If there is a free courier, the order is delivered on the day of the order or the next day. You can specify the delivery time of the order cialis australia Clear work, respectful attitude to the buyer, timely delivery.
Microsoft word - experiment3.doc
THIN LAYER CHROMATOGRAPHY
A. PRELAB ASSIGNMENT
Prepare a Table of Physical Constants for the following compounds.
(Consult the Merck Index as well as the Handbook of Chemistry and Physics).
B. THEORETICAL BACKGROUND
The theory of chromatography will be discussed in the laboratory lecture.
However, a few points do require emphasis. The most often used stationary
phases, silica gel and alumina, are polar materials. Consequently, the least polar
compound will have the highest Rf value. Increasing the polarity of the eluting
solvent will increase the Rf values of all compounds, but will not
order of the Rf values. If the solvent is too polar, all compounds will follow the
solvent front, Rf = 1.0. Too low a solvent polarity will leave all compounds at the
origin, Rf = 0. Neither of these situations is useful in analyzing mixtures. Usually,
the polarity of the eluting solvent can be adjusted by mixing solvents of different
polarities to yield Rf values between 0.1 and 0.9. Under all conditions, spots of
identical compounds should yield identical Rf values. However, in some
situations, different compounds can also yield identical Rf values. Therefore,
confirmation of the identity of a compound by TLC requires that the compounds
be analyzed against standard compounds under several sets of conditions.
A. GENERAL PROCEDURES
1. Application of Sample
It is best for samples to be applied as small (2-3 mm diameter) spots.
During development of the plate, the spots tend to diffuse, so that the final
spot is considerably larger than the original spot. The larger the final spot,
the more difficult it is to resolve a mixture into completely separate spots.
Small spots can be obtained by using fine capillary tubes as applicators.
Compounds or mixtures are applied approximately 1 cm apart in an even
line about 1 cm from the bottom of the TLC plate.
2. Development of the Plates
The eluting chamber must be saturated with the eluting solvent vapor. To
assure vapor saturation, a piece of solvent-saturated filter paper is used
as a liner for the chamber. The level of solvent in the bottom of the
chamber must be below the positions of the spots on the plate. Too high a
solvent level will wash the spots off the plate, preventing any meaningful
analysis. The plate is placed into the chamber and the solvent is allowed
to rise up to within 0.5 cm of the top of the silica gel coating. When this
occurs, the plate is removed from the developing chamber and the extent
of solvent migration (solvent front) is marked on the plate right away.
3. Visualization of Spots
Colored compounds are easily located on TLC plates. However, the vast
majority of organic molecules are white or colorless, and thus they are
invisible on TLC plates. In such analyses, some method must be used to
make the spots visible. Although several techniques are available, one of
the simplest and most used methods is the I2 (Iodine) visualization
technique. In this process, the TLC plate is placed in an iodine chamber.
Most organic compounds adsorb yellow-brown I2 molecules. Thus, spots
of chromatographed compounds appear as yellow brown spots after the
plate has been treated with I2. When the plate is removed from the I2
chamber, the I2 will gradually diffuse away from the spots, so the plate
should be immediately marked to indicate the positions of spots. Another
method used to visualize the spots is to observe the plate under UV light.
Most conjugated compounds appear as violet or purple spots under UV
light, if the coating on the plates contains a UV absorber.
4. Rf Values
Rf values for many compounds are published for specific sets of
conditions (sample quantity, solvent system, temperature and coating
material). Often these conditions are difficult to duplicate, so the best way
to identify a compound in a mixture is to compare the behavior of the
mixture and a standard on the same plate. Even then, variations in the
thickness of the coating can cause significant variation of Rf values on the
1. Analysis of a Mixture of Colored Compounds
Transfer small samples (several drops) of the four standard colored
Choose the lightest colored standard solution for this part of the experiment.
Spot this standard on the TLC plate in increasing order of applications (e.g.
1 for position 1, 2 for position 2, etc.). Develop the plate using a 3:2 mixture
of hexane and acetone. From the appearance of the final spots, choose the
appropriate number of applications for your subsequent plates with the
For your initial analysis, spot your unknown and the colored standard
compounds on one plate. Be sure to note which spot corresponds to which
Develop the plate using a 3:2 mixture of hexane and acetone. Your
unknown may contain two or three of the standard compounds. Draw a
replica (to scale) of this plate in your Results section. Calculate the Rf
values and record in a table (See sample on the next page).
