Formal Report On Exp 5

Formal Report on Exp 5
COLUMN AND THIN LAYER CHROMATOGRAPHY
Mark Paul P. Pastrana, Mariah Ericka M. Patawaran, Princess Juneire M. Peligro,
Francisco Q. Pua III, Rose Anne L. Quyo and Janille P. Ragpa
Group 8 2B Medical Technology Organic Chemistry Laboratory
ABSTRACT
The main objectives were to separate the colored components of malunggay leaves by means of
column chromatography, as well as to determine the purity of the components using thin layer
chromatography (TLC) and measure the Rf values of the colored components obtained herein. For
column chromatography, the sample prepared was loaded into a Pasteur pipette plugged with cotton
and uniformly packed with silica gel. The eluents used were 7 mL hexane:acetone (7:3), 5 mL
hexane:acetone (1:1), 5 mL ... Show more content on Helpwriting.net ...
Without letting the column run dry, hexane:acetone (1:1) was introduced into the column, and in the
same manner the eluates were collected. This was the same for the succeeding eluents, and went on
until no more colored eluates could be obtained from the column. Cotton
Cotton
Silica gel
Silica gel
Pasteur pipette
Pasteur pipette
Iron clamp
Iron clamp
Iron stand
Iron stand
Figure 2 Column Chromatography 3. Thin Layer Chromatography The eluates obtained from
column chromatography were applied on a TLC plate pre–coated with silica by spotting it seven
times per color using a capillary tube. Each spot was dried before applying the next. A developing
chamber was prepared by placing an amount of the solvent system, hexane:acetone (7:3), into a
beaker. Filter paper was used to line the walls of the beaker, and was then covered with a watch
glass to equilibrate the chamber. Once the filter paper was saturated with the solvent system, the
TLC plate was carefully placed in the beaker to develop. When the solvent system had reached
about a centimeter from the upper end of the TLC plate, the plate was removed, and before allowing
it to air–dry, the solvent front was marked. Once air–dried, the plate was placed under a UV lamp to
visualize the components to determine any additional colors that were invisible without UV light.
Solvent system

Solvent system
Beaker
Beaker
Watch Glass
Watch Glass
Figure 3.1 Thin Layer Chromatography
Figure 3.2 Thin Layer
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Thin Layer Chromatography ( Tlc )
Thin layer chromatography (TLC) is a technique used to identify components within a compound
and determine the compounds purity. The purpose of this experiment is to determine what solvent
works the best at separating the different pigments of the paprika extract. The solvents used to
develop the plate are hexane and methyl tert–butyl ether (MTBE), as well as three mixtures of the
two solutions (5% MTBE in hexane, 10% MTBE in hexane, and 30% MTBE in hexane). Hexane is
a non–polar alkane that is used as a solvent for many different things and MTBE is a polar ether that
is used as a fuel additive. In this experiment, five TLC plates were set up and ran in each of the five
prepared solvents. There are four steps when performing TLC which include: spot (touch the
capillary to the plate briefly so the compound runs out and forms a small spot on the plate), develop
(choose a solvent to develop the plate, then place the slide in the developing chamber), visualize
(when the solvent reaches the stop line, remove and visualize the data), and calculate (measure the
distance between the spot and start line to calculate the Rf values). Rf= Distance traveled by spot
Distance traveled by solvent Procedure Extraction of carotenoids from paprika powder. First weigh
out 0.5 g of paprika powder and place it in a 50 mL beaker. Then add a magnetic stir bar to the
beaker and 5 mL of methyl tert– butyl ether (MTBE), allowing it to stir for 5–10 minutes. Vacuum
filter the solution

Thin Layer Chromatography Lab Report
The Thin Layer Chromatography of Drugs lab taught me how chromatography could be used to
identify the compounds of an unknown drug. This procedure relied on using compounds that had a
known travel distance, which served as the control, and examining how far the compounds in the
unknown drug traveled. The results of this experiment indicated that drugs are chemical molecules
with a diverse amount of compounds i.e.: caffeine, acetaminophen, asprin, etc. The essentials of this
lab relied on two factors that allow for the separation of different compounds from the drug. The
first factor is how soluble the compound is in the given solvent. The more soluble the substance, the
farther it will travel due to the bond strength between it and the solvent. ... Show more content on
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This is due to DCM being able to effectively dissolve organic substances. In this experiment DCM
and water were both mixed with an impure organic compound in a test tube. DCM was able to
isolate the organic compound from the water. This was due to the organic compound dissolving in
the DCM rather than in the water. Additionally, what makes DCM a good organic solvent is the fact
that it is denser than water. This allows for the distinction between the DCM layer and water layer,
since the DCM layer will collect at the bottom of the test tube. It can then be transferred out of the
test tube and collected, thus separating the organic compound from the

Paper Chromatography Lab
The objective of this "Chromatography Lab" is to determine the number of substances and their Rf
values present as dyes in different markers/inks.
Chromatography deals with physically separating a mixture into it's very own components.
Chromatography is used for keeping compounds from mixtures. The process that is involved in it, is
separating out a mixture of chemicals, that are most likely either in a liquid or gas form. This
process allows the chemicals/substances, to past each other, which is either a liquid or solid. The
main type of chromatography process is paper chromatography. Paper chromatography process was
what we used for/during this lab. Paper chromatography is used to separate the components of ink,
dyes, plant compounds (chlorophyll), make–up, and other substances. The other chromatography
processes include: Liquid, Thin–Layer, and Gas. Liquid chromatography is used for identifying
unknown plant pigments and other compounds. Thin–layer chromatography uses thin plastic or
glass trays to identify the composition of pigments, chemicals, and other unknown substances. Gas
chromatography is used to figure out the chemical composition of unknown substances, like the
different compounds that are in gasoline. ... Show more content on Helpwriting.net ...
The procedures for this "Chromatography Lab" include eight steps, which are talked about down

What Is Thin Layer Chromatography
FUNDAMENTAL UNITS OF LIFE ARE found in molecules consisting of amino acids. In the
center of every amino acid is an α–carbon with an amino group, a carboxy group, a hydrogen, and a
side chain group (R–group) attached to it. The difference in side chain groups (R–groups) is what
distinguishes the different amino acids. If an amino acid has a positively charged (+) side–chain, it
is a basic amino acid. If an amino acid has a negatively charged (–) side–chain, it is a acidic amino
acid. If an amino acid has a polar side–chain its categorized as a polar amino acid, and if it had a
nonpolar side–chain is is categorized as a nonpolar amino acid. There are two chiral formations for
every amino acid : D–formation and L–formation. The formation of the ... Show more content on
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It can give the polarity of that solution, reactivity of the solution, the phase of the reaction of the
solution and the hydrophobicity of the solution. With this, one can determine the type of bonds in a
solution relative to the other solutions as well as relative to the developer. There are three steps to
the process of Thin Layer Chromatography; spotting, development (mobile phase), and
visualization. During the spotting process solution(s) is/are dabbed with a capillary tube on the
origin line of the TLC sheet to the required amount to give a starting point. TLC sheets have a polar
silica gel on them which will react with the dabbed solution(s). The more polar the solution the
more it will react with the polar silica gel. Once placed in the developer tank the mobile phase
begins, as the somewhat polar developer (less polar than the silica gel) will move up the the TLC
sheet through capillary action. The amino acids that are more hydrophobic will move along with the
developer upward and away from the silica gel whereas the more hydrophilic amino acids will be
more attracted to the polar silica and will move less than hydrophobic amino acids which are more
attracted to the mobile phase caused by the developer. The solvent front is determined when the
developer has almost reached to top of the TLC sheet. It is important to avoid it reaching to top,
because it can cause a skew in data if allowed. The amino acids that are first to reach the top will sit
at the top while the other less hydrophobic amino acids will get closer causing a skewed perspective
in polarities between the amino acids at the top and the rest. After this the TLC sheet is removed, the
solvent front is drawn, and the sheet now needs to dry. The next step in the development phase is
helpful when identifying differences in amino acids. The TLC sheet is sprayed with ninhydrin which
catabolizes the amino acids, then

