It is instrumental analytical technique. it is one of the major type of chromatography technique. its basic principle is adsorption. it has many applications in various fields
chromatography, principle, adsorbent of TLC, mobile phase of TLC, techniques in TLC, preparation of TLC plate, standards for TLC, advantages, disadvantages of TLC, Application of TLC.
In this slide contains principle, instrumentation, methodology, and application of gel chromatography.
Presented by: SATHEES CHANDRA (Department of pharmaceutical analysis).
RIPER, anantapur
chromatography, principle, adsorbent of TLC, mobile phase of TLC, techniques in TLC, preparation of TLC plate, standards for TLC, advantages, disadvantages of TLC, Application of TLC.
In this slide contains principle, instrumentation, methodology, and application of gel chromatography.
Presented by: SATHEES CHANDRA (Department of pharmaceutical analysis).
RIPER, anantapur
This presentation contains all the topics related to column chromatography. That includes introduction, principle,apparatus, experimental aspects of column chromatography, application of column chromatography, advantage and disadvantage of column chromatography with reference.
High Performance Thin Layer Chromatography (HPTLC) instrumentationMadhuraNewrekar
HPTLC is an advancement of TLC. It is a high performance liquid chromatography with automation compared to Thin Layer Chromatography(TLC).Speed, Efficiency and Accuracy are important advantages. Evaluation time is less due to updated automation in instrumentation.
Steps involved in HPTLC and the materials and instruments required in those steps are described in brief.
HPTLC- Principle, Instrumentation and Software (Abhishek Gupta)Abhishek Gupta
HPTLC is the improved method of TLC which utilizes the conventional technique of TLC in more optimized way
It is also known as planar chromatography or Flat-bed chromatography.
In this slide contains principle of IR spectroscopy and sampling techniques.
Presented by: R.Banuteja (Department of pharmaceutical analysis).
RIPER, anantpur.
introduction, history, principle, experimental techniques, evaluation on chromatogram, adv. & dis-adv., common problems, comparision, applications and analysis of drugs through TLC(2000-2017)
This presentation contains all the topics related to column chromatography. That includes introduction, principle,apparatus, experimental aspects of column chromatography, application of column chromatography, advantage and disadvantage of column chromatography with reference.
High Performance Thin Layer Chromatography (HPTLC) instrumentationMadhuraNewrekar
HPTLC is an advancement of TLC. It is a high performance liquid chromatography with automation compared to Thin Layer Chromatography(TLC).Speed, Efficiency and Accuracy are important advantages. Evaluation time is less due to updated automation in instrumentation.
Steps involved in HPTLC and the materials and instruments required in those steps are described in brief.
HPTLC- Principle, Instrumentation and Software (Abhishek Gupta)Abhishek Gupta
HPTLC is the improved method of TLC which utilizes the conventional technique of TLC in more optimized way
It is also known as planar chromatography or Flat-bed chromatography.
In this slide contains principle of IR spectroscopy and sampling techniques.
Presented by: R.Banuteja (Department of pharmaceutical analysis).
RIPER, anantpur.
introduction, history, principle, experimental techniques, evaluation on chromatogram, adv. & dis-adv., common problems, comparision, applications and analysis of drugs through TLC(2000-2017)
Thin-layer chromatography (TLC) is a chromatography technique used to separate non-volatile mixtures. Thin-layer chromatography is performed on a sheet of glass, plastic, or aluminium foil, which is coated with a thin layer of adsorbent material, usually silica gel, aluminium oxide (alumina), or cellulose.
This is about on TLC. and I hope it will helpful for you.
In this describe about their introduction, principle, application, procedure, methodology, RF value, and their advantage, disadvantage
Thank you😊
Thin layer chromatography technique - easier, cheaper.
