principle: Microwave absorption by unpaired electrons in a strong magnetic field. Provides information about electronic structure.

- Steps: 1) Place sample in magnetic field, 2) Irradiate sample with microwaves, 3) Detect microwave absorption as function of magnetic field.

- Uses: Study free radicals, identify paramagnetic species, examine properties of materials.

7. Emission Spectroscopy

- Principle: Excited atoms and molecules emit electromagnetic radiation at characteristic wavelengths as they relax.

- Steps: 1) Excite sample using heat/electrical energy, 2) Measure intensity of emitted light at different wavelengths.

- Uses: Identify elements, quantify analyte concentration, flame tests.

8. Energy Dispersive X-ray Spectroscopy

- Principle: X-rays emitted from a sample due to bombardment by electron beam are measured by energy. 

- Steps: 1) Bombard sample with electron beam, 2) Detect emitted X-rays, 3) Measure X-ray energy spectra.

- Uses: Elemental analysis and chemical characterization of a sample.

9. Fluorescence Spectroscopy

- Principle: Fluorescent light is emitted from a sample after excitation by a light source. 

- Steps: 1) Excite sample with light source, 2) Measure intensity and wavelengths of emitted fluorescent light.

- Uses: Analyze organic compounds, biochemical analysis, medical diagnostics.

10. Fourier Transform Infrared Spectroscopy

- Principle: Infrared light passing through a sample is measured by an interferometer to obtain an interferogram, which is decoded by Fourier transform.

- Steps: 1) Pass IR light through sample, 2) Measure interferogram, 3) Apply Fourier transform to get spectrum.

- Uses: Identify organic, polymeric, and some inorganic materials.

11. Gamma-ray Spectroscopy

- Principle: Nuclear transitions in atomic nuclei result in gamma-ray emissions at characteristic energies.

- Steps: 1) Excite sample to induce nuclear reactions, 2) Measure energy spectrum of emitted gamma-rays. 

- Uses: Identify isotopes, analyze nuclear reactions.

12. Infrared Spectroscopy

- Principle: Infrared radiation excites molecular vibrations which absorb at specific wavelengths based on chemical structure.

- Steps: 1) Expose sample to IR radiation, 2) Measure IR absorption or transmission spectrum.

- Uses: Identify functional groups, analyze organic compounds, quantify components.

13. Magnetic Resonance Spectroscopy

- Principle: Atomic nuclei absorbed in a magnetic field absorb and re-emit electromagnetic radiation. Frequency depends on the nucleus and chemical environment. 

- Steps: 1) Place sample in magnetic field, 2) Irradiate sample with radio waves, 3) Detect emitted radiation.

- Uses: Analyze molecular structure and chemical environment.

14. Mass Spectrometry 

- Principle: Ions formed from a sample are separated based on mass-to-charge ratio.

- Steps: 1) Ionize analyte molecules, 2) Separate ions based on m/z, usually with electric/magnetic fields, 3) Detect ions.

- Uses: Identify unknown compounds, quantify molecules, determine molecular structure.

15. Molecular Spectroscopy

- Principle: Interaction of molecules with electromagnetic radiation at varying wavelengths is measured to derive structural and dynamic information.

- Steps: 1) Expose sample to EM radiation source, 2) Measure interaction (absorption, emission, scattering, etc.) as function of wavelength.

- Uses: Determine molecular structure, identify functional groups, analyze molecular motions/transitions.

16. Mössbauer Spectroscopy

- Principle: Based on recoil-free emission and absorption of gamma rays by atomic nuclei bound in a solid. 

- Steps: 1) Irradiate sample with gamma rays, 2) Measure absorption spectrum of gamma rays.

- Uses: Study chemical environment and magnetic properties of iron-containing samples.

17. Nuclear Magnetic Resonance Spectroscopy

- Principle: Nuclei in a magnetic field absorb and re-emit electromagnetic radiation at characteristic frequencies. 

- Steps: 1) Place sample in magnetic field, 2) Irradiate sample with radio waves, 3) Detect emitted radiation. 