The objective of experiment 6 was to learn how to separate an unknown mixture by gas
chromatography in order to analyze the data about the unknown compounds in the mixture.
The objective of experiment 7 was to utilize a thin–layer chromatography to test the purity and
contents of each vial from the previous experiment.
Introduction The process of separating unknown mixtures with similar polarity to its counterparts
pose a problem for most. One method to separate a mixture based on interactions, in the gaseous
phase, is through a process known as gas chromatography. Gas chromatography consists of a
mixture going through a mobile phase, being immersed in a gas, and a stationary phase, being
physically separated through a non–volatile liquid. A gas chromatograph with a flame ionization
detector consists of several components. The components of a gas chromatograph are a heated
injection port (to inject an ... Show more content on Helpwriting.net ...
Thin–layer chromatography (TLC) consists of a TLC plate (stationary phase) that is partially
immersed in a solution (mobile phase) to separate compounds on it, based on polarity. When an
unknown compound, with distinct polar components, is placed on the TLC plate, the components
with low polarity will have traveled the farthest from the start line it was set on. A compound with
low polarity also has a large Rf value () on a TLC plate. The Rf value is a ratio that depicts how
much a compound interacted with the TLC plate. However, the compounds from experiment 5 were
separated with an adsorption chromatography column. These compounds will only have one band
on the TLC plate, since they should be pure compounds. The comprehension of TLC
chromatography and how to properly separate compounds with an adsorption chromatograph with
an adsorption chromatograph column were essential to produce quantitative data in experiment

Thin Layer Chromatography (TLC) is an extremely useful technique for monitoring reactions. It is
also used to determine the proper solvent system for performing separations using column
chromatography. TLC uses a stationary phase, usually alumina or silica, that is highly polar
(standard) or non–polar (reverse phase), and a mobile phase, some solvent whose polarity you will
choose. In 5.301, and in most lab applications, you will use standard phase silica plates. You will
apply your reaction mixture in solution to the plate then "run" the plate by allowing a solvent (or
combination of solvents) to move up the plate by capillary action. Depending on the polarity of the
components of the mixture, different compounds will travel different distances up the plate. More
polar compounds will "stick" to the ... Show more content on Helpwriting.net ...
3) Fill TLC chamber with 1–2 mL of the desired solvent system. Place a large piece of cut filter
paper in the chamber as well. 4) Spot the compound on the baseline of the TLC plate. We will use
commercial spotters, but spotters can be pulled from hot Pasteur pipets –you may see this in your
UROP. If you are monitoring a reaction, make sure to spot the starting material, the reaction
mixture, and a co–spot of both. 5) Run the TLC. Let the solvent go about 90% of the way up the
plate. 6) Remove the plate from the chamber and mark the solvent front immediately with a pencil –
you will use this to calculate the Rf. 7) Let the solvent dry off of the

PRACTICAL REPORT ON THE ISOLATION AND
IDENTIFICATION OF...
PRACTICAL REPORT ON THE ISOLATION AND IDENTIFICATION OF CODEINE AND
PARACETAMOLPRACTICAL REPORT ON THE ISOLATION AND IDENTIFICATION OF
CODEINE AND PARACETAMOL
AIM: To extract codeine and paracetamol from its tablet by solvent extraction and tentatively
identify in comparison to standards using Thin Layer Chromatography.
INTRODUCTION:
Codeine or methyl morphine, an alkaloid, was first isolated in 1832 from raw opium. It
concentration ranges from 0.2% to 0.8%. Mostly used for its analgesic, anti–tussive and anti–
diarrheal capabilities (Tremlett, Anderson and Wolf, 2010). Paracetamol also known as
acetaminophen (n–acetyl–p–aminophenol, APAP) on the other hand, is a useful non– steroidal anti–
inflammatory drug (NSAID). It is commonly ... Show more content on Helpwriting.net ...
On a thin chromatography plate, five spots were placed ( as shown in table 2) and the plate was
developed using chloroform/methanol. This was later visualized with dragendorff's reagent under
the UV light. All separated components were observed, identified and recorded.
RESULTS:
Table of observed pH
SOLUTION Initial pH Final pH
Basified sample 10 12 TABLE 1
Table of Retention factor (RF value)
Rf = Distance travelled by the substance (cm) Distance travelled by the solvent (cm)
SUBSTANCE Distance travelled by substance (cm) Distance travelled by Solvent (cm) Retention
factor value (Rf)
Chloroform extract 3.0 4.0 0.75
Codeine positive control 3.0 4.0 0.75
Paracetamol positive control 4.0 4.0 1.00
Chloroform (negative control) 3.5 4.0 0.86
Diluted sample 4.0 4.0 1.00 TABLE 2
DIAGRAM:
Fig 3: The Developed Chromatographic Plate.
DISCUSSION:
Running the chloroform extracts and diluted sample together with two positive controls and a
negative control on a single chromatographic plate simultaneously, the retention factor(Rf) of five

different samples were determined. The RF value of the chloroform extract(0.75) tallied with that of
the codeine positive control and that of diluted sample(1.00) with the paracetamol positive control.

Acetaminophen Chromatography
Introduction The purpose of this experiment is to analyze the following analgesics: acetaminophen,
aspirin, caffeine, and ibuprofen. This analyzes is to be done using thin layer chromatography (TLC).
Also, in this experiment we isolate β–carotene from spinach using column chromatography (CC).
Chromatography is a separation technique. It separates a mixture in one state of matter by moving it
through another substance in another state of matter (Woodford, 2016). Chromatography consists of
a compound that does not move in the entire experiment (stationary phase) and another compound
that runs through the stationary phase (mobile phase).
Thin layer chromatography uses a thin sheet of silica gel as the stationary phase. The
chromatography plate is then to be placed vertically inside a chromatography chamber (Thin layer
chromatography, NA). Column chromatography works very similar to TLC, but instead of using a
thin layer of silica gel, in this case we fill a column with this gel (University of Toronto, NA).
Finally, we are looking to calculate the value of Rf for every substance. Rf is defined as the distance
travelled by the compound divided by the distance travelled by the solvent in the TLC experiment
(Weldegirma, 2016). ... Show more content on Helpwriting.net ...
The first part of this experiment uses thin layer chromatography (TLC) while the second part
involves column chromatography

