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Unit 33

EXPERIMENT 33
Identification of an Unknown ___________________________________________
E33.1 OVERVIEW In this experiment you will identify an unknown organic liquid by a combination of analytical tools: IR spectrum, chemical tests, refractive index and 1H-NMR. Your unknown will be a liquid with no more than nine carbon atoms and will have only one of the following functional groups: alcohol, alkyl halide, aldehyde, ketone, carboxylic acid or ester. In addition, your unknown may have an ether group. This experiment is different from other experiments you have performed so far. The amount of experimental work is minimal, but you will be required to fully interpret your data and solve the unknown before leaving the lab. As always, the TA will be there to assist you and to clarify any questions, especially about 1H-NMR. You should come to the lab very well prepared with a good working knowledge of IR and, above all, 1HNMR. E33.2 EXPERIMENTAL PROCEDURE
Background reading: 11.1-11.3; Unit 31; Unit 33 Estimated time: 3-3.5 hr

Obtain an unknown liquid sample from the stockroom. Write down the unknown’s number in your lab notebook. Obtain the IR of your unknown. Your IR should be “nice looking” with the bands ending in sharp peaks rather than being flat at the bottom. These flat signals result when too much sample is used. To avoid this, use a very small drop of liquid: touch your liquid with a Pasteur pipet and then just touch the IR plate with the pipet. This will deposit a very small amount of liquid on the plate, enough to obtain a good IR. Cover the plate with the other half and spread the liquid by rotating the plates. By analyzing the IR spectrum you should be able to tell if your unknown contains any of the following functional groups: alcohol, carboxylic acid, aldehyde, ketone, ester. You should also be able to determine if your compound is aliphatic or aromatic. The lack of IR bands for OH and C=O will mean that your compound is an alkyl halide. If you suspect a carboxylic acid, you should further confirm your suspicion by checking the pH of a solution of your unknown in water (one drop of unknown in 1 mL of water). If the pH is clearly acidic (pH < 3) your unknown is likely to be a carboxylic acid. You should also perform the silver-nitrate-in-ethanol test. Most carboxylic acids produce a precipitate of a silver carboxylate when treated with silver nitrate.

4-dinitrophenylhydrazine. perform the sodium-iodide-in-acetone test and the silver-nitrate-in-ethanol test. Tertiary alcohols soluble in the Lucas reagent (ZnCl2 in HCl(c)) react immediately and give an insoluble tertiary alkyl chloride that separates as an immiscible layer or as an emulsion. but it may also be helpful in anchoring the integration. Aromatic aldehydes and ketones do not react.1-11. Aliphatic aldehydes give a positive test (the formation of a brown-reddish precipitate of cuprous oxide) with the Fehling’s reagent. For example if your spectrum shows two triplets and no other multiplet (doublet. you should try to anchor the integration by making a reasonable assumption about the number of hydrogens under a certain peak. to your sample. • Interpret the multiplicity of the peaks following the n+1 rule. Esters do not react. Primary alcohols do not react with the Lucas reagent at room temperature. These tests will allow you to distinguish between primary and tertiary alkyl halides. if you suspect an aldehyde. If you suspect an alkyl halide. it’s very likely that . The TA will show you how to use the refractometer.2 Unit 33 If you suspect an alcohol.3 and measure the index of refraction of your unknown liquid. The interpretation of the NMR should be done in the lab. After you have identified your unknown. such as water. If you are on the right track the TA will give you the 1H-NMR to interpret.14 for a list of commonly used solvents and their NMR signals. the CRC and any other reliable handbook or online source can be used for this. a ketone or an ester perform the 2. If you suspect an aldehyde. otherwise he/she will help you reorient. Make sure that you estimate the fourth decimal place. If your 2.3000-1. quartet. Since you do not know the molecular formula. performed the chemical tests and measured the index of refraction.4-nitrophenylhydrazine test was positive. Example. triplet. Aldehydes and ketones react almost immediately and give a yellow-orange (sometimes red) precipitate when treated with 2. The Lucas test is limited to alcohols soluble in the Lucas reagent (monofunctional alcohols with fewer than six carbon atoms). compare the index of refraction measured with the literature value as further confirmation of its identity. your integration will be in relative terms. you should perform the Lucas test.). Your assumption should be based on the functional group present in your molecule and knowing that the unknown is a relative small molecule with 9 or fewer carbons. These are the steps to follow: • Notice the solvent used to obtain the NMR and make sure that you do not assign solvent or impurity signals. The Merck Index. Secondary alcohols soluble in the Lucas reagent react more slowly than tertiary alcohols and cloudiness or an immiscible layer forms within 5-10 minutes. Read sections 11. including water. perform the Fehling’s test as well. That peak should integrate for only 1H. they give a negative test as evidenced by the lack of a precipitate. The index of refraction of your unknown will be in the range 1. p. check your results with your TA.4dinitrophenyldrazine test. • Integrate the 1H-NMR following the steps discussed in lab lecture (section 33. 732). This will tell you not only the number of neighbors for each type of hydrogen. After you have run the IR spectrum. etc.5500. Check Table 33. However.9. locate the aldehyde hydrogen.