Handling is easy. Used as an identification test also purity test. It comprises of stationary and mobile phase. There are various types of chromatography technique. TLC consists of three steps - spotting, development, and visualization. The Rf value is used to quantify the movement of the materials along the plate. Rf is equal to the
distance traveled by the substance divided by the distance traveled by the solvent. Its value is
always between zero and one. A TLC analysis might be summarized something like, "Using a silica
gel plate and ethyl acetate as the development solvent, unknown mixture X showed three spots
having Rf's of 0.12, 0.25, and 0.87". CThere are three components in TLC:
(1) the TLC plate (stationary phase), the development solvent (mobile phase), and the sample to be
analyzed (solute). In our experiment the TLC plate consists of a thin plastic sheet covered with a
thin layer of silica gel.
Chromatography : A seperation techniqueSHIVANEE VYAS
Chromatography is a method of seperating mixture of components into individual components through equlibrium distribution between two phases.
Each chromatographic method essentially consists of 2 phases a staionary phase and a mobile phase.
Stationary phase : solid or liquid
Mobile phase : liquid or gas
Selection of an animal model is one of the most important steps in any of the experimental pharmacological study.
Animal model preferred for the study must be producing similar disease profile as in the human.
Screening models for evaluation of anti ulcer activitySIVASWAROOP YARASI
A sore that develops on the lining of the oesophagus, stomach or small intestine.
Ulcers occur when stomach acid damages the lining of the digestive tract. Common causes include the bacteria H. Pylori and anti-inflammatory pain relievers including aspirin.
Upper abdominal pain is a common symptom.
Treatment usually includes medication to decrease stomach acid production. If it is caused by bacteria, antibiotics may be required.
Irritable bowel syndrome - diagnosis, pathophysiology and pharmacologySIVASWAROOP YARASI
irritable bowel syndrome (IBS) is a common disorder that affects the large intestine. Signs and symptoms include cramping, abdominal pain, bloating, gas, and diarrhoea or constipation, or both. IBS is a chronic condition that you'll need to manage long term.
ROLE OF IMMUNE CELLS IN CANCER AND TARGETING IMMUNE CELLS FOR CANCER THERAPYSIVASWAROOP YARASI
Cancer immunotherapy is a therapy used to treat cancer patients that involves or uses components of the immune system. Some cancer immunotherapies consist of antibodies that bind to, and inhibit the function of, proteins expressed by cancer cells. Other cancer immunotherapies include vaccines and T cell infusions.
Separation techniques are those techniques that can be used to separate two different states of matter such as liquids and solids.
Separation is an important asset to purify component of interest from a mixture.
Adulteration is a practice of substituting original crude drug partially or whole with other similar looking substances but the latter is either free from or inferior in chemical and therapeutic properties. Adulteration in simple words is the debasement of an article. OR Adulteration is broadly defined as admixture or substitution of original or genuine article/ drug with inferior, defective or otherwise useless or harmful substances.
ADULTRANT : The adulterant must be some material which in both cheap and available in fairly large amounts.
The growth and development of plants is regulated by a number of
chemical substances which together exert a complex interaction to
meet the needs of the plant. Five groups of plant hormones are well
established; they are the auxins, gibberellins, cytokinins, abscisic acid
and its derivatives, and ethylene. These substances are of wide distribution
and may, in fact, occur in all higher plants. They are specific
in their action, are active in very low concentrations, and regulate cell
enlargement, cell division, cell differentiation, organogenesis, senescence
and dormancy. Their action is probably sequential. Other hormones
concerned with flower formation and reproduction, but as yet
uncharacterized, have also been envisaged. The essential role of these
substances is illustrated by cell and tissue cultures; without the addition
of suitable hormones no development or cell division occurs. The effects of these very active substances on the production of
secondary metabolites, particularly with a view to producing plants
containing an enhanced proportion of active constituent, are of interest
to pharmacognosists. In such studies the manner in which the results
are recorded is all-important, particularly as the treatment may also
influence the size of the test plant compared with the controls. For
commercial purposes yield per hectare is an obvious criterion, whereas
for biosynthetic studies yield per plant or per cent fresh weight may be
of more significance. For final drug evaluation per cent dry weight is
the most likely requirement.
