RATIFICATION PAGE Complete report of Organic Chemistry I with the title Column Chromatography and Thin Layer Chromatography

which made by: Name : Sri Agustiani Basir

Reg. Number : 101304159 Class Group Department : ICP Chemistry : IV : Chemistry

After checked by assistant and assistant coordinator. So, this report accepted.

Makassar, June Assistant coordinator Assistant

2011

(Ahmad Fudhail Majid, S.Pd ) Known by, Lecturer of Responsibility

( Humaerah Azahra )

( Iwan Dini, S.Si, M.Si )

A. Title of experiment The title of experiment is Column Chromatography and Thin Layer Chromatography.

B. Purpose of experiment In the end of experiment, the student excepted understands about: 1. The student to know basic techniques column chromatography and thin layer chromatography. 2. The student to know basic principles of chromatography. 3. The student to know basic principles from the influence of substituent to the electrofil in aromatic compound.

C. Preview of literature Chromatography is an extremely sensitive technique for purification and separation of substances. Chromatography possesses two important features: a. Only a very small sample is required for analysis, and b. It is capable of effectively separating very complex mixtures (Henry: 1981). Chromatography was first introduced by Michael Tswet (1906), a botanist from Rusia. In this experiments he managed to separate the chlorophyll and other color pigments in plant extracts using calcium carbonate powder is filled into glass columns and petroleum ether as solvent. Understanding chromatography involves separation method based on the differential distribution of sample components between two phases. According to this definition always involves two phases chromatography, the stationary phase and mobile phase. Stationary phase can be either solids or liquids that are bound to the solid surface (paper or an absorbent). Whereas the mobile phase to a liquid called the eluent or solvent, or an inert carrier gas. The movement of this mobile phase resulted in differential migration of components in the sample.

Based on the separation mechanism known as four different types of chromatography, chromatography, namely: c. ion a. adsorption chromatography, and b. d. partition exclusion

exchange

chromatography,

chromatography. In adsorption chromatography, a solid stationary phase and phase motion can be either liquid or gas. Solute adsorbed by the surface of solid particles. An example of this type of chromatography is thin layer chromatography (TLC). Basically, thin layer chromatography is very similar to paper chromatography, particularly on how to do it. The apparent difference in the separation media, which used a thin layer of fine adsorbent on board glass, aluminum or plastic instead of paper. A thin layer of this adsorbent in the separation process serves at the stationary phase. Adsorbent material can be used as stationary phase silica gel, alumina and cellulose powder. Silica gel particles containing hydroxyl groups on its surface which will form a hydrogen bond with polar molecules. Water is absorbed in the gel to prevent the polar molecules. From reaching the surface. To fix the gel is activated by heating so that the absorbed water can be issued. Alumina can separated weak polar compounds (Subagyo:2000). Organic compounds analytic interaction model with silica gel:

All types of chromatography involves a process of equilibrium dynamic molecules and rapidly between 2 phase (stationary and mobile). Equilibrium between the two phases is dependent on 3 factors: 1. Polarity and molecular size tube separated 2. Polarity stationary phases 3. Polarity mobile phase Molecular polarity is determined by its structure. Therefore, the more polar molecules will be separated, the stronger interaction with the stationary phase, resulting molecule it stays longer in the stationary phase. In contrast, non polar molecules are smaller affinity of the stationary phase will tend to be in the mobile phase longer and will be eluted first. The more polar compounds will be separated, so if you use a polar stationary phase such us silica gel, the compound will be bound strongly to the stationary phase and will be separated in order last. In general, if the chromatography used a polar stationary phase, first choose non polar solvents as the mobile phase to elute the components in the mixture (Anonym:2011). Factory equipment maker to chromatography from year to year race improve accuracy, sensitivity an ease of operations, because it is not surprising when a device is often quickly become outdated. Similarly, in the field of purification of a product such as chemicals and pharmaceuticals, preparative chromatography plays a fairly important. Fields that use services such as preparative chromatography preparation enzymes, purification of drugs results biosynthesis, the isolation of bioactive materials from natural materials, the manufacture of standard compounds in the petroleum industry, purification of drinking water and water for laboratory, industrial waste water cleaning/ household etc other (Chairil:1994). Phenol also known as carbolic acid, is an organic compound with the chemical formula C6H5OH.

OH

OH

Or

Phenol is appreciably soluble in water, with about 8,3 g dissolving in 100 mL (0,88 M). Phenol is highly reactive toward electrophilic aromatic substituent as the oxygen atoms electrons donate electron density into the ring (Rasyid:2009).