Avoid skin contact and inhalation. Perform these tests in the fume hood. If you leave a peak without interpretation. • Keep in mind that contrary to IR. Chemical tests Interpret your IR spectrum and decide which one(s) of the following tests you will perform. Use the correlation tables (Tables 33.5-0. separation of an immiscible layer or emulsion. Gloves should be changed often and removed immediately after completion of the chemical operations. In a small test tube place 0. the time it takes for the change to occur and whether it happens at room temperature or after heating.12) and compare them with the expected values (Table 33. every peak of the NMR spectrum should be accounted for (except for very small peaks that may belong to impurities). • Before you leave the lab you must present the name and structure of the unknown to your TA and explain your reasoning.2). Tertiary alcohols react very fast and form an immiscible layer almost immediately. Primary alcohols do not react. • • • • • • Lucas test If you suspect an alcohol. Record your observations. The match between your calculations and the experimental values should be acceptable.3 Unit 33 each triplet integrates for 2H (the 2H that split the neighbor’s signal into a triplet). • Measure the coupling constants (section 33. 2. it’s very likely that your proposed structure is wrong.7 mL of Lucas reagent (ZnCl2 in concentrated HCl) add about seven drops of your unknown and mix well. Silver nitrate produces unsightly stains. a change in color. Secondary alcohols form an immiscible layer within 5-10 minutes. If the unknown doesn’t dissolve.4-dinitrophenylhydrazine is a skin irritant and may cause skin allergies. • Pay attention to the chemical shifts to determine the environment of each hydrogen (shielded or deshielded). check first its solubility in water: In a small test tube add 1 mL of water and then add a drop of the unknown. go ahead and perform the Lucas test below. Chemical tests are based on observable changes such as the formation of a precipitate. Note the time it takes to form an emulsion or an insoluble layer. precipitate formation. consider it insoluble and do not perform the Lucas test below.3-33. Hydrochloric acid is corrosive and may cause severe burns. Avoid skin contact.8) to calculate the chemical shifts of all the hydrogens. Record your observations carefully. mix well. If the unknown dissolves in water. Avoid skin contact and inhalation. Safety First Wear gloves when performing the reactions. Its insolubility is an indication that the molecule likely has six or more carbons. . Tartrates are irritant. etc. describe changes in color.

ketones react more slowly and may take a few minutes before they produce a yellow-orange precipitate. The product alkyl chloride is insoluble and separates as an immiscible layer or as an emulsion. Thus. R R R OH C R + HCl (c) ZnCl2 R C R Cl + H2O soluble 3o > 2o >> 1o insoluble The role of ZnCl2 is to act as an acid catalyst (Lewis acid). a nucleophilic substitution reaction catalyzed by acids.4-dinitrophenylhydrazine reagent (2. The reaction takes place by an SN1 mechanism and that’s why tertiary alcohols are the most reactive. This test is based on the reaction of alcohols with HCl. The test is useful only for alcohols soluble in the Lucas reagent (small alcohols with fewer than six carbons).5-0.4-Dinitrophenylhydrazine test In a disposable 13x100 mm test tube. Record your observations. .7 mL of 2. Esters give a negative test. no colored precipitate forms.4 Unit 33 Cleaning up • With the aid of a Pasteur pipet transfer the contents from the Lucas test tubes to a container labeled “Lucas Test Waste. followed by secondary alcohols. Primary alcohols do not react with Lucas reagent at room temperature. the formation of an emulsion or immiscible layer is considered a positive test.4-dinitrophenylhydrazine in solution with H2SO4 and ethanol/water) and add five drops of the unknown and mix thoroughly with the help of a glass rod. Aldehydes react almost immediately giving a yellow-orange precipitate. place 0. In a negative test. R R Zn2+ OH R R Zn O H C R C R R R C R + (ZnOH)+ HCl R R C R Cl + Zn2+ + H2O 2. that leads to the formation of alkyl chlorides.” Rinse the test tubes with a little ethanol and dispose of the rinses in the same container.

