Chemical and biological analysis: Lets go fishing molecules Ajeet - - PowerPoint PPT Presentation

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Chemical and biological analysis: Lets go fishing molecules Ajeet - - PowerPoint PPT Presentation

May 26, 2023 13 likes •205 views

Chemical and biological analysis: Lets go fishing molecules Ajeet Singh and Venkat Panchagnula Bioseparations Group Chemical Engineering Division, National Chemical Laboratory Pune 411008 A simple test to distinguish between similar looking liquids

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Chemical and biological analysis: Lets go fishing molecules

Ajeet Singh and Venkat Panchagnula

Bioseparations Group Chemical Engineering Division, National Chemical Laboratory Pune 411008

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A simple test to distinguish between similar looking liquids

  • Water / Isopropyl alchohol / detergent water
  • Water – Won’t spread on wax paper
  • Alcohol and detergent water will spread
  • Alcohol mark has a smooth edge and detergent will have

an irregular edge

  • Alcohol evaporates faster, detergent evaporates the

slowest

Many similar experiments can be found on the American Chemical Society Website www.acs.org

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Forensic Science – Finger prints with ninhydrin reaction

Live demo of ninhydrin revealing fingerprints

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Amino acids, peptides and proteins

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www.langara.bc.ca/.../Assets/aminoAcidStruc.jpg http://en.wikipedia.org/wiki/File:Protein‐primary‐structure.png

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Milk

  • 87% H20
  • True Protein 3%
  • Crude protein 3.1%
  • Casein 2.6%

Protein content in Milk determined by Kjeldahl method that measures Nitrogen present

http://chem.vander‐lingen.nl/articles/The_melamine_milk_scandal/id/126/itemid/380

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Tainted milk trial opens in China Tainted milk made thousands of Chinese babies ill Six men have gone on trial in China accused of making and selling the chemical at the centre of the tainted milk scandal.The tainted milk is blamed for killing at least six children and making 300,000

  • ther people ill.The men are accused of adding melamine to raw

milk to make it appear high in protein. Friday, 26 December 2008

Melamine found in Cadbury goods The recall affects mainland China, Hong Kong, Taiwan and Australia Preliminary tests have found melamine in Cadbury's Chinese‐made chocolates, the company says.At least 50,000 Chinese babies have fallen ill and four been killed by milk tainted with the industrial chemical.Cadbury had earlier recalled 11 chocolate types from mainland China, Hong Kong, Taiwan and Australia because of fears of contamination.It remains unclear how much melamine was in the recalled products, a Cadbury spokesman told the BBC.

Monday, 29 September 2008 Source for the news items: http://news.bbc.co.uk

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Melamine

  • Used in plastics and fertilizers
  • Scrap melamine is unusable left over product

containing 70% melamine and other derivatives including cyanuric acid

  • In China, melamine scrap and low cost soya

protein powder were added

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Chromatographic separation of melamine in milk

Melamine standard Milk with melamine Milk without melamine + + + + + + + +

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Need for a separation ‐ Chromatography

  • Principle of chromatography & TLC
  • Chlorination – the amine gps will be

chlorinated; In the presence of starch, iodine replaces Cl and gives color (WHY?) ‐

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Methods to analyze melamine in milk sample

  • Thin layer chromatography (movie )
  • HPLC (movie)
  • Mass spectrometry
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Mass Spectrometric Analysis of Milk and Contaminated Milk Uncontaminated Milk Contaminated Milk with Melamine

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Mass Spectrometric Analysis of Melamine Melamine Molecule

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Combiflam

Paracetamol Ibubrufen Combiflam + + + + + + + + ` Combiflam separation by TLC – Live demo

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Human Proteome: ~22,000 Genes ------ ~1 million proteins

Figure Adapted from Biochemistry, Mathews, Addison Wellesley

Genomics to Proteomics

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http://www.scandfish.com/gfx/clpost_namericanfish_large.jpg

Proteomics: Fishing the ‘Unknown Fishes’

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Gel electrophoresis

Gel acts as a sieving media Separation based on mass

  • Electrophoretic mobility

+ + + +

  • +
  • +
  • +
  • +

+ +

Applied potential ‘E’

  • +
  • +
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What am I working on?

  • Protein separation, isolation and detection
  • Agricultural applications
  • Biomarker discovery and medical diagnostics
  • Fundamental understanding
  • Protein encapsulation and delivery
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Take home messages…

  • Analytical science – can involve

– Simple chemical reactions – Complicated physiochemical phenomenon – Separation, purification, isolation – Simple color detection to complicated signal generation and detection [Electrochemical, mass spectral analysis] – Discovery of analytes to tackle problems relating to human diseases

  • Covers wide range of applications
  • Encompasses many areas – Science, Engineering and

Medicine

  • Known fish can be caught (from a fish tank)
  • Plenty of scope to fish in the ‘high seas’ – rich

dividends to reap