2. Materials and Method
All the chemicals were obtained from commercial sources and used without further purification.
Melting point was measured in digital melting point apparatus (Veego, VMP–DS) model. 1H
nuclear magnetic resonance (NMR) spectral was recorded at room temperature on a 300MHz liquid
state NMR spectrometer in (Brüker Biospin, Switzerland) using tetramethylsilane as internal
standard. The reactions were monitored by thin–layer chromatography (TLC) using precoated plates
(Merck). All solvents utilized in thin layer chromatography were distilled before use.
2.1. Experimental
General Procedure for Thia–Michael Addition of Thiols to 2–nitro–3–phenyl–3–
(phenylthio)propan–1–ol and its derivatives (6a–6f)
Thiol 2.45g of 1 equiv (22.33 mmol) was added to a mixture of Nitroolefins 4g of 1 equiv (22.33
mmol) in DAPCO 2 equiv, and the mixture was vigorously stirred at room temperature until the
nitroolefin was consumed completely (5 ... Show more content on Helpwriting.net ...
Elemental Analysis for C15H15NO3S: Calculated: C 62.26; H, 5.23; N, 4.84; O, 16.59; S, 11.08;
Found: C 62.26; H, 5.23; N, 4.84; O, 16.59; S, 11.08.
3–(4–ethylphenyl)–2–nitro–3–(phenylthio)propan–1–ol (6b):
Yield: 88%; Colourless solid; mp: 92–93 ˚C; 1H NMR (300 MHz, CDCl3): δ 1.19 ( t, J=7.8 Hz,
3H), 2.24 (bs, 1H), 2.58 (q, J= 7.8 Hz, 2H), 4.26–4.36 (m, 2H), 4.65 (d, J=10.2, 1H), 4.98–5.05 (m,
1H), 7.08–7.27 (m, 9H); 13C NMR (75 MHz): δ 15.26, 28.44, 52.55, 62.39, 92.22, 127.85,
128.20, 128.57, 129.13, 132.22, 133.71, 133.84, 144.48. Elemental Analysis for C17H19NO3S:
3
Calculated: C 64.33; H, 6.03; N, 4.41; O, 15.12; S, 10.10; Found: C 64.33; H, 6.03; N, 4.41; O,
15.12; S,

Spinach Paper Chromatography
In this experiment, the process of chromatography was taken place in order to identify the different
types of pigments present in spinach leaf. This was done by carrying out two types of experiments
which are thin layer chromatography (TLC) and paper chromatography. Both of the experiments
were done using a similar procedure except that they both used a different stationary phase. Paper
chromatography used paper, whilst, TLC used a silica plate. Propanone was used as the extraction
solvent for spinach leaves and chromatography solvent was used as the mobile phase in the
experiment. The results portray that the pigments has rose to different heights through the stationary
phase as they separated into different coloured pigments. The only pigments ... Show more content
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A line was drawn with a pencil approximately 1cm from, and parallel to the bottom of the plate.
Plant material was placed into a mortar with a pinch of sand and was grinded with a pestle, to help
the plant material break down into a liquid form. 1–2cm of propanone was added into the mortar
along with the plant material and was grinded until the liquid appeared to be a dark green colour. A
micropipette tip was dipped into the liquid and a tiny drop of the extract was transferred onto the
middle of the pencil line on the TLC plate. Care was taken to ensure that the spot got no bigger than
3mm diameter. More drops were added each time after the spot dried properly. The drops were
added until the spot turned dark green which took about 5–10 drops. 10ml of running solvent was
added into the chromatography tank (made up of 5 parts cyclohexane: 3 parts propanone: 2 parts
petroleum ether). The chromatography plate was placed into the beaker so that the plate dips into the
solvent making sure the pigment spot was above the surface of the solvent. The beaker was covered.
The chromatogram was left inside the beaker for about 5–10

Thin Layer Chromatography
TITLE : Reactions and separation of sugars by TLC.
AIM : Thin–Layer Chromatography can show many different characteristics of a mixture. It is
recognized for isolation , separation ,identification, and anaylsis of the mixture's components. The
purpose of this experiment is to separate carbohydrates into its pure components such as mixtures of
monosacrides by TLC. TLC is used to identify sugars in normal and pancreatic disease urine, the
procedure is easy and reproducible .
INTRODUCTION: technique and applications for sugar crhomatography have continued to advance
steadily since publication of previous edition in 1969. Though thin layer chromatography has
become most popular for rapid identification.
Sugars exist in solution as an equilibrium mixture of open chain and closed ring(orcyclic), strutures.
sugars that can be oxidised by mild oxidising agents are called reducing sugars because oxidising
agent is reduced in the reaaction. A non reducing sugar is not oxidised by oxidising agent . All
common monosaccrides are reducing sugars. there are several test forr sugars which are based on
composition or specific group. Hence sucrose is not a reducing agent due to bonding of oxygen with
carbon to form glycosidic bonds between monosacrrides. Therefore they are not free to let go so
cant be reduced , some of test includes seliwanoff 's test which distinguish between aldose and
ketose sugars.Other test for sugars is benedicts test which commonly used to detect presence of

Abstract. Thin Layer Chromatography is a simple procedure that allows you to determine how many
and what kind of compounds are in a mixture. By understanding the properties of the TLC system
and how it combines with the functional groups of amino acids the retardation factor (Rf) values can
be calculated and compared to specific hydropathy values of amino acids. Thin layer
chromatography is used to separate and recognize a certain compound. The silica gel that is located
on the outside of the TLC sheet functions as the stationary phase and the solvent mixture functions
as the mobile phase. Amino acid solution is added to the thin layer chromatography sheet and is then
placed in a developing tank that contains developer/solvent; only the bottom of the sheet is touching
the solvent. This solvent acts as the mobile phase, and the amino acids slowly will rise up the TLC
sheet by capillary action. A very important component of TLC is Rf value. Which measures the
distance a substance has moved ... Show more content on Helpwriting.net ...
By separating mixtures and identifying how different compounds can work together by moving
within the developer. The amino acids used interact in two phases, the mobile and the stationary
phase. Thin layer chromatography works mostly on capillary attractions. Due to surface tension
interactions the mobile phase is forced upward to the stationary phase because of the capillary
attractions. A–amino acids are the 20 common amino acids. They are made up of a carboxyl group
and an amino group, which is bonded to a carbon atom. Their side chains, or R groups is what
makes each amino acid unique, the R groups vary in size, electrical charge, and structure. Ninhydrin
is used to detect the amino acids, because they are otherwise colorless. When the ninhydrin reacts
with the amino acids it gives off a purple colored spot. The amino acids can be identified once the
Rf is calculated and compared with reference

Thin Layer Chromatography Lab
Thin layer chromatography is the separation of substances using different solvents that run up the
TLC plate to find different materials and colors in the sample. Three solvents in this test were one
hundred percent acetone, one hundred percent isopropyl, and distilled water. In this test we timed
the speed of each solvent and recorded the best color content.
PACE Introduction
Thin Layer Chromatography is a commonly used experiment in forensics, and is the separation of
substances using different solvents. What I want to find out is what the fastest solvent is, but keeps
the best color on the TLC plate. The three solvents in the experiment are one hundred percent
acetone, one hundred percent isopropyl, and distilled water. Acetone is nail polish remover and
isopropyl is rubbing alcohol. In the experiment, it required distilled water because it is purified and
is always the same substance in the container. ... Show more content on Helpwriting.net ...
TLC was invented in 1941 for forensics, but wasn't perfected on paper until 1944(chromatography–
online.org). Schaiber was the one who invented TLC and he was a German scientist. Some other
facts are that there are many other ways to do TLC and that it's not a very old process.
Forensic scientists use this in cases dealing with a lot of drugs or explosives. They use much more
complicated equipment though to perform their tasks. Many explosives can be tested and most
common drugs can be tested too. Some reasons to test these substances are, one the government
doesn't want people to have drugs or explosives, and two, and people might find out and get really
concerned. All in all this is very important to society. The hypothesis is what would be the fastest
solvent that will keep the best color

