The document discusses thin layer chromatography (TLC), a technique used to separate mixtures based on differential partitioning between a mobile phase and stationary phase. TLC involves preparing a TLC plate by coating it with an adsorbent stationary phase. Samples are applied as spots and the plate is developed in a mobile phase, allowing separation. Visualization identifies the components, which are used to analyze samples and identify unknown compounds. TLC is useful for separating polar compounds like amino acids, sugars, and natural products.

1. (I.S.F. COLLEGE OF PHARMACY MOGA)
Presented by-
Atul Chaudhary
M.PhaRm. First Year
PHARMACEUTICS

2.  Chromatography is a laboratory technique for
the separation of a mixture. The mixture is dissolved in a
fluid called the mobile phase, which carries it through a
structure holding another material called the stationary
phase. The separation is based on differential
partitioning between the mobile and stationary phases
Subtle differences in a compound's partition coefficient
result in differential retention on the stationary phase
and thus affect the separation2/7/2020 2T.L.C. ASSIGNMENT

3.  Thin-layer chromatography (TLC) is a widely
employed laboratory technique used to separate
different biochemicals on the basis of their relative
attractions to the stationary and mobile phases.
 TLC is very versatile; multiple samples can be
separated simultaneously on the same layer, making
it very useful for screening applications such as
testing drug levels and water purity.
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6.  The slurry, which is a mixture of stationary phase and
water is prepared by using the ratio mentioned earlier.
After preparing the slurry, the TLC plates can be
prepared by using any one of the following techniques.
 Pouring
 Dipping
 Spraying
 Spreading
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7.  Normal thickness of 0.25cm is used for analytical
purpose and 2mm thickness for preparative purpose
 Then the spreader is rolled only once on the plate.
 The plate are allowed for setting . This is done to
avoid cracks
 The plates are activated by keeping in an oven at 100
degree centegrade to 120 degree centigrade for 1
hour
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9.  2 – 5 of 1% solution of sample or standard is spotted
using a capillary tube or micropipette. The spots
should be kept at least 2cm above the base of plates
and the spotting area should not be immersed in
mobile phase in a developing chamber.
 The sample is applied on the narrow bands
 The width of the band must be as narrow as possible
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11.  Step 1: Prepare the developing container
 Step 2: Prepare the TLC plate
 Step 3: Spot the TLC plate
 Step 4: Develop the plate
 Step 5: Visualize the spot
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13.  UV Light: For aromatics + conjugated systems.
Iodine: Visualizes ~half the time.
 Anisaldehyde: For many aldehydes, ketones, and
alcohols.
 Vanillin: For many aldehydes, ketones, and
alcohols.
 Permanganate: For alkenes, alkynes, or oxidizable
groups (aldehydes, alcohols).
 Phosphomolybdic Acid (PMA): For alcohols,
phenols, alkenes, and many carbonyl compounds.
 Iron(III) Chloride: For phenols.
 Bromocresol Green: For acidic compounds
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14.  TLC is more useful for the analysis of polar compounds
like Amino acids, Sugars, Natural products etc.
 The Different types of applications are listed below
1. Seperation of mixture of drugs of chemical or biological
origin, plants extracts etc,
2. Seperation of carbohydrates (sugar) Vitamins, proteins,
alcohals, glycosides, aminoacids etc
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15. 1. Identifications of drugs
2. Identifications of related compounds in drugs
3. To detect the presence of foreign substance in
drugs
4. To detect decomposition products in drugs
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17.  Bernard Fried and Josef Sherma
 Prem kumar Jaiswal
 Egon Stahi
 Chatwal
 Google Wikipidia
 Slideshare
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