2 (© desired compound (© desired compound ‘© impurities © impurities Figure 1 Figure 2 Extraction occurs when the desired Washing occurs when impurities change compound changes layers, leaving layers, leaving the desired compound behind impurities behind however, extraction almost always refers to the transfer of compounds — from one liquid solvent to another liquid solvent. eer ‘A compound can be separated from impurities in a on ‘ ‘extracting the compound from the original or first solvent into a solvent. The compound must be more soluble in the second solvent than in the first solvent, and the impurities must be insoluble in the second solvent. ‘Also, to effect the extraction, the two solvents selected must immiscible, or not soluble in one another, so that they produce separate solvent layers. After dissolving the mixture in the first solvent, th solution is added to the second solvent. The two layers are vigorously to maximize the surface area between them. This mixing facilitates transfer of a dissolved compound from one layer to another. Once tt transfer process is complete, the layers are again allowed to form, as in Figure 1. Separation of the two layers then completes the separation of t desired compound from the impurities. Washing is the reverse process, in which the impurities are removed the second solvent, leaving the desired compound in the original sol ‘The amount of compound to be extracted determines whether macros ‘or microscale techniques should be employed for the extraction. The chemical principles associated with the extractions are identical, but techniques are somewhat different. " Extractions using larger quantities of solvents, tens or hi milliliters, require a separatory funnel, as shown in Figure 3. The sob layers are mixed by shaking the separatory funnel. Then the layers allowed to reform. The bottom layer is drained through the stopcock; top layer is poured from the top of the separatory funnel. Microscale extractions can be conducted using a test tube or a tube. Mixing and separating the layers can be done using a Pasteur p{© 2004 Cengage Leaning ‘TECHO705: Separating Acids and Neutral Compounds by Solvent Extraction Figure 3 A separatory funnel used for ‘macroscale extractions Figure 4 Ethers and hydrocarbons are less dense than woater Dichloromethane is more dense than water chlorinated solvent, such as dichloromethane. When the two immiscible solvents are placed into a container, two liquid layers result. The more dense solvent is always the bottom layer. It is important to identify the solvent in each layer. Hydrocarbons and ethers are less dense than water or the dilute aqueous solutions used in extractions. When one of these nonpolar solvents is used, the water layer is the bottom layer, as shown in Figure 4 However, dichloromethane is more dense than water. When dichloro- methane is used as the nonpolar solvent, the water layer will be the top layer, as shown in Figure 5. Although the identity of each layer can be established from the density of each solvent, their identities should be confirmed. To confirm the identities of the layers, one or two drops of water are introduced just below the surface of the top layer. If the drops of water mix with the top layer, then the top layer is the water layer. If the drops of water fall through the top layer to the layer below, then the water layer is the bottom one. It is a good practice to save all layers in labeled containers until the experiment is complete and the desired product is isolated. Often the two solvents will not completely separate after shaking, due to the formation of an emulsion at the interface between them. An emulsion is a suspension of small droplets of one liquid in another liquid. Emulsions are generally opaque or cloudy in appearance and are often mistaken as a third layer. The small size of the droplets in an emulsion causes the separation of the two solvents to take place very slowly. Several procedures may be helpful to facilitate this separation. For example, gentle swirling of the container, addition of a few drops of saturated aqueous sodium chloride (NaC) or ethanol, or addition of more solvent to dilute the solutions may help. In particularly difficult cases, it may be necessary to filter the mixture to remove small solid particles that promote emulsion formation.“ “TECHO70S: Separating Acid and Neural Compounds by Soren Exracon A simple, but useful, guide to solubility is like dissolves like. That is, nonpolar compounds, including most organic compounds, are more soluble in nonpolar solvents than in polar solvents. On the other hand, ionic and polar compounds are more soluble in polar solvents, such as water. These solubility differences can be exploited to separate nonpolar compounds from ionic or polar compounds. For example, synthetic reactions often produce ionic, inorganic salts a8 by-products of the desired nonpolar organic product. In such cases, these salts are removed by washing the nonpolar solvent with water. The organic compound remains dissolved in the nonpolar solvent. Some organic compounds are sufficiently polar to be quite soluble in water. Extraction of such polar compounds into a nonpolar solvent is often difficult: The process can be facilitated by using the technique called salting out. Inorganic salts, such as NaCl, are dissolved in water to reduce the solubility of the organic compound in the aqueous layer. Under these conditions, the organic compound preferentially dissolves in the nonpolar layer. Extraction is a particularly effective means of separating organic ‘compounds if one compound in the mixture can be chemically converted to an ionic form. The ionic form is soluble in an aqueous layer and can be extracted into it. Other non-ionized organic compounds in the mixture will remain dissolved in the nonpolar solvent layer. Separation of the two layers results in the separation of the dissolved compounds. Tonic forms of some organic compounds can be produced by reacting them with aqueous acids or bases (see Figure 6 below). Reacting organic ‘acids with bases such as sodium hydroxide (NaOH) converts these acids to water-soluble anions. Reacting basic amines with dilute aqueous acid solutions such as hydrochloric acid (HCD converts the amines to water- soluble cations. ‘The extent to which an acid-base reaction proceeds to completion depends upon the relative acidity and basicity of the reactants and products. Reactions occur so that stronger acids and bases react to produce weaker conjugate bases and acids. Recall that the pK, is a measure of the acidity of an acid, as shown in Equation 1. PK, = —log K,base, OH”, removes a 3. The stronger * from p-toluic acid to form the salt, p-toluate. The polar Y ‘aqueous solution. Both OH” and p-toluate are bases. The 0 6 indicates that OH is a stronger base than p-toluate, with a pK, of ‘The stronger base takes H* from the weaker base. ‘Similarly, OH” is a stronger base than p-tert-butylphenoxide ion, with 4 pK, of 102. So OF takes I from per butyiphenol to form the water soluble p-teri-butylphenoxide ion. Sodium hydrogen carbonate (NaHCO), with a pK, of 6.4, is a weaker base than p-tert-butylphenoxide ion, so HCO3~ will not take H* from p+tert-butylphenol, as shown in Equation 4. As a result, p-tert-butylphenol is not converted to a salt in aqueous sodium hydrogen carbonate and does not become water soluble. Although aqueous NaHCO; is not sufficiently basic to react with p-tert- butylphenol, NaHCO; will react with p-toluic acid to form the water soluble p-toluate, as shown in Equation 5. Oy dpceear = ay ke + m0 42 poluic acid (pK, =42) Piotuaearion P16 cay) + or = ct O)j-o so 4.3) plert-butylphenol (PK, = 102) ptert-butylphenoxide anion PAr= 16 cw {(O)—on +HCO; 4" — NoREACTION 4.4) perbutylphenol (pK, = 102) i cy C—OH + Hoo > a O-bo- +#4,00, 4.5) poluic acid (pK, = 42) ‘poisan ieica a ia F agu Te postion of an acid-tse determined by the relative acidity ofthe reactant acid and the product acid $Microscale Extraction ‘TECHO705: Separating Acids and Neutral Compounds by Solvent Extraction ‘The p-toluic acid and the p-tet-butylphenol can be recovered by adding HCI to the aqueous solutions. The p-toluate and /Iphenoxide ions are stronger bases than is Cl-, so each one takes H” from HCl. The acid forms are not water soluble, so they precipitate from solution. ‘The procedure you will use in this experiment exploits the differences in these reactions to separate p-toluic acid and p-tert-butylphenol from the nonpolar solvent in which they are dissolved. First, you will extract only prtoluic acid into NaHCOs solution. Then, you will extract p-tert= butylphenol into NaOH solution. Next, you wili add HCl to each of the extracts to precipitate the water-insoluble p-toluic acid and p-tert- butylphenol. You will isolate the precipitates from the solutions by vacuum filtration, then air dry them. A flowchart for the separations is shown in Figure 8 on the next page. A third compound, acetanilide, does not react with either NaOH or NaHCO, and remains dissolved in the nonpolar solvent. To recover acetanilide, you will dry the nonpolar layer with anhydrous sodium sulfate (NazSO,) and evaporate the solvent in a fume hood. You will recrystallize the acetanilide in an ice bath. After you dry the compounds, you will measure the mass of each isolated compound. Finally, you will measure the melting point of each compound and assess its ‘purity by comparing the experimentally ‘measured melting point with the literature value. Equipment 3 beakers, 50-mL hot plate 2 beakers, 250-mL” melting point capillary tubes 15-mL centrifuge tube, 5 Pasteur pipets, with latex bulb with plastic cap pH test paper filter paper, to fit sand bath’ Hirsch filter funnel thermometer, ~10 to 110 °C glass stirring rod 2 watch glasses 10-mL graduated cylinder weighing paper Hirsch filter funnel, with 50-mL filter flask and gasket “one for the ice bath, the other to support the centrifuge tube ‘sand in crysalizing dish on electric ot plate or sand in electric heating well with hea controller|© 2004 Cengage Learning CgHgNHCOCH, P-CHs—CgHy—COO™ P-CiHy—CoH,OH Hen as allt B= e208 sotution eit oe ether aqueous filtrate residue (extraction) (filtration) discard P-CH;—CgH,—COOH ‘CgHsNHCOCH, P-CyHy—CgH,O~ 8 |-— HCI solution a (jensiees te residue (anhydrous) (filtration) decantate residue | discard P-C,He—GgH,0H (decantation) tiie H=NHCOCH,, discard vapor (evaporate) discard (CgHgNHCOCH, Figure 8 Flowchart for separations using microscale ecnigues“a ‘TECHO705: Separating Acids and Neural Compounds by Solvent Extraction Reagents and Properties ‘Molar mass Substance Quantity (g/mol) mp (°C) — bp CC) acetanilide 60 mg 135.16 113-115 tert-butyl methyl ether 5 mL 88.15 55-56 p-tert-butylphenol 100 mg 15022 98-101 3M hydrochloric acid = 2mL. 05M sodium hydrogen carbonate 6mL 05M sodium hydroxide 6 mL sodium sulfate, anhydrous 05g p-toluic acid water, 100 mg 136.15 180-182 distilled or deionized Preview * Dissolve acetanilide, p-toluic acid, and p-tert-butylphenol in t-butyl methyl ether (see Figure 8) ‘+ Extract p-toluic acid from the ether layer with NaHCO; solution, separating the layers + Extract p-tert-butylphenol ffm the ether layer with NaOH solution, separating thedayers * Isolate p-toluic acid by adding HCI solution to the aqueous NaHCO, layer + Isolate p-tert-butyiphenol by adding HCI solution to the aqueous NaOH layer + Isolate acetanilide by drying the ether solution, then evaporating t-butyl methyl ether in a fume hood * Dry the isolated compounds and measure their masses * Measure the melting point of each compound, and compare to literature value PROCEDURE 1. Preparing the Extraction ‘Wear departmentally approved safety goggles at all times while in the Z\ 3 chemistry laboratory. Always use caution in the laboratory, Many chemicals are potentially harmful, Prevent contact with your eyes, skin, and clothing. Avold ingesting any of the reagents. CAUTION Acetanilide is toxic and irritating. terButyl methyl ether is flammable and Inritating.(© 204 Cengage Learning ‘TECHO705: Separating Acs and Neutral Compounds by Solvent Extraction 2. Extracting p-Tolule Acld 3. Extracting p-tert- Butyiphenol Weigh 50-70 mg of acetanilide and 80-120 mg each ot vatyiphenel: Record the exact mass ofeach sold. Place 5 mL of tert ether into a 15-mL centrifuge tube. Add the three solids to the Tee Seah? Oa cage Fas nod ia to Goslre CULE Reactions between sodium hydrogen carbonate (NaHCO,) and acids produce carbon dioxide (CO,) gas, which can result in foaming. ‘Add 2 mL of 0.5M aqueous NaHCO; to the ether solution in the centrifuge tube. Gently and thoroughly mix the two layers in the centrifuge tube. NOTE: To ensure complete reaction, it is important to mix the layers well. If you are Using a centrituge tube with a tightly fited cap that does not leak, vigorous sheking can achieve this mixing, An alternative technique is to repeatedly draw the mixture into a Pasteur pipet and then to forcefully discharge the mixture back into the tube. Once any initial reaction has subsided, place the plastic cap on the centrifuge tube and shake gently. Remove the cap to allow any CO. gas to escape. Repeat this process several times, gradually increasing the intensity with which you shake the tube. Shake vigorously, because thorough mixing of the layers is essential. Support the centrifuge tube in a beaker or flask and allow the liquid layers to separate. Confirm the identity of the layers by using a Pasteur pipet to introduce one or two drops of water just below the surface of the top layer. Make certain no air isin the pipet tip. Closely observe what happens to the drops. Using a Pasteur pipet, carefully remove the aqueous layer and transfer it to a labeled 50-mL. beaker. To remove any toluic acid remaining in the ether layer, add a second 2-mL NaHCO; portion to the tube containing the ether mixture. Shake the tube vigorously. Remove the aqueous layer and combine it in the beaker with the first extract. Repeat with a third 2-mL NaHCOs portion. Add 1 ml of distilled or deionized water to the centrifuge tube and mix. Remove the aqueous layer and combine it with the three NaHCO, solution extracts in the 50-mL beaker. Save this aqueous solution for Part 4. ORI iia acai neracemcemee | peeeeeceereme ores (NEON) (6 torso ened Gorrosive,d. 1) 8) cue Add 2 mL. of 0.5M NaOH to the ether solution remaining in the centrifuge tube, Shake the tube vigorously. Using a Pasteur pipet, remove the aqueous layer and transfer the layer {nto a clean, labeled, 50-mL beaker. Repeat the extraction of the ether layer with a second 2-mL NaOH portion. Remove the second NaOH layer and combine it with the first. Repeat with a third 2-mL NaOH portion. Add 1 ml of water to the ether remaining in the centrifuge tube and mix, Remove the aqueous layer and combine it with the three NaOH extracts. Save the NaOH extracts in the 50-mL. beaker for Part 5. Save the ether layer remaining in the centrifuge tube for Part 6.4. Isolating p-Toluic Acid 5. Isolating p-tert- Butylphenol 6. Isolating Acetanilide ‘TECHO705: Separating Acids and Neutral Compounds by Solvent Extraction ‘3M hydrochloric acid (HCI) is toxic. Adding 3M HCI to the NaHCO; solution will produce C02, causing a large amount of foaming. Select the 50-mL beaker containing the NaHCO, extracts from Part 2. Add 3M HCI dropwise to the NaHCO; solution to precipitate the p-toluic acid. Notice that foaming occurs and a precipitate of p-toluic acid forms. Continue to add the 3M HCI, dropwise with stirring, until no more solid is produced and the solution tests acidic (pH < 3). NOTE: To test for acidity, remove a drop ofthe Solution with a string rod and place the drop on a small piece of pH test paper. Weigh a filter paper and record its mass. Using the weighed filter Paper, separate the crystals from the solution using vacuum filtration with a Hirsch funnel. Support the crystals and paper on a watch glass and allow the crystals to air dry. Select the 50-mL beaker containing the NaOH extracts from Part 3. To remove any remaining traces of tert-butyl methyl ether that might inhibit the crystallization of the phenol, heat the NaOH solution to about 60 °C on a hot plate in a fume hood. Remove the beaker from the hot plate and allow the solution to cool To precipitate p-tert-butylphenol, add 3M HCI dropwise to the cooled solution until it tests acidic. If the phenol separates as an oil, cool the ‘mixture in an ice bath to facilitate crystallization. Weigh a filter paper and record its mass. Using the weighed filter paper, separate the p-tert-butylphenol crystals from the solution by filtration with a Hirsch funnel. Support the crystals and paper on a watch glass and allow the crystals to air dry. Anhydrous sodium sulfate (NazSO,) is Irritating and hygroscopic. A Select the centrifuge tube containing the ether layer from Part 3. Add approximately 0.5 g of anhydrous Na,S0O, to the centrifuge tube to remove any traces of water from the ether-acetanilide solution. Cap the tube, shake it, and allow it to stand for 5 min, NOTE: After anhydrous NagSQ, adsorbs water, it wil look like salt or sugar. Weigh a 50-mL beaker and record its mass. Decant the dried ether acetanilide solution into the 50-mL beaker, leaving the Na;SO, in the centrifuge tube. Evaporate the ether, in a fume hood, by warming the beaker on a 50 °C sand bath while gently blowing air or nitrogen over the solution, Avoid overheating. pe be ee eee NOTE: Too much heat causes acetanilide to sublime. When the ether has evaporated, ‘8 small amount of oll will remain, and the container wil fee! hot tothe touch.