In this experiment, the objective was to use thin layer chromatography to identify the major active
ingredients in commercial analgesic preparations. TLC is a method to separate compounds and to
see how many compounds are present in the mixture. The separation into components is also
dependent on the solvent used. When TLC is performed, the Rf values are determined for the
sample and they are compared to the Rf values of the standards. Similar Rf values help identify the
standards that might be present in the sample. I used TLC to identify the major active ingredient in
Aleve, a pain medication. Six standards were used to determine which was the main ingredient in
Aleve, but only three standards had similar Rf values to the Rf values of Aleve. The standards and
their Rf values are summarized below. Rf Values S2: 0.67, 0.83 B1: 0.66 S5: 0.55 B2: 0.65 S6: 0.39
... Show more content on Helpwriting.net ...
Comparing Rf values, S2, Ibuprofen, has a similar Rf value to Aleve, but because it separated into
two components, one having a higher Rf value than Aleve, it is not likely that the main ingredient in
Aleve is ibuprofen. Standard S5, naproxen sodium, has a smaller Rf value than Aleve, but it is the
next closest one. Comparing both S2 and S5, I believe S5, naproxen sodium, is present in my
sample. S6, aspirin, has an Rf value that is too small to be the major ingredient in

Recrystallisation and Chemical Separations Essay
Introduction:
Recrystallization is used for the purification of solid compounds. The recrystallization process relies
on the fact that majority of compounds are more soluble in hot solvent than in cold. The hot
saturated solution containing the compound will have unwanted impurities and will be filtered out
and cooled to produce the pure crystal constituents of the compound.
Thin layer chromatography can be used as a physical method to segregate compounds from natural
sources. E.g. Spinach leaves are visibly green, but consist of a variety of components that have more
colour than others. This experimental procedure uses compounds from spinach leaves that are
exposed to chromatography, TLC plate to indicate the different pigments ... Show more content on
Helpwriting.net ...
Part C:
Clear silica gel turned visibly green once green food dye was added. The first band within column
(yellow) was collected after Nacl was added; methanol was added to the column to start the second
mobile phase which extracts the second band of (blue) liquid from the remaining silica gel + green
food dye solution. The column chromatography produced two beakers of blue and yellow from the
green food dye. Wavelengths of maximum absorption were calculated: yellow = 428nm; blue =
629nm; food dye green = 629 & 426nm.
Discussion:
Part A:
The warm water was added to the dehydrated mixture of table salt, sand and the copper sulphate.
The solution changed to a visibly blue homogenous colour. The sand and some of the copper
crystals that did not dissolve remained at the bottom of the beaker. A small amount of copper
sulphate residue was left in the collection funnel. The temperature of the solution was too high for
the mixture to bind and recrystallize; Ethanol was added to the mixture to lower solubility. The
beaker was then placed in a cooler for duration of 10mintues to decrease temperature and increase
the rate of recrystallization. When the beaker was removed from the cooler it was still visibly blue
indicating in was not a complete recovery. Recovered copper sulphate pentahydrate crystals were
solid, multi edged and uniform in assembly. High level of purity.

Part B:
The chromatogram involved

Experiment 2a Adsorption Chromatography ( Tlc )
NAME: ____Amy Hua_______________________
Experiment 4a Adsorption Chromatography (TLC)
Summary of Points for Experiment 4a:
Item Possible Points Actual Points
Pre–Lab 2 Notebook: N/A N/A
Purpose/Table of Reagents 2
Corrections 2
Blank Spaces 2
Signatures 2
TLC data (4–in notebook) 8
Coherent 2
Conclusions (absent here) 1
Sub–Total = 21 multiply Sub–Total x 2= 42 Report: N/A N/A
Introduction 2
Data and Calculations 8
Less Points–Missing Data N/A N/A
Unknown Identity 10
Data Analysis / Conclusions 5 TOTAL 67 minus any page overage 0 minus for late reports 0 minus
for TA points 0
FINAL TOTAL POINTS 67
INTRODUCTION
In this experiment, the technique of thin layer chromatography is used to identify the identity of
unknown. To aid in seeing the spots from the thin layer chromatography, UV light and iodine
adsorption is used.
DATAAND CALCULATIONS
PART I:
Below are two depictions of the developed TLC's for 2– and 4–hydroxy acetophenone. The first was

visualized under UV light and the second by iodine adsorption.
PART II:
Below are two depictions of the developed TLCs for an unknown analgesic versus known standards.
Both TLC's were visualized by UV light.
UNKNOWN ID (10 points):
UNKNOWN #: ________5_______________
IDENTITY OF UNKNOWN ANALGESIC: ___caffeine and most likely acetylsalicylic acid but
possibly ibuprofen___________
DATAANALYSIS AND CONCLUSIONS
In

Experiment 5
Title : Thin Layer Chromatography
Objectives:
i. To distinguish polar and non–polar solvents. ii. To familiar with the analysis technique by using
the thin layer chromatography. iii. To differentiate the retention factor, Rf for different compounds.
[pic]
Result:
|Compound |Distance traveled by the compound |
|o–nitroanaline |2.45 |
|p–nitroaniline |1.70 |
|Unknown sample ... Show more content on Helpwriting.net ...
(Or, more likely, given the level you are probably working at, someone else has already done all the
hard work for you, and you just use the solvent mixture you are given and everything will work
perfectly!)
Chromatography is a word used to encompass a range of techniques in which mixtures of pure
substances are separated into the individual substances by using a mobile phase (usually a liquid or
gas) to push the mixture along a stationary phase (usually a solid or liquid coated on a solid).
Because the individual substances have different molecular structures, they interact differently with
both the stationary and mobile phases, and consequently are "pushed" at different rates by the
mobile phase.
Thin–Layer Chromatography (TLC) is a simple and inexpensive technique that is often used to
judge the purity of a synthesized compound or to indicate the extent of progress of a chemical
reaction. In this technique, a small quantity of a solution of the mixture to be analyzed is deposited
as a small spot on a TLC plate, which consists of a thin layer of silica gel (SiO2) or alumina
(Al2O3) coated on a glass or plastic sheet. The plate constitutes the stationary phase. The sheet is
then placed in a chamber containing a small amount of solvent, which is the mobile phase. The
solvent gradually moves up the plate via capillary action, and it carries the deposited

Thin Layer Chromatography Of Amino Acid
Thin– Layer Chromatography (TLC) of Amino Acids
Introduction:
Thin layer chromatography (TLC) separates compounds, through migration, from a mixed solution
with the assistance of a solvent and an absorbent strip of cellulose. The purpose of this lab is to
allow students to determine an unknown amino acid by comparing results to six known amino acids
(slowest to fastest: lysine, glycine, alanine, tyrosine, phenylalanine, and leucine) and properly
separate a combined solution of amino acids using the TLC method. But to fully understand the lab,
two terms must be understood: amino acid and protein. Amino acids are building blocks of proteins
linked by peptide bonds that contain both carboxyl and amino functional groups.
The six amino acids used in this experiment: ... Show more content on Helpwriting.net ...
Rf values for known and unknown amino acids. Lanes 2 and 4 were used to identify the unknown
(Unknown T) amino acid with the slight assistance of Lane 1.
Discussion:
Based on my results from Lanes 2 and 4, I predict Unknown T to contain both lysine and glycine
based on the slow reaction/movement the unknown experienced once the solvent was introduced
and the shortest distance it underwent. My prediction was supported with the development of Lane
2. As I analyzed Lane 2, it became obvious that the six Rf values represented the six known amino
acids; the lowest Rf value was lysine and the highest Rf value was leucine. Once the Rf values for
the unknown was calculated it was easy to identify the two amino acids present in the unknown
sample due to corresponding values in both lanes. Although one value (Rf = 0.515) in Lane 4 did
not completely match with the second value (representing glycine) in Lane 2, I can conclude that the
Rf value in Lane 4 was still glycine because the values are similar and close in range, and Lane 1
helps support my theory as

