Silica Gel | Aluminium Oxide Column chroamtography


Published on

Buy Silica Gel Powder for Silica Gel, Aluminium Oxide, Paper and Flash Column Chromatography us in Pharmaceutical Industries – Bulk Drugs & API, Nutraceuticals, Herbal Extracts products manufacturers, Research Laboratories, Laboratories Chemical Repackaging, Contract Research Laboratories. Column Chromatography is one of the most useful methods for purification & separation (Isolation) of individual desire compound from mixture of unwanted compounds.

Published in: Business
  • Be the first to comment

No Downloads
Total views
On SlideShare
From Embeds
Number of Embeds
Embeds 0
No embeds

No notes for slide

Silica Gel | Aluminium Oxide Column chroamtography

  1. 1. Column Chromatography PRODUCTS: SILICA GEL ALUMINIUM OXIDE
  2. 2. What is Chromatography?Chromatography has been developed into a new method of separation of mixture of compounds mainly when they are available in small quantities.This method is very useful when the components of a mixture have almost the same physical and chemical properties and hence can’t be separated by other usual methods of separations.The term chromatography means writing in colour (Chroma = Colour & Graphy = To write).
  3. 3. Types of Chromatography• Paper Chromatography• Gas Chromatography• Thin Layer Chromatography• Solid - Liquid Chromatography (Column Chromatography) 1. Gravity Chromatography 2. Flash Chromatography 3. High performance Liquid Chromatography
  4. 4. What is column chromatography?• Column chromatography is one of the most useful methods for purification & separation (Isolation) of individual desire compound from mixture of unwanted compounds.• It is often used for preparative applications on scales from micrograms up to kilograms• It is a solid - liquid technique in which the stationary phase is a solid & mobile phase is a liquid.• The stationary phase or adsorbent in column chromatography is a solid. The most common stationary phase for column chromatography is Silica Gel, followed by Alumina Oxide.• The mobile phase or eluent is a liquid. It is either a pure solvent or a mixture of different solvents.• It can be used for molecules whose molecular weight is < 2000 g/mol
  5. 5. High Performance Liquid Chromatography
  6. 6. Thin Layer Chromatography Thin layer chromatography, or TLC, is a method for analyzing mixtures by separating the compounds in the mixture. (All types of Chromatography works on same principle.) Uniform layer of Silica Gel coated onto a piece of glass, aluminium or rigid plastic plates are being used for analysis. The mobile phase moves through the stationary phase and carries the components of the mixture with it.  Different components travel at different rates. Application:  To determine the number of components in a mixture,  Identification of compounds, and  Purity of a compound.
  7. 7. Thin Layer Chromatography It is used in Analytical study to analyzed microgram (0.000001 g) quantity samples and it takes around 5-10 minutes for result. Size available: 1. Silica Gel G for TLC (With Binder) 2. Silica Gel H for TLC (Without Binder) 3. Silica Gel GF 254 – with fluorescent indicator 4. Silica Gel HF 254 with fluorescent indicator. In Thin Layer Chromatography fluorescent is added in stationary phase to analyze substances which are colourless. This we can not see with our naked eyes. When plate will expose to UV light then it will Glow except the spots of substances & spot will look like a dark patch.
  8. 8. Selection of Stationary Phase & Mobile Phase• Removal of impurities • It requires balancing act • Columns are available in glass tubes, between solvent & stainless steel & vary in length & diameter.• No. of components to be compound’s polarity. • Resolution of column depends on both separated • The compound must also diameter & length of adsorbents packed in• Length of the column used be soluble in water so they column.• Affinity differences are not permanently • Resolution improves with increase in length between components adsorbed. & reduces with increase in diameter.• Quality of adsorbent used • For e.g. 25 gms of adsorbents will provide a better separation in a 1 cm diameter column than 2 cm diameter column. • Column dimensions - length & diameter ratio (10:1, 30:1 or 100:1)Stationary Mobile Phase ColumnPhase (Silica (Solvents)Gel or Alumina)
  9. 9. How Scale Up take place• Analytical Scale: Column Inner Dia - 4,5,6 mm & it is done at milligram scale.• Semi Preparative: Column Inner Dia - 10, 20, 30 mm & it is done at milligram scale.• Preparative: Column Inner Dia – 40, 60, 80, 80 mm & it can be done at gram Scale• Pilot scale: Kg scale (1, 2, 4, 10, 12 Kg)• Commercial scale: Bulk Quantity (50, 100, 200, 400 Kgs or more)
  10. 10. Methods of Column Packing Dry Method : Add dry silica / Alumina to the column and apply to the bottom of the column. This will compress the silica gel and keep it compressed for the next steps. Packing can be improved by tapping the column. While applying vacuum; pour solvent in it. Allow the solvent to move though the column until reaches to the bottom. At this stage vacuum is not require. Allow 5–6 columns value of solvent to flow through the column to make sure it is complete packed. Drain the solvent till the solvent level is just even with the surface of the stationary phase