- Uses: Determine molecular structure, identify compounds, quantify analytes.

18. Photoelectron Spectroscopy

- Principle: High energy photons eject electrons from a sample. Kinetic energy measured provides information about elemental composition and chemical environment.

- Steps: 1) Irradiate sample with X-rays/UV rays, 2) Measure kinetic energy of emitted electrons.

- Uses: Determine elemental composition, chemical bonding, electronic structure. 

19. Raman Spectroscopy

- Principle: Based on inelastic scattering of monochromatic light by molecules which provides vibrational info.

- Steps: 1) Illuminate sample with monochromatic source, 2) Measure frequency shifts in scattered light.

- Uses: Identify molecules, investigate sample composition.

20. UV Spectroscopy

- Principle: Samples absorb ultraviolet light at wavelengths corresponding to allowed electronic transitions.

- Steps: 1) Expose sample to UV light source, 2) Measure absorption spectrum in UV region.

- Uses: Quantify analytes, identify organic compounds, determine purity.

21. UV/Vis Spectroscopy

- Principle: Molecules containing π-electrons or non-bonding electrons can absorb UV/Vis light resulting in electronic transitions.

- Steps: 1) Expose sample to UV/Vis light, 2) Measure absorption spectrum.

- Uses: Quantify analytes using Beer-Lambert law, identify analytes, determine kinetics.

22. X-ray Photoelectron Spectroscopy 

- Principle: X-ray irradiation causes photoelectron emission, with binding energies characteristic of each element.

- Steps: 1) Irradiate sample with X-rays, 2) Measure kinetic energy of emitted photoelectrons. 

- Uses: Determine elemental composition, chemical states, electronic structure.

References:

- Skoog, Holler and Crouch - Principles of Instrumental Analysis 

- Banwell and McCash - Fundamentals of Molecular Spectroscopy

- Hollas - Modern Spectroscopy

- McQuarrie - Quantum Chemistry 

- Wilson and Walker - Principles and Techniques of Practical Biochemistry

- Günzler and Gremlich - IR Spectroscopy: An Introduction

How does the Suzuki coupling reaction work?

The Suzuki coupling is a cross-coupling reaction used to form new carbon-carbon bonds between an aryl or vinyl boronic acid and an aryl or vinyl halide catalyzed by a palladium complex. 

The Suzuki coupling reaction couples aryl halides and arylboronic acids to form biaryl compounds. This versatile C-C bond forming reaction has been widely studied to expand its scope and improve efficiency.

Various catalytic systems have been explored, including photocatalytic coupling using palladium catalysts, Suzuki-Miyaura coupling of fluoroarenes, and nickel-catalyzed variants as a more earth-abundant option. Solvent selection and greener processing conditions have also been evaluated.

These diverse Suzuki coupling approaches have enabled numerous applications from protein modification to the synthesis of complex polyaromatic structures. Ongoing work continues to optimize Suzuki coupling for different substrates and reaction conditions to enhance the sustainability, efficiency, and selectivity of this important cross-coupling reaction.

The key steps are:

1. Oxidative addition - The palladium catalyst undergoes oxidative addition with the aryl/vinyl halide, inserting itself between the carbon-halogen bond. This forms an organopalladium complex.

2. Transmetalation - The organopalladium complex then undergoes transmetalation with the boronic acid, replacing the halide with an aryl/vinyl group from the boronic acid. This forms a new organopalladium complex.

3. Reductive elimination - Reductive elimination of the palladium catalyst from the complex then occurs, forming the new carbon-carbon bond between the two aryl/vinyl groups originally from the halide and boronic acid. The active palladium catalyst is regenerated. 

The Suzuki reaction works well for joining aryl or vinyl units together efficiently under relatively mild conditions. It requires a base, usually an aqueous hydroxide, to activate the boronic acid. 

The Suzuki coupling is a very useful tool in organic synthesis and pharmaceutical research for building biaryls and styrenes.