Oral controlled drug delivery systems - Various Approaches SIVASWAROOP YARASI
these are the drug delivery systems which are given orally and the drug release is such that it releases at a controlled way at a predetermined rate for a particular period of time.
this is an act that comes under Indian judiciary. it deals about the cultivation, supply and proper usage of narcotic substances. it has its own committee that regulates the activities according to the act.
liposomes are novel drug delivery dosage systems, where the drug is entrapped in phospholipid bilayered vesicles. the release of drug from the vesicles can be controlled or sustained.
the follwing presentation contain structure, classification and preparation methods, characterization and applications of liposomes.
the following document contains various diagnostic test for screening liver function. and interpretation of results, which may confirm the presence of a disease or disorder
Autacoids - pharmacological actions and drugs related to them. SIVASWAROOP YARASI
Autacoids or "autocoids" are biological factors which act like local hormones, have a brief duration, and act near the site of synthesis. The word autacoids comes from the Greek "autos" (self) and "acos" (relief, i.e. drug).
it is a method of miscellaneous instrumental analytical technique. it is one of the thermal analytical techniques used. it also has wide applications in the field of pharmacy.
The Art Pastor's Guide to Sabbath | Steve ThomasonSteve Thomason
What is the purpose of the Sabbath Law in the Torah. It is interesting to compare how the context of the law shifts from Exodus to Deuteronomy. Who gets to rest, and why?
Ethnobotany and Ethnopharmacology:
Ethnobotany in herbal drug evaluation,
Impact of Ethnobotany in traditional medicine,
New development in herbals,
Bio-prospecting tools for drug discovery,
Role of Ethnopharmacology in drug evaluation,
Reverse Pharmacology.
2024.06.01 Introducing a competency framework for languag learning materials ...Sandy Millin
http://sandymillin.wordpress.com/iateflwebinar2024
Published classroom materials form the basis of syllabuses, drive teacher professional development, and have a potentially huge influence on learners, teachers and education systems. All teachers also create their own materials, whether a few sentences on a blackboard, a highly-structured fully-realised online course, or anything in between. Despite this, the knowledge and skills needed to create effective language learning materials are rarely part of teacher training, and are mostly learnt by trial and error.
Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
This webinar will introduce you to my framework, highlighting the key competencies I identified from my research. It will also show how anybody involved in language teaching (any language, not just English!), teacher training, managing schools or developing language learning materials can benefit from using the framework.
The Indian economy is classified into different sectors to simplify the analysis and understanding of economic activities. For Class 10, it's essential to grasp the sectors of the Indian economy, understand their characteristics, and recognize their importance. This guide will provide detailed notes on the Sectors of the Indian Economy Class 10, using specific long-tail keywords to enhance comprehension.
For more information, visit-www.vavaclasses.com
Welcome to TechSoup New Member Orientation and Q&A (May 2024).pdfTechSoup
In this webinar you will learn how your organization can access TechSoup's wide variety of product discount and donation programs. From hardware to software, we'll give you a tour of the tools available to help your nonprofit with productivity, collaboration, financial management, donor tracking, security, and more.
2. Introduction:
• Thin layer chromatography was first discovered
by Izmailov and Schreiber in 1938. Further Stahl
(1958) perfected the method and developed
equipment and standardized adsorbents for the
preparation of uniform layers on glass plates.
And this technique, chromatography using thin
layers of an adsorbent held on a glass plate or
other supporting medium is called as thin layer
chromatography.
3. Definition:
• Thin layer of chromatography is a method if
analysis in which the stationary phase (a finely
divided solid) is spread as a thin layer on a rigid
supporting plate and mobile phase.
• TLC is often named by other names such as drop,
strip, spread layer, surface chromatography and
open column chromatography.