  11. 11. Methods of Column Packing Wet Method: Fill the column about one third with solvent In a beaker, measure out the required amount of silica / alumina. In another beaker, take solvent approximately one and a half times the amount of silica / alumina.  Add silica/alumina to the solvent while swirling in small quantity at a time. Use a glass rod to mix the slurry. Pour some of the slurry into column & allow solvent to drain to avoid overflowing. Tap the column carefully to encourage bubbles to rise and the silica to settle Continue to move the slurry to the column until all the silica or alumina is added. Wash the inside of the column by pouring solvent down the inside edge. Drain the solvent till the solvent level is just even with the surface of the stationary phase
  12. 12. How does separation take place?
  13. 13. Types of ColumnsGravity Column Chromatography: Flash Chromatography:Solvent is allowed to move down the Solvent is pushed down the column bycolumn by gravitational forces. positive air pressure
  14. 14. ApplicationSeparation of mixture of compoundsPurification processPurification of PhytochemicalIsolation of metabolites i.e. Small moleculesEstimation of drugsProcess DevelopmentPurify Natural compoundsTo separate active component from Plant materialHerbal Extraction
  15. 15. Adsorbents used in chromatography method• Silica gel (SiO2) and alumina (Al2O3) are two adsorbents commonly used for column chromatography.• These adsorbents are sold in different mesh sizes such as 60-230 mesh, 100-200 mesh, 200-400 mesh & tailor made.• Adsorbent particle size affects how the solvent runs through the column.• Smaller particles (higher mesh size i.e. 230-400 mesh) are used for flash chromatography & larger particles (lower mesh size i.e. 60- 120/60-200) are used for gravity chromatography.
  16. 16. Difference between Normal Phase & Reverse Phase Chromatography Normal Phase Chromatography Reverse Phase Chromatography • It uses a polar stationary phase and a • It uses a non polar stationary phase and non-polar (low Polarity Solvents) mobile a polar mobile phase. phase. • Non-polar compounds elute faster than • Polar compounds elute faster than non polar compounds. polor compounds. • When we increase polarity of mobile • When we increase polarity of mobile phase elution time will increase. phase elution time will decrease. • It can not be reused / reproducible • It Can reused / reproducible • Mobile phase are non polor i.e. IPA, • Mobile phase are polor compounds hexane, dichloromethane, chloroform, ethyl ether, and isopropyl alcohol (IPA). such as water, acetonitrile, methanol
  17. 17. Advantages & Disadvantages of column chromatography Advantages DisadvantagesIt can be used in both analytical and preparative applications. Time consuming ProcessIt is used to identify the number of components of a mixture.It is also used to separate and purify important quantities of those More amounts of Mobile Phasecomponents for subsequent evaluation (Solvents) requiredAny type of mixture can be separated Scale up process will take a long time to properly prepare & useAny quantity of mixture can be separatedThere is wider choice of Mobile Phase (Solvents) Automation makes the techniques more complicated & expensive It is low cost process and disposability of the stationary phase once it is used in the processProcess can be scale up form lab scale to commercial scaleAutomation is possible
  18. 18. Types of Company we need to focus Types of Department we need to contact• Pharmaceutical Industries – Bulk • R & D – Research & Development Drugs & API I. Organic Synthesis Lab, II. Medicinal Chemistry lab,• Nutraceuticals III. Novel Drug Discovery,• Herbal Extracts products IV. Clinical Research, manufacturers V. Pilot Scale lab, VI. Preparative Lab,• Research Laboratories VII. Semi Preparative Lab• Laboratories Chemical Repackers • Q.A. / Q.C.• Contract Research Laboratories • Process department / Production Department • Purchase – At Last
  19. 19. Silent FeaturesManufacturing since 1973 – Consistence suppliesThe product offered is highly active materialOur products has got higher surface areaThe product is having better & controlled pore volumesThe Pore diameter is strictly between 50-60AThe bulk density is lower, thus you require less qty of material on column.The product does not offer hydrolysis of your drugs after separations.We offer batch to batch reproducible results.Selectivity & kinetics are maintained constant ( better values)Higher theoretical plates counts.Manufactured under strict GMP normsISO 9001 accredited manufacturing firm
  20. 20. Comparison Between Silica Gel & Alumina OxideSilica Gel Alumina OxideChemical For mula SiO2 (Silicon Dioxide). Chemical Formula Al2O3 (Aluminium Trioxide ).it is acidic in nature which we are making it to It can be Acidic, Basic & NeutralneutralSilica Gel has Higher Surface area as compare Alumina’s Surface Area is less than Silica Gel Alumina i.e. 350-550 m2/gm 140-160 m2/gmSizes available: 35-70, 60-120, 70-230, 100-200, Size Available: 100-300 mesh & 200-400 mesh230-400 meshBulk Density: 040-0.65 gm/ml Bulk Density: 0.90-1.2 gm/mlPore Dia: 20, 60, 100, 300, 1000A Pore Dia: 50-60A