D. Apparatus and reagents 1. Apparatus a. Thermometer 1100C b. Volumetric glass 10 mL c. Erlenmeyer flask 50 mL d. Balance e. Stir bar f. Stopwatch g. Beaker glass 100 mL h. Separating funnel 250 mL i. Spray bottle j. Column glass (contain the cotton) k. Erlenmeyer flask 250 mL l. Elution bottle m. Beaker glass 50 mL n. Statif and clamp o. Spiritus, asbes, and tripod 2. Reagents a. Silica gel TLC b. TLC plat c. Concentrated Nitride acid (HNO3) 1 piece 2 pieces 1 piece 1 piece 1 piece 1 piece 4 pieces 1 piece 1 piece 1 piece 1 piece 1 piece 7 pieces 1 piece 1 piece

d. Phenol (C6H5OH) e. Methylen Chloride (CHCl3) f. H2O g. Sodium Sulfate Inhidrate (Na2SO4) h. Benzene (C6H6) i. Iod Crystal j. Ice k. Tissue l. Palm leaf rib

E. Work procedure 1. Phenol nitration a. 1,5 ml of saturated HNO3 to mix 3,5 ml of water, to cool until 5oC b. To add the mixture between HNO3 and water into erlenmeyer which is contain 1,5 gram of phenol c. To shake, and then to arrange the mixture temprature 20 oC - 5oC for 15 minutes and then between 30o - 5oC for 15 minutesto cool it in water d. To add 3,5 ml of cool water into the HNO3 solution and to extract for 2 timeswith 5 ml methylenchloride e. To wish mixture with 5 ml of aquadest f. To dry with sodium sulphate inhidrate , then vapor its solvent in water evaporator, then to filtrate it. 2. Column Chromatography a. To prepare glass column which contain cotton and fill into it Methylen chloride b. To add alumina into column c. To add gel silica and put the mixture of phenol nitration and did fraction for three fraction and to observe what happen

3. Making of thin layers chromatography (TLC) a. To drop each fraction on plat of gel silica by using palm leaf rib b. To set the plat on eluent bottle that contain of benzene c. To vapor with iodium concentration d. To calculate of Rf value

F. Observation result a. Nitration of phenol 3,5 mL H2O + 1,5 mL HNO3 + 3,5 mL ice water + 1,5 gr phenol brown solution

brown oil + 5 mL Methylen chloride

form two layers; up layer (brown) down layer (black brownish) brown solution + 5 mL Methylen chloride two layers; up layer

(brown) down layer (black brownish) Nitrophenol + Na2SO4 anhydrate b. Column Chromatography

black brownish + H2O

black brownish solution.

1 gram silica gel + 15 drops Nitrophenol green powder result 12 fractions. c. Thin layer chromatography The solutions from the fraction drop into the plat into the benzene and dry with iodium solid. So, we get data: Fraction I and II dont show dot in TLC plat Fraction III, Rf = Fraction IV, Rf = Fraction V, Rf = Fraction VI, Rf = = 0,21 = 0,24 = 0,21 = = 0,21

 Fraction VII, Rf = Fraction VIII, Rf = Fraction IX, Rf = Fraction X, Rf = Fraction XI, Rf = Fraction XII, Rf = Fraction XIII, Rf = Fraction XIV, Rf = Fraction XV, Rf = Fraction XVI, Rf =

= 0,21 = 0,21 = 0,23 = 0,20 = 0,20 = 0,20 = 0,20 = 0,23 = 0,23 = 0,23

Note: Rf 0,2 it is 2,4 dinitrophenol

G. Analysis data From observation result, we get data: III. Spot distance Eluen distance Rf = IV. Spot distance Eluen distance Rf = V. Spot distance Eluen distance Rf = = 0,8 = 3,8 = = 0,21

= 0,8 = 3,8 = = 0,21

= 0,9 = 3,8 = = 0,24

VI.

Spot distance Eluen distance Rf =

= 0,8 = 3,8 = = 0,21

VII.

Spot distance Eluen distance Rf =

= 0,8 = 3,8 = = 0,21

VIII.

Spot distance Eluen distance Rf =

= 0,8 = 3,8 = = 0,21

IX.

Spot distance Eluen distance Rf =

= 0,9 = 3,9 = = 0,23

X.

Spot distance Eluen distance Rf =

= 0,8 = 3,9 = = 0,20

XI.

Spot distance Eluen distance Rf =

= 0,8 = 3,9 = = 0,20

XII.

Spot distance Eluen distance Rf =

= 0,8 = 3,9 = = 0,20

XIII.

Spot distance Eluen distance Rf =

= 0,8 = 3,9 = = 0,20

XIV.

Spot distance Eluen distance Rf =

= 0,9 = 3,9 = = 0,23

XV.

Spot distance Eluen distance Rf =

= 0,9 = 3,9 = = 0,23

XVI.