4. Superiority of TLC over other
chromatographic technique:
• Simple equipment. Low cost.
• Short development time.
• Separation of micro gram of the substances can be
achieved.
• Any type of compound can be analyzed.
• Wide choice of stationary phase. May b employed for
adsorption, partition (reversed phase also) or ion
exchange chromatography.
• Early recovery of separated components.
• Easy visualization.
• Sensitivity is better 10 to 100 times paper
chromatography.
• Variable thickness of thin layers.
• Chemically inert stationary phase.
5. Principle:
• The principle for separation is adsorption.
• One or more compounds are spotted on a thin layer
of adsorbent coated on a chromatographic plate.
• The m/p flows through because of capillary action.
• The components move according to their affinities
towards the adsorbent.
▫ More affinity towards the s/p – travels slowly
▫ Lesser affinity towards the s/p – travels faster.
• Thus the components are separated on thin layer
chromatographic plate based on the affinity of the
components towards the s/p.
6. Steps involved in TLC:
• Selection of adsorbent.
• Selection of glass plate.
• Coating of the adsorbent on to the glass plate.
• Activation of adsorbent.
• Purification of layer.
• Selection of mobile phase.
• Spotting of the sample.
• Development
• Visualization
• Quantitative and qualitative analysis.
7. Selection of adsorbent:
• A large no. of coating materials are available.
• Adsorbents classified into:
8. Adsorbent:
Acidic:
E.g. silica
For acidic analytes
Basic:
E.g. alumina
For basic analytes
Neutral:
E.g. keisleguhr,
Cellulose powder.
For neutral analytes
Adsorbent:
active
Has more active
sites for binding to
the analyte.
inactive
Has less active
sites for the
binding of analyte.
9. • As stationary phase a special finely ground matrix is
coated on the glass plate, a metal or a plastic film as
a thin layer ( approx 0.25mm).
• Adsorbent doesn’t adhere to plate properly. To
overcome this problem binders are used like
gypsum, starch, hydrated silicon dioxide etc are
added to the adsorbent. E.g. Silica gel G 60, where G
indicates gypsum and 60 is particle size.
• A fluorescent indicator like zinc silicate are added to
the adsorbent to simplify visualization of spot. The
advantage is non fluorescent U.V absorbing analytes
can be detected on a thin layer containing
fluorescent additive.
▫ E.g. silica gel GF
• Such substances show up as dark spots as green
fluorescent indicator.
10. Glass plates:
• Glass plates which are specific dimensions like
20cm × 20cm (full plate), 20cm × 10cm (half
plate), 20cm × 5cm (quarter plate).
• Microscopic slides can also be used for some
applications like monitoring the progress of
chemical reaction. The development time is
much shorter like 5mins.
• They should withstand temperatures used for
drying the plates.
11. Coating of adsorbent on glass plate:
• Main aim is to get uniform thickness of the layer.
• Many techniques are used:
• Pouring:
▫ Measured amount of slurry is poured and plate is tipped
back and forth to spread uniformly.
▫ Disadvantage – uniform thickness cannot be ensured.
• Dipping:
▫ Whole plate is dipped into slurry.
▫ Disadvantage – backside of plate is also coated and more
amount of slurry is required even to prepare fewer plates.
• Spraying:
▫ Suspension of adsorbent is sprayed onto plate.
▫ Disadvantage – uniform thickness cannot ne achieved.
12. • Spreading:
▫ Widely used.
▫ The slurry after preparation poured into a TLC
spreader and the thickness of layer is adjusted by
adjusting a knob in the spreader.
▫ Now the spreader is rolled on the plate or the plate is
moved while applying the slurry .
▫ Thickness of 0 – 2mm (0.25 for analytical purposes
and 2mm for preparative purpose).
• Pre coated plate:
▫ Ready to use thin layers are now available pre coated.
▫ These are expensive
▫ Thickness varies from 0.1 to 0.2mm.