Sugar content in organic products versus natural products tested with thin layer chromatography
Carbohydrates are one of the most important components for food sources. Carbon based molecules
are also called organic compounds. Large organic molecules have elaborate shapes. The chain of
carbon atoms is called a carbon skeleton. Other monosaccharides carbon skeletons may have 3 to 7
carbons. 5 carbon sugars are called pentoses and six carbon sugars are called hexoses, which are the
most common. The formula for a Monosaccharide Carbohydrate is CH2O. Carbohydrates refer to a
class of molecules ranging from small sugar molecules to large polysaccharides (long chain of
monosaccharides).(Reece 2012) These sugars behave differently and have different ... Show more
content on Helpwriting.net ...
(n.d.). Retrieved July 9, 2015, from Sciencelab website: http://www.sciencelab.com/msds.php?
msdsId=9927062 Material safety data sheet aniline [Fact sheet]. (n.d.). Retrieved July 9, 2015, from
sciencelab.com website: http://www.sciencelab.com/msds.php?msdsId=9927435
Material safety data sheet phosphoric acid, 85% [Fact sheet]. (n.d.). RetrievedJuly 9, 2015, from
sciencelab.com website: http://www.sciencelab.com/ msds.php?msdsId=9927393
Material safety data sheet ethyl acetate [Fact sheet]. (n.d.). Retrieved July 9,2015, from
Sciencelab.com website: http://www.sciencelab.com/msds.php?msdsId=9927165
Material safety data sheet isopropyl alcohol [Fact sheet]. (n.d.). Retrieved July 9, 2015, from
Sciencelab.com website: http://www.sciencelab.com/msds.php?msdsId=9924412
Material safety data sheet pyridine [Fact sheet]. (n.d.). Retrieved July 9,2015, from
fscimage.fishersci.com website: https://fscimage.fishersci.com/msds/19990.htm
Stereoisomerism optical isomerism [Lecture notes]. (2000). Retrieved June 25, 2015, from
chemguide.co website: http://www.chemguide.co.uk/basicorg/

Thin–layer chromatography (TLC) is a chromatography technique used to separate the components
of a mixture. It can be used to monitor the progress of a reaction, determine the purity of a
substance, and identify compounds present in a given mixture. TLC is performed on a sheet of glass,
plastic, or aluminum foil, which is coated with a thin layer of adsorbent material, usually silica gel,
aluminum oxide, or cellulose. This layer of adsorbent is known as the stationary phase. After the
sample has been applied on the plate, a solvent or solvent mixture is drawn up the plate via capillary
action (known as the mobile phase). The presence of hydroxyl groups in the adsorbent renders the
surface of silica gel highly polar. Thus, polar functionality ... Show more content on Helpwriting.net
...
Because different analytes ascend the TLC plate at different rates due to polarity, separation is
achieved. Caffeine was found to be the most polar compound out of all the other analgesics with the
lowest Rf factor of 0.33 due to its xanthine core which contains two fused rings, a pyrimidinedione
and imidazole. Out of the four amine groups present in caffeine, the pyrimidinedione in turn
contains two amide functional groups that exist predominately in a zwitterionic resonance, the
location from which the nitrogen atoms are double bonded to their adjacent amide carbons atoms.
The nitrogen atom is also capable of forming a hydrogen bond and participating in dipole–dipole
interactions, along with its london–dispersion forces, thereby making it an extremely polar
molecular compound and allowing it to form coordination bonds with silica gel in the TLC. The
second most

By: Salien Mansi
Student ID: Smansi2
[TLC Separation of Amino Acids]
Objective:
Separation and identification of the amino acids in a mixture by thin layer chromatography.
Experiment was done to understand the effects of Thin layer Chromatography using various
concentration mixtures of solvents.
Thin layer chromatography [TLC]:
The importance of Chromatography is for the separation of compounds in a mixture. The separation
happens between the stationary and the mobiles phases. This experiment itself is based on the non–
covalent interactions between the stationary phase, the different compounds and the solvent.
Thin Layer Chromatography is a mechanism used in order to monitor the progress of a chemical
reaction and the purity of a compound in order to identify those mixtures.
In some cases, a separation may fully divide the mixture into its pure constituents but it mostly
converts a mixture of a chemical substance into two or more product mixtures.
Thin layer chromatographic (TLC) is performed on a sheet of glass which is coated with a very thin
layer of adsorbent such as silica gel. The silica gel acts as the stationary phase and the solvent
mixture acts as the mobile phase.
In the perfect solvent system, the compounds are soluble to different degrees.
Uses of TLC:–
1– Purity of any sample
2– Identification of compounds
3– Examination of reactions
4– Biochemical analysis
5– One of the most important application of TLC is in separation of multicomponent pharmaceutical

formulations.
6– In food and cosmetic industry, TLC method is used for separation and identification of colours,
preservatives, sweetening agent, and various cosmetic products.
The separation is based on different factors:
1– Solubility: The more soluble a compound is a solvent, the faster it will accelerate up the plate.
2– Attraction: The attraction between the compound and the silica, the more the compound combine
with silica the lesser movement arise.
3– Size of the compound: The bigger the compound the slower it accelerates.
RF value is considered as a very important characteristic used in thin layer chromatography.
Individual components can be measured by an RF value or a Retaining factor value. This zero to
one value

Thin Layer Chromatography And Column Chromatography
Thin Layer Chromatography and Column Chromatography
By Maggi Shelton
Under the Supervision of Dr. Mills Chavonda
The Department of Chemistry and Physics
Milledgeville, GA 31061 Abstract:
Thin layer chromatography and column chromatography are two different methods that allow for
the separation of two miscible solvents. Through column chromatography, a mixture of nonpolar
fluorine and polar fluorenone was successfully separated. Thin layer chromatography was then used
in order to determine the success of the separation based upon the calculated Rf values, which if the
Rf value fell below zero or was above one, then the separation was not successful.
Introduction: Methods used to separate miscible solvents include thin layer chromatography (TLC)
as well as column chromatography. A method used to separate a mixture of miscible solvents is
column chromatography, which is used to purify and separate compounds. The particular speed of
the solvents used in the experiment depended upon the properties, being polar or nonpolar, as well
as the properties of the prepared column (INSERT SOURCE HERE). With column chromatography,
there are two phases that include mobile and stationary phases. In the experiment performed, the
glass column used was first packed with a piece of cotton followed by a layer of sand in order to
keep the silica gel in place and also to prevent the gel from flowing out of the tube when the
particular solvent was added. Based upon whether the

Thin-Layer Chromatography Lab
Introduction Thin–layer chromatography, also known as TLC, is a principle that describes how
various compounds travel multiple distances when placed as a thin layer on a plate. TLC is a
technique that can be used to determine how many components are in a mixture. TLC can also be
used to determine a specific compound in a mixture. After performing TLC, the retention factor (Rf)
can be used to determine a specific compound in a mixture. The retention factor (Rf) is During TLC,
there is a step called elution. In elution the components on the TLC plate will have different
solubilities and different adsorption strengths. The liquid at the bottom of the TLC chamber is called
eluent. The purpose of this experiment was to use TLC to separate and identify ... Show more
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If the solutions are placed to close together on the TLC plates, then is possible that after being place
in the TLC chamber, they will end up close together and it will be difficult to distinguish which spot
belongs to a certain solution. Another way to improve this experiment would be to make sure that
the bottom of the TLC plate is parallel to the bottom of the jar in the TLC chamber. If it is not
parallel, then the solvent front will not develop properly and it will make determining the spots or
comparing the spots more difficult. The last way to improve this experiment would be to make sure
that the .5% glacial acetic acid in ethyl acetate inside of each chambers does not evaporate. If it does
evaporate this could affect the development of the solvent