5 Unit 33 Cleaning up • • With the aid of a Pasteur pipet transfer the contents from the 2. Fehling’s test In a small test tube place 0. The mechanism for this reaction is complex and not fully understood and we will not discuss it here. Record your observations.4-dinitrophenylhydrazone insoluble absorbs blue appears orange Esters do not react with 2.4-dinitrophenylhydrazine (absorbs violet) resulting in a product that appears orange (the complementary color of blue).5 mL of Fehling’s reagent A (copper sulfate in water) and 0. The mechanism for this reaction (a type of nucleophilic addition to the carbonyl) is discussed on page 419 in the lab book. can be easily oxidized with Cu2+.4-dinitrophenylhydrazine test in the container labeled “2.4-dinitrophenylhydrazine absorbs violet appears yellow 2. Aromatic aldehydes and ketones do not react. The aldehyde gets oxidized to a carboxylic acid (or a carboxylate under the basic conditions of the test) and the copper gets reduced to Cu+ (in the form of Cu2O) and occasionally to Cu0 (metallic copper). Aliphatic aldehydes give a positive test. Dispose of the test tubes from the 2. The end result is the formation of an insoluble 2. A positive test is the formation of a reddish precipitate of cuprous oxide (Cu2O) or metallic copper deposited on the walls of the tube (copper mirror).” Rinse the test tubes with a little ethanol and dispose of the rinses in the same container. especially aliphatic aldehydes.4dinitrophenylhydrazone which has a more extended system of conjugation than the original hydrazine and absorbs visible light at longer wavelengths (blue) than the original 2. . Cleaning up • With the aid of a Pasteur pipet transfer the contents from the Fehling’s test tubes to a container labeled “Fehling’s Test Waste.4-DNPH test.4-dinitrophenylhydrazine. R C R O + H2N-NHO 2N NO2 HA R C R N-NHO 2N NO2 + H2O 2.liquid waste.” This test is based on the nucleophilic addition of ammonia derivatives. such as hydrazines (H2N-NH-R).” Rinse the test tubes with a little ethanol and dispose of the rinses in the same container.solid waste.4-DNPH test. This test is discussed on pages 420 and 429 of the lab book. Add two drops of the liquid sample and heat the liquid in a boiling water bath for about 10 minutes. Aldehydes.4-dinitrophenylhydrazine test tubes to a container labeled “2. to the carbonyl group of aldehydes and ketones.5 mL of Fehling’s reagent B (sodium hydroxide and sodium potassium tartrate in water).

O HO O O OH O O O Cu2+ + 2 HO O O 2 HO- O Cu O O O HO O O + 2 H2O tartrate blue complex Silver nitrate in ethanol test In a small and perfectly dry test tube place 1 mL of a 0. The tartrate forms a complex with the Cu2+ and keeps it in solution where it can react with the aldehyde. In general. If no precipitate is observed within five minutes at room temperature. 276 and 285 of the lab book. alkyl iodides react faster than alkyl bromides and alkyl bromides faster than alkyl chlorides. is a reddish precipitate. This test is also discussed on p. The tartratecopper complex is deep blue. Carboxylic acids produce an insoluble silver carboxylate.6 Unit 33 H R C O + 2 Cu2+ blue + 5 HO- tartrate heat O R C O + 3 H2O + Cu2O reddish precipitate an aliphatic aldehyde a carboxylate The reaction. Tertiary alkyl halides and allylic halides react very fast. To prevent this from happening. and the product of the reaction. Alkyl halides react with silver nitrate in ethanol and give silver halide which separates as a precipitate. Aryl halides and vinylic halides give a negative test even upon heating. . Carefully record your observations. The reaction is considered positive if a precipitate forms. In the presence of strong base (NaOH) needed for the redox process. Chelating agents are important in industry. medicinal chemistry and biological process for their ability to form stable complexes with metals (as an example.1 M solution of silver nitrate in ethanol. The formation of this precipitate is considered a positive test. secondary alkyl halides react slower and primary alkyl halides only after heating. is accompanied by a drastic change in color. Cu2+ under basic conditions is deep blue. think of the heme group in hemoglobin as a chelating agent for the Fe2+ ion found in its center). Tartrate is what we call a chelating agent. usually Cu2O rather than metallic copper. place the solution in a water bath at approximately 80oC and observe if a precipitate forms. a redox process. Add two-three drops of your unknown and mix well. Cu2+ would precipitate out in the form of insoluble Cu(OH)2 and would not react with the aldehyde. Observe the formation of a precipitate and notice how long it takes to form. tartrate is added along with the NaOH. A chelating agent is an organic compound that can form two or more coordinate bonds with one central metal ion. The reaction is performed in the presence of tartrate to keep the Cu2+ in solution under basic conditions.

