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.

1. Column Chromatography
PRODUCTS:
SILICA GEL
ALUMINIUM OXIDE

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. 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. 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. High Performance Liquid Chromatography

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. 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. 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 Column
Phase (Silica
(Solvents)
Gel or Alumina)

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. 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. 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. How does separation take place?

13. Types of Columns
Gravity Column Chromatography: Flash Chromatography:
Solvent is allowed to move down the Solvent is pushed down the column by
column by gravitational forces. positive air pressure

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. 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. 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. Advantages & Disadvantages of column chromatography
Advantages Disadvantages
It can be used in both analytical and preparative applications.
Time consuming Process
It 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 Phase
components for subsequent evaluation (Solvents) required
Any type of mixture can be separated Scale up process will take a long
time to properly prepare & use
Any quantity of mixture can be separated
There 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 process
Process can be scale up form lab scale to commercial scale
Automation is possible

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. Silent Features
Manufacturing since 1973 – Consistence supplies
The product offered is highly active material
Our products has got higher surface area
The product is having better & controlled pore volumes
The Pore diameter is strictly between 50-60A
The 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 norms
ISO 9001 accredited manufacturing firm

20. Comparison Between Silica Gel & Alumina Oxide
Silica Gel Alumina Oxide
Chemical 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 & Neutral
neutral
Silica Gel has Higher Surface area as compare Alumina’s Surface Area is less than Silica Gel i.e.
to Alumina i.e. 350-550 m2/gm 140-160 m2/gm
Sizes available: 35-70, 60-120, 70-230, 100-200, Size Available: 100-300 mesh & 200-400 mesh
230-400 mesh
Bulk Density: 040-0.65 gm/ml Bulk Density: 0.90-1.2 gm/ml
Pore Dia: 20, 60, 100, 300, 1000A Pore Dia: 50-60A