Beta-lactams

Beta-lactams are an essential antibiotic drug class produced by both fermentation and synthetic methods. Their unique reactivity enables the generation of novel compounds for pharmaceutical applications.

Beta-lactams are an important class of compounds in biological and synthetic chemistry. They are commonly used as antibiotics to treat bacterial infections. Beta-lactams work by inhibiting penicillin-binding proteins that are crucial for bacterial cell wall biosynthesis. Most beta-lactams are produced by fermentation or modification of fermented intermediates, except for carbapenems and aztreonam which require synthetic routes.

The reactivity of the beta-lactam ring has been widely studied, making it a useful substrate in synthetic organic chemistry. The ring can be opened through various reactions to generate new biologically relevant compounds. Efficient synthesis of new beta-lactams can be achieved from amidines using promoters like bismuth, indium, and copper salts. Diverse C4-N-substituted beta-lactams can also be prepared by nucleophilic reactions. A commonly used method in beta-lactam chemistry is the Ketene-Imine Staudinger Reaction.

Bibliography:

Zerong, Daniel, Wang. (2023). The Chemistry and Biology of Beta-Lactams.   doi: 10.1201/9781003330288

(2023). Enzymatic biosynthesis of β-lactam antibiotics.   doi: 10.1016/b978-0-443-19059-9.00007-4

Japheth, O., Ombito, and, Girija, S., Singh. (2019). Recent Progress in Chemistry of β-Lactams. Mini-reviews in Organic Chemistry,  doi: 10.2174/1570193X15666180914165303

Bimal, K., Banik., Alberto, Boretti. (2021). Hypotheses for synthesis of novel chiral beta-amino-beta-lactams through amidines.   doi: 10.1016/J.RECHEM.2021.100158

Rajneesh, Kaur., Divya, Tripathi., Kuldeep, Singh., Raman, Singh. (2018). Recent advances in β-lactam chemistry.  J. Integr. Sci. Technol., 2018, 6(2), 46-51

Adrian, Saura-Sanmartin., Laura, Andreu-Ardil. (2023). Stereoselective synthesis of β-lactams: recent examples.. Organic and Biomolecular Chemistry, 21(16):3296-3306. doi: 10.1039/d3ob00309d

Anusandhan National Research Foundation Act 2023

 

Anusandhan National Research Foundation Act 2023

An Act to provide for the registration and regulation of a financially research ecosystem, and open scientific research activities for the private sector in the India in accordance with the National Education Policy guideline.
Enacted by	Parliament of India
Assented to	15 August 2023
Commenced	1 December 2023
Bill citation	No. 25 2023
Status: In force (5th Feb 2024)

https://dst.gov.in/sites/default/files/NRF.pdf 

AI Tools for Scientists & Teachers

 Here are some of the top AI tools that can be useful for scientists and teachers:

For Scientists:

- AI Lab Tools - Enables scientists to easily build, train, and deploy machine learning models without coding.

- Transcriptic - Provides robotic cloud laboratories for remote biological experimentation. Enables automation of pipetting, instrumentation, and data collection.

- Atomwise - Uses deep learning for structure-based small molecule drug discovery. Can predict novel drug candidates. 

- Arterys - FDA-cleared medical imaging analytics platform that automates analysis of MRIs and CT scans.

- Benchling - Cloud-based software platform that helps manage laboratory data and automate research workflows.

- Sophia Genetics - Leverages AI to help diagnose genetic diseases, cancers, and complex disorders.

- SciBite - Text analytics to help extract insights from unstructured scientific data like research papers and reports.

For Teachers:

- Socratic - AI tutoring app that provides personalized learning recommendations and micro-explanations for STEM subjects.

- Quill - NLP technology to provide grammar and writing feedback for students' essays. Helps teach skills.

- Aleks - Adaptive learning platform using AI to individualize math instruction for K-12 students.

- Century - AI platform that generates personalized learning paths in math and English for each student.

- Thinkster Math - Virtual AI math tutor that analyzes students' weaknesses and provides customized lessons.

