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Review on Thin Layer Chromatography

Preethi J1*, Harita B2 and Rajesh T3

1Acharya Nagarjuna University, Guntur, Andhra Pradesh, India

2ASN College of Pharmacy, Tenali, Andhra Pradesh, India

3Bhavan’s Vivekananda College, Osmania University, Hyderabad, Telangana, India

*Corresponding Author:
Preethi J
Acharya Nagarjuna University
Guntur, Andhra Pradesh, India
Tel: 7794080768
E-mail: [email protected]

Received Date: February 18, 2017; Accepted Date: March 21, 2017; Published Date: March 27, 2017

Citation: Preethi J, Harita B, Rajesh T (2017) Review on Thin Layer Chromatography. J Formul Sci Bioavailab 1: 107.

Copyright: © 2017 Preethi J, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Thin-layer chromatography (TLC) is a technique which is used to distinct non-volatile mixtures. This Thin-layer chromatography is can be executed on a sheet of glass, plastic, or aluminium foil, which is covered with a thin layer of adsorbent material, usually silica gel, cellulose or aluminium oxide (alumina). In this, the layer of adsorbent is called as the stationary phase. Chromatography is a division procedure that each natural scientist and organic chemist knows about.


Chromatography; Adsorbent; Stationary phase; Exploration


Chromatography is a partition strategy that each natural scientist and organic chemist knows about. I, myself, being a natural scientist, have routinely completed chromatographic partitions of an assortment of blend of mixes in the lab [1-3]. In fact, I was leafing through my exploration slides and ran over a pictorial portrayal of a genuine chromatographic partition that I had done in the lab.

To start with, as appeared in the Figure 1, I ran a thin layer chromatography (TLC) plate. This is essentially a rectangular bit of glass plate, covered with a thin layer of silica. I connected a spot of the response blend simply over the base of the plate (indicated with a strong line), and set the plate in a jug that contained a suitable natural dissolvable (for this situation, 1:1 volume by volume blend of hexane: ethyl acetic acid derivation was utilized), with quite recently enough volume to plunge the lower edge of the plate [4-6]. Bit by bit by slender activity, the dissolvable began ascending the silica plate, and as should be obvious the response blend isolated into 3 spots with unmistakable hues when the dissolvable had achieved the dissolvable front check [7,8] (Figure 1).


Figure 1: Thin layer chromatography.

Principle of Separation of Different Components

Differential affinities (quality of attachment) of the different parts of the analytic towards the stationary and portable stage bring about the differential division of the segments. Fondness, thus, is managed by two properties of the atom: 'Adsorption' and 'Dissolvability'.

We can characterize adsorption as the property of how well a part of the blend adheres to the stationary stage, while dissolvability is the property of how well a segment of the blend disintegrates in the versatile stage [9-11].

Higher the adsorption to the stationary stage, the slower the particle will travel through the segment (Figure 2).


Figure 2: A TLC plate with silica gel coating which is stationary phase.

Higher the solvency in the versatile stage, the speedier the atom will travel through the segment (Figure 3).


Figure 3: A TLC plate in the mobile phase within a TLC chamber.

Thin layer chromatography (TLC), is regularly found in research facility tests [12-14]. Thin layer chromatography utilizes a glass, metal, or plastic plate that is covered with the stationary stage, more often than not silica gel. A little drop of the blend that is being investigated is put a short separation from the base of the TLC plate. The TLC plate is then put into a chamber or tank with the versatile stage, similar to water, ethanol, (CH3)2CO, or a blend of solvents [15-19] (Figure 4).


Figure 4: A TLC experiment showing the separation of black ink into different parts.

Calculating Rf Values

In this experiment that all you need to know what is the number of various colors made up the blend, you could simply stop there [20-23]. In any case, estimations are regularly taken from the plate with a specific end goal to help distinguish the mixes introduce. These estimations are the separation went by the dissolvable, and the separation went by individual spots.

At the point when the dissolvable front draws near to the highest point of the plate, the plate is expelled from the measuring glass and the position of the dissolvable is set apart with a different line before it has an opportunity to vanish [24-27].

The Rf value for each dye is then worked out using the formula:


The Rf value can be utilized to recognize mixes because of their uniqueness to each compound [28-33]. When contrasting two distinct mixes under similar conditions, the compound with the bigger Rf value is less polar in light of the fact that it doesn't adhere to the stationary stage the length of the polar compound, which would have a lower Rf value.

Rf values and reproducibility can be influenced by various distinctive variables, for example, layer thickness, dampness on the TLC plate, vessel immersion, temperature, profundity of versatile stage, nature of the TLC plate, test size, and dissolvable parameters. These impacts ordinarily cause an expansion in Rf values. In any case, on account of layer thickness, the Rf value would diminish on the grounds that the portable stage moves slower up the plate.