©2004 Cengage Leaming “TECHO7OS: Sepang Acids and Neural Compounds by Solvent Exuacion » Figure 9 ‘Mixing solutions in a separatory funnel Add 10 mL of 05M aqueous NaHCO; to the ether solution in the separatory funnel. Place the glass stopper in the top of the separatory funnel, and invert the funnel while holding the stopper in place, as shown in Figure 9, Gently mix the two layers by rocking the separatory funnel bback and forth. With the funnel inverted, open the stopcock to vent any gas that is generated. Listen for the gas as it exits through the stopcock. Continue this ‘mixing process, gradually increasing the force of the mixing, until the funnel can be shaken quite vigorously with no gas being produced upon venting. Place the funnel in the support ring and allow the layers to separate, Confirm the identity of the layers by using a Pasteur pipet to introduce one or two drops of water just below the surface of the top layer. Make certain no air is in the pipet tip. Closely observe what happens to the drops. Remove the glass stopper from the top of the funnel, and open the stopcock to allow the aqueous layer to drain into a clean, labeled 100-mL beaker. When the interface between the layers just reaches the bottom of the funnel (top of the stopcock), close the stopcock to retain the ether layer in the funnel. NOTE: It you open the stopcock while the glass stopper is in the top of the separatory funnel, a slight vacuum wil be created, and the bottom layer will not drain from the funnel Add a second 10-mL NaHCO; portion to the funnel to remove any p+toluic acid remaining in the ether layer. Mix with frequent venting. After the layers have separated, drain the aqueous layer into the beaker with the first extract, Repeat with a third 10-mL NaHCO; portion. Add 5 mL. of distilled or deionized water to the separatory funnel and mix. Drain the water layer into the beaker containing the three NaHCO solution extracts. Save this aqueous solution for Part 4.3. Extracting p-tert- 4, Isolating p-Toluic Acid ‘TECHO705: Separating Acids and Neutral Compounds by Solvent Extraction (0.5M Sodium hydroxide (NaOH) is toxi Add 10 mL of 0.5M NaOH to the ether solution remaining in the separatory funnel. Mix the layers as before so that the NaOH and the p-tert- butylphenol can react. Remember to mix cautiously at first with frequent venting through the stopcock. Allow the layers to separate, and drain the aqueous NaOH layer into a clean, labeled 100-mL beaker. Repeat the extraction of the ether layer with a second 10-mL NaOH portion. Drain the NaOH layer from the separatory funnel into the 100-mL beaker containing the first NaOH extract. Repeat with a third 10-mL NaOH portion. Add 5 mL of water to the ether remaining in the separatory funnel and mix. Allow the layers to separate. Drain the water layer into the 100-mL. beaker containing the three NaOH extracts. Save the NaOH extracts for Part 5. Save the ether layer remaining in the separatory funnel for Part 6. ‘3M Hydrochloric acid (HCI) is toxic. Adding 3M HCI to the NaHCO; solution ZA will produce CO,, causing a large amount of foaming. Select the 100-mL beaker containing the NaHCO, extracts from Part 2. To precipitate the p-toluic acid, carefully add 3M HCI to the NaHCO; solution. Notice that foaming occurs, and a precipitate of p-toluic acid forms. Continue to add the 3M HCI, dropwise with stirring, until no more solid is produced and the solution tests acidic (pH < 3). NOTE: To test for aciity, remove @ drop ofthe solution with a stiring rod and place the drop on a small piece of pH test paper. Se Weigh a filter paper and record its mass. Using the weighed filter Paper, separate the crystals from the solution using vacuum filtration with 4 Biichner funnel. Support the crystals and paper on a watch glass and allow the crystals to air dry. Select the 100-mL beaker containing the NaOH extracts from Part 3. To remove any remaining traces of tert-butyl methyl ether that might inhibit the crystallization of the phenol, heat the NaOH solution to about 60 °C on hot plate in a fume hood. Remove the beaker from the hot plate and allow the solution to cool. ‘To precipitate crystals of p-tert-butylphenol, carefully add 3M HCl to the ‘cooled solution until itis acidic. If the phenol separates as an oil, cool the mixture in an ice bath to facilitate crystallization. a filter paper and record its mass. Using the weighed filter Paper, separate the p-feri-butylphenol crystals from the solution filtration with a Buchner funnel. Support the crystals and paper on. ‘watch glass and allow the crystals to air dry.{© 2004 Cengage Leaning -TRCHO705: Separating Acid and Neutral Compounds by SolvencExraction| 6. Isolating Acetanilide 8. Cleaning Up ‘Anhydrous sodium sulfate (Na,SO,) is Irritating and hygroscopic. Select the separatory funnel containing the ether layer from Part 3. Transfer the ether-acetanilide solution from the separatory funnel to a clean 125-mL Erlenmeyer flask. Add approximately 1g of anhydrous Na;SO, to the flask to remove any traces of water from the solution. Stopper the flask and allow it to stand for 5 min with occasional swirling. i a tS IRN mata ee NOTE: After anyhydrous NazSO, adsorbs water, it wil ]0ok tke salt or sugar. Weigh a 100-mL beaker and record its mass. Decant the clear, dried ether-acetanilide solution into the 100-mL, beaker. Evaporate the ether, in a fume hood, by warming the beaker on a hot plate while a stream of air passes over the solution. Avoid overheating. NOTE: Too much haat causes acetanilide to sublime. When the ether has evaporated, ‘small amount of oil will remain, and the container will fee! hot to the touch. Crystallize the oil residue, the acetanilide, by cooling the beaker in an ice bath. If necessary, scratch the bottom of the beaker with a glass stirring rod, or add a seed crystal, to induce crystallization. Allow the acetanilide crystals to dry. When all of the samples are dry, measure the mass of each compound. Measure the melting point of each compound, and assess its purity by comparing the measured melting point with the literature value. Use the labeled collection containers provided by your laboratory instructor. Clean your glassware with soap or detergent. thoroughly with soap or detergent before leaving the“TECHO70S: Separating Acids and Neutral Compounds by Solvent Extraction Pre-Laboratory Assignment 1. Briefly describe the hazards you should be aware of when you work with: (a) tert-butyl methyl ether (b) 3M HCL 2. Briefly explain or describe the following: (2) What is the difference between extraction and washing? (b) How would you determine which layer is the aqueous layer after you add NaHCO; solution to the ether solution of your compounds? (© Why is the NaOH extract heated before acidification? (a) What two visible evidences of reaction will you see when you acidify the NaHCO; extract with HCI solution? (e) In which layer would p-toluic acid be more soluble if p-toluic acid were added to a two-layer mixture of tert-butyl methyl ether and water? .© 2004 Cengage Leaning‘TECHO708: Separating Fetrocene and AceHferrocene by Adsorption Column Chromatography Adsorption column chromatography is a technique that uses a solid stationary phase, the adsorbent, packed in a glass column, and a solvent, the mobile phase, that moves slowly through the packed column. A solvent used as a mobile phase is called an eluent. In an adsorption column chromatography experiment, a mixture of ‘compounds is added to the eluent. As the eluent moves through the column, the stationary phase and the mobile phase interact with the compounds in the mixture, The differences in attraction of the compounds to the stationary and mobile phases result in the compounds moving at different rates through the packed column, separating from one another. A compound attracted more strongly by the mobile phase will move rapidly through the column, and elute from, or come off, the column dissolved in the eluent. In contrast, a compound more strongly attracted to the stationary phase will move slowly through the column. Alumina (AlO;) and silica gel (SiO-xH,O) are the most commonly used adsorbents for adsorption column chromatography. Alumina is generally suitable for chromatography of less polar compounds. Silica gel Rives good results with compounds containing polar functional groups. Even within a mixture of relatively low polarity compounds, the more polar compounds of the mixture bind tightly to the adsorbent; less polar ones bind more loosely. Separation occurs when an eluent of low to moderate polarity is passed through the column. Less polar compounds of the mixture readily dissolve in the eluent and move through the column. More polar compounds haye a stronger attraction to the adsorbent than to the moving eluent. When differences in attraction are great enough, the compounds can be separated, A mobile phase consisting of a single eluent may be sufficient to separate and elute each compound of a mixture. However, ifthe mixture to be separated. includes compounds with a wide range of polarities, two eluents may have to bbe used, In such a case, the eluent of lower polarity is used first. Table 1 shows the relative polarity of various eluents used as mobile phases. Eluents more polar than ethers should not be used with alumina Table 1 Relative polarities of various eluents hexane tetrachloroethane benzene toluene trichloroethane dichloroethane increasing diethylether polarity tert-butyl methyl ether ethyl acetate acetone T ethanol methanol ‘Water Y‘© 1007 Cengage Leaming scale chromatography column Preparing Adsorption Chromatography because such eluents either react on the alumina surface or they dissolve the alumina. Figure 1 shows the two most common types of columns employed in microscale column chromatography. The packed adsorbent is called the column bed. A typical column has a bed height ten times the column diameter. There are two satisfactory methods for packing adsorption chromato- graphy columns, the dry pack method and the slurry pack method. Using the Dry Pack Method The dry pack method, shown in Figure 1(a), uses a short-stem Pasteur pipet asa column. A small cotton or glass wool plug at the pipet bottom, layered with a small amount of sand, provides a level base for the adsorbent. Dry adsorbent is added. A sample mixture containing the compounds to be separated must also bbe added to the column in a dry form. Typically, the mixture is dissolved in a minimum volume of a moderately polar eluent, the solution is mixed with a small amount of adsorbent, and the eluent is removed by evaporation. The dry sample-adsorbent powder is added to the top of the column bed. Finally, a small amount of pure adsorbent is placed on top of the sample to prevent any disruption of the sample layer with the of eluent.Detecting the Separated Compounds Figure 2 (a) A vertical column allows ‘good separation of compounds, ‘while (b) a nonvertical column ‘causes non-horizontal bands ‘and poor compound separation ‘TECHO70S: Spaating Herocene and Aceercen by Adorson Column Chromatography Separation of the sample mixture compounds is achieved by adding eluent to the top of the column bed. Eluent is collected as it elutes from the bottom of the column, Using the Slurry Pack Method A microscale chromatography column, shown in Figure 1(b), contains a stopcock to control eluent flow and a glass frit to support the adsorbent. Although such a column could be packed using the dry pack method, the slurry pack method is more commonly used. The slurry used is a mixture of adsorbent and the chosen eluent. The column is packed by closing the stopcock, filling the column half full with the eluent, and adding the adsorbent-eluent slurry. During the addition of the slurry, the stopcock is opened to allow eluent to drain slowly through the column as the adsorbent bed packs. Air bubbles must not be allowed to form in the bed. Such bubbles cause the bed to be irregular and interfere with the uniform movement of the mixture compounds through the column. The mixture sample is added to the column either dissolved in a minimum volume of a moderately polar eluent or as a dry sample-adsorbent powder, as described in the dry pack method. Separation of the mixture compounds is achieved by adding eluent to the top of the column bed. Eluent is collected as it elutes from the bottom of the column, If the separation requires more than one eluent, the column is packed using the less polar eluent. For example, hexane is less polar than tert-butyl methyl ether (TBME), as shown in Table 1. If both hexane and TBME are to be used, the column is packed with hexane. Each compound forms a band, or area of concentration, when a mixture is placed on the column. As a compound is attracted to the eluent, that band moves through the column. The column must be vertical at all times, as shown in Figure 2(a) If the column is not kept upright, Figure 2(b), bands may elute unevenly, preventing effective separation. band 9,(© 1007 Cengage Leeming Using the Dry Pack Method To calculate the percent recovery of the mixture compounds, the recovered TaMia eh co STs DA MRNA se Facet recovery for exch ‘compound can be calculated, as shown in Equation 1. " _ SS ee) oo) Percent recovery = \ mass of compound in original mixture, (Eq. 1) Equipment 2 beakers, 50-mL 2 Pasteur pipets, with latex bulbs 3-mL conical vial 9-mm Pasteur pipet cotton or glass wool sand bath, with thermometer" 4 Erlenmeyer flasks, 50-mL #1 stopper fine sand support stand labels utility damp ‘marking pen 20-cm wire microspatula ‘sendin cyslising dh ond nati hing wal with hen conor Reagents and Properties ‘Molar mass Substance Quantity — (gimol) mp CC) BCC) acetylferrocene 35mg «2881-83 alumina 33g 102 2054 tert-butyl methyl ether 20 mL. 88 55-56 _ ferrocene 35 mg 186 174.176 "hexane 20 mL 86 6Poe TECHO708: Separating Ferrocene and Acerylferocene by Adsorption Column Chromatography Preview ‘+ Prepare dry pack column + Prepare ferrocene-acetylferrocene sample + Apply sample to column ‘+ Elute ferrocene + Blute acetylferrocene ‘+ Evaporate eluents from ferrocene and acetylferrocene ‘+ Measure masses of separated compounds PROCEDURE A Chemical Alert 1. Labeling and Weighing Collection Containers 2, Preparing a Dry Pack Column 3. Preparing a Sample Pea etn fert-butyl methyl ether—flammable and irritant hexane—flammable and irritant Wear departmentally approved safety goggles at all times while in th chemistry laboratory. 4 Label four 50-mL Erlenmeyer flasks “hexane”, “hexane eluent”, “TBME”, and “TBME eluent’, respectively. Label two 50-mL beakers “ferrocene” and “acetylferrocene”, respectively. Weigh each beaker and record the masses. Prepare a column in @ short-stem, wide-bore, 9-mm Pasteur pipet by placing a small plug of cotton at its tip. Use a wire to push the cotton into place. Do not make the cotton plug too tight or eluent flow will be restricted. Using a utility clamp, attach the column to a support stand, making certain the column is vertical. Ifthe utility clamp is too large to firmly hold the column, use paper towels or a split stopper to hold the column in the clamp. Pour approximately 50 mg of sand on top of the cotton plug. Then slowly pour 3 g of alumina on top of the sand. Tap the side of the pipet to pack the column as you add the alumina. tert-Butyl methy! ether (TBME) is flammable and irritating. Keep TBME away from flames and other heat sources. Use TEME under a fume hood. ‘Acetylferrocene is highly toxic. Prevent eye, skin, and clothing contact. Avoid ingesting and inhaling these compounds. Weigh 70 mg of a 50:50 mixture of ferrocene and acetylferrocene. Place the ‘mixture into a 3-mL vial. Add 800 uL of TBME to the vial and swirl the vial(© 1097 Cengage Leaning “TECHO708: Separating Frrocene and Aezyfrrocene by Adsorption Column Chromatography o 4, Applying the Sample to the Column 5. Eluting Ferrocene 6. Eluting Acetylferrocene to dissolve the mixture. Add 100 mg of alumina to the solution and mix. ‘Note that the alumina does not dissolve. Evaporate the mixture to dryness by placing the vial in a 60 °C sand bath under a fume hood. Use a gentle stream of air or nitrogen to speed the ‘evaporation. ‘Add the dry alumina-sample mixture to the top of the column. Add an additional 200 mg of alumina on top of the sample to protect the sample layer from disruptions when adding eluent. Hexane is flammable and irritating. Keep hexane away from flames and other hheat sources. Use hexane under a fume hood. Prevent eye, skin, and clothing contact. Avoid ingesting and inhaling this compour Half-fill the 50-ml. flask labeled “hexane” with hexane. Use this hexane as your working stock of eluent. Refill if necessary. Place the flask labeled “hexane eluent” under the column. Using a Pasteur pipet, slowly add hexane, in 1-mL increments, to the top of the column. Allow the liquid to flow down the side of the column, taking care not to disturb the alumina bed. Collect the hexane as it elutes from the column. NOTE: Do not alow the column to go dry. necessary, place a #1 atopper in the top. (of the column to stop the eluent flow for short time periods. Continue to add hexane to the top of the column until the bottom of the yellow ferrocene band is at the bottom of the column bed. NOTE: When the batom edge of the colored band reaches the bottom edge of the alumina (the top edge ofthe sand), the ferrocene is ready to elute, Elution ofthe band is ‘complete when the top edge ofthe colored band has eluted from the bottom tip ofthe column, Remove the hexane eluent flask from under the column, and replace it with the preweighed 50-mL beaker labeled “ferrocene”. Collect the hexane containing the ferrocene, using this beaker. After all the ferrocene has eluted from the bottom of the column, remove the beaker from under the column. Place the hexane eluent flask under the column and proceed immediately to Part 6. Half-fill the 50-mL flask labeled “‘TBME” with TBME. Use this TBME as your working stock eluent. Refill if necessary. Allow the hexane above the column bed to flow into the bed. When the top of the bed just begins to appear dry, use a Pasteur pipet to carefully add 1 mL of TBME to the top of the column. Remove the hexane eluent flask from under the column, and replace it with the flask labeled “TBME eluent’. Collect the eluent in this flask.6 ‘TECHO708: Separating Ferrocene and Acetyfesrocene by Adsorption Column Chromatography Continue adding TBME, in I-mL increments, until the bottom of the orange acetylferrocene band is at the bottom of the column bed. Remove the TBME eluent flask from under the column. Replace the flask with the preweighed 50-mL beaker labeled “acetylferrocene”. Collect the TBME containing the acetylferrocene, using this beaker. If crystals of acetylferrocene form at the tip of the column, fill a Pasteur pipet with TBME. Use the TBME to rinse the acetylferrocene into the labeled beaker. After all the acetylferrocene has eluted from the column, remove the acetylferrocene beaker from under the column. Collect any additional eluent in the TBME eluent flask. 