Application of chromatographic technique
- 2. Presented BY Dr Devraj K.C. PG 1st year Guided By:- Dr Poornima B S Assistant Professor Dravyaguna Department
- 4. Chromatography Is a technique used to separate and identify the components of a mixture The separation of a mixture by distribution of its components between a mobile and stationary phase over time ◦ mobile phase = solvent ◦ stationary phase = column packing material
- 5. A mixture of two different compound passing it through mobile phase to stationary phase that retains some compounds longer than others in stationary phase. In stationary phase the Retention factor (Rf) value is calculated where Rf value of each and every compound is different.
- 6. Rf = distance moved by substance distance moved by solvent front Some terminology in chromatograph
- 7. Type of chromatography Solvent Paper chromatography Air, alcohol Thin Layer Chromatography Hexane, ether petroleum, alcohol. Gas chromatography He, Ar, N2 High Performance Liquid Chromatography Cyclohexane, n-hexane, carbon tetrachloride, ethanol, methanol, air
- 10. Type of chromatography Material Paper chromatography Filter paper, cellulose Thin Layer Chromatography Silica gel, alumina, polyamide Gas chromatography Squalene, apezion, carbowax M High Performance Liquid Chromatography C-8, C-18, Licosorb, Silicone
- 11. Retention Time. Retention time (RT) is a measure of the time taken for a solute to pass through a chromatography column.
- 13. • Paper Chromatography – separates dried liquid samples with a liquid solvent (mobile phase) and a paper strip (stationary phase) • Thin-Layer Chromatography – separates dried liquid samples with a liquid solvent (mobile phase) and a glass plate covered with a thin layer of alumina or silica gel (stationary phase) • Liquid Chromatography – separates liquid samples with a liquid solvent (mobile phase) and a column composed of solid beads (stationary phase) • Gas Chromatography – separates vaporized samples with a carrier gas (mobile phase) and a column composed of a liquid or of solid beads (stationary phase) •
- 14. 1. Pharmaceuticals industry -To identify the drug, avoid impurities ,determine stability, doses form and to find the quantity of drugs - Heavy metal analysis of drug i.e. Cadmium, Arsenic, Mercury. -Pesticidal residue and micro chemical analysis of drugs. 2. Analysis of natural contamination - Phenol & Mercury from sea water 3. Forensic test - Determination of steroid in blood, urine & sweat. - Detection of psychotropic drug in plasma
- 15. 4. Clinical test - Monitoring of hepatic cirrhosis patient through aquaporin 2 in the urine. 5. Food and essence manufacture - sweetener analysis in the fruit juice - preservative analysis.
- 17. i) paper chromatograph can be used for isolating known and unknown substances. ii) Identifying pure substances from mixtures. iii) Determination of decomposed products.
- 20. TLC vido
- 21. i)Qualitative and quantitative analysis of the herbs and herbal drugs. ii) Analysis of crude samples containing multiple components. iii) The separation process is easy to follow especially with colored compounds for this Specific and sensitive colour reagents can be used to detect separated spots (Dragendroff reagent/Kedde reagent) iv) Several samples can be separated parallel to each other on the same plate resulting in a high output, time saving, and a rapid, low-cost analysis.
- 22. High-performance Thin layer chromatography(HPTL)
- 24. Preparative and analytical HPLC are widely used in pharmaceutical industry for isolating and purification of herbal compounds. There are basically two types:- i) Low pressure HPLC (typically under 5 bar) ii) High pressure HPLC (pressure>20 bar)
- 25. i) LC-MS has become method of choice in many stages of drug development. I.e. Chemical standardization of an aqueous extract of the mixture of the compound herbal formulation. ii) LC-MS is useful in analysis of amino glycosides their solubility in water, plasma protein binding capacity, and their excretion.
- 29. Where w=width and h=height of peak
- 32. .Identification and quantifying of aromatics and volatile substances of medicinal plants.
- 35. SFC permits the separation and determination of a group of compounds that are not conveniently handled by either gas or liquid chromatography. It is useful for the separation of much less polar compounds such as many natural products, including fat soluble vitamins, carotenoids, and lipids along with separation of each individual components of herbal drugs.
- 40. i) Complex mixture of herbal drug can be separated by chromatography passing through mobile to stationary phase. ii) Mobile phase is either gas or liquid but stationary phase is ever solid. iii) Component in mobile and stationary phase depends upon the test drug. Iv) Chromatographic technique is compulsory in the field of herbal drug standardization, authentication and manufacturing.
- 41. v) For identification, authentication, quantifying, preparation and standardization of ayurvedic herbal drugs many physical, chemical, biological, and instrumental technique are used among them chromatography is physicochemical method. vi) Paper chromatography is used to identify and separate simple compound of herbal drugs. vii)High-performance liquid chromatography (HPLC) and High-performance thin layer chromatography (HPTLC) are backbone of ayurveda for drug standardization and Authentication.
- 42. vii) Gas Chromatography technique is widely use to detect and quantify volatiles and aromatics of herbal drugs. viii) Supercritical fluid chromatography is the hybrid production of gas and liquid chromatography for extraction from crude herbs.
- 43. Sharma A., Shanker C., Tyagi L.K., Singh M., Rao V. Herbal Medicine for Market Potential in India: An Overview. Academic J. Plant Sci. 2008; 1 (2): 26-36. 2 Kadam P.V., Yadav K.N., Shivatare R.S., Pande A.S., Patel A.N., Patil M.J. Standardization of Gomutra Haritaki Vati: An Ayurvedic Formulation. Int. J. Pharm. Bio. Sci. 2012; 3(3): 181 – 187. 3. Mukharjee P.K., Wahile A. Integrated approaches towards drug development from Ayurveda and other Indian system of medicines. J. Ethanopharmacol. 2006; 103 (1): 25-35. 4. Choudhary N., Sekhon B.S. An overview of advances in the standardization of herbal drugs. J. Pharm. Educ. Res. 2011; 2 (2): 55-70. 5. Amit J., Sunil C., Vimal K., Anupam P. Phytosomes: A revolution in herbal drugs. The Pharma Review 2007; 11- 13. 6. Jena A., Saha D., Biswal B., Jana S.B., Koley A., Sur D., Battacharya A. Pharmacognostic Studies of leaves of Pterospermum Suberifolium. Int. J.
- 44. Thank you