1. What was the main aim of the practical? (5–marks) The aim of this practical is to isolate lipid
extracts of amniotic liquid through thin layer chromatography (TLC). This method is based on the
separation of substances that form an analytical mixture because of the different affinity on the two
phases (mobile and stationary). This experiment will concentrate on measuring the quantity of
Phosphatidylglycerol, Sphingomyelin and Phosphatidylcholine from the amniotic liquid of two
patients. These are substances present in surfactants and can provide a general profile of lung
maturation and also provide information about the infant chances of developing respiratory distress
syndrome (RDS). Surfactants are fundamental as they keep the air sacs ... Show more content on
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The first spot represents the Phosphatidylcholine; which structure is characterised by a charged
polar head. This spot has not migrated very much due to a minimal attraction of the solute for the
solvent. The stationary phase used in this experiment was formed by silica gel. The covalent
bonding present on this adsorbent surface creates a very polar stationary phase, explaining why the
Phosphatidylcholine did not migrate very further. The second spot was formed by Sphingomyelin
and Phosphatidylcholine. Both compounds contain a phosphoryl choline polar head group. The spot
containing these two components managed to travel a bit further compared to the first spot, thanks
to the presence of wide range of chain lengths on the tail of the sphingomyelin molecule; that
succeeded in slightly interact with the mobile phase. (Mitnick M, 2016). The third spot represents
the Phosphatidylglycerol. Its structure is formed by a neutral glycerol group that allows it to interact
with a polar mobile phase providing an increased migration. (RD, 2016) The size of the compounds
can also affect the solubility in the respective mobile phase. For example, Phosphatidylglycerol is
the substance that travelled the most also because of its small size. (Mitnick, DeMarco and Gibbons,

Chromatography techniques are one of the most useful methods available for the separation or
isolation of organic compound from a mixture. The most common used chromatographic technique
includes column chromatography, thin –layer chromatography (TLC), etc. The purpose of this lab
exercise is using the gel filtration chromatography to separate different proteins from a mixture.
Collecting elutes (run off), and calibrating a curve to show how the column separates molecules by
molecular weight, and identify the molecular weight of the proteins. The sample mixture used in this
lab contained Blue Dextran that has a size of 2x106 Da, and has blue in color. Cytochrome C has a
size of 13,000 Da, pink in color. Potassium Chromate has a size of 192 ... Show more content on
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It is very important not to let the bed run dry during any part of the experiment because the pockets
of air in the bed will cause uneven flow of molecules through the column. Next, 13 clean, dry
cuvettes were collected and labeled 0 to 12. 1mL of water was measured by the P–1000 cuvette and
was transferred into the cuvette that labeled 0 in order to measure the amount of fraction that we
will collect. The sample mixture that included Blue Dextran, Cytochrome of was obtained and was
measured 250 μL by using the P–1000 pipette. Then, the 250 μL was carefully transferred into the
column that already packed. Add the mixture by placing your pipette against the wall of the column
so as not to disturb the bed. The mixture was allowed to enter the bed, close to the bottom the bed
(about 1 cm) by opening the stop cock . Then, about 15 mL of buffer was added into the column.
3mL of buffer added slowly each time to prevent the overflow and avoid disturb the bed. 1 mL
fractions were collected into each cuvette until there is no color present in the last fraction. 1 mL of
fraction was measured by keep the #0 cuvette contained water next to the cuvette that you want to
measure. After, collecting the fraction, the fraction was analyzed and recored the color and the
intensity. Each fraction was given the number based on the scale 0–5 based their color. The bigger
number the darker color they have and more intense. All the waste was discarded to appropriate
waste

Lab #1
Extraction and Thin Layer Chromatography
Kaya Gaudet 6784928
January 9, 2013
Line Structure References
BenzophenoneBiphenylBenzoic Acid
Extraction: A process used to separate different compounds in a mixture based on their solubility in
an immiscible substance. Usually an aqueous and organic phase are used. During extraction the
desired compound moves one phase and leaves the unwanted substance behind. Extraction normally
has to be done a few times to leave all of the impurities behind. TLC plating can be used to mark the
progress of the extraction.
Thin Layer Chromatography (TLC) Plating: This is a common method that is used to monitor the
progress of a reaction and determine when it is complete. In our ... Show more content on
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The blue, on the other hand, would probably not benefit form salting because it showed no affinity
towards the non–polar organic compound and the increase in polarity in the water would just make
it more attracted to the substance it was already in.
Part B: This introduction to thin layer chromatography challenged us to identify an unknown
compound by comparing it to the TLC plates of two other known compounds. Using a mixture of
2:8 ethyl acetate and hexanes gave us a mix of polar and non–polar solvent and gave more accurate
results. The Rh values for our unknown matched most closely with those of benzophenone but
weren't an exact match with values of 0.57 for benzophenone and 0.54 for the unknown compared to
0.78 for biphenyl and 0.59 for the unknown. In the lab questions we are asked to compare the
structures and polarity of three molecules; benzophenone, biphenyl, and benzoic acid. After
evaluating those it seems plausible that benzoic acid was our unknown salt because it would have a
very similar polarity to benzophenone while airing on the slower/more polar side.
Part C: This part just allowed us to see the effects of using an entirely polar or entirely non–polar
mobile phase and measure its effectiveness. It's obvious from just glancing at the plates that there is
much less obvious separation in these two mixtures and either the solvent was too polar and almost
all of the compounds reached the top (ethyl acetate) or it wasn't

Thin Layer Chromatography Lab
Brianda Mendez
Lab 05 Report
Introduction:
Thin–layer chromatography (TLC) is a very commonly used technique in synthetic chemistry for
identifying compounds, determining their purity and following the progress of a reaction. Thin–
layer chromatography or TLC, is a solid–liquid form of chromatography, it involves the distribution
between two phases. The stationary phase is a polar adsorbent, it is coated on a glass slide or plastic
sheet creating a thin layer of the particular stationary phase. The mobile phase, a liquid or a gas
flows through the stationary phase and carries the components of the mixture with it. In
chromatography, the retardation factor, is the fraction of a chemical constituent that undergoes
analysis in the mobile phase of the chromatographic system.1 The Rf is defined as the ratio of the ...
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The bigger the Rf, the further the spot moved and that the Rf should be the same for a component
regardless of how far the solvent moves.
Results & Discussion: Comparing the efficiency between the plates with the unknown was quite
effective, the first plate contained the unknown, Aspirin, Acetaminophen and Ibuprofen. The
outcomes came out very clear, Aspirin had a measure of 2.9cm, Acetaminophen had 2.6cm ,
Ibuprofen had 3.5cm and my unknown had 3cm. It was clear that Ibuprofen had too high of a
distance, Acetaminophen was too low of a distance, while Aspirin was right around the length my
unknown traveled. With that being said, I kept aspirin as one of my options to test later during the
KCU plate. In the second plate, It contained Salicylamide, Caffeine, and my unknown. The
Salicylamide traveled at a rate of 2.9 cm , Caffeine only traveled .8 cm and my unknown traveled 3
cm once again. It was visually clear that both Aspirin and Salicylamide had a distance .1 cm away
from my