13.
14. Activation of adsorbent:
• Coated plates are kept in air for 30 min and
then in hot air oven at 110°C for another 30 min.
• The dried plates can be stored in
thermostatically controlled oven or in
desiccators and can be used when required.
15. Purification or washing of plates:
• Silica gel contains iron as impurity which causes
a considerable distortion of chromatograph.
• To purify the air dried plates are given a
preliminary development with methanol-conc.
Hcl (9:1, v/v). The iron gets migrated with
solvent front to the upper edge of the plate.
• The plates are again dried and activated.
• Washing can be done for precoated TLC plates
also.
16. Selection of mobile phase:
• If the chemical nature if the sample that is to be
separated is known then it is possible to know a
suitable solvents by using original stalh’s triangle
which is inter-relating adsorbent activity, nature of
the solute and nature of the solvent.
• If the triangle is rotated so that at corner M points to
the type of mixture to be separated, this specifies at
corners S and E respectively, the necessary activity
of the adsorbent and the optimum polarity of the
eluent.
17.
18. • Suppose a mixture has hydrocarbons and
ketones. Then from stahl’s triangle, it is found
that an active adsorbent is required, together
with a non polar solvent.
• Mixtures of two or more solvents of different
polarity often give better separation than
chemically homogenous solvents.
• They should be as pure as possible.
19. Application of sample:
• Concentration of the sample or standard solution should
has to be minimum.
• The spots should be at least 2cm above the base of the
plate and spotting area should not be immersed in m/p.
• Agla micro syringe is used for quantitative work,
however capillary tubes can be used for qualitative work.
• To spot the plate, simply touch the capillary tube end to
the coated side of the plate. The solvent should quickly
evaporate leaving mixture behind on the plate.
• Excessive spotting leads to smearing, smudging and spot
overlap will result making identification of separated
components difficult.
20. Development:
Development tank:
• Different chambers of different sizes are used to hold
TLC plates
• Different development tanks are available
▫ Flat bottom chambers
▫ Twin trough chambers – it require less solvent and two
plates can be developed.
▫ Cylindrical tanks
• Chamber saturation should be done.
• Tank should be lined inside with filtered paper
moistened with m/p so as to saturate with atmosphere.
• If chamber saturation is not done edge effect occurs.
• Edge effect: where the solvent front moves faster in
middle of the plate than that of the edges. Therefore
spots are distorted and not regular.
21.
22. Development techniques:
• One dimensional development or vertical :
▫ Conventional method
▫ Solvent flows against gravity, because of capillary action. (bottom
to top).
• Horizontal TLC:
▫ plate is kept in horizontal manner.
▫ Spotting is done in middle of the plate and m/p is added slowly
through sides.
• Descending TLC:
▫ Flow of solvent is assisted by gravity and hence development is
faster.
▫ Solvent holder is on top.
• Multiple development TLC:
▫ Similar to vertical TLC.
▫ After developing once, the plate is dried and then again kept in
same mobile phase (same composition) and in same direction.
▫ Without detection multiple developments are done.
▫ It is done to separate some complex mixture ( more no. of
compounds).
23. • Step wise TLC:
▫ Plate size is bigger 30cm plate.
▫ Usually done when development distance is long.
▫ Development:
Allow first m/p to travel upto 15 to 18cm and then it is stopped and
dried.
Now it is again kept in another m/p and development is done.
• Two dimensional TLC:
▫ First the plates are developed in one axis and the plates after drying are
developed in other axis.
▫ When large number of compounds or complex mixtures are need to be
separated this method can be followed.
▫ Either same solvent or different solvent system can be used.
• Gradient TLC:
▫ Isocratic – same composition of m/p used throughout development .
▫ Gradient – ratio of m/p changed.
• Fractionating TLC:
▫ Spotting is done in the middle at top of the plate and the m/p is forced
though the edges of the plate.
▫ Different fractions are collected at different times based the affinity of
the analyte towards the adsorbent.