7. Drying and Weighing Evaporate the eluents from the ferrocene and acetylferrocene beakers by ‘Compounds placing them in a 60 °C sand bath under the fume hood. Use a gentle stream of air or nitrogen to speed evaporation. When the eluents have evaporated, allow the beakers to cool. Weigh each beaker. Record the masses. Subtract the masses of the respective ‘empty beakers. 8. Cleaning Up Use the labeled collection containers provided by your laboratory instructor. Transfer the contents of the hexane and hexane eluent flasks to the container labeled “Recovered Hexane’. Transfer the contents of the TBME and TBME eluent flasks to the container labeled “Recovered tert-Butyl Methyl Ether’. Scrape the recovered ferrocene and acetylferro- cene from their respective beakers, and place the compounds in the con- tainers labeled “Recovered Ferrocene’ and “Recovered Acetylferrocene’, respectively. Remove the column from the utility clamp and place the column in the container labeled “Used Columns”. Place the Pasteur pipets into the container labeled “Used Pasteur Pipets’. Clean your glassware with soap or detergent. Wash your hands thoroughly with soap or detergent before leaving the laboratory. Using the Slurry Pack Equipment 3 beakers, 50-mL microspatula 3-mL conical vial 2 Pasteur pipets, with latex bulbs chromatography column sand bath* 4 Erlenmeyer flasks, 50-mL support stand labels utility clamp marking pen “sand in crystallizing dish or sand in electric heating well with heat controller(© 1067 Cengage Learning ‘TECHO708: Separating Ferocene and Aceyferrocene by Adsorption Column Chromatography 2 Reagents and Properties ‘Molar mass ‘Substance Quantity (g/mol) mp CC) bp °C) acetylferrocene 35 mg 28 81-83 alumina 33g 102 2054 tert-butyl methyl ether 20 mL 88 55-56 ferrocene 35 mg 186 174-176 hexane 50 mL 86 0 Preview * Prepare slurry pack column + Prepare ferrocene-acetylferrocene sample + Apply sample to column + Elute ferrocene + Elute acetylferrocene ‘+ Evaporate eluents from ferrocene and acetylferrocene ‘+ Measure masses of separated compounds PROCEDURE A Chemical Alert 1. Labeling and Weighing Collection Containers 2. Preparing a Slurry Pack Column einen Cee ee era ee eee hexane—flanimable and irritant ‘Wear departmentally approved safety goggl chemistry laboratory. Label four 50-mL Erlenmeyer flasks “hexane”, “hexane eluent’, “TBME”, and “TBME eluent’, respectively. Label two 50-mL beakers “ferrocene” and “acetylferrocene”, respectively. Weigh each beaker and record the masses. Hexane is flammable and irritating. Keep hexane away from flames and a ‘other heat sources. Use hexane under a fume hood. Prevent eye, skin, and clothing contact. Avoid ingesting and inhaling this compoun Using @ utility clamp, attach a chromatography column equipped with a stopcock and a glass frit to a support stand, making certain the column is vertical.3. Preparing a Sample ‘4, Applying the Sample to the Column 5. Eluting Ferrocene ‘TECHO706: Separating Feerocene and Aceryierocene by Adsorption Column Chromatography Fill the 50-mL flask labeled “hexane’’ with hexane. Use this hexane as your working stock of eluent. Refill if necessary. Close the stopcock, and half-fill the column with hexane. Mix 3 g of alumina with approximately 20 mL of hexane in a 50-mL. beaker. Swirl the beaker to form a slurry. Pour the slurry into the column. At the same time, ‘open the stopcock slightly to allow hexane to drip from the bottom of the column. As the column bed packs, tap the sides of the column to ensure good adsorbent packing and to dislodge any air bubbles that may be present. Continue adding the slurry, with tapping, until all of the alumina is in the column. Rinse the beaker with hexane, and add the rinses to the column. Do not allow the column to go dry. After the bed has packed, close the stopcock, making certain that hexane is present above the top of the column bed. Wf possible, use tert-butyl methyl ether (TBME) under a fume hood. TBME is flammable and irritating. Keep TBME away from flames and other heat sources. Acetylferrocene is highly toxic. Prevent eye, skin, and clothing contact. Avoid ingesting and inhaling these compounds. Weigh 70 mg of a 50:50 mixture of ferrocene and acetylferrocene. Place the mixture into a 3-mL vial. Add 800 wL of TBME to the vial, and swirl the vial to dissolve the mixture. Add 100 mg of alumina to the solution and mix. Note that the alumina does not dissolve. Evaporate the mixture to dryness by placing the vial in a 60°C sand bath under a fume hood. Use a gentle stream of air or nitrogen to speed the evaporation. Open the column stopcock to allow the hexane level to reach the top of the alumina bed. When the top of the bed just begins to appear dry, close the stopcock. NOTE: No excess hexane should be visible above the column bed. No air bubbles should be present in the bed. Only the very top of the bed should appear dry. ‘Add the dry alumina-sample mixture to the top of the column, Add an ‘additional 200 mg of alumina on top of the sample to protect the sample layer from disruptions when adding eluent. Place the flask labeled “hexane eluent” under the column. Open the stopcock and collect the hexane as it elutes from the column. Using a Pasteur pipet, carefully add hexane, in 1-mL. increments, to the top of the column, taking care not to disturb the alumina bed. NOTE: Do not allow the column to go dry. When air bubbles are allowed to form in the ‘lumina bed, elvent wil flow around the bubbles, leaving sample onthe alumina. Large air bubbles may stop the eluent flow completely. Continue to add hexane to the top of the column until the bottom of the yellow ferrocene band is at the bottom of the column bed.a iti‘—s J “TECHO706: Separating Ferrocene and Acerferocene by Adsorption Column Chromatography n NOTE: When the bottom edge of the colored band reaches the botiom edge of the alumina (the top edge of the glass fr), the ferrocene is ready to elute. Elution of the. ‘band is complete when the top edge ofthe colored band has eluted from the bottom tip. ‘ofthe column. Remove the hexane eluent flask from under the column, and replace ‘under the column and proceed immediately to Part 6. 6. Eluting Acetylferrocene Allow the hexane above the column bed to flow into the bed. When the top of the bed just begins to appear dry, close the stopcock. Remove the hexane ‘eluent flask from under the column, and replace it with the flask labeled “"TBME eluent’. Half-fill the 50-mL flask labeled “TBME” with TBME. Use this TBME as your working stock eluent. Refill if necessary. Use a Pasteur pipet to carefully add 1 mL of TBME to the top of the column. Open the stopcock and collect the eluent in the flask. Continue adding the TBME, in 1-ml, increments, until the orange acetylferrocene band is at the bottom of the column bed. Remove the TBME eluent flask from under the column. Replace the flask with the preweighed 50-mL beaker labeled “acetylferrocene”. Collect the TBME containing the acetylferrocene, using this beaker. If crystals of acetylferrocene form at the tip of the column, fill a Pasteur pipet with TBME. Use the TBME to rinse the acetylferrocene into the labeled beaker. After all the acetylferrocene has eluted from the column, close the stopcock. Remove the acetylferrocene beaker from under the column. Place the TBME eluent flask under the column. Open the stopcock, and collect any additional eluent in the flask. 7. Drying and Weighing Evaporate the eluents from the ferrocene and acetylferrocene by placing Compounds the beakers in a 60°C sand bath under the fume hood. Use a gentle stream of air or nitrogen to speed evaporation. When the eluents have evaporated, allow the beakers to cool. Weigh each beaker. Record the masses. Subtract the masses of the respective empty beakers. 8. Cleaning Up Use the labeled collection containers provided by your laboratory instructor. Transfer the contents of the hexane and hexane eluent flasks to the container labeled “Recovered Hexane”. Transfer the contents of the TBME and TBME eluent flasks to the container labeled “Recovered tert-Butyl Methyl Ether’. Scrape the recovered ferrocene and acetylferro- cene from their respective beakers, and place the compounds in the containers labeled “Recovered Ferrocene” and “Recovered Acetylferro- cene”, respectively. Remove the column from the utility clamp and place the column in the container labeled “Used Columns”. Place the Pasteur pipets into the (© 1097 Cengage Learringn ‘TECHO706: Separating Ferrocene and Aceylertocene by Adwrption Column Chromatography container labeled “Used Pasteur Pipets”. Clean your glassware with soap or detergent. ‘Wash your hands thoroughly with soap or detergent before leaving the laboratory.ume) 2886000 1861 ©3. Briefly explain why the chromatography column should be kept absolutely vertical while it is being packed and used. 4. How will you know when you should begin to collect the acetylferrocene sample as it elutes from the column? 5. What is the purpose of adding alumina on top of the sample?at kone Nd dl Fone oe “Te eth Nel Tin lan al Le {© 1988 Congage Losing 6 REAC Dehydrating Cyclohexanol Prepared by Carl T. Wigal, Lebanon Valley College PURPOSE OF THE EXPERIMENT Dehydrate cyclohexanol to prepare cyclohexene. Characterize cyclohexene by i ammonium cerum (V) nitrate test, bromine test, infrared spectroscopy and/or refractive index. EXPERIMENTAL OPTIONS ‘Semi-Microscale Dehydration Microscale Dehydration Using Glassware with Elastomeric Connectors Using a Hickman Still Assembly Product Characterization BACKGROUND REQUIRED You should be familiar with basic laboratory techniques for measuring volumes and masses. You should know how to conduct a simple distil- lation. For product characterization, you should know how to measure refractive index and /or obtain and interpret an infrared spectrum. BACKGROUND INFORMATION Elimination reactions involve the loss of a small molecule (H-X) from adjacent carbon atoms, resulting in pi-bond formation. Consequently, elimination reactions are good synthetic methods for producing alkenes or alkynes. These reactions occur through a process called heterolytic bond cleavage. Heterolytic bond cleavage occurs when one atom leaves a compound with both electrons of the original bond, resulting in the formation of ions. For example, elimination of H-X from an organic molecule involves the loss of a proton (H*) and a leaving group (x), as shown in Figure 1 on the next page. The leaving group departs with both electrons from the original C-X bond. The electrons in the adjacent CH bond form the new pi bond of the alkene, with loss of the proton. Sr ee ere es LAs, CENGAGE © mcegge usm ALOFT PESNED ou coed yb py He eb ace Learning” svt sonto unt ay tm yay msrp dee ma eg ee pa ‘tg xe dry ey hun lars tan pent oot prt Sn Oe ea ep wtp apeB REACO712: Dehydrating Cycloheranol Figure 1 Elimination of HX from an organic molecule The elimination of water (H-OH) from alcohols was one of the earliest organic reactions studied. This reaction, still widely used, is called a dehydration reaction. In many cases, alcohol dehydration is an acid- catalyzed reaction that proceeds by an elimination mechanism called El. ‘The E1 mechanism for the dehydration of 2-methyl-2-butanol is shown in Figure 2. The first step of dehydration is a proton transfer from the acid catalyst to the oxygen atom of the alcohol. This protonation forms an oxonium ion, the conjugate acid of the alcohol. Weak bases are good leaving groups, so changing the leaving group from hydroxide to water favors the reaction Step 1 HC poets ue HC poets By soo ree Be Sar ie Hyc~ ‘ee H 2methy2-butanol ees Step 2 ee phe 10 ane Pou cH, pte HO ie a CH;CH, carbocation Step 3 sachs HCC, ™“, i cH CHy CH, ik 2emethyt 2 buene methyl Luteo (major product) (minor product} 2 ET mechanism for the dehydration of 2-methyl-2-butanot(© 1098 Cengage Leaning Qualitative Tests [REACH712: Dehydrating Cylhexanol a The second step of the dehydration reaction is loss of water from the eninge Wocstia'n pod catia cuvocas, Tesl@eaar a mechanism is rate-determining. Not all alcohols dehydrate at the same rate. Alcohols are classified according to the number of alkyl groups attached to the carbon bearing the hydroxyl group. The terminology used to describe the degree of substitution is tertiary (3°), secondary (2°), and primary (1°). Experi- mental evidence shows that the ease of alcohol dehydration follows the trend 3° > 2° > 1°. This reactivity directly relates to the stability of the carbocation intermediate formed during the rate-determining step of the reaction. In the third and final step, a proton is released from a carbon atom adjacent to the positively charged carbon. The electrons previously com- prising the C-H bond form the new carbon-carbon pi bond. The formation of two isomeric alkenes is possible in elimination reactions where a proton can be lost from either of two different carbon atoms. Saytzeff’s rule states that the orientation of the double bond favors the more thermodynamically stable alkene; that is, the alkene with the greatest number of alkyl groups bonded to the carbons of the double bond. Thus, dehydrating 2-methyl-2-butanol produces primarily 2-methyl- 2-butene, a trisubstituted alkene, rather than 2-methyl-1-butene, a disub- stituted alkene. In this experiment, you will dehydrate cyclohexanol to form cyclohexene. Because cyclohexene has a lower boiling point than cyclohexanol, the cyclohexene can be distilled away as it forms. You will isolate and characterize the cyclohexene by performing qualitative tests for alcohols and alkenes. Your laboratory instructor will tell you whether to further characterize the cyclohexene by measuring its refractive index and/or by generating infrared spectra for both cyclohex- anol and cyclohexene. The presence of the hydroxyl group of an alcohol can be determined by observing the reaction of an alcohol with ammonium cerium(IV) nitrate, (NH:Ce(NO;)e. A positive test for an alcohol is indicated as the yellow (NH9;Ce(NO}), solution turns red when complexed with an alcohol, as shown in Figure 3. Even small contaminating amounts of alcohol can cause a slight color change. The presence of a carbon-carbon double bond of an alkene can be determined by observing the reaction between bromine and an alkene, as shown in Figure 4 on the next page. Bromine is a reddish-brown color. A Positive test is indicated by the decolorization of the bromine solution. (NHg2Ce(NO3), + R-OH —® [alcohol + reagent] yellow alcohol red complex Figure 3 Reaction of ammonium cerium(IV) nitrate with an alcohol0 REACO712: Dehydeating Cyeloheranal R R R Seat, + Beng noe ie RR RoR OBF iss SA rstanonn eae Figure 4 Reaction of bromine with an alkene SEMI-MICROSCALE DEHYDRATION Equipment 100-mL beaker magnetic stir bar distillation apparatus 2 Pasteur pipets, with latex bulb condenser, with tubing 5-ml. sample vial distilling head sand bath’ 10-mL round-bottom flask 13 x 100-mm test tube! receiver flask* support ring thermometer, 10 to 260°C, 2 support stands or equivalent, with adapter 3 utility clamps ‘vacuum adapter wire gauze 10-mL graduated cylinder ““0-ml. vial or 10-ml.round-bottom flask ‘stirring hot plate with crystallizing dish filled with sand or magnetic stirrer and electric flask heater fled with sand ‘or enti tbe Reagents and Properties ] : ‘molar mass bp a substance quantity (gimol) CC) (ght ‘ calcium chloride 025g 11099 ‘ anhydrous if cyclohexanol 284 g 100.16 160 0948 cyclohexene” 8215 83 osit ; . phosphoric acid, 85% 40 mL 98.00 1.685 sulfuric acid, 02 mL 98.08 1.840 concentrated product Preview Assemble the distillation apparatus ‘Add cyclohexanol, sulfuric acid, and phosphoric acid to the flask Distill the reaction mixture and collect the distillate in a receiver Transfer the distillate to a test tube Remove the bottom layer of the distillate Dry the top layer with anhydrous calcium chloride(© 1008 Cengage Learning REACO712: Dehydrating Cyelohexanol a PROCEDURE 1. Using Distillation to 2, Isolating Cyclohexene ‘anhydrous calclum chloride—irritant and hygroscopic cyclohexanol—irritant and hygroscopic ‘eyclohexene—flammable and irritant phosphoric acld—corrosive i t : i i Wear departmentally approved safety goggles at all times while in the chemistry laboratory. | ncenttrated sulfuric acid (H,S0,) is toxic and an oxidizer. Phosphoric acid (HsPO,) is corrosive. Prevent eye, skin, clothing, and combustible materials contact. Cyclohexanol is an irritant and hygroscopic. Avoid inhaling vapors and ingesting these compounds. Use a fume hood. Assemble the distillation apparatus shown in Figure 5 on the next page. If necessary, use substitute glassware as directed by your laboratory instructor. Remove the 10-mL round-bottom flask from the apparatus. Place 4.0 mL of 85% HPO, and 2.84 g (3.0 mL) of cyclohexanol in the round-bottom flask. Add 5 drops of concentrated H,S0, to the flask and add a magnetic stir bar. Reattach the round-bottom flask to the distillation apparatus. Start the flow of water through the condenser. Tur on the magnetic stirrer. Heat the reaction mixture while stirring until the product starts to distill. Continue the distillation, collecting the product in the receiver flask until no more liquid distills or until the temperature of the thermometer rises above 85 °C. ‘Tum off he heat. Allow the apparatus to cool. Turn off the magnetic stirrer. Remove the receiver flask from the distillation assembly. Use a Pasteur pipet to transfer the distillate into a centrifuge tube or small test tube. i | Cyclohexene is flam: y from flames or other heat sources. Prevent eye, skin, and clothing contact. Avold inhaling vapors. Use a fume hood. Sameer SSS a tas Ce Eh Notice that as the distillate in the tube cools, two layers form. Use the Pasteur pipet to