instrumental analysis of poison part 1 - Dr Pooja Prasad.pptx

the ppt is made in the simplest way to understand the functionality of poison. The instruments that are required to identify the poison or poison components at the microlevel have been described. Also deals with their uses and functions.

1. Instrumental Analysis of Poison Dr Pooja Prasad PG Scholar

2. To detect poisons ,drugs and different chemicals Chromatography - process of separating components of mixture Electrophoresis – movement of charged particles in a fluid under electric field. Spectrometry – measurement of the interactions between light and matter, its radiation intensity and wavelength

3. Chromatographic techniques Column chromatography Paper chromatography Thin layer chromatography Gas chromatography

4. " Chromato graphy " is a technique for separation, “ Chromato gram " is the result of chromatography, “Chromato graph " is the instrument used to conduct chromatography.

5. It is a tube like structure made up of glass or plastic. Generally Chromatography has always two phase components Stationary phase – solid/liquid Mobile phase- liquid/gas In column Chromatography S. phase has solid and M. phase has liquid This procedure can be run manually or automatic. for automatic – a machine called AKTA system Materials used for column chromatography- silica gel, aluminium Solvent s used as mobile phase – ethanol, acetone, water, acetic acid, pyridine.

6. Steps to do manual column chromatography 1. pick a stationary phase (silica , aluminium) Dry packing Wet packing 2 .Column packing

7. Column Chromatography Applications Isolate active ingredients Separating compound mixtures Determine drug estimation from drug formulations Remove impurities Isolate metabolites from biological fluids.

8. Paper chromatography Normal/ cellulose paper - Whattman filter paper (31ET) coarser and faster. Modified paper Hydrophillic paper - modified with methanol, glycerol, formamide Hydrophobic paper- modified with acetylation of OH groups leads to hydrophobic in nature, also be said as reverse phase chromatography Impregnation of silica, alumna, ion exchange resins can also be made

9. Principle used Partition seen in liquid to liquid Adsorption- solid- liquid

10. Thin layer chromatography Instead of paper, silica gel and aluminium gel is used in glass plate. technique used to isolate non-volatile mixtures

11. Thin layer chromatography…. cont … experiment is conducted on a sheet of aluminium foil, plastic, or glass which is coated with a thin layer of adsorbent material. The material usually used is cellulose, or silica gel TLC is one of the fastest, least expensive, simplest and easiest chromatography technique.

12. to apply sample spots, thin marks are made at the bottom of the plate with the help of a pencil. Apply sample solutions to the marked spots. Place the plate in the TLC chamber and close it with a lid. It is kept in such a way that the sample faces the mobile phase. Thin layer chromatography…. cont …

13. Immerse the plate for development. Remember to keep the sample spots well above the level of the mobile phase. Do not immerse it in the solvent. Wait till the development of spots. Once the spots are developed, take out the plates and dry them. The sample spots can be observed under a UV light chamber. Thin layer chromatography…. cont …

14. If sample is invisible then UV visualization – polyphenol Acid treatment- dip- air dry- hot plate- brown spot Iodine vapour treatment- compound C=C ,double bond carry atom Vallinium stain – Alcohol, aldehyde, ketone Ninhydraine treatment- Amino acid if sample forms streak then it shows sample has applied in large quantity. If solvent is not moved equally then it shows that the gel is not applied properly in uniform thickness. pencil is used instead of pen , because pen contains blue ink or black ink. It will spread along with silica gel.

15. Thin Layer Chromatography Applications…. cont …   qualitative testing of Various medicines such as sedatives, local anesthetics, anticonvulsant tranquilizers, analgesics, antihistamines, steroids, hypnotics is done by TLC. TLC is extremely useful in Biochemical analysis such as separation or isolation of biochemical metabolites from its blood plasma, urine, body fluids, serum, etc. TLC can be used to identify natural products like essential oils or volatile oil, fixed oil, glycosides, waxes, alkaloids, etc.

16. It is widely used in separating multicomponent pharmaceutical formulations. It is used in the food industry, to separate and identify colours , sweetening agent, and preservatives It is used in the cosmetic industry . Thin Layer Chromatography Applications

17. Disadvantages Of Thin Layer Chromatography: Thin Layer Chromatography plates do not have longer stationary phase. When compared to other chromatographic techniques the length of separation is limited. The results generated from TLC are difficult to reproduce. Since TLC operates as an open system, some factors such as humidity and temperature can be can affect the final outcome of the chromatogram. The detection limit is high and therefore if you want a lower detection limit, you cannot use TLC. It is only a qualitative analysis technique and not quantitative.

18. Gas chromatography Mobile phase- Gas Stationary phase – liquid/ Solid Used to analyse the DNA, Protein, Mixture molecule

19. Gas chromatography (GC) combined with mass spectrometry (MS) gives the best possible results on sample analysis. Gas chromatography separates the components of a mixture in an extremely powerful separation method. Gas chromatography

20. The identity of an unknown compound cannot be established on the basis of its retention time. Gas chromatography is coupled with a mass spectrometer for such types of analysis. The gas chromatography (GC) separates the components of the mixture while the mass spectrometry gives structural information of such components . Gas chromatography

21. It can be used during the Olympic games for stimulants, narcotics, and anabolic tests. analysis of polymeric compounds and complex petroleum products like paraffin, cycloparaffin, olefin, cycloolefin, and alkyl benzenes. gas chromatography-mass spectrometry combinations are used to characterize the properties of fuels and lubricating oils. Gas chromatography

22. Nitrogen , helium

23. Low boiling point will move fast High boiling point will move slow In case boiling point is same then it determines with their size of molecules.

24. Mass spectrometry Powerful micro-analytical technique Requires few nanomoles of the sample Used to Quantify known materials Identify known compounds within a sample To elucidate the structure and chemical properties of molecule . Technique studies the effect of ionizing energy of molecules

25. CHEMICAL FINGERPRINT Mass spectrum of each compound is always unique determine mass to charge ratio (m/z) by a mass analyzer. Uses- Isotopes isolation- C12, C13, C14 Molecular fragments Atom Mass spectrometry cont..

26. Gas   chromatography or liquid  chromatography  combined with mass spectrometry (MS) gives the best possible results on different types of sample analysis. The mass spectroscopy principle is used to measure the exact molecular weight and molecular formula of organic compounds. The presence of certain structural units in the organic compound can be recognized by mass spectrometry. Mass spectrometry cont..

27. testing and discovery of drugs, food contamination detection, pesticide analysis, isotope ratio determination, protein identification, and carbon dating. characteristics of proteins and protein complexes and the sequencing of peptides. drinking water tests, pesticide screening, soil contamination analysis, pollution monitoring, and different types of heavy metal analysis. Mass spectrometry cont..

28. drug discovery and analysis, pharmacokinetic and pharmacodynamics analysis, and metabolite screening. drug development, clinical tests, disease screening, drug therapy monitoring, cancer screening, and diagnosis. 3. In forensic applications , fingerprinting analysis, trace evidence analysis, confirmation of drug abuse, and explosive residues identification or bombing investigation. Mass spectrometry cont..

29. High Performance Liquid Chromatography Works to Separates Identify Quantify Mobile phase- liquid Resolution power of this test is good because it has ability to distinguish between the components Identification is done on the basis of polarity Sample – blood, bacterial toxins, fungal toxins etc.

30. Next Session