From your results, choose the standards that appear to make up your
unknown. Next, spot a plate with the standards that you have chosen, your
unknown, a mixture of the standards prepared by spotting them one over
the other, and a mixture of unknown and the selected standards. Develop
the plate and draw a replica of it in your Results section. When you are sure
of the identities of the compounds in your unknown mixture, calculate Rf
values for all spots on the last plate. Record your values in a table similar to
Note: To obtain unequivocal results you may require more plates than those
described above. If so, record information only for those plates that run
properly and contribute to your determination of the identity of the
components of your unknown mixture. Some compounds may streak when
run on TLC plates. If they do so consistently, this can be used to help
identify the compounds. Use an I2 chamber to visualize any questionable
2. TLC Analysis of Analgesic Drugs
Analgesics, compounds that relieve pain, range from aspirin to morphine
and related narcotics. In addition to aspirin, several other chemically similar
compounds are widely used in non-prescription analgesic tablets. Among
these are salicylamide and acetaminophen. Caffeine is sometimes added to
these formulations to overcome drowsiness. Other ingredients may be
added for different therapeutic effects, such as antispasmodic or slight
sedative actions. In addition to the active ingredients, the tablets contain
other substances which act as binders or enhance dissolution. In this
experiment, you will obtain as an unknown an over-the-counter analgesic
drug. You will identify the unknown drug by TLC comparison with these
known reference compounds: aspirin (acetylsalicylic acid), acetaminophen
(4’-acetamidophenol or 4’-hydroxacetanilide), caffeine and salicylamide.
Your unknown will be one of those listed below; the structures of the
Obtain your unknown analgesic drug and samples (a few drops) of
solutions of the four reference compounds. Record the number of your
unknown. Dissolve the analgesic in 1 mL of 1:1 CH2Cl2: methanol. Stir the
mixture well and be sure all solid dissolves. Identify your unknown by TLC
vs. four samples of the known ingredients, using the same method as you
did with your first unknown. Be sure to determine Rfs of all compounds
involved. With this sample, develop the plates using a solvent mixture
having the composition 15:1 CH2Cl2: acetic acid. Observe the developed
plates under ultraviolet (UV) light in the hood. Then, develop the plates in
an iodine chamber, as demonstrated. Draw replicas of useful plates in
your Observations section. Calculate Rf values of all spots on the first
plate and on your confirmatory plate and record the results in tabular
format, as you did with the colored compounds.
An unknown compound, X, was analyzed by TLC using two different
eluting solvent mixtures. Three standard compounds, G, H, and I, were
used. The Rf values for the analyses are given below. Which of the three
standards is most likely to be compound X? Explain your reasoning in two
An unknown mixture was analyzed using TLC ped on a silica gel plate and
developed in 1:1 acetone:hexane, and gave 2 spots with Rf values of 1.0
Can you say for sure how many compounds are contained in the
mixture? If so, how many are there? Explain.
If not, what further experiments might be performed to determine
the number of components in the mixture? Explain.
On the basis of the results of your TLC analysis, list the analgesic standard
compounds and colored standard compounds in order of decreasing
polarity. Explain your reasoning. Keep it brief.
F. DISCUSSION AND CONCLUSIONS
GHS LABORATORY OUTREACH TEST CODES AND SPECIMEN REQUIREMENTSneonatal is in test description 17HP Separate serum after clotting and custody collection procedure. Refrigerated. LAVENDER. Centrifuge and separate plasma specimens from cells. Transfer 1.5 mLs to a plastic, transport tube. Label Sample with "Plasma" sticker. Do not thaw. Frozen. GHS LABORATORY OUTREACH TEST CODES AND SPECIM
CCD4A-2220-EU Care Call Mini Magnetic Door Monitor Part of the Care Call Paging System for Carers Getting Started 1. Install a 9 volt PP3 Alkaline or Lithium battery. ONLY USE ALKALINE OR LITHIUM TYPE BATTERIES. 2. When a battery is first fitted the indicator light flashes red then green several times, there is a brief pause and it then flashes green several times. The number o