remove the majority of the bottom layer. Place the3. Cleaning Up REACO712: Dehydrating Cyeloheranol Figure 5 Distillation apparatus for semi-microscale dehydration bottom layer into the container labeled “Recovered Acid Layer”, provided by your laboratory instructor. ‘Anhydrous calcium chlorice is irritating and hygroscopic. Avoid inhaling dust. Winiss tke Day the top organic layer by placing about 0.25 g of anhydrous calcium chloride into the test tube. Let the test tube stand for 5 min. Weigh a clean 5-mL sample vial. Using a clean dry Pasteur pipet, remove the liquid from the test tube, and transfer the liquid to the tared sample vial. Weigh your product Characterize your cyclohexene using the methods in the Product Characterization section designated by your laboratory instructor. Place your recovered materials in the appropriate labeled collection containers as directed by your laboratory instructor. Clean your glassware with soap or detergent. Wash your hands thoroughly with soap or detergent before leaving the laborator| a substance quantity (ghmol) eo) (glmL) ‘calcium chloride, 025g 11099 anhydrous cyclohexanol 1422 g 100.16 160 0.948 cyclohexene 8215 8 git phosphoric acid, 85% 15 mL. 98.00 1.685 sulfuric acid, 0.12 mL, 98.08 1.840 ‘concentrated i Assemble the distillation apparatus ‘* Add cyclohexanol, sulfuric acid, and phosphoric acid to the flask © Distill the reaction mixture and collect the distillate in a receiver © Transfer the distillate to a test tube ‘* Remove the bottom layer of the distillate © Dry the top layer with anhydrous calcium chloride © Tare a sample vial ‘* Transfer the cyclohexene to the sample vial * Weigh the cyclohexene PROCEDURE ‘anhydrous calcium chloride—irrtant and hygroscopic aNDehydrate Cyclohexanol ee Distillation to REACO712: Dehydrating Cyclohexanol phosphoric acid—corrosive ulturie acid—toxic and oxidizer ‘Wear departmentally approved safety goggles at all times while in the chemistry laboratory. Concentrated sulfuric acid (H,S0,) is toxic and an oxidizer. Phosphoric acid (HsPO,) Is corrosive. Prevent eye, skin, clothing, and combustible materials contact. Cyclohexanol is an irritant and hygroscopic. Avoid inhaling vapors and ingesting these compounds. Use a fume hood. Assemble the distillation apparatus shown in Figure 6. Remove the 5-mL round-bottom flask from the apparatus. Place 1.5 mL of 85% H3PO, and 1,422 g (1.5 mL) of cyclohexanol into the flask. Add 3 drops of concentrated H,SO, to the flask and add a magnetic stir bar. Reattach the round-bottom flask to the distillation apparatus. Turn on the magnetic stirrer. Heat the reaction mixture while stirring until the product starts to distill. Continue the distillation, collecting the product in the receiver vial until no more liquid distills or until the temperature of the thermometer rises above 85°C. ‘Tum off the heat. Allow the apparatus to cool. Turn off the magnetic stirrer. Remove the receiver vial from the distillation assembly. Use a Pasteur pipet to transfer the distillate into a centrifuge tube or small test tube. thermometer — thermometer adapter Figure 6 Distillation apparatus for microscale dehydration using glassware with elastomeric connectors© 1098 Cengage Learring 2, Isolating Cyclohexene 3. Cleaning Up 85 REACO712: Dehydrating Cycloheranol Gyclohexene Is flammable and irritating. Keep away from flames or other heat sources. Prevent eye, skin, and clothing contact. Avoid inhaling vapors. Use a fume hood. (icine: oo Sanaa aD ii ‘Use the Notice that as the distillate in the tube cools, two layers form. Pasteur pipet to remove the majority of the bottom layer. Place the bottom layer into the container labeled “Recovered Acid Layer’, provided by your laboratory instructor. ‘Anhydrous calcium chloride is irritating and hygroscopic. Avoid inhaling dust. Dry the top organic layer by placing about 0.25 g of anhydrous calcium chloride in the test tube. Let the test tube stand for 5 min : Weigh a clean 5-mL sample vial. Using a clean dry Pasteur pipet, remove the liquid from the tube and transfer the liquid to the tared sample vial. Weigh your product. Characterize your cyclohexene using the methods in the Product ‘Characterization section designated by your laboratory instructor. Place your recovered materials in the appropriate labeled collection con- tainers as directed by your laboratory instructor. Clean your glassware with soap or detergent. ‘Wash your hands thoroughly with soap or detergent before leaving the ZN laboratory. Peres ame MICROSCALE DEHYDRATION Using a Hickman Stilt ‘Assembly Equipment S-mL conical vial support stand 10-mL graduated cylinder 10 x 75-mm test tube’ Hickman still thermometer, —10 to 150°C, magnetic spin vane with adapter* 2 Pasteur pipets, with latex bulb thermometer, -10 to 260°C, 3-mL sample vial or equivalent sand bath* 2 utility damps aan rang tes With and or magnetic ster and electric flask heater or centrifuge tube oft Hickman sl86 REACO712; Dehydeating Cylohexanol Reagents and Properties molar mass bp a substance quantity (gimol) CO) (g/ml) calcium chloride 025g 110.99 anhydrous cyclohexanol 1422 g 100.16 160 0.948 cyclohexene 8215 83 ost phosphoric acid, 85% 15 mL. 98.00 1.685 sulfuric acid, 0.2 mL. 98.08 1.840 concentrated “produat Preview * Assemble the distillation apparatus * Add cyclohexanol, sulfuric acid, and phosphoric acid to the vial ‘© Distill the reaction mixture into the still collar © Transfer the distillate to a test tube © Remove the bottom layer of the distillate © Dry the top layer with anhydrous calcium chloride © Tare a sample vial © Transfer the cyclohexene to the sample vial '* Weigh the cyclohexene PROCEDURE CHEMICAL ALERT N anhydrous calcium chloride—irritant and hygroscopic cyclohexanol—irritant and hygroscopic ‘cyclohexene—flammable and irritant phosphoric acid—corrosive Sulfuric acid—toxie and oxidizer Cin as /\ ‘Wear departmentally approved safety goggles at all times while in the chemistry laboratory. 1. Using Distillation to Dehydrate Cyclohexanol Concentrated sulfuric acid Hz80,) Is toxic and an oxidizer. Phosphoric acid (HPO,) is corrosive. Prevent eye, skin, clothing, and combustible materials ‘contact. Cyclohexanol is an irritant and hygroscopic. Avoid inhaling vapors and ingesting these compounds. Use a fume hood,‘© 1088 Cengage Learning Dilation epprtu for micocle dhydraton using Hickman stil Assemble the distillation apparatus shown in Figure 7. Remove the 5-mL conical vial from the apparatus. Place 1.5 mL of 85% HPO, and 1.422 g (1.5 mL) of cyclohexanol into the vial. Add 3 drops of concentrated H,SO, to the vial and add a magnetic spin vane. Reattach the vial to the distillation apparatus, ‘Tum on the magnetic stirrer. Heat the reaction mixture while stirring until the product starts to distill. As the product starts to collect in the still, use a Pasteur pipet to remove the liquid from the still. [Nove 1] Transfer the liquid into a centrifuge tube or a small test tube. Continue the distillation and collection until no more liquid distills or until the temperature of the thermometer rises above 85°C. ‘Tum off the heat: Allow the apparatus to cool. Turn off the magnetic stirrer. Cyclohexene yy heat sources. Prevent eye, skin, and clothing contact. Avoid inhaling vapors. Use a fume hood. ear ae ‘Notice that as the distillate in the test tube or vial cools, two layers form. Use the Pasteur pipet to remove the majority of the bottom layer. Place the bottom layer into the container labeled “Recovered Acid Layer’, provided laboratory instructor.REACD712: Dehydrating Cyeloheanol ‘Anhydrous calcium chloride is irritating and hygroscopic, Avold inhaling dust. Dry the top organic layer by placing about 0.25 g of anhydrous calcium chloride into the test tube. Let the test tube stand for 5 min. Weigh a clean 3-mL sample vial. Using a clean dry Pasteur pipet, remove the liquid from the test tube and transfer the liquid to the tared sample vial. Weigh your product. Characterize your cyclohexene using the methods in the Product Characterization section designated by your laboratory instructor. 3. Cleaning Up Place your recovered materials in the appropriate labeled collection containers as directed by your laboratory instructor. Clean your glassware with soap or detergent. Wash your hands thoroughly with soap or detergent before leaving the laborator PRODUCT CHARACTERIZATION Equipment 5 Pasteur pipets, with latex bulb white spot plate 3 test tubes, 10 x 75-mm Reagents and Properties ‘molar mass bp substance quantity (gimol) eo ammonium cerium(V) 06 mL nitrate test reagent bromine test reagent 024 mL cyclohexanol 0416 mL. 100.16 160 cyclohexene 0.16 mL 8215 83 dichloromethane 24 mL 8493 40 14-dioxane 0.36 mL 88.11 100-102 Preview © Test standards and product for presence of alcohol using ammonium cerium(IV) nitrate reagent © Test standards and product for presence of alkene using bromine reagent ‘* Compare cyclohexanol and cyclohexene product using infrared analysis © Measure refractive index of cyclohexene productREACD712: Dehydrating Cycohexanol PROCEDURE ‘ammonium cerium(IV) nitrate—irritant and oxidizer ‘bromine—highly toxic and oxidizer ceyclohexanol—initant and hygroscopic ‘cyclohexene—flammable and irritant dichloromethane—toxic and iritant 11,4-dloxane—flammable and suspected carcinogen A-dloxane—flammable and suspected carcinogen ‘Wear departmentally approved safety goggles at all times while in the chemistry laborator CRYO a egg 1. Using Ammonium Cerium(tV) Nitrate to Test fcAlotels, ‘Ammonium cerium(V) nitrate, (NH,)2Ce(NOs)6, Is irritating and an ‘oxidizer. 1,4-Dioxane s flammable and a suspected carcinogen. Keep 1,4-dioxane away from flames or other heat sources. Prevent eye, skin, and clothing contact. ‘Avoid inhaling vapors. Use 1,4-dioxane in a fume hood. ‘Cyclohexanol is irritating and hygroscopic. Cyclohexene is flammable and irritating. Keep cyclohexene away from flames or other heat sources. Prevent eye, ‘skin, and clothing contact. Avoid inhaling vapors. Place 5 drops of the (NH,),Ce(NO3). test reagent and 3 drops of 14-dioxane ome i400 iswedssasovert. in each of three wells of a white spot plate. [Note 2] Add 2 drops of cyclohexanol to the first well and stir. Observe any color change and record your results. Add 2 drops of cyclohexene from the reagent bottle to the second well and stir, Observe any color change and record your results. Add 2 drops of your cyclohexene product to the third well and stir. Observe any color change and record your results. 2. Using Bromine toTest for Alkenes CITY Bromine (Bris highly toxic. Dichioromethane is toxic and iritating. ‘Cyclohexano! is irritating and hygroscopic. Cyclohexene is flammable and irritating. Keep cyclohexene away from flames or other heat sources. Prevent eye, skin, and clothing contact. Avoid inhaling vapors. Use these reagents in a fume hood. Label three small test tubes “cyclohexanol”, “cyclohexene”, and “product”. Place 2 drops of cyclohexanol into the tube labeled “cyclohexanol”. Place 2 drops of cyclohexene into the tube labeled “cyclohexene”. Place 2 drops of Your product into the tube labeled “product” Add 20 drops of dichloromethane into each test tube, and stir. [Note 3]. Add 2 drops ofthe Bra test reagent to each tube and stir. Observe any color change. Record your results sm & Dichicrmthane 1 used os a : ; ice g3, Using Infrared Analysis to Compare Cyclohexanol and Your Gyclohexene Product rt & Sat pates are tragic and rygtoncopc. Donat use water ome tbe ‘plate, Even moisture tom your tages ‘nll aac the plates. Use gloves nd ‘ai handle he pats by te edges 4, Using Refractive Index to Characterize Your Cyclohexene Product 5. Cleaning Up aT & The reactive inex at 20°C aloud by using he folowing eque ‘ion. where T's the ambient temperate ‘in degrees Cols and ns the vtrac- the Inder measured ot ambient tom perature 1B = + 000085(7-20°C) REACO712: Dehydrating Cyelohesanol Table 1 Refractive indices (20°C) Meee TP tle 20h ssiemnainbatene aia eal wet 1.3329 cyclohexanol 1.4650 cyclohexene eo Obtain the operating instructions for using the infrared spectrometer from your laboratory instructor. Obtain a set of KBr, NaCl, or AgCI salt plates and a holder. [Nore 4] Place 1 drop of your cyclohexanol between the salt plates. Gently press the plates together to remove any air bubbles. Place the plates in the holder and secure the plates. Run and plot the IR spectrum according to your operating instructions. Repeat this procedure for your cyclohexene product. Examine the region of the spectrum above 1500 cm~'. Assign the bonds that give rise to these absorptions. Obiain the operating instructions for the refractometer from your laboratory instructor. Measure the refractive index for your cyclohexene product, Measure the laboratory temperature in °C. Make temperature corrections, if necessary. [Nore 5] Compare your refractive index to the literature values shown in Table 1. Place your recovered materials in the appropriate labeled collection containers as directed by your laboratory instructor. Clean your glassware with soap or detergent. Le Te /\, ‘Wash your hands thoroughly with soap or detergent before leaving the laboratory. Kbibbheagiiartes tance‘© 1908 Congage Leeming on REAOU712: Dehydrating Crloheranal Post-Laboratory Questions 1. Calculate the percent yield that you obtained from this reaction. 2. Did your product cause a color change with (NH,);Ce(NOs), test reagent? Explain your results. 3. Did your product cause a color change with the Br test reagent? Explain your results, 4. Compare the IR spectra for cyclohexanol and your product. What IR evidence do you have that your product is cyclohexene and not cyclohexanol? Briefly explain. 5. (a) Calculate the refractive index of your product at 20°C. (b) Compare the refractive index of your product to the data of Table 1. Does the result indicate that your product is pure? Briefly explain. (©) If the refractive index of your product differs from the listed value, what is the most likely contaminant in your product, as indicated by the refractive index? Briefly explain. 6. What would be the major product obtained from the El dehydration of 2-methylcyclohexanol? 7. Outline a mechanism for the dehydration of I-methyl-1-cyclohexanol. Would you predict this reaction to be faster or slower than the reaction you performed?© 1008 Cengage Leming [REACO712: Dehydrating Cyclohexanol 2 Tame Pre-Laboratory Assignment 1. What safety precautions must be observed when using concentrated H2SO, and HsPO,? 2. (a) Write the chemical equation for the dehydration of cyclohexanol. (b) Using the following information, calculate the theoretical yield for the dehydration of 3.0 mL of cyclohexanol ‘molar mass a substance (imol) (int) cyclohexanol 100.16 0.948 cyclohexene 8215 osit 3. When 2-butanol undergoes E1 dehydration, three alkenes are obtained. Draw the structures for these alkenes. Which alkene would you predict to be formed in greatest abundance?‘Sy2 Mechanism REACO714: Studying SN and SN2 Reactions: Nucleophilic Substicution Acid-base reaction: AcHY +. 'BT) gm Ate op strong acid strong base weak base weak acid Nucleophilic Substitution: L—C— + Nu me LI + = —C—Nu | organic nucleophile leaving onan reactant POU prnduct Figure 1 Comparison of an acid-base reaction and a nucleophilic substitution reaction substitution reactions share several characteristics with acid-base reactions, as shown in Figure 1 Groups that are good leaving groups in nucleophilic substitution Teactions are weak bases in acid-base reactions. Strong bases are typically 800d nucleophiles in substitution reactions. In an acid-base reaction, a Proton is transferred from the conjugate acid of a weak base to a strong base. In a similar fashion, nucleophilic substitution reactions often involve the transfer of a carbon group from a weak base, the leaving group, to a stronger base, the nucleophile. Nucleophilic substitution reactions may occur by one of two common mechanisms, designated $y2 and Sy1 The Sy2 mechanism derives its designation from the fact that two chemical species—the organic reactant and the nucleophile—participate in the rate- determining step of the reaction. This mechanism has only one step, as shown in Equation 1 oy me “ re 2s EL Nu a (Eq. 1) ay \ Nu The reaction is initiated by an attack of the nucleophile on the carbon bonded to the leaving group. This back-side attack produces a product in Which the configuration of the carbon atom is inverted Some factors that can have a significant effect on the rate of Sy2 reactions include: (@) The leaving group: Leaving groups are invariably weak bases. The Presence of a good leaving group in the organic compound is essential for a nucleophilic substitution reaction to occur. 