- Duolingo - Language learning app using AI to tailor courses based on users' proficiency. 

- DreamBox - Adaptive K-8 math education platform driven by AI that adjusts lessons to each learner's level.

- Metacog - AI tutoring tool focused on improving reading comprehension skills for students.

Crafting an Effective Research Article

Writing and Publishing a Research Article

Publishing research articles is critical for scientists, scholars, and academics to advance knowledge and their careers. But writing a compelling research paper that clearly communicates your study and findings is easier said than done. In this blog post, I'll share tips on how to craft an impactful research article.

Choose the Right Journal

Selecting the optimal journal to submit to is an important first step. Consider the aim, readership, and types of articles published in potential journals. Choosing a journal that aligns with your research topic and approach signals relevancy to editors and reviewers. Identifying the right outlet sets your article up for success.

Structure with IMRAD

Organize your article using the standard IMRAD format - Introduction, Methods, Results, and Discussion. This structure helps logically convey key parts of your study. The introduction orients readers to the research area, gaps in knowledge, and your specific aims. The methods section provides sufficient detail for others to reproduce your experiment. Results presents analyzed data and discoveries. Finally, the discussion interprets findings, compares them to other studies, notes limitations, and explores implications.

Write a Strong Introduction 

The introduction is crucial for framing your entire study. After reviewing pertinent literature, clearly state the research question, objective, or hypothesis. Succinctly explain why your study is needed based on gaps in understanding. The final paragraph should outline the article's structure and summarize the contents. This "roadmap" helps readers navigate the key points.

Explain the Methods

The methods section should provide enough specifics that competent researchers can replicate your study. Describe the experimental design, subjects, equipment, procedures, and analysis techniques. Cite previously published methods. Using subsections can help organize more complex endeavors. The goal is to give readers a clear picture of how the study was conducted.

Present the Results

The results section objectively presents key quantitative and qualitative findings without interpretation. Use text, figures, and tables to summarize analyzed data and illustrate important outcomes. Figures should have descriptive captions and tables should be formatted cleanly. Only include displays that enhance understanding of results - not raw data.

Discuss and Contextualize 

This section allows you to interpret implications of results within the field. Compare your outcomes to prior studies stated in the introduction. Explore possible mechanisms behind the observations. Acknowledge limitations and reasonable alternative explanations. End by highlighting the study's overall contributions and suggesting next steps for future research. 

Refine and Review

With the main sections drafted, refine the article to ensure it meets submission requirements. Format citations and references consistently. Seek feedback from colleagues. Review the article carefully to confirm it is coherent, flows logically, and follows the target journal's style guidelines. Finally, choose a concise, descriptive title and create an abstract that summarizes the key points.

Writing an effective research article takes time and effort. But clearly communicating your work to advance science and knowledge in your field is worth the investment. What strategies have you found helpful for writing high-quality papers? Let me know in the comments!

Practical Books

 

S.No	Medium	Class	Subject with description	Link
1	11	English	Automobile Practicals-Whole book
2	11	English	Electronic Engineering Practicals-Whole book
3	11	English	Mechanical Engineering Practicals-Whole book
4	11	English	Zoology-Zoology Practicals
5	11	English	Bio-Zoology Practicals
6	11	English	Statistics-Statistics Practicals - 10.09.2018
7	11	English	Home science-Practicals
8	11	English	Nutrition & Dietetics-Practicals
9	11	English	Micro biology-Practicals
10	11	Tamil	Automobile Engineering Practicals-Whole book
11	11	Tamil	Electronic Engineering Practicals -Whole book
12	11	Tamil	Mechanical Engineering Practicals -Whole book
13	11	Tamil	Zoology -ZoologyPracticals
14	11	Tamil	Bio-Zoology -Bio-Zoology Practicals
15	11	Tamil	Botany -Botany Practicals - 10.09.2018
16	11	Tamil	Home science - Practicals
17	11	Tamil	Nutrition and dietics - Practicals
18	11	Tamil	Micro biology - Practicals

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