Thin Layer Chromotography Discussion
Thin–Layer Chromotography Discussion and Conclusion
Easton Montgomery
Discussion:
In thin–layer chromatography a liquid is pumped across a bed of particles. The liquid that is pumped
across is called the mobile phase and the particles are the stationary phase. A mixture of the
molecules that will be separated is put into the mobile phase. Thin–layer chromatography tells
you/helps you determine the number of compounds in a mixture, the purity of a compound, and the
identity of compounds if you have examples to pull information from. Thin–layer chromatography
is used to separate nonvolatile mixtures. The dye that was the most polar was the color red and pink
which was Rhodamine B and the least polar was the light pink color which was Sudan IV. Our first
TLC plate had five ... Show more content on Helpwriting.net ...
You can use thin–layer chromatography for quantitative analysis to some extent. If you use solutions
of known concentrations and compare the unknown to the standards, then you can gain some
quantitative information. You can also calculate Rf values taking the distance traveled divided by
the total distance of the eluent. After we chopped up our spinach leaf we used hexane and ethanol to
separate out compounds in our leaf. After adding water, we removed the top layer and spotted a
TLC plate to separate out our compounds. We also used a mixture of hexane/ethyl acetate (1:1) and
hexane/ethyl acetate (20:1) to use as our developing solution. The (1:1) solution gave use six
compounds separated out on a TLC plate with xanthophyll being the most polar and carotene being
the least polar. On the other hand, the (20:1) solution gave us one compound that moved through our
TLC plate. Looking at the plate it seems like it is the carotene that moved while everything else
remained near the polar side of the plate. The most polar molecule was Rhodamine B (bright pink–
red) then Fast Green (blue) then we had a clear compound show up, then Bismark Brown (yellow–
orange) and the least polar compound was Sudan IV

Thin Layer Chromatography Essay
The isolation and purification of phosphatidylcholine from egg yolk is successfully achieved
through aluminum oxide chromatography. The chromatography–purified fractions of PC is
concentrated by doing thin layer chromatography. The TLC plate from first two fractions showed
indication of PC, which describes the separation is very faster than the typical experiment. Usually
fractions 5–9 contains the highly concentrated PC ( Clingman ).The net weight of the PC obtained
after all the purification is 1.14grams.The TLC analysis between crude PC and purified PC is
compare with standard lyso PC.The purified PC and crude PC has same Rf value but crude PC has
additional spots that are not present in purified run. Since the purified PC has one prominent ...
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Another factor that influence micelle formation is , the salt concentration and pH. The membrane
phospholipids are disrupted by mild nonionic detergents to isolate them but the isolated lipids
should be functional to react with the enzyme phospholipases. If pH and salt concentration is not
maintained at biological range, the micelles formed could be nonfunctional or denatured. This will
make the phospholipases reactivity with lipids impossible.
TLC is used this experiment is discontinuous method to analyze phospholipase reaction.
Spectrophotometry can be used to monitor enzymatic reaction activity continuously without
interrupting. However, TLC analysis more suitable method than spectrophotometry for following
phospholipase reactions because the products of the reaction does not have light absorption qualities
such as aromatic or conjugated pi system. However, products have polarity properties, which can be
exploited to measure the reaction using TLC.
This experiment has successfully demonstrated the isolation efficiently by TLC plate analysis. In
addition, site specific cleavage of phospholipases and reaction sensitivity to detergent also evident
from this

Essay On Thin Layer Chromatography
The procedure shown demonstrated how to determine the composition of unknown compounds,
based off of information that's known from given analgesics, by using thin–layer chromatography
(TLC)2. With some silica gel, a coated plate, and developmental solvent, it was possible to
distinguish unique measurements and characteristics of each unknown compound3. This was all
possible due to chemistry and the bonds that exist within the different compounds and materials.
Polarity and non polarity played a big part in how the results were achieved, as well as the chemical
structure and shape of the compounds1. The chemical structures of each compound are what allows
for them to be either polar or non polar. Within the experiment, depending on the polarity ... Show
more content on Helpwriting.net ...
While the iodine composition wasn't equal, caffeine was the closest to unknown one. Unknown two
was found to be most similar to Aspirin due to the low Rf values and and lack of appearance
underneath an UV light. Neither of them presented any color or distinction under the lights and
aspirin only lightly showed any trace of iodine components. Unknown three seemed most similar to
Acetaminophen and caffeine. This is because of the similarities in distance, Rf value, dark green or
gray color, and the strong presence under UV lights in the second spotting Lastly, the fourth
unknown once again seemed very similar to acetaminophen, and additionally caffeine as well. The
distance, Rf value, and the darkness of the second spot matches with acetaminophen. The first spot
seems to match up the most to caffeine, once again due to Rf values, distance in cm, and basic color
schemes. The most polar seems to be the Aspirin and the Salicylamide, this was indicated by their
lack of movement during the experiment. The most non polar seemed to be the Acetaminophen and
caffeine, you can tell by noting how far the distances were compared to the other

Evaluation Of The Practical -you Work As An Analyst For A...
Assignment 3.3
The aim of the practical –You work as an analyst for a snack food manufacturer.You have just
changed to a new supplier of soy sauce but are worried that the taste is not as strong and you suspect
that some of the key ingredients have been changed. Explain how you are going to do this.
Chromatography is a method used mainly in labs in order to isolate organic compounds from
inorganic compounds in order to be observed. Via this technique, scientists are able to differentiate
between the substances which create a particular compound. Chromatography can be used to
separate mixtures of coloured compounds. Mixtures which are suitable for separation by
chromatography are inks, colouring agents in foods and dyes. You can ... Show more content on
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Chromatography is also used in food analysis to detect microorganisms in drinking water. Scientists
whom observe the environment also use this method in order to measure pollution levels within the
area. Forensic scientist use it in order to analyse substances for DNA such as blood alongside to
discover what substances are such as drugs. Alongside this, chemists can use it to differentiate rates
of progression of chemicals alongside to observe what substances are present in a given sample.
https://uk.answers.yahoo.com/question/index?qid=20090125083047AA4IQ73
http://www.discoveriesinmedicine.com/Bar–Cod/Chromatography.html
http://www.pharmainfo.net/pharma–student–magazine/application–and–importance–thin–layer–
chromatography–analysis–and–research––0
Equipment
–Ruler
–Pencil
–TLC Plate
–Capillary tubes
–250ml Beaker
–Watch Glass / Petri dish
–Butanol
–Acetic acid
–Water
–Amoy soy sauce , Kikkoman soy sauce and Sainsbury's soy sauce.
–Hairdryer

–Petri Dish
–Wool
–Ninhydrin
–Latex gloves.
How to prepare the sample for analysis–
Prepare a solution of Butanol, acetic acid and water within the ratio 4:1:2.
Mix each soy sauce sample with an equal volume of butanol, in a 50:50 ratio, shake and leave
prepared mixture for 15 minutes in order for the different layers to become clear.
Method
1) Draw a faint pencil line 1cm below a TLC plate.
2) Use a capillary tube to put a

Component Analysis of Common Analgesic Tablets by Thin–Layer Chromatography
Objective
The objective of this lab was to use Thin Layer Chromatography to separate the different
compounds found in four over–the–counter drugs. TLC uses the polarities of the different
compounds to separate them and the uses their Rf value to determine the exact compound(s)
present. Compounds with higher polarity will stay closer to the baseline because they have a
stronger connection to the silica.
Method
For this experiment the analgesic tablet and ethanol mixture was already made prior to lab, so that
was not done. The eluent was poured into the beakers and covered to allow he air to also get
saturated, and to ensure none evaporated. The TLC plates were label ... Show more content on
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Shaking the plate could have moved around compounds before they dried leading to incorrect data.
In addition, trying to distinguish between a streak and two overlapping spots was difficult for some
compounds. Specifically Excedrin during the second trial, there was a big streak with some indents
on the side, but because of the size it was concluded that they were not separate spots; after looking
up the true active ingredients and then reviewing the plates again, they should have been marked as
separate