2) The carbon group: An Sy2 reaction occurs fastest when the approach of the nucleophile to the carbon is unhindered by the presence of bulky groups. For example, the reaction occurs faster at primary carbon atoms than at secondary carbon atoms. (3) The nucleophile: For nucleophiles in which the attacking atoms are of comparable size, nucleophilicity parallels basicity. That is, stronger© 1908 Cengage Leaning REACO714: Studying SN1 and SN2 Reactions: Nucleophilic Substitution a bases are better nucleophiles than weaker bases. When nucleophiles differ in size, however, the larger, more polarizable atoms are more nucleophilic, even though they are weaker bases. ; (@) The solvent: Reactions involving negatively charged nucleophiles occur much more rapidly in polar-aprotic solvents; that is, solvents in which anionic nucleophiles are poorly solvated. Dimethylsulfoxide (DMSO) is an example of a polar-aprotic solvent. Meet leophili itt i mechanism, in ants Nucleophilic substitution reactions may occur via an Sy mect eee which only the organic reactant is involved in the rate-determining step, as shown in Equation 2. The nucleophile reacts in a fast step. INH ng wets | ee lot eC Nu. + Eg.2) \ it mp = | slow estes i Swi reactions occur when conditions favor ionization of the organic reactant. Such conditions include: (2) The leaving group: The leaving group (L) must be a weak base. 2) The carbon group: The rate-determining step involves production of a carbocation. This step will occur faster for those compounds that yield more stable carbocations. For example, tertiary compounds react faster than secondary compounds. Primary compounds react extremely slowly. Carbocation intermediates are also stabilized by dispersal of the positive charge through delocalization of electrons. Sy reactions that produce such resonance-stabilized carbocation intermediates are also quite fast. For example, Sw1 reactions occur readily with molecules that form allyl or benzyl carbocations, (3) The solvent: Ionization processes are facilitated by polar solvents. Because the rate of an Syl reaction is directly dependent on such an ionization, the reaction occurs faster in more polar solvents. In the first part of this experiment, you will convert I-butanol to ‘-bromobutane as an example of an Sy2 reaction. Because the OH group of the alcohol is nota weak base and, therefore, not a good leaving group, you will conduct the reaction in a strongly acidic solution. In an acidic environment, the alcohol exists as its conjugate acid, and the leaving group is a water molecule, a weak base, as shown in Equation 3 ” ib ote CHsCHiCH:CHs—OH —*- CHsCH;,CH,CHj—OH; —> CHsCH,CH,CH;—Br + H,0 (Eq.3) butanol sa ‘-oromobutane The relatively uncrowded primary carbon atom is open to back-side attack by an appropriate nucleophile. Most nucleophiles are strong bases and could not exist in this acidic environment because they would be tapidly protonated. Bromide ion, however, is a vey weak base that is strongly nucleophilic due to its very large size. Bromide ion is an effective nucleophile even in a very acidic medium, In the second part of this experiment, you will study factors affecting the rates of Sy1 reactions. In Sw1 reactions, solvent molecules often serve ac: ; i : ; “TECHO70S: Separating Acids and Neutral Compounds by Solvent Extraction Post-Laboratory Questions 1, Calculate the percent recovery of each of the compounds recovered from the original mixture. 2. If you mistakenly extracted the ether solution first with NaOH solution and then with NaHCO3 solution, no crystals would be produced upon acidification of the NaHCOs layer. Briefly explain. 3. What product would you obtain if you evaporated the water from the NaOH layer prior to acidifying the layer? 4. Suppose that you used dichloromethane instead of tert-butyl methyl ether as the nonpolar solvent in this experiment. What changes in the procedure would you make in view of the fact that dichloro- ‘methane is more dense than water? 5. Benzoic acid (CoHs-COOH) is a weak acid (pK, = 4.2) and naphthalene is neutral, neither acidic or basic. Prepare a flowchart for the separation and recovery of benzoic acid and naphthalene. ‘Molar mass (gimol) mp (°C) bp CO) PK, benzoic acid 122.12 124 249.2 42 naphthalene 128.16 80.2 217.9 neutral 6, After comparing the melting points of each of your compounds to their respective literature values, comment on the purity of each compound.(© 1008 Cengage Learring trated. concent 1.04 sodium hydrogen 2 mL ‘carbonate, 5% | Place a boiling chip, 1-butanol, and HBr solution into a test tube Slowly add concentrated H.SOx Heat at reflux for 1 hr Allow the reaction mixture to cool, then separate the acid layer ‘Wash the organic layer with water, aqueous NaHCOa, then water again Dry the organic layer with anhydrous CaCl. Purify the product by distillation Characterize the product by boiling point, refractive index, density, or IR spectroscopy PROCEDURE 1. Conducting the Reaction ‘1-bromobutane—flammable and irritant ZN ‘1-butanol—flammable and irritant 48% hydrobromic acid—toxic and corrosive sulfuric acid—toxic and oxidizer ‘Wear departmentally approved safety goggles at all times while in the chemistry laboratory. Pe a AR a Hydrobromic acid (HBr) is toxic and corrosive. Concentrated sulfuric acid ZN (H:S0,) is toxic and oxidizing. HBr and H,S0, can ca case . scan cause severe bi ‘pill, notify your laboratory instructor immediately. ahaa ‘-Butanol is flammable and irritating. Do not use ; ear flames or other Sources. Prevent eye, skin, and clothing contact. Avold inhaling fumes and =umttAs te rscton occu, you shou ‘ose production of «second ba ‘Gyr One lye conta fw wate and (tte ce entae F-bemo-blare produced byte reacon. ae 2 Once te quid he test de feachos 4 eeaay refx conden, pro- ‘ood wae acon “Factors Meng tho ates ofc Ranctone ter ‘mode 2. Washing the Reaction Mixture TE As he sold anhyecus Cac: ‘emoves wal: rom he uid Vxomo. bone, the Saud layer wit become ler. Suing the mit wil facstate Ihe process. ‘3. Distilling the Product (optional) REACO714: Studying SN1 and SN2 Reactions: Nucleophilic Substicution Insert an 18 x 150-mm test tube into a 125-mL Erlenmeyer flask. Place the flask on a balance and tare the balance. Add 1.00 g of 1-butanol to the test tube. Add 2.0 mL of 48% HBr and_an acid resistant boiling chip. Then slowly add, with mixing, 1.0 mL of concentrated H2SO. Remove the test-tabe from the Erlenmeyer flask. Clamp the test tube vertically to a support stand with only the very bottom of the tube in a sand bath. Heat the mixture to boiling using a low setting, ‘Avoid loss of HBr; do not allow the condensation ring to rise more than. 2 em in the test tube. Reflux the solution for 1 hr. (Nove 1]. [Nore 2} +-Bromobutane is flammable and irritating. Do not use near flames or other heat sources. Prevent eye, skin, and clothing contact. Avoid inhaling fumes and ingesting 1-bromobutane, Label two 100-mL. beakers ““Acid Layer” and “Washes”, respectively, Place 25 mL of distilled or deionized water into the beaker labeled “Acid Layee’ abel ierdlins complete, remove the test tube from the sand bath. Allow the test tube to cool Transfer the contents of the test tube to a 15-mL. centrifuge tube. Use a Pasteur pipet to remove the bottom acid layer. Place the acid layer into the beaker labeled “Acid Layer” To wash the 1-bromobutane layer, add 2 mL of water to the centrifuge tube and mix well. Then use a Pasteur pipet to remove the upper water layer. Place the water into the beaker labeled “Washes”, Wash the 1-bromobutane again, first with 2 mL of 5% aqueous sodium hydrogen carbonate (NaH1CO;), then with a second 2 mL of water. Each time, use the Pasteur pipet to remove the aqueous layer and place the layer into the beaker labeled “Washes”. After the final washing with water, allow the two layers to completely separate, Use a clean Pasteur pipet to transfer the 1-bromobutane layer to a clean, dry 5-mL vial. Add 4-5 granules of anhydrous CaCl, to the vial to dry the product. Allow the product to dry for 5-10 min. [ore 3] If you do not conduct the optional distillation in Part 3, measure and record the mass of your product. Tare a 5-mL vial. Use a clean, dry Pasteur pipet to transfer the product to a clean, dry 13 x 100-mm test tube. Add a boiling chip. Clamp the test tube vertically in a sand bath and heat the product until the liquid refluxes in the test tube. Submerge the tip of a clean, dry Pasteur pipet into the 1-bromobutane vapors, as shown in Figure 2. Draw the vapors into the pipet, where they will condense. Transfer the condensed 1-bromobutane to the tared 5-mL vial. Repeat this process until no more vapors are obtained. Measure and record the mass of the 1-bromobutane collected.© 1998 Cengage Learning REACO714: Seudying SII and SN2 Reactions: Nucleophilic Substzuion 101 4. Characterizing the Product [uore 4] ITE 4: Use the product characteriza. tion techniques designated by your ‘aboratry etucton 5.Cleaning Up art 5: The ttractve Index at 20°C calculated by using te folowing ea ton, where Tis the amber operate in degrees Clue, and ny tthe Totacine index measured at ambient temperatu, 8 = n+ 0.00045 (T-20°C) SS) / Figure 2 Test-tube distillation apparatus Characterize the product by measuring its boiling point, density, or refractive index. Record the temperature when measuring the refractive index. Make a temperature correction for the refractive index. [Nott 5] ‘Compare your experimental values with the literature values given under Reagents and Properties. Obtain an infrared spectrum of your product. Compare your product spectrum with a spectrum of the I-butanol used as the reagent. Pay particular attention to the regions at 3500 cm~' and 1050 cm". Place your recovered materials in the appropriate labeled collection containers as directed by your laboratory instructor. Clean your glassware with soap or detergent. (QUT iar rama) your hands thoroughly with soap or detergent before leaving the laboratory. SYNTHESIZING 1-BROMOBUTANE Equipment 50-mL beaker 10-mL graduated cylinder 400-mL beaker* 2 Pasteur pipets, boiling chip, acid-resistant with latex bulbt 15-mL centrifuge tube product vial" 100-mL. flask heater, rubber stopper, one-hole with heat controller rubber tubing.standard taper glassware: support ring pa condenser, with tubing 2 support stands ' distilling head 18 X 150-mm test tube po 10-mL round-bottom flask’ 2 utility clamps =" 100-mL round-bottom flask Y-tube or T-tube! ' thermometer, ~10-260 °C, bt with adapter \a vacuum adapter i ior ice bath {esteaeqlpment for optional procedure, 4. Distifling the Product a Nae ee ee 1s Reagents and Properties z _ molar mass bp density ‘substance quantity (gimol) = (°C)_— (gh) "B a ‘1-bromobutane* 1370 102 1.276 1.4390 " V-butanol 505 74d 1180810 1.390 calcium chloride, 10g ‘ anhydrous hydrobromic acid, 10.0mL_——80.9 149 18% sulfuric acid, 40mL 981 1841 concentrated sodium hydrogen 5,0 mL 1.04 carbonate, 5% d aqueous . Sprodact ce Preview - * Place a boiling chip, 1-butanol, and HBr solution into a roundbottom flask * Slowly add concentrated H;SO, to the cooled reaction mixture ‘© Heat the mixture at reflux for 1 hr © Codistill the product with water © Wash the organic layer with water, 5% NaHCO;, then water again © Dry the organic layer with anhydrous CaCl, © Purify the product by distillation © Characterize the product by boiling point, refractive index, density, or IR spectroscopy Hii A a \, ‘1-bromobutane—flammable and irritant ‘1-butanol—flammable and irritant(© 1008 Cengage Learning REACO714; Studying SNI and SN2 Reactions: Nuceophilic Substitution 103 1. Conducting the Reaction ITE: As the reaction occu, you sho- id ne the production of a s8cond Saud layer. One eer contains the water ‘and acid; the ether contains 1-bromo- butane produced bythe reaction {att Once the qui inthe ask roa: hes a steady refx condion, procees wth the secon "Factors Acting the Fates of S.1Reactons”on the page 1, 48% hydrobromic acid—toxic and corrosive sulfuric acid—toxic and oxidizer | Wear departmentally approved safety goggles at all times while in the ‘chemistry laboratory. | Hydrobromic acid (HBr) Is toxic and corrosive. Concentrated sulfuric acid (H_S0,)is toxic and otidizing. HBr and H,SO, can cause severe burns. In case of a ‘pill, notity your laboratory instructor immediately. 4-Butanol is flammable and irritating. Do not use near flames or other heat ‘sources. Prevent eye, skin, and clothing contact. Avoid inhaling fumes and ingesting these compounds. Use the 400-mL beaker to prepare an ice-water bath. Place an acid resistant boiling chip, 5.0 g (6.2 mL) of 1-butanol, and 10 mL of 48% HBr into a 100-mL round-bottom flask. Cool the mixture in the ice bath. Slowly add to the round-bottom flask, with stirring, 4 mL of concentrated HSOx, Fit the flask witha condenser for reflux, as shown in Figure 3. Fit the top of the condenser with a rubber stopper equipped with a Y-tube. Use rubber tubing to connect one arm of the Y-tube to the water aspirator. Leave the other arm open. Tum on the water to the aspirator to draw the acid fumes generated by the reaction. j Heat the mixture to boiling, Reflux the mixture gently for 1 hr. [Note 6] [Nore 7] Reflux apparatus for macro-scale preparation104 2.Codistillation of the Product ‘nr You wt observe wo ayers nthe fost ibe because the +-bromebulane ‘Mil cogs wih water Ones you collet YO-18 of cla, ony water shouts remain in ho pot ‘3.Washing the Product ‘REACO714: Studying SN1 and SN2 Reactions: Nucleophilic Substitution 1-Bromobutane is flammable and irritating. Do not use near flames or other heat sources. Prevent eye, skin, and clothing contact. Avoid inhaling fumes and ingesting 1-bromobutan« Once the reflux is complete, remove the heater from the flask and allow the flask to cool. Add 10 mL. of distilled or deionized water and an additional boiling chip. ‘Arrange the apparatus for distillation, as shown in Figure 4. Distill the mixture into an 18% 150-mm test tube until you collect 10-15 mL of distillate. [wore 8] Place the pot residue in the container labeled “Recovered Pot Residue”, provided by your laboratory instructor. Label a 50-ml. beaker “Washes”. Use a Pasteur pipet to remove the upper aqueous layer from the distillate. Place this layer into the labeled beaker. To wash the 1-bromobutane layer, add 5 mL of water to the test tube ‘and mix well. Use a Pasteur pipet to remove the water layer and transfer the water to the labeled beaker. ‘Wash the 1-bromobutane again, first with 5 mL of 5% aqueous sodium hydrogen carbonate (NaHCO;), then with a second 5 mL of water. Each és “ « i io a ic yt cc " a a oh { of z :EE ESN eee eee are ee {© 1998 Cengage Learing REACD714: Studying SNI and SN2 Reactions: Nuleophic Subscuton 105 aT: Toe Si layer becomes clear 38 the sod atycrous Cas xtcts waar trom te bau -bremebutee, 4, Distilling the Product (Optional) 5. Characterizing the Product [uore 10] Tw: Use the procuct characteriza. tion tectiques ‘destnated by your laboratory into. 6. Cleaning Up ITE The rotactve index at 20°C it calculated by using he following equ: ten, where Tis he ambien temperature in degrees Catius, and nb the ‘etractvo index messed at ambient temperature, 1B = nl, +0.00085(T-20 C) time, use the Pasteur pipet to remove the aqueous layer and place the layer {nto the labeled beaker. ‘After the last water wash, remove as much of the water as possible with the Pasteur pipet. Transfer the contents ofthe test tube to a centrifuge tube to facilitate the removal of the water. Again, remove as much water as possible. ‘Add approximately 1 g of anhydrous CaCl, to dry the product. Allow the product to dry for 5-10 min. [Nore 9] If you do not conduct the optional distillation in Part 4, measure and record the mass of your product. Tare a product vial. Use a clean, dry Pasteur pipet to transfer the dried ‘[-bromobutane into a 10-mL round-bottom flask. Assemble a distillation apparatus using the 10-mL round-bottom flask. Make certain the distilla- tion apparatus is clean and dry. Distill the product into the tared vial, collecting the portion that distills from 99-103°C, Measure and record the mass of the 1-bromobutane collected, Characterize the product by measuring its boiling point, density, or refractive index. Record the temperature when measuring the refractive index. Make a temperature correction for the refractive index. [Note 11] Compare the experimental values with the literature values given under Reagents and Properties. Obtain an infrared spectrum of your product. Compare your product spectrum with a spectrum of the I-butanol used as the reagent. Pay particular attention to the regions at 3500 cm™' and 1050 cm=! Place your recovered materials in the appropriate labeled collection containers as directed by your laboratory instructor. Clean your glassware with soap or detergent. your hands thoroughly with soap or detergent before leaving the ZN laboratory. a a FACTORS AFFECTING THE RATES OF Sy1 REACTIONS Equipment 2 Erlenmeyer flasks, 125-mL. 50-uL micropipet 50-mL graduated cylinder 100-:L micropipet* 100-mL graduated cylinder timer marking pen “or adjustable micropipet set to 200iREACO714: Seudying SN1 and SN2 Reactions: Nucleophilie Substitution 106 a Reagents and Properties i eee ‘ molar mass bp density, , substance quantity (gma) (°C) glen) 4250 Dbromopropane 50 yl. 1230 «58 114i 2-propanol 150 mL. oo 82 079 1.3770 (05M sodium 12mL | hydroxide phenolphthalein 30 drops 2bromo-2-methyl 200 uL 137 7m 11914279 propane 2-chloro-2-methyl 50 yl. 26 52 085 1.3848 propane Preview Measuring the Effect of the Leaving Group on Reaction Rate ‘© Prepare a 50% 2-propanol/water solvent mixture * Divide the mixture into two portions * Add phenolphthalein indicator and NaOH solution ‘* Add alkyl halides and measure the time required for discharge of the indicator color Measuring the Effect of the Alkyl Group Structure on Reaction Rate ‘© Prepare a 50% 2-propanol/water solvent mixture ‘* Divide the mixture into two portions ‘© Add phenolphthalein indicator and NaOH solution © Add alkyi halides and measure the time required for discharge of the indicator color Measuring the Effect of Solvent Polarity on Reaction Rate © Prepare 40% and 60% 2-propanol/water solvent mixtures | ‘* Add phenolphthalein indicator and NaOH solution j © Add 2-bromo-2-methylpropane and measure the time required for discharge of the indicator color \ { PROCEDURE 2-bromopropane—flammable and irritant AN 2-bromo-2-methylpropane—flammable 2-chloro-2-methylpropane—flammable ae :(© 1998 Cengage Learning [REACO714: Snadying SN1 and SN2 Reactions: Nucleophilic Substiution 107 1. Measuring the Effect of the Leaving Group on Reaction Rate tate 1: The phenopprnain snouc be bight pink nth slighty Basic sluion 2. Measuring the Effect of the Alkyl Group Structure on Reaction Rate 2-propanol—flammable and irritant ‘sodium hydroxide—corrosive and toxic en ee ery ce ee ee ‘Wear departmentally approved safety goggles at all times while in the chemistry laboratory. 