If no precipitate is observed after three minutes at room temperature. Dispose of the unused unknowns in the bag labeled “Unknowns-waste. Alkyl chlorides and alkyl bromides react with sodium iodide in acetone and give an alkyl iodide and sodium chloride or sodium bromide. aryl halides and vinyl halides do not react. R X + NaI acetone R I + NaX X = Cl.7 Unit 33 R X + AgNO3 ethanol R ONO2 + AgX The reaction takes place by an SN1 mechanism and therefore tertiary alkyl halides react faster than secondary. place the solution in a water bath at approximately 50oC for about five minutes and observe if a precipitate forms. therefore. Record your observations.” EXPERIMENT 33 REPORT . primary and secondary alkyl chlorides and secondary and tertiary alkyl bromides react upon heating at 50oC. Carboxylic acids react with silver nitrate in ethanol and produce a precipitate of silver carboxylate.” Rinse the test tubes with a little ethanol and dispose of the rinses in the same container. Primary alkyl halides react slowly and give a precipitate only upon heating.1 M solution of a 15% (w/v) solution of sodium iodide in acetone. aryl halides and vinylic halides do not react even after heating. R-COOH + AgNO3 ethanol R-COOAg + HNO3 Sodium iodide in acetone test In a small and perfectly dry test tube place 1 mL of a 0. Cleaning up • • With the aid of a Pasteur pipet transfer the contents from the silver nitrate and sodium iodide test tubes to a container labeled “Alkyl Halides Tests Waste. 277 and 285 of the lab book. Br This test is based on the limited solubility of sodium chloride and sodium bromide in acetone. Add two-three drops of your unknown and mix well. Primary alkyl bromides react at room temperature. The reaction takes place by an SN2 mechanism and. Tertiary alkyl chlorides. respectively. which are insoluble in acetone. Tertiary alkyl chlorides. while sodium iodide (the reagent) is soluble in acetone. primary alkyl bromides and chlorides react faster than secondary ones. Bromides react faster than chlorides. sodium chloride and sodium bromide (the possible side products) are not. This test is also discussed on p. The formation of a whitish precipitate of sodium chloride or bromide is considered a positive test. The reaction is considered positive if a precipitate of sodium chloride or sodium bromide forms.

O O O O 13) Using the concept of critical angle (show a drawing).4-dinitrophenylhydrazones are more conjugated than 2. What is considered a positive test? Which alkyl halides react faster? 11) Write a chemical reaction for 1-bromobutane and sodium iodide in acetone. Interpret and discuss the results of such tests. What is considered a positive test? Which alkyl halides react faster? 12) How would you distinguish these two compounds by 1H-NMR? Use chemical shift arguments.2). 6) What is considered a positive 2. Present the important bands in the usual table format. Show all your calculations. Calculate the chemical shifts of all the hydrogens using correlation tables. What is considered a positive result for the Lucas test? 3) Which alcohols react faster with the Lucas reagent? Why? 4) Write a chemical reaction for acetone and 2. 8) What is considered a positive Fehling’s test? Do aromatic aldehydes react? 9) Using chemical reactions.8 Unit 33 Pre-lab 1) How would you distinguish each compound in the following pairs by IR spectroscopy? Give the bands (with their assignments) that you would use to tell each compound apart. coupling and chemical shift. Account for coupling according to the n+1 rule.4-nitrophenylhydrazine test? 7) Write a chemical reaction for formaldehyde and the Fehling’s reagent. Which band(s) did you use to identify the functional group? 3) Report the results of your chemical tests. Measure the coupling constants and compare them with table values (Table 33. Show your integration calculations. If you do not know their structures check them out in a textbook or online: a) 1-pentanol and 1-pentanal b) benzoic acid and phenol c) cyclohexanone and benzaldehyde d) methyl acetate and acetic acid 2) Write a chemical reaction for 3-methyl-3-pentanol and the Lucas reagent. explain the role of tartrate in the Fehling’s test. What is a chelating agent? 10) Write a chemical reaction for 2-methyl-2-chlorobutane and silver nitrate in ethanol. if applicable.4dinitrophenylhydrazine. 5) Explain how 2. Show the mechanism for the reaction. Briefly explain your reasoning. 4) Report the index of refraction of your unknown. Present the data in a table format. 2) Interpret the IR of your unknown. Account for integration. In-lab 1) Identify your unknown. including pH determination. How does it compare with the index of refraction reported in the literature? 5) Interpret the 1H-NMR. . give the definition of the index of refraction in the form of an equation. Draw its structure and name it. Include calculation of chemical shifts using correlation tables.4-dinitrophenylhydrazine.