Comparing Polarity Of Caffeine And Acetaminophen
In this experiment, thin layer chromatography (TLC) was used to identify and compare polarity of
two molecules, caffeine and acetaminophen. Chromatography is defined as the separation of a
mixture of chemicals as they flow at different rates over a stationary phase based on their relative
polarity. Caffeine, the more polar molecule had a greater affinity for the polar silica gel stationary
phase causing it to consistently have a lower retention factor regardless of the mobile phase. This
methodology can be effectively used to distinguish and analyze the polarity various of chemical
mixtures such as within medicines, inks, etc.
In thin layer chromatography a stationary phase, silica gel with a glass backing, is dotted on a pencil
drawn

Chemical Components Of Anacin And Tylenol, Using Thin...
Abstract
The goal of this experiment was to find out active chemical components in Anacin and Tylenol,
using Thin Layer Chromatography technique. This technique uses the difference in the
intermolecular forcer and polarity to separate mixtures. Comparing Rf values were then used to
determine the active chemical components in the two analgesics. The overall result was that
Acetaminophen exists in Tylenol and Acetylsalicylic Acid exists in Anacin.
Introduction
The main goal of this lab was to determine which of the acetaminophen and acetylsalicylic acid
exists in Anacin and which exists in Tylenol. In order to achieve this goal, Thin Layer
Chromatography, an approach that uses the difference in the strength of intermolecular forces and
polarity in molecules to separate mixtures, was used In this investigation, a TLC plate and ethyl
acetate (solvent) were used to measure the Rf values of four different solutions of Tylenol, Anacin,
Acetaminophen, and Acetylsalicylic acid. The value of Rf depends on the strength of the
intermolecular forces that exists in molecules. The stronger the intermolecular forces are, the harder
the solvent moves the molecule up, resulting a small Rf value. In contrast, molecules with weak
intermolecular forces tend to have a high Rf value.
Additionally, Rf values are also dependent on the polarity of molecules. The more polar the
molecule is, the smaller Rf value, and the less polar the molecule is, the lager the Rf value.
Therefore, the outcome

Thin Layer Chromatography In Drug Screening
Drug screening, which is generally used in criminal situations, health care and the workplace has
recently become more common. The ease of use and fast results have increased the use of
immunoassays; however, these can cause false positive results. This can lead to severe
consequences if they are not confirmed by secondary analysis, such as gas chromatography–mass
spectrometry (GCMS). The drugs screened during this experiment were ketamine, methadone,
diazepam, codeine and imipramine. This report discusses the use of thin layer chromatography as a
drug screen technique. The results obtained from this are compared to the secondary analysis from
GCMS which show that the drugs from the unknown sample were ketamine, methadone and
codeine. The main techniques are examined to gain a better understanding of the methods used.
Potential false positive and false negative results that can occur are also discussed.
Introduction
A drug screen is an analysis of a physiological fluids such as urine, blood, sweat, or saliva to
determine the presence or absence of specified drugs or their metabolites. The main applications of
drug testing include police officers testing for the presence and concentration of alcohol in the
blood, detection of the presence of performance enhancing drugs in sport and employers screening
staff for illegal drugs.
When there are a large number of samples for drug screening, immunoassay techniques are favoured
as they are cheaper and faster. This is

The Preparation of Acetaminophen (Paracetamol) with Thin...
Experiment 3: The preparation of acetaminophen (paracetamol) with thin layer chromatography
(TLC) to monitor the reaction.
Abstract:
This experiment is to demonstrate the preparation of paracetamol and its properties. Reflux and
filtration of 4–aminophenol and acetic anhydride formed the crude sample. Further analysis of dry
white crystals were used to give quantitative measurements and a percentage yield of 46% was
obtained. The overall conclusion is that the acetic anhydride reacted with the –NH2 group.
Materials and Methods:
1. We weighed out (on a top–pan balance) 4–aminophenol (about 11.0g) in a weighing boat and
transfer the powder to a 250cm3 round–bottomed flask (RBF) using a powder funnel. 2. Then
recorded the amount ... Show more content on Helpwriting.net ...
–Sample + Ethanol= Colourless solution
FeCl3 was then added to this solution and a positive colour change was seen, as it turned green
–Sample + Ethanol + FeCl3 = Dark Green solution
This was then repeated using Salicyclic Acid instead of the sample to give a reference material as it
will give a positive test for a phenol.
–Salicyclic Acid + Ethanol = Colourless solution
–Salicyclic Acid + Ethanol + FeCl3 =Purple solution
The positive colour change from colourless to dark green of the sample means that the phenol group
is present in the sample. Therefore indicating that the acetic anhydride has reacted with the amino
group.
3. Determine the melting point and record the infrared spectrum of the dry solid.
Melting Point:

The melting point of sample obtained is: 1680C
Literature Value: [1]: 169–1720C
Fig 1.1–Infrared Spectra of Paracetamol:
The infrared spectrum of paracetamol shows the appearance of a new peak at 1561 – 1650cm–1 this
represents the carbonyl group from the acetic anhydride, meaning that paracetamol has been
formed. The –OH peak is still present at 3109 – 3319cm–1 this shows that the phenol group is still
attached to paracetamol. From the infrared spectra you can also see that the –NH2 group has
disappeared. This is as it has been selectively acetylated by acetic anhydride to form paracetamol.
4. TLC Plate analysis and Rf values of important spots.
Fig 1.2–Thin Layer Chromatography of Student samples 1 and

OBJECTIVE The purpose of this experiment is to analyze mixtures of compounds prior to, during
and after a separation scheme. This experiment also allows monitoring reactions of organic
molecules, and determines the identity of a mixture of compounds. STRUCTURES AND
PHYSICAL PROPERTIES OF REACTANTS [1] SOLVENTS | a. Hexane1Molecular Molarity:
86.18 g/molBoiling Point: 69 ºCMelting Point: –95ºCDensity: 0.659 g/mL at 25ºCWater Solubility:
Insoluble in waterColor/Texture: Colorless/LiquidHazardous Information: May cause irritation to
the eyes, skin and respiratory system. May cause drowsiness and/or dizziness. Suspected of
damaging fertility or unborn child. May cause damage to organs. Toxic to aquatic life. Highly
flammable liquid and ... Show more content on Helpwriting.net ...
The distance traveled by Benzophenone was 2.9cm, which gives it an Rf value of 0.67. The third
spot contained Biphenyl, which traveled a distance of 3.6cm, which gives it an Rf value of 0.84.
Unknown B had two spots: the first spot had a traveled distance of about 2.3cm which gives it an Rf
value of about 0.53 and the second had a traveled distance of about 2.9cm, which gives it an Rf
value of about 0.67. Judging from the Rf values of unknown B, the two compounds present are
Benzhydrol and Benzophenone. DISCUSSION Separation of compounds is a result of competition
by the stationary phase (adsorbent) and the mobile–phase (developing solvent). Generally, a non–
polar developing solvent should be used for non–polar compounds and a polar developing solvent
for polar compounds. Selecting a solvent is a trial–and–error process. Solvents that are not polar
enough does not cause the original spot to move far enough, as well as, solvents that are very polar
causes all spotted material to move along the solvent front. As a result, both give no to

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