2-Bromo-2-methylpropane, 2-chloro-2-methylpropane, and 2-propanol are ‘flammable and irritating. Do not use near flames or other heat sources. Prevent eye, skin, and clothing contact. Avoid breathing fumes and ingesting these compounds. To prepare a 50% mixture of 2-propanol in water, place 50 mL of 2-propanol into a 100-mL graduated cylinder and add enough distilled or deionized ‘water to make 100 mL. Mix well. Place 50 mL of the mixture into each of two 125-mL Erlenmeyer flasks. ‘Add 5 drops of phenolphthalein indicator and exactly 200 pL of 0.5M NaOH to each of the flasks. Mix well. [Nore 12] ‘Add 50 iL of 2-bromo-2-methylpropane, with swirling, to one of the flasks. Measure and record the time required for the solution to become colorless. ‘Add 50 iL of 2-chloro-2-methylpropane, with swirling, to the second flask. Again measure and record the time required for the solution to become colorless. mopropane, 2-bromo-2-methylpropane, fopanol are flam- ‘mable and irritating. Do not use near flames or other heat sources. Prevent eye, skin, and clothing contact. Avoid breathing fumes and ingesting these compounds. Prepare 100 mL of a 50% mixture of 2-propanol in water as described in Part 1. Divide the mixture equally between two 125-mL Erlenmeyer flasks. ‘Add 5 drops of phenolphthalein and exactly 200 ul of 0.5M NaOH to each of the flasks. Mix well ‘Add 50 pL of 2-bromo-2-methylpropane, with swirling, to one of the flasks. Measure and record the time required for the solution to become colorless. ‘Add 50 uL of 2-bromopropane, with swirling, to the second flask. Again measure and record the time required for the solution to become colorless. If the solution does not become colorless within 15 min, stop the reaction and record the time as >15 min.3, Measuring the Effect of Solvent Polarity on Reaction Rate 4,Cleaning Up ure you conduct ti section of the experiment cing ther ime for Section eI ret to that secton REACO714: Studying SN} and SN2 Reactions: Nucleophilic Substitution ‘2-Bromo-2-methylpropane and 2-propanol are flammable and irritating. Do not use near flames or other heat sources. Prevent eye, skin, and clothing ‘contact. Avoid breathing fumes and ingesting these compounds. To prepare 50 mL of a 40% mixture of 2-propanol in water, place 20 ml. of 2-propanol into a 50-mL graduated cylinder and add enough water to make 50 mL. Place the mixture into a labeled 125-mL Erlenmeyer flask. To prepare 50 mL of a 60% mixture of 2-propanol in water, place 30 mL. of 2-propanol into a 50-mL graduated cylinder and add enough water to make 50 mL. Place the mixture into a labeled 125-mL Erlenmeyer flask. ‘Add 5 drops of phenolphthalein and exactly 200 iL. of 0.5M NaOH to ‘each flask. Mix well. ‘Add 50 iL of 2-bromo-2-methylpropane, with swirling, to each flask. Measure and record the time required for each solution to become colorless. Place all alcohol-water reaction mixtures in the container labeled “Recovered Alcohol-Water Mixtures’, provided by your laboratory instructor. Clean your glassware with soap or detergent: [Nove 13] Wash your hands thoroughly with soap or detergent before leaving the laboratory.{© 1908 Cengage Learning REACO714: Studying SN1 and SN2 Reactions Nucleophilic Substation 109 Post-Laboratory Questions 1. Calculate the percent yield of I-bromobutane obtained in your experiment. 2. What experimental evidence can you provide that the product isolated in your synthetic experiment is 1-bromobutane? 3. Which compound, 2-bromo-2-methylpropane or 2-chloro-2-methyl-propane, reacted faster in your ‘Syl experiment? What were the relative rates of the two reactions? 4. Based on your answer to question 3, which is the better leaving group, Br or CI? Are these results consistent with the relative basicities of these two ions? Briefly explain. 5. Which compound, 2-bromo-2-methylpropane or 2-bromopropane, reacted faster in your Sul experiment? How are the reactivities of 2-bromo-2-methylpropane and 2-bromopropane related to the stabilities of the carbocations produced as intermediates in the reaction? Briefly explain. 6. Which of the two solvent mixtures, 40% 2-propanol or 60% 2-propanol, is more polar? Briefly explain. 7. In which of the two solvent mixtures did 2-bromo-2-methylpropane react faster? Account for your results in terms of the effect of solvent polarity on the rate-determining step in this Syv1 reaction. ‘8 Use your results to explain which variable—leaving group, alkyl structure, or solvent polarity—has the greatest impact on the rate of an Sy1 reaction.© 1908 Cengage Learning 3, Why should reflux of the microscale reaction mixture be gentle, with the condensation ring remaining close to the surface of the liquid in the test tube? 4, By observing the reaction mixture, what visual evidence can be gained to indicate that 1-butanol is being converted to 1-bromobutane? 5. What is the function of anhydrous CaCl, in this experiment?12 REACD714: Studying SN and SN2 Reactions: Nucleopilic Substcuion 6. The limiting reagent in the production of 1-bromobutane is I-butanol. Calculate the theoretical yield of 1-bromobutane for your experiment. Record your results here and in your laboratory notebook. 4 7. How does the Syl reaction in this experiment cause the acid-base indicator, phenolphthalein, to change color? Briefly explain. ‘8. Why is it important that the volume of 0.5M NaOH be measured exactly?8 Isolating Caffeine from Tea Prepared by Robert Silberman, SUNY, Cortland, and William M. Loffredo, East Stroudsburg University PURPOSE OF THE EXPERIMENT Extract caffeine from instant tea, purify the crude extract using recrystalli- zation ot sublimation, and determine the mass percent of caffeine in the instant tea. EXPERIMENTAL OPTIONS Extracting Caffeine Purifying Caffeine ‘A. By Recrystallization B. By Sublimation ‘You should be familiar with distillation, extraction, evaporation of solvents, recrystallization, and sublimation. D INFORMATION Caffeine is probably the most commonly used addictive drug. It is a stimulant consumed daily by millions of people as they drink coffee, tea, and soft drinks. The discovery that caffeine is a stimulant is generally attributed to the Chinese and has been known for thousands of years. Pierre Jean Robiquet first isolated the pure compound in 1821. Chemically, caffeine belongs to a large class of organic compounds called alkaloids, which vary widely in structure and reactivity. All alkaloids have a nitrogen atom that allows most of them to accept a proton (H*) and act as a base, or alkaline substance. The name “alkaloid” comes from this characteristic. The basic properties of most alkaloids make them easy to extract from plant materials by using aqueous acid solutions. Therefore, alkaloids were among the first compounds isolated from plants. CENGAGE © 280Copu any AL RTS RSEMED opto vo ey Be yt en mb ma . Learning nts own hm yay mans pai de ere hgh tie ng sary We etn ara rs «aa ie oe perc Secen al WE Be Sop eta epr ebngeseelbepdihe. 1200 Cengage Leaning© CHy N N’ > ANON SYN'T0732: Isolating Caffeine from Tea CH, ctfine Structural formulas of some xanthine alkaloids Examples of alkaloids range from important medicinal drugs such as quinine, used in the treatment of malaria, and morphine, a powerful painkiller, to commonly abused drugs such as cocaine and nicotine. The search for new plant alkaloids continues. Caffeine is not the only alkaloid stimulant found in beverages. Tea also contains theophylline and cocoa contains theobromine. Theophylline has been used as a bronchial dilator for asthmatics and as a heart stimulant. Theobromine is sometimes used as a diuretic. All three compounds are chemically related and belong to a subclass of alkaloids known as xanthines. Figure 1 shows the structures for xanthine and its derivatives, caffeine, theophylline, and theobromine. They differ only in the number and the placement of methyl groups, CH. In medicinal chemistry, structurally similar compounds, such as the xanthines, frequently exhibit related physiological activities. Thus, in their search for new and better drugs, medicinal chemists often synthesize a large number of compounds that are structurally similar to one that exhibits beneficial medicinal properties. For example, barbiturates, penicillins, and antihistamines are different families of useful drugs. Many methods for separating natural products from plants have been devised. These methods include extractions, steam distillation, and sublimation of pure substances from crude extracts, In this experiment, you will extract caffeine from tea. You will purify the caffeine either by recrystallization or by sublimation. You will calculate the mass percent of caffeine in the sample by using Equation 1. oy as of purified caffeine, g ‘mass of instant tea, g 100%) (Eq. 1) Equipment 25-mL beaker’ 25-mL filter flask, with #2 neoprene 50-mL beaker adapter and vacuum tubing’ 400-mL beaker* 125-mL filter flask, with vacuum 15-mL centrifuge tube! tubing condenser funnel, general purpose conical centrifuge tube 10-mL graduated cylinder distilling head Hirsch funnel, with adapter! 50-mL Erlenmeyer flask, hot plate with stopper microburner! 125-mL. Erlenmeyer flask microspatula*(© 1909 Cengage Learning ‘$xNVT0732:llating Caffeine from Tea 15 Pasteur filter pij support ring Pasteur pipet, with latex bulb’ —_ support stand ‘1-mL transfer pipet, with bulb test tube, 13 x 100-mm' phase separation paper test tube, 16 x 150-mm® juct vial thermometer, —10 to 110°C 50-mL round-bottom flask 3 utility clamps sor hot-water bath “for recrytalization or sublimation Sf phase separation paper snot available Reagents and Properties ade re ee ‘molar mass a substance quantity —(glmol) bp CC). mp (°C) (ghml) eee eee caffeine” 234-236 dichloromethane 10mL 84.93 40 1.325 diethyl ether/hexene, 1:1 2mL hexane ImL 86.18 6 0.659 instant tea 10g 2-propanol -2mL 60.10 82 sodium hydroxide 2M 5 mL sodium sulfate, 05g anhydrous* product "if phase separation paper is not available Preview Mix instant tea, 2M NaOH, and dichloromethane, and shake for 10 min © Separate the aqueous layer from the organic layer * _Distill the dichloromethane on an 80°C water bath to reduce its volume tolmL A. Purifying by Recrystallization ‘© Evaporate the remaining dichloromethane from the crude caffeine © Dissolve the crude caffeine crystals in a minimal amount of hot 2-propanol, cool, and add hexane © Filter the purified caffeine and wash with 1:1 diethyl ether/hexane solution © Weigh the crystals B, Purifying by Sublimation Evaporate the remaining dichloromethane from the crude caffeine Assemble the apparatus for vacuum filtration Apply vacuum to the flask containing the crude caffeine Carefully add the ice-water mixture to the centrifuge tube while the flask is under vacuumus ‘SYNNTO732; elating Caffeine fom Tes © Carefully and gently heat the outside of the flask to cause the caffeine to sublime ‘© Allow the flask to cool to room temperature while still under vacuum Pour the ice-water mixture out of the centrifuge tube and remove any water around the rim © Disassemble the apparatus and carefully scrape the caffeine crystals off of the tube into a weighed container © Weigh the crystals PROCEDURE Extracting Caffeine tart: Phase separaton paper ows ‘organic soon lo pass trough while ‘aoping aqueous solons ine cone ‘Wear departmentally approved safety goggles at all times while in the chemistry laboratory. ‘Always use caution in the laboratory. Many chemicals are potentially harmful. Prevent contact with your eyes, skin, and clothing. Avoid ingesting any of the reagents, Dichloromethane is toxic and irritating. Use a fume hood when working, with dichloromethane. Sodium hydroxide (NaOH) is corrosive. Tare a clean dry 50-mL Erlenmeyer flask. Place 1.0 g of instant tea into the flask. Add 5 mL of 2M NaOH and 10 mL of dichloromethane. Stopper the flask. Make certain that the stopper used with the 50-mL, Erlenmeyer flask fits tightly to prevent leakage during shaking. Gently shake the mixture for 10 min by repeatedly inverting the flask. Loosen the stopper periodically to release any pressure build-up. Avoid vigorous shaking, which will lead to the formation of emulsions. Allow the contents of the flask to settle. To separate the layers, pour the mixture through phase separation paper into a 125-mL. Erlenmeyer flask. [Nore 1]. Place the aqueous layer in the labeled collection container, provided by your laboratory instructor. ‘Sodium sulfate is irritating and hygroscopic. Minimize air contact. If phase separation paper is not available, place the mixture into a conical centrifuge tube. Allow the contents to settle. Use a Pasteur filter pipet to remove the bottom dichloromethane layer, which contains the crude caffeine. Place the dichloromethane layer into a 16 x 150-mm test tube. ‘Add 0.5 g of anhydrous sodium sulfate. Allow the layer to dry for 10 min(© 1900 Cengage Learning ‘SYNTO732: loating Calin fom Tea u7 ‘Transfer the dichloromethane into a 50-mL round-bottom flask. Assemble a simple distillation apparatus using the round-bottom flask. Heat a water bath to 80°C. Using the hot-water bath, distill the dichloromethane from the crude crystals until approximately 1 mL of rmathane souion containing the cue dichloromethane remains in the flask [Note 2]. case Disassemble the distillation apparatus. Place the 80°C water bath in a fume hood. 2. Purifying Caffeine [wore3] A. By Recrystallization oT 2: Use eit pureaton rocedre ‘Aor B as ected by your laboratory ineructor, ‘2-Propanol and hexane are flammable and irritating. Diethyl ether is highly flammable and toxic, Use a fume hood. Keep these compounds away from flames, or other heat sources. ‘Transfer the dichloromethane remaining in the flask to a 25-mL beaker. Heat the beaker on the water bath to remove the remaining dichloro- methane, leaving the crude caffeine crystals in the beaker. Place approximately 2 mL of 2-propanol into a clean, dry 13 x 100-mm_ test tube and place the tube in the hot-water bath. Heat the 2-propanol to 80 °C. Do not boil the 2-propanol away. If necessary, add more 2-propanol to the test tube. ‘Add hot 2-propanol dropwise to the crude dry crystals until the crystals just dissolve. Keep the beaker containing the crude crystals near the boiling point of 2-propanol, using the hot-water bath. Do not exceed 2 mL of 2-propanol total volume. Allow the solution to cool to room temperature. Slowly add, with stirring, 1 mL of hexane to the cooled 2-propanol solution. Let the mixture stand for 3 min. Collect the pure caffeine crystals by vacuum filtration using a Hirsch funnel. Wash the crystals carefully with 1 mL of 1:1 diethyl ether/hexane solution. Weigh the crystals and record the mass. Place the crystals in a labeled product vial By Sublimation Always inspect all glassware for cracks or chips before use. Cracked glassware can implode under vacuum. Discard any damaged glassware in the container provided by your laboratory instructor. Transfer the dichloromethane remaining in the flask to a 25-mL filter flask. Heat the flask on the hot-water bath to remove the remaining dichlor- ‘omethane, leaving the crude caffeine crystals in the flask. Clamp the filter flask containing the crude caffeine to a support stand, with the bottom of the flask approximately 10 to 12.5 cm from the base, as shown in Figure 2 on the next page.us sar & Be sure 0 place the ico-water fue into the carfuge hie far 2 ‘ecuim le elabiehed inthe fk terse, war vapor rom the ak may ‘condense on he ouside fhe oat ube ‘Any wate present onthe ouside of te tea ibe wil dsove the catene 3.Cleaning Up ‘SYNNT0732: Isolating Caffeine from Tes Figure 2 Apparatus for purifying caffeine by sublimation Insert a clean, dry 15-mL centrifuge tube through a #2 neoprene adapter. Position the tube in the filter flask so that the bottom of the tube is within 1 cm of the flask bottom. Connect the side arm of the filter flask to a trap and aspirator. Turn on the aspirator and check to see that the centrifuge tube assembly is held tightly, indicating that a vacuum is being produced in the flask. Apply the vacuum to the flask. After 1-2 min, carefully fill the tube with the crushed ice-water mixture [Note 4]. Light a microburner. Adjust the flame cone to a low heat intensity. Use the microburner flame to gently heat the outside of the filter flask. Keep the burner moving in order to evenly heat the outside of the flask. Warm the flask just enough to sublime the caffeine without melting it. Check to see that the sublimed caffeine is condensing onto the outside of the tube. ‘When no more caffeine condenses onto the outside of the tube, remove the burner. Allow the flask to cool to room temperature. Do not disconnect the vacuum source at this time. Carefully pour out the remaining ice-water mixture from the tube. Use a paper towel to absorb any water around the top of the flask and the stopper. When the flask opening is dry, break the vacuum. Remove the tubing from the filter flask and turn off the aspirator. ‘Tare a product vial. After the centrifuge tube has come to room temperature, disassemble the apparatus. Carefully scrape the caffeine from the outside of the tube into the tared product vial Weigh the caffeine crystals and record the mass. Label the product vial. Use the labeled collection containers provided by your laboratory instructor for disposal of all solutions and the purified caffeine. Clean your glassware with soap or detergent. ‘Wash your hands thoroughly with soap or detergent before leaving the ZN laboratory. — Ss© 1990 Cengage Leaming ‘SYNTTO732: Isolating Caffeine from Tea ug POST LABORATORY QUESTIONS 1. Calculate the mass percent of caffeine in your instant tea sample. 2, On the average, people use one teaspoon (2.5 g) of instant tea to make an Boz glass of iced tea. The average glass of iced tea contains 10 mg of caffeine per oz. (a) Calculate the mass percent of caffeine in a glass of iced tea. (b) Are your experimental results consistent with these facts? Briefly explain. (©) Ifnot, explain why your experimental results differ from these data.‘SYNTO732: Isolating Caffeine from Tea 121 Pre-Laboratory Assignment 11. Why should you use a fume hood when working with dichloromethane? 