Advantages of TLC

TLC is extremely easy to utilize and reasonable. Students can be shown this method and apply its comparative standards to other chromatographic systems [34-37]. There are little materials required for TLC (chamber, watch glass, slender, plate, dissolvable, pencil, and UV-light). Therefore, once the best dissolvable is discovered, it can be connected to different procedures, for example, High execution fluid chromatography.

TLC can be utilized to guarantee immaculateness of a compound. It is anything but difficult to check the virtue utilizing an UV-light. Recognizable proof of most mixes should be possible basically by checking Rf writing values. You can adjust the chromatography conditions effortlessly to build the improvement for determination of a particular segment.

Disadvantages of TLC

TLC plates don't have long stationary stages. Consequently, the length of partition is constrained contrasted with other chromatographic methods. Additionally, as far as possible is a ton higher. On the off chance that you would require a lower identification restrict, one would need to utilize other chromatographic procedures. TLC works as an open framework, so elements, for example, moistness and temperature can be outcomes to the consequences of your chromatogram [38-41].

The effect of saturation material

The RF values of all the metal ions were found to be zero on carbamide- formaldehyde layer impregnated with acidic admixture, ICF6 [NaDDC (20%)+H3PO4 (4%)], in the mobile phases. These perceptions bolster the way that metal dithiocarbamate buildings are shaky in acidic blends. The alkaline impregnation material, ICF5 [NaDDC (20%)+NaOH (4%)] was observed to be the best as it gives minimal spots for all the metal particles. Consequently, it appears that NaDDC go about as a complexing specialist and additionally adsorbent. The part of water solvent salts is smothered because of the overabundance of NaDDC (20%).

The effect of mobile phase

The RF values were found to be zero for the metal ions on thin layer of carbamide – formaldehyde impregnated with any of the six impregnation materials in the mobile phase of highest dielectric constant (ε=78.54) such as water. It is on line with the way that the dithiocarbamates of metal particles of nuclear number more than 20 are water insoluble [42-45]. The differential RF values have been acquired for the metal particles in low dielectric consistent versatile stage, for example, carbon tetrachloride (ε=2.24), benzene (ε=2.27), acetone (ε=20.7), ethanol (ε=24.5) and methanol (ε=32.7). Consequently unmistakably the RF estimations of metal dithiocarbamates rely on upon its dissolvability in the portable stage, the adsorption partiality and pH of the impregnation materials [46]. Along these lines, the most extreme quantities of detachments have been accomplished in least dielectric consistent portable stage that is carbon tetrachloride. Henceforth carbon tetrachloride is by all accounts better portable stages for metal particles chromatography on NaDDC utilizing carbamideformaldehyde polymer layer.

Chromatographic conditions

A basic, exact, fast, particular, and financial elite thin-layer chromatography (HPTLC) strategy has been set up for concurrent investigation of Domperidone (DMP), Paracetamol (PCM) and Tramadol Hcl (TMD) in tablet measurements shapes. The chromatographic divisions were performed on precoated silica gel 60254 plates with toluene-ethylacetate-butanol-smelling salts 5:4:1:0.2 (v/v) as portable stage [47-52]. The plates were produced in a 7.0 cm at encompassing temperature. The created plates were examined and measured at their single wavelength of greatest assimilation at roughly 278 nm for DMP and PCM, individually. Test conditions, for example, chamber measure, chamber immersion time, movement of dissolvable front, opening width, and so forth was basically examined and the ideal conditions were chosen. The medications were palatably settled with Rf 0.18 ± 0.02 for DMP, Rf 0.25 ± 0.02 for PCM and for TMD Rf 0.50 ± 0.02. The technique was approved for linearity, exactness, accuracy, and specificity [53-55]. The adjustment plot was straight between 100-600 ng/band for DMP, 3250-19500 ng/band based for PCM and 375-2250 ng/band based for TMD. The cut off points of location and evaluation for DMP were 9.95 and 30.16 ng/band, individually; for PCM they were 64.30 ng and 194.87 ng/band and for TMD 5.51 and 16.70/band. This HPTLC technique is financial, touchy, and less tedious than other chromatographic strategies. It is an easy to use and significance instrument for examination of consolidated tablet measurement frames.


The Thin layer chromatography is used in many applications such as identification, purification, testing and determination of active ingredient of pharmaceutical drugs. TLC helps in Separation of multicomponent pharmaceutical formulations, Qualitative analysis of alkaloids, cosmetology. It is the simple technique to separate the amino acids, it is a normal laboratory technique and also less time and less money consuming technique.


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