24. Visualization:
• Colored spots can be visually detected. But for
detecting colorless spots, the following techniques
are used:
• Non specific methods: no. of spots can be
detected but not exact nature or type of compound.
▫ Iodine chamber method : where brown or amber
colored spots are observed when the paper are kept in
a tank with few iodine crystals at the bottom.
▫ UV chamber for fluorescent compounds: when
viewed under UV chamber at 254 or at 365 nm,
fluorescent compounds can be detected.
25. • Specific methods: specific spray or detecting or
visualizing agents are used to find out the nature of
compound or for identification purposes.
▫ Eg.
Fecl3 for phenolic and tannin compounds
ninhydrin for amino acids.
• The detecting techniques can also be categorized as:
▫ Destructive technique: when specific spray agents
are used the samples are destroyed before detection.
Eg. Ninhydrin reagent
▫ Non destructive technique: methods like UV
chamber, iodine chamber, densitometry method
doesn’t destroy the sample even after detection.
26. Qualitative analysis:
• Rf value:
▫ Rf= dist travelled by solute/dist travelled by solvent
front
▫ value ranges from 0-1
▫ Ideal values are 0.3-0.8.
▫ It is characteristic to each compound in a particular
combination of sp and mp.
▫ The unknown compound can be identified by
comparing its rf values with standard’s.
▫ if rf = 1 – analyte move along with m/p without
separating.
▫ If rf = 0.1 – analyte is adsorbed to adsorbent and not
moving.
27. • Rx value:
▫ Distance travelled by sample/dist travelled by
standard.
▫ It is always closer to 1.
▫ m/p allowed to travel throughout the plate.
• Rm values:
▫ To find whether compounds belong to a
homologous series.
▫ It is a combined value.
▫ Delta Rm = log (1/Rf – 1).
28. Quantitative analysis:
• Can be carried out in two ways:
• Direct method: quantitative determination is
undertaken directly on the layer.
• Indirect method: the substances are removed from
the adsorbent and then determined after elution.
• Direct methods:
▫ Visual assessment of chromatogram.
▫ Determination by measurement of spot area – this method
is based on a relation between spot and amount of the
substance present.
▫ Quantitative TLC incorporating densitometer.
▫ Direct spectrophotometry on thin layer-
By evaluation of wavelength of maximum absorbance.
Characterization of chromatogram zones by reading the
absorption or fluorescence curves directly from TLC is done by
chromatogram spectrophotometer introduced by zeiss, stahl
and jork.
29. • Indirect method:
▫ Done after eluting the individual spots with
suitable solvent and then filtering off the
stationary phase.
▫ The exact quantity of compounds can be
determined by conventional methods like
colorimetry, uv spectrophotometry, fluorimetry,
flame photometry, electrochemical methods of
analysis.
▫ The spotted area is scooped using certain
apparatus.
30. Kips apparatus:
• Also a kind of destructive method.
• It has two arms, one connected to vacuum and the
other collects the spots.
• It has a sintered glass filter in arm that is connected
to vacuum.
• It has a bulb with extracting solvent in between
these two arms.
• Vacuum tries to suck the particles at the spot and
these particles fall into bulb and sintered glass filter
prevent the particles getting into the vacuum pump.
• The particles fall into the blub with extracting
solvent and the s/p is filtered off if present.
• Now certain conventional methods like colorimetry,
uv are used.
31. Applications:
• TLC is very commonly used technique in synthetic
chemistry for identifying compounds, determining
their purity and the progress of a reaction.
• Separation of vitamins, antibiotics, proteins,
alkaloids, glycosides.
• Identification of drugs e.g. amino caproic acid,
digitoxin, levodopa.
• Detecting the decomposition products in drug.
• Identification of organic compounds
▫ Even no. alcohols from decanol through
hexacosanolcan be separated on keiselguhr G with
cyclohexane as a developing solvent