Spot distance Eluen distance Rf =

= 0,9 = 3,9 = = 0,23

So, Rf is 0,2 it is 2,4 dinitrophenol

H. Discussion 1. Nitration of phenol This experiment concentrated HNO3 put into water produce colorless yellow solution. The purpose of water added HNO3 so that not happen explosion or fully because concentrated HNO3 very reactive and have a high concentration. When the added HNO3 into water is cooled before, this purpose is concentrated HNO3 not occur spontaneous solution. And then a solution added phenol produce brown solution. A solution arrange of temperature 200 50C function for the temperature is not hot so the reaction can occur completely, if through of temperature 200 50C the solution form not nitration phenol but other solution. After that a solution added ice water, function of the ice water for neutralization temperature and then done the extraction with way filled HNO3 + H2O and phenol into separating funnel and added methylen chloride, the function for separate organic compound (phenol) with water. When extract formed two layers are up layer: brown solution and down layer: brown like black solution. Form two layers caused different of polarity and density of solution, water to have a characteristic polar and organic

compound have a characteristic non polar. Density of phenol bigger than water (density of water = 1 gr/ mL, phenol = 1,07 gr/ mL) so can known up layer is water and down layer is organic compound. Then organic compound layer washed with water. The function of washed with water is to wash and bond of acid for not residue so that produced Nitrophenol. Nitrophenol added Na2SO4 is to bond of water which residue at Nitrophenol compound after that vaporized so that gets pure Nitrophenol. Purpose of nitration of phenol for knows that phenol can nitration or not, so that to know phenol can nitrate. 2. Column chromatography Basic a principle of chromatography is separated mixture of compound that the components base the different each speed of migration of component between two phases is stationary and mobile phase. At experiment, result from nitration into the column had contained silica gel, function of the adding easy is elution. After nitration result out we must to wait until yellow layer to appear and yellow drop out from column. The fraction which contained there are 16 fraction. The function of column chromatography is to separate of compounds which found in nitration phenol. This column chromatography which function as stationary phase is silica gel, silica gel is absorber which versatile and silica gel can bounded with hydrogen with other solution and a solvent if there is water. And which function as mobile phase is methylen chloride. 3. Thin layer chromatography This experiment result from fraction of chromatography column tested which use thin layer chromatography, purpose from tested is to identify compounds which contain at fraction of column chromatography. Media which used in thin layer chromatography is thin plate. Then to dip a palm leaf rib in each fraction and put 1 drop result of fraction to each dot has been determine in layer plate. After that put in elution bottle contained benzene, function of benzene is to

mediator in process absorbent of spot. Than vaporizing in iod solution which to function is to show a spot can see so that to make easier identification process. According to result of experiment obtained Rf is 0,2. Can concluded according to Rf obtained is 2,4 dinitrophenol. The mechanism reaction:
OH OH NO 2 OH NO 2

+ HNO 3
I. Closing a. Conclusion

+HNO 3
NO 2

From the observation, we can get conclusion as follow: 1. Basic techniques of column chromatography are applied

chromatography or adsorption base kind of phase which use. - Basic techniques of thin layer chromatography are chromatography which use TLC adsorbents (smooth) depend in glass of board. 2. Basic techniques of chromatography are separated mixture of compound that the component bases the different each speed of migration of component between two phases is stationary and mobile phase. 3. Result of experiment 2,4 dinitrophenol.

b. Suggestion 1. For the practicant, we must have a great take care when observed the change of color or reaction that was occur when done the experiment. 2. For the laborant, must attention for the reagent that need to done the experiment.

BIBLIOGRAPHY Anonyma.

2011.

Penuntun

Praktikum

Kimia

Organik

(K12051)

Farmasi.

http://diaharrazy.files.wordpress.com/2010/12/praktikumkoki2051fa2.pdf. Online. Accessed on May 26th 2011. Anwar, Chairil, dkk. 1994. Pengantar Praktikum Kimia Organik. Yogyakarta: Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Gajah Mada. Henry,N.W. 1981. Matter Under Investigation. Hongkong: Jacaranda Kimia, FMIPA, UNM. Rasyid, Muhaidah. 2009. Kimia Organik I. Makassar: Badan penerbit Universitas Negeri Makassar. Subagyo. 2005. Kimia Analitik II. Malang: Penerbit Universitas Negeri Malang.

Answer The Question

1.

With see the Rf value from derivate of phenol and known the thin layer chromatography (TLC) principle is partition,how tol level or series the polarity of phenol fraction ? Answer : The levels or series the polarity of phenol fraction are : a. 2,4,6-trinitriphenol b. 2,4-dinitrophenol c. p-nitrophenol d. o-nitrophenol

2.

Suggest a technique of thin layer chromatography used for getting pure compound (preparatif)? Answer: The method tthat we can use for getting pure compound in a thin layer is preparation of nitrophenol because nitrophenol is first step to add mixture which will be used in chromatography coulumn and will continued thin layer chromatography.