2. Why is gentle shaking necessary during the extraction of caffeine from tea? 3. Why is it important to not overheat the dichloromethane while extracting the caffeine? EE eee ee ewe eee eee eee ee ee ee eS oe 0st Cinna ni12 ‘S¥INTO732: loatng Caffeine fom Tea 4, Why should the 2-propanol be kept hot while dissolving the crude crystals? eh lh eGWAL bo ‘ Hite siren Mee(© 1008 Cengage Learning 9 SYNT Brominating Alkenes Prepared by Carl T, Wigal, Lebanon Valley College PURPOSE OF THE EXPERIMENT ‘Synthesize vicinal cihalides by brominating alkenes. Characterize vicinal dihalides by using the silver nitrate test and by using melting point mea- ‘surement to determine the relative stereochemistry. EXPERIMENTAL OPTIONS Using Semi-Microscale Techniques to Brominate Alkenes Cinnamic Acid cis-Stilbene trans-Stilbene Using Microscale Techniques to Brominate Alkenes Cinnamic Acid cis-Stilbene trans-Stilbene BACKGROUND REQUIRED You should consult your textbook for the Cahn-Ingold-Prelog System for assigning the configuration of a chiral center. You should be familiar with techniques for reflux, for vacuum filtration, and for melting point mea- surement. BACKGROUND INFORMATION The halogenation of alkenes is an important reaction in the chemical industry. For example, over 8 million tons of 1,2-dichloroethane per year are produced by the addition of chlorine (Cl,) to ethylene. This product is used both as a solvent and in the preparation of polyvinyl chloride, PVC, a common organic polymer used in household plumbing. The products obtained from alkene halogenation are called vicinal dihalides because the two halogen substituents are attached to adjacent carbon atoms. When the halogen used is either bromine (Br) or chlorine (Cl,), halogenation of alkenes occurs rapidly at room temperature, and the ee Ne CENGAGE © ®:8cmoe tering AL RTS RESEND Nop wk cn epee ay man Learning: temtsont caine nay mana ec ore aug as an cm smi pang i Vt tan oma an oan ee yet eri nt Scena 7 Unde ayia ep weeps14 ‘SYNTO719: Brominating Allens resulting vicinal dihalides are stable. Fluorination is a violent reaction that is difficult to control and is accompanied by several side reactions. Jodination is an endothermic process, resulting in vicinal diiodides that tend to revert to alkenes. Consequently, the most common applications of alkene halogenation are chlorination and bromination. Typically, alkenes undergo reactions through electrophilic addition, a process in which the alkene pi (n) bond is replaced with two sigma (0) bonds. The general mechanism of electrophilic addition involves two steps, as shown in Figure 1. The first step involves reaction of an electron deficient species, called an electrophile (E*), with the electron-rich x bond of the alkene. The two electrons of the x bond shift toward the electrophile, forming a new carbon— electrophile abond. This step results in formation of a positively charged intermediate. In most instances, the positive charge centers on a carbon atom, so the electron-deficient intermediate is called a carbocation. In the second step, the electrophilic carbocation reacts with an electron- rich species called a nucleophile (Nu”). The nucleophile donates an electron pair to the positively charged intermediate forming a carbon-nucleophile bond. Alkene bromination follows the same general mechanism with a few important modifications, as shown in Figure 2. In the first step, the proximity of the 1 electrons of the alkene to Br, polarizes the bromine bromine bond. This polarization induces a bond dipole that allows Br. to act as an electrophile. Electrons flow from the x bond to the polarized Bra, forming a carbon-bromine bond and breaking the bromine-bromine bond, producing a bromide ion (Br-). The positively charged intermediate in bromination is not a carbocation, but a bromonium ion. This cyclic intermediate results from a nonbonding electron pair from bromine that ae or carbocation Figure 1 Electrophilic addition to an aikene1908 Cengage Learning HO,C Figure 3 maleic acid fumaric acid SYNTO719: Brominating Alkenes 125 stabilizes the positive charge on carbon. The bromonium ion is more stable than a simple carbocation because all atoms of the bromonium ion have an ‘octet of electrons. In the second step, Br~ acts as a nucleophile, attacking the electron- deficient bromonium ion. The second carbon-bromine 6 bond of the vicinal dihalide forms in this step. An important feature of this step is the resulting stereochemistry. Bromide ion adds to the side opposite the carbon-bromine bonds of the bromonium ion. This process is called anti-addition. Anti- addition of Br- occurs because Br- is blocked from one face of the bromonium ion by the bromine atom. The consequence of anti-addition of Br, to cyclic alkenes is the formation of trans vicinal dihalides. Chiral carbon atoms, or chiral centers, are generated in many organic reactions. A chiral carbon is a carbon atom that is bonded to four different substituents. As a consequence, two different configurations are possible for a chiral center: rectus (R) or sinister (S). In the case of alkene bromination, the formation of the two carbon- bromine obonds could result in two new chiral centers in the vicinal dihalide. Using the 2" rule, where n is the number of chiral centers in a molecule, a maximum of four stereoisomers could result from alkene bromination. However, not all stereoisomers are formed during a single bromination. Bromonium ion formation preserves the stereochemical integrity of the starting material. Therefore, trans alkenes form trans bromonium ions and cis alkenes form cis bromonium ions. Consider the bromination of maleic and fumaric acids, as shown in Figure 3. When maleic acid, the cis isomer, is brominated, the bromo- nium ion formed has cis configuration. The product is a mixture of two HO,C , e ae Ge Br a \" COH HOC, (sCOH ne g 2 aes oe Rar HH Nee Neape con p . SoH Br HO,C"Y bie 253. Pr © Bromination of maleic acid and fumaric acid126 SYNTO719: Brominating Alkenes stereoisomers. These stereoisomers are enantiomers having the absolute configurations of 2R,3R and 25,38. Enantiomers are molecules that contain chiral centers and are non-superimposable mirror images. Anti-addition of Br~ to the cis bromonium ion can take place by either path a or path b in Figure 3. Addition by path a results in 2R,3R, while addition via path b results in 25,3. Addition occurs at the same rate by either path; therefore, the two enantiomers are produced in equal amounts. A mixture containing equal amounts of a pair of enantiomers is called a racemic mixture, A racemic mixture is often designated by placing (+) at the front of the name. Unlike bromination of maleic acid, bromination of fumaric acid results ina single stereoisomer. The bromination of fumaric acid results in a trans bromonium ion intermediate. Addition of Br” to the trans bromonium ion by either path a or path b yields the same compound. The absolute stereo- chemistry of the product is 2R,3S, which is identical to 2S,3R. This compound is an example of a meso compound, which is a molecule that contains chiral centers, but also contains an internal plane of symmetry. As a result, meso compounds have superimposable mirror images. Meso dibromides result from the bromination of symmetrically disubstituted trans alkenes. Ifthe alkene is not symmetrically substituted, a pair of enantiomers will result. The relative stereochemistry of the enantiomers could have both chiral centers having the same configuration (R,R or S,S) or the opposite configuration (R,S or $,R). The prefixes erythro and threo are used to differentiate these stereoisomers. Erythro refers to a pair of enantiomers having a configuration similar to the sugar erythrose. The erythro form is often described as “‘meso-like” because the molecule would have a plane of symmetry if the two dissimilar groups were equivalent. Threo refers to a pair of enantiomers having a configuration similar to the sugar threose. ‘These configurations are shown in Figure 4. In this experiment, you will brominate an alkene using pyridinium tribromide, a comparatively safe, convenient source of bromine. You will characterize your product by measuring its melting point and by conducting a silver nitrate test, Vicinal dihalides react with alcoholic silver nitrate within five minutes to form a precipitate of the corresponding silver halide. This reaction can serve as a simple test for the presence of bromine or chlorine atoms. You will determine the relative stereochemistry of your Product by melting point measurement because the stereoisomers’ melting points differ significantly. CHO CHO H7-OH HO--H H--OH HO--H CH;OH = CHOH (RR) (ss) (ererythrose @ Figure 4 Configurations of (a) erythrose and (b) threose(© 1998 Cengage Leaming SYNTO719: Brominating Alkenes a7 USING SEMI-MICROSCALE TECHNIQUES TO BROMINATE ALKENES Equipment 250-mL beaker 2 Pasteur pipets, with latex bulb 25-mL filter flask, reflux assembly ‘with vacuum tubing condenser, with tubing filter paper 25-mL round-bottom flask 10-mL graduated cylinder thermometer, —10 to 260°C Hirsch funnel, with adapter sand bath* magnetic stir bar spatula magnetic wand 2 support stands ‘melting point capillary tubes 13x 100-mm test tube micropipet, 100 to 1000-uL. 2 utility clamps * string hotplate with crystallizing dish filled with sand or magnetic stirer and electric ask heater filled with sand Reagents and Properties molar mass bp = mpd substance quantity (gimol) = °C)——PC) (gm) acetic acid’, glacial 6 mL. 60.05 118 1.049 cinnamic acid 450 mg 148.16 133 1,2-dibromo- 340.07 1,2-diphenylethane’ 2,3-dibromo-3-pheny 30797 propanoic acid’ ethanol", 95% 05 mL pyridinium tribro 06-1.155g 319.84. mide* silver nitrate’, 2% in 0.5 mL ethanol cis-stilbene 300 pL. 180.25 145mm 1011 trans-stilbene 300 mg 180.25 124 amount for one brominatin * product Preview * Assemble the reflux apparatus * Add the alkene, pyridinium tribromide, and acetic acid © Reflux the reaction mixture Allow the reaction mixture to cool to room temperature Remove the stir bar Add water and cool the reaction mixture in an ice-water bath Use vacuum filtration to isolate the product Dry and weigh the product128 SYNTO719: Brominating Alkenes © Measure the melting point of the product © Test the product with silver nitrate reagent ‘1.Assembling the Apparatus ‘2. Brominating the Alkenes [nore 1] {WM You abort nstructor wil \iesgrate wach akenes you wl Brom: AIT & The sci matrls ae rt sou ‘at rom emperare acetic aoc, ZAIDI. a ommmay N, acetic acid—corrosive ‘innamic acid—irritant ethanol—flammable and irritant pyridinium tribromide—corrosive and lachrymator silver nitrate—toxic and oxidizer times while in the ‘Wear departmentally approved safety goggl chemistry laboratory. Assemble the reflux apparatus shown in Figure 5. Acetic acid is corrosive. Pyridinium tribromide is corrosive and a ZN lachrymator. Prevent eye, skin, and clothing contact. Avoid inhaling or ingesting these compounds. Use a fume hood to dispense these reagents. ee eae rE a SRO aT Cinnamic Acid Cinnamic acid is irritating. Prevent eye, skin, and clothing contact. WN —— a Remove the 25-ml. round-bottom flask from the apparatus. Place 450 mg of cinnamic acid, 6.0 mL of acetic acid, and 1.155 g of pyridinium tribromide in the round-bottom flask. Add a magnetic stir bar. [Note 2] Proceed to Part 3. cis-Stilbene Remove the 25-mL round-bottom flask from the apparatus. Place 300 uL of is-stilbene, 6.0 mL of acetic acid, and 600 mg of pyridinium tribromide in the round-bottom flask. Add a magnetic stir bar. [Nort 2] Proceed to Part 3. trans-Stilbene Remove the 25-mL round-bottom flask from the apparatus. Place 300 mg of trans-stilbene, 6.0 mL of acetic acid, and 600 mg of pyridinium tribromide in the round-bottom flask. Add a magnetic stir bar. [Nore 2] Proceed to Part 3. Lake. i ‘=(© 1008 Cengage Leeming SYNTO719:Brominating Akenes ae Hrs tunnel ic aspirator 25m tor flask Figure 6 Semi-microscale reflux apparatus Vacuum filtration apparatus 3. Refluxing the Reaction 4.Collecting, Washing, and Drying the Crystals 5. Identitying the Product 6.Cleaning Up Reattach the round-bottom flask to the reflux apparatus. Start the flow of water through the condenser. Heat the reaction mixture to reflux while stirring. Reflux for 20 min. After 20 min, remove the flask from the heat. Allow the reaction mixture to cool for 5 min. Turn off the water. Remove the condenser and use a magnetic wand to remove the stir bar. Add 8.0 mL of distilled or deionized water to the flask. Prepare an ice-water bath by half filling a 250-mL beaker with equal volumes of ice and water. Place the flask in the ice-water bath for 15 min. While the reaction mixture is cooling in the ice bath, assemble a vacuum filtration apparatus, as shown in Figure 6. Turn on the water to the aspirator and moisten the filter paper with a few drops of water. Filter the crystalline solid using the vacuum filtration apparatus. Wash the crystals with 3.0 mL. of water. Allow the crystals to dry in the Hirsch funnel by pulling air through the funnel for 15 min. Weigh your dried product and record its mass. Ethanol is flammable and irritating. Keep away from flames or other heat ZN Sources. Silver nitrate is toxic and oxidizing. Prevent eye, skin, and clothing contact. Avoid inhaling fumes and ingesting these compound: Measure and record the melting point of the product. Using a small test tube, dissolve approximately 10 mg of the product in 0.5 mL of 95% ethanol To this test tube, add 0.5 mL of 2% ethanolic silver nitrate. Allow the test tube to stand for 5 min. Record the presence or absence of a Precipitate. Place your recovered materials in the appropriate labeled collection containers as directed by your laboratory instructor. Clean your glassware with soap or detergent.130 SYNTO719: Brominating Alkenes Wash your hands thoroughly with soap or detergent before leaving the laboratory. 7 USING MICROSCALE TECHNIQUES TO BROMINATE ALKENES Equipment t 250-mL beaker 10-mL graduated cylinder conical vial reflux assembly* Hirsch funnel, with adapter condenser, with tubing magnetic stir bar or spin vane 5.0-mL conical vial melting point capillary tubes elastomeric connector 100-4L micropipet reflux assembly* 2 Pasteur pipets, with latex bulb condenser, with tubing sand bath elastomeric connector spatula 5.0-mL round-bottom flask 2 support stands 25-ml filter flask, 13 x 100-mm test tube with vacuum tubing thermometer, ~10 to 260°C filter paper 2 utility clamps forceps" * use reflux assembly indicated by your laboratory instructor ora magnetic wand * stirring hot plate with crystallizing dish filled with sand or magnetic stirrer and electric flask heater filled with sand Reagents and Properties ’ molar mass mpd substance quantity (gimol) bp CC) CC) (g/mL) acetic acid*, glacial = 2 mL. 60.05 118 1.049 cinnamic acid 150mg 148.16 133 ! 12-dibromo- 340.07 12-diphenylethane’ 2,3-dibromo-3-pheny/- 307.97 propanoic acid’ ‘ ethanol", 95% 05 mL pyridinium, 200-385 mg 319.84 tribromide* silver nitrate’, 2% in 0.5 mL. ethanol cis-stilbene 100 uL 180.25 1454" trans-stilbene 100 mg 180.25 124 : * tmnounts for one bromination * product Preview Assemble the reflux apparatus ‘© Add the alkene, pyridinium tribromide, and acetic acid© 1008 Cengage Learning ‘SYNTO719: Brominating Alkenes 135, Pre-Laboratory Assignment 1. What safety precautions must be observed when using (@) pyridinium tribromide? (b) acetic acid? 2. Calculate the theoretical yield for the bromination of both stilbenes and cinamic acid, assuming the presence of excess pyridinium tribromide. Note the theoretical yields here and in your laboratory notebook. 3. Draw the mechanism, including the intermediate bromonium ion, generated in the bromination of trans-2-butene. 4. (@) Look up and draw structures for cinnamic acid, cis-stlbene, and trans-stlbene. (©) Predict the relative stereochemistry of each product and draw the predicted structures.( 132 SYNTO719: Brominating Alkenes ‘ d 2. Brominating the Alkenes [vere] tun You weessery never wi Ab@te acid Is corrosive, Pyridinium tribromide is corrosive and a ‘erate which akenvs you wil trom Jachrymater. Prevent eye, skin, and clothing contact. Avoid inhaling or ingesting nae these compounds. Use a fume hood to dispense these reagents. Remove the 5.0-ml. conical vial (or round-bottom flask) from the apparatus. Place 150 mg of cinnamic acid, 20 mL. of acetic acid, and 385 mg of pyridinium tribromide in the conical vial (flask). Add a magnetic stir bar. [nore 2] Proceed to Part 3. cis-Stilbene Remove the 5.0-mL conical vial (or round-bottom flask) from the apparatus. Place 100 uL of cis-stlbene, 2.0 mL of acetic acid, and 200 mg of pyridinium tribromide in the conical vial (flask). Add a magnetic stir bar. [Note 2] Proceed to Part 3 trans-Stilbene Remove the 5.0-mL conical vial (or round-bottom flask) from the apparatus. Place 100 mg of rans-stilbene,2.0 mL of acetic acid, and 200 mg of pyridinium tribromide in the conical vial (ask). Add a magnetic stir bar. [Nore 2] Proceed to Part 3. / 3.RefluxingtheReaction _Reattach the conical vial (flask) to the reflux apparatus. Start the flow of water through the condenser. Heat the reaction mixture to reflux while 4 stirring. Reflux for 15 min. After 15 min, remove the vial (flask) from the heat. Allow the reaction mixture to cool for 5 min. Remove the condenser and use forceps or a magnetic wand to remove the stir bar. Add 2.5 mL of distilled or deionized water to the conical vial (flask), Prepare an ice-water bath by half-filling a 250-mL beaker with equal volumes of ice and water. Place the vial (flask) in the ice-water bath for 15 min. 4,Collecting, Washing, and While the reaction mixture is cooling in the ice bath, assemble a vacuum Drying the Crystals filtration apparatus, as shown in Figure 8. Turn on the water to the aspirator and moisten the filter paper with a few drops of water. Filter the crystalline solid using the vacuum filtration apparatus. Wash the crystals with 3 mL of water. Allow the crystals to dry in the Hirsch funnel by pulling air through the 2 aia a funnel for 15 min. Weigh your dried product and record its mass.© 1098 Cengage Leeming 5. Identifying the Product 6. Cleaning Up ‘S¥NTO719: Brominating Alkenes Hirsch tunnel SS 10 aspirator | —25-mt. ter task Figure 8 Vacuum filtration apparatus Ethanol is flammable and Irritating. Keep away from flames or other heat sources. Silver nitrate is toxic and oxidizing. Prevent eye, skin, and clothing contact. Avoid inhaling fumes and ingesting these compounds. Measure and record the melting point of the product. Using a small test tube, dissolve approximately 10 mg of the product in 0.5 mL of 95% ethanol. To this test tube, add 0.5 ml of 2% ethanolic silver nitrate. Allow the test tube to stand for 5 min. Record the presence or absence of a precipitate. Place your recovered materials in the appropriate labeled collection containers as directed by your laboratory instructor. Clean your glassware with soap or detergent. | Wash your hands thoroughly with soap or detergent before leaving the laboratory. pesenungererm: wesruueen eer14 SYINTO719; fron POST-LABORATORY QUESTIONS 1. (a) Compare the melting point of your product(s) to the data provided. In each case, identify the product you produced, compound mp CC) Grimans pha incam aaa (threo 4 (Lrerythro 203, (£)-1,2-dibromo-1,2-diphenylethane 110 meso-1,2-dibromo-1,2-diphenylethane 238 (b) Draw your product in its correct stereochemical configuration. (© Compare your results with your predictions for Pre-Laboratory Assignment question 4. 2.Calculate the percent yield that you obtained from your alkene brominations, 3. (a) When silver nitrate solution was added to your product, what did you observe? (b) Explain your observations. 4, Write reactions for the brominations you performed, in each case showing the intermediate bromonium ion that formed© 1008 Cengage Learning ‘SYNTO719: Brominating Alkenes 135, Pre-Laboratory Assignment 1. What safety precautions must be observed when using (@) pyridinium tribromide? (b) acetic acid? 2. Calculate the theoretical yield for the bromination of both stilbenes and cinamic acid, assuming the presence of excess pyridinium tribromide. Note the theoretical yields here and in your laboratory notebook. 3. Draw the mechanism, including the intermediate bromonium ion, generated in the bromination of trans-2-butene. 4. (@) Look up and draw structures for cinnamic acid, cis-stlbene, and trans-stlbene. (©) Predict the relative stereochemistry of each product and draw the predicted structures.© 1098 Cengage Leaming 10 TECH Isolating Clove Oil from Cloves Using Steam Distillation Prepared by Joseph W. LeFevre, SUNY Oswego PURPOSE OF THE EXPERIMENT Isolate clove oil from cloves by steam distillation and extraction. Use reactions with bromine, potassium permanganate, and iron(II) chloride to characterize the product. Analyze the product purity by thin-layer chromatography. EXPERIMENTAL OPTIONS Semi-Microscale Steam Distillation Microscale Steam Distillation Using Glassware with Elastomeric Connectors Using the Hickman Still Characterizing the Product BACKGROUND REQUIRED You should be familiar with distillation, extraction, drying organic solvents, speeding evaporation of organic solvents, thin-layer chromato- graphy, and general microscale techniques. BACKGROUND INFORMATION Simple and fractional distillations are carried out on miscible mixtures. Ideal mixtures follow Raoult's law: The total vapor pressure of the system is determined by adding together the products of the vapor pressure and the respective mole fraction of each compound. For a two-compound system, this relationship is shown in Equation 1, where P+ is the total vapor pressure, P;° and P,° are the vapor pressures of pure compounds and 2, and X; and X> are the respective mole fractions. rr ee ae eee GUS CENGAGE © 108 coe ewig AL A RESEND No pref newt comedy ag he my bmp © Learning: — min stecnt say omarhyuy nas gp to radar) reg amg png bY son oar ero ern Srp el pra mee rs Sc YT of 7 de Sates Cpt At wh pr wie pie petFigure 1 Structures for (a) eugenol and ©) eugenol acetate TECHO722: solating Clove Ol from Cloves Using Steam Distillation Py = PPX: + PPX: (&q-1) Distillation can also be performed on mixtures in which the two compounds are not miscible. This process is called codistillation. When ‘one of the compounds is water, the process is called steam distillation. When two immiscible liquids are distilled, the total vapor pressure Pr above the liquid is equal to the sum of the vapor pressures of each compound. This relationship, known as Dalton’s law, is shown in Equation 2. Pr=Pp+P? (Eq. 2) ‘The respective mole fractions are not included in this equation because, in ‘an ideal situation, each liquid vaporizes independently of the other. When Py is equal to atmospheric pressure of 760 torr, compounds 1 and 2 begin to codistill, with each compound contributing to Pr. Consider water as compound 1. The vapor pressure of pure water at its boiling point of 100 °C is 760 torr. Because compound 2 also contributes to Pz, the mixture will-wistill at a temperature less than 100 °C. The actual distillation temperature will depend on the vapor pressure of compound 2. Steam distillation offers an advantage in that volatile compounds that are unstable or have high boiling points can codistill with water at relatively low temperatures. This process avoids decomposition that might occur at the normal boiling point of the compound of interest. For example, eugenol, the ‘major compound of clove oil, boils at a relatively high temperature of 254 °C. Steam distillation avoids this high temperature and results in the distillation of eugenol at a temperature slightly less than 100 °C. In practice, steam distillation is usually carried out by one of two methods. In the first method, an excess of water is added to the compound of interest in a distilling flask, The mixture is then heated to the boiling, point. The resulting vapor is condensed and collected in a receiving flask. The compound of interest is then separated from the water, often by extraction. In the second method, steam is bubbled into the compound of interest to effect the distillation. In this experiment, you will use the first method because it is easier to set up. Clove oil belongs to a large class of natural products called the essential oils. Many of these compounds are used as flavorings and perfumes and, in the past, were considered to be the “essence” of the plant from which they were derived. Cloves are the dried flower buds of the clove tree, Eugenia caryophyllata, found in India and other locations in the Far East. Steam distillation of freshly ground cloves results in clove oil, which consists of several compounds. Eugenol is the major compound, comprising 85-90 percent. Eugenol acetate comprises 9-10 percent. These structures are shown in Figure 1. (CH;CH=CH @ es CH;CH=CH, HO’ HyC a{© 1098 Cengage Leaning 139 -TeCHH0722 lating Clove Oil om Cowes Using team Disillaion igenol hydroxyl contains a carbon-carbon double bond and an aromatic group called a phenol. These functional groups provide the basis for Simple chemical tests used to characterize the clove oil. A solution ‘bromine (Brz) in dichloromethane decolorizes as Bra reacts with the double bond to form a colorless compound, as shown in Equation 3. A positive test is the disappearance of the red Br, color. Qualitative Tests Tar Br pe yes a een Nae w, Ea. 3) ee Br [edie couprund (colores A potassium permanganate (KMnO,) solution can oxidize a double etree earns to form a 1,2-diol with the simultaneous reduction of Mn”* in KMnO, to Mn** in manganese dioxide (MnO2), as shown in Equation 4. A positive test is the disappearance of the purple KMnO, and the appearance of MnO; as a muddy brown precipitate. 3 A +2 Keno, + 4 HO — 3 JK + 2MnO, + 2 KOH He (Eq. 4) (purple) 363 rowed - Phenols (Ar-OH) react with the Fe** ion in iron(II) chloride (FeCls) to give complexes that are blue, green, red, or purple, as shown in Equation 5. The color may last for only a few seconds or for many hours, depending on the stability of the complex. Sates Oar 4 ArO,,, | sOAr 6ArOH + Fe+ <== Fe" + 6H aro” | Noar oat Oar (Eq. (yellow) (blue, green, red, or purple) In this experiment, you will steam distill clove oil from freshly ground cloves. Following the distillation, clove oil and water will be present in the receiving flask. Because clove oil will be a minor fraction of the distillate, the clove oil must be extracted from the water into an organic solvent such as dichloromethane. Removing the dichloromethane layer leaves clove oil as the product.vo Semi-Microscale Steam Distillation vive vale ‘TECHO722: isolating Clove Oil from Cloves Using Steam Distillacion Equipment boiling chips ‘sand bath’? Bunsen burner 125-mL separatory funnel® cotton* standard-taper glassware electric flask heater Claisen connecting tube 50-mL Erlenmeyer flask, condenser, with stopper with adapter and tubing glass stirring rod distilling head 10-mL graduated cylinder 100-mL round-bottom flask 50-mL graduated cylinder 2 round-bottom flasks, 50-mL ‘marking pen thermometer, ~10 to 260 °C, microspatula with adapter mortar and pestle support ring 3 Pasteur pipets, with latex bulb 2 support stands powder funnel 3 utility clamps “for Pasteur iter pipet ‘For hot-water bath ‘Ysand in crystallizing dish on electric hot plate or sand in electric heating well with heat controller Salso use as addition funnel Reagents and Properties Molar mass Quantity (ghmol) bp °C) 58 21 mL 84.93 40 164.20 254 10 mL 32.04 64.7 10 mL 58.44 sodium sulfate, anhydrous 058 142.04 ‘product ‘Preview ‘+ Grind the cloves with a mortar and pestle * Place the ground cloves and water in the distilling flask + Assemble the steam distillation apparatus +» Distill the mixture + Extract the clove oil into dichloromethane ‘+ Dry the dichloromethane layer with anhydrous Na;SO, ‘+ Remove the dichloromethane from the clove oil by distillation « Weigh the clove oil(© 1998 Cengage Learning ‘TECHO722; lsolaing Clove Oil from Cloves Using Steam Distillation ua PROCEDURE A eee ChemicalAlet MRQaRE Cee cia ee Wear departmentally approved safety goggles at all times while in the chemistry laboratory. 4. Conducting Steam Weigh 5 g of whole cloves. Grind them to a coarse powder, using a mortar Distillation and pestle. Reweigh the powder and record the mass. Use a powder funnel to transfer the ground cloves to a 100-mL round- bottom flask. Add 40 mL. of deionized or distilled water and a boiling chip to the flask. Mix well with a glass stirring rod. Mark the level of the mixture on the side of the flask. ‘Add 30 mL of water to a 50-mL round-bottom flask. Mark the level of the water on the side of the flask. Then discard the water from the flask. Assemble the steam distillation apparatus shown in Figure 2. Use the 100-mL round-bottom flask as the pot. Use the 50-mL round-bottom flask as the receiver. If a vacuum adapter is not used, make certain there is an opening to the atmosphere. Pour 100 mL of water into the addition funnel. Start the flow of water through the condenser. Adjust a Bunsen burner flame to lessen the hot central cone. Heat the pot by waving the flame back and forth under the pot. Maintain a distillation rate of approximately one drop every 3-5 s. — thermometer Figure 2 Semi-microscale steam distillation apparatus142 2. Extracting the Clove Oil ‘TECHO722: lsolating Clove Oil from Cloves Using Steam Disillaion NOTE: Do not heat the mature too rapidly. The clove mixture tends to foam when rapidly heated. The burner tlame can easily be added and withdrawn to control the heating rate, Add water to the pot at 10-min intervals to keep the water level at the mark. Stop the distillation when approximately 30 mL of distillate has been collected. Dichloromethane is toxic and irritating. Use a fume hood. Clove oil (eugenol) is irritating. Prevent eye, skin, and clothing contact. Allow the receiver to cool to room temperature. Carefully pour the distillate from the receiver into a 125-mL separatory funnel. Add 10 mL of saturated NaCI solution. Using a Pasteur pipet, carefully rinse the condenser and the inside neck of the receiving flask with 5 mL of dichloromethane. Swit] the flask gently to dissolve the remaining clove oil. Add this dichloromethane to the distillate in the separatory funnel. NOTE: Significant amounts of clove oll will adhere to the condenser and the sides and eck of the receiving tlask. Cap the separatory funnel and gently swirl the contents for several seconds. Vent the separatory funnel frequently. After the pressure has been vented, shake the contents vigorously to thoroughly mix the two layers. ‘Swirl the separatory funnel. At the same time, gently tap the outside of the separatory funnel with your index finger to force into the bottom layer any droplets of dichloromethane that are adhering to the sides of the funnel. Allow the layers to separate. Drain the lower dichloromethane layer into a 50-mL Erlenmeyer flask, making certain that none of the aqueous layer is transferred to the flask. Rinse the condenser and the receiver with a second 5-mL portion of dichloromethane. Transfer the rinsing to the separatory funnel. Repeat the extraction of the aqueous layer. Drain the second dichloromethane extract from the separatory funnel and combine it with the first one in the 50-mL. Erlenmeyer flask. Repeat the rinsing and extraction process with a third 5-mL portion of dichloro- methane. Combine the third extract in the same 50-mL Erlenmeyer flask. Anhydrous sodium sulfate (Na,SO,) is irritating and hygroscopic. Do not inhale and ingest this compound. Add approximately 05 g of anhydrous Na;SO, to the flask containing the dichloromethane extracts. Stopper the flask. Allow the extracts to dry for 5 min. Weigh a clean, dry 50-mL round-bottom flask to the nearest 0.001 g and record the mass. Using a Pasteur filter pipet, transfer the dried dichloro- ‘methane into the flask, making certain that no Na;SO, is transferred with the(© 1008 Cengage Learing “TECHO722;llating Clove Ol from Cloves Using Steam Dilation 43 3. Cleaning Up solution. Use three additional 2-mL portions of dichloromethane to rinse the 'NaSO, and ensure complete transfer of the clove oil to the beaker. ‘Assemble a simple distillation apparatus using the 50-mL round- bottom flask as the pot. Add a boiling chip. Use a 40 °C sand bath or a hot- water bath to distill the dichloromethane away from the product. When all of the dichloromethane has been distilled, cool the flask. Weigh it to the nearest 0.001 g and record the mass. Subtract the mass of the empty flask to obtain the mass of the clove oil ‘Methanol is flammable and toxic. Keep away from flames or other heat sources. Prevent eye, skin, and clothing contact. Use a fume hood. Dissolve the clove oil in 10 mL of methanol. Proceed to the Characterizing, the Product Section later in this module. Place your recovered materials in the appropriate labeled collection containers as directed by your laboratory instructor. Clean your glassware with soap or detergent. Wash your hands thoroughly with soap or detergent before leaving the laboratory. MICROSCALE STEAM DISTILLATION Using Glassware with Elastomeric Connectors Equipment 25-mL beaker 10-mL graduated cylinder boiling chip marking pen 15-mL. centrifuge tube, with cap microburner copper metal sponge microspatula cotton* mortar and pestle 10-mL Erlenmeyer flask, 3 Pasteur pipets, with latex bulb with stopper 1.0-mL pipet 125-mL Erlenmeyer flask sand bath’ glass stirring rod support ring glassware, support stand with elastomeric connectors thermometer, ~10 to 260 °C, distilling head/air condenser _ With adapter distilling tube, 2 utility clamps with syringe port wire gauze 10-mL round-bottom flask IemL syringe “for Pasteur Site pipet ‘sand in crystallizing dish on electric hot plate or sand in electric heating well with heat controllerM4 ‘TECHO722; Isolating Clove Oil from Cloves Using Steam Distillation Reagents and Properties ee ee Molar mass Substance Quantity (gimo) bp CC) cloves 05 dichloromethane 4mL- 84.93 40 eugenol” 164.20 254 ice ‘methanol 1 mL. 32.04 47 sodium chloride, sat. solution 1 mL 58.44 sodium sulfate, anhydrous 50 mg 142.04 ‘product Preview + Grind the cloves with a mortar and pestle + Place the ground cloves and water in the distilling flask + Assemble the steam distillation apparatus + Distill the mixture + Extract the clove oil into dichloromethane + Dry the dichloromethane layer with anhydrous Na,SO, + Remove the dichloromethane from the clove oil « Weigh the clove oil PROCEDURE A ‘Chemical Alert 1. Conducting Steam Distillation eee urea eee Siren erie Cu eee me ceeree Wear departmentally approved safety goggles at all times while in the a chemistry laboratory. Fe ee ereecene ee eR YO UO Grind 10 whole cloves to coarse powder, using a small mortar and pestle. Weigh 0.400-0.500 g of the powder and record the mass to the nearest 0.001 g. Use a microspatula to carefully transfer the ground cloves to a 10-ml. round-bottom flask. Add a boiling chip and 4 mL. of deionized or distilled water. Mix well with a glass stirring rod. Mark the level of the mixture on the side of the flask. Add 3 mL of water to a 15-mL centrifuge tube. Mark the level of the water on the side of the tube. Then discard the water from the tube.(© 1008 Cengage Learing ‘TECHO722: lolting Clove Oil from Cloves Using Star Distillation M5 Figure 3 Microscale steam distillation ‘apparatus using elastomeric connectors 2. Extracting the Clove Oil Place a small plug of copper metal sponge in the distillation head to help prevent the mixture from foaming over into the centrifuge tube when the distilling flask is heated. Fill a 125-mL Erlenmeyer flask three-quarters full with crushed ice. Place the centrifuge tube in the flask. Assemble the remainder of the distilling apparatus, as shown in Figure 3. Start the flow of water through the condenser. Adjust a microburner flame to lessen the hot central cone. Heat the pot by waving the flame back and forth under the pot. Heat the mixture to maintain a distillation rate of approximately one drop every 5 s. NOTE: Do not heat tha mixture too quickly. Rapid heating may cause the mixture to foam violently. A microbumner flame can easily be added and withdrawn to control the heating rate. Draw 1.0 mL of water into the syringe. Add water dropwise to the pot every 5 min to keep the water level to the mark. Refill the syringe with water as needed. Add more ice, as needed, to the Erlenmeyer flask containing the centrifuge tube. Stop the distillation when 3 mL of distillate has been collected in the centrifuge tube, | Dichloromethane is toxic and irritating. Use a fume hood. Clove oi (eugenol) is irritating. Prevent eye, skin, and clothing contact. Remove the centrifuge tube from the flask. Add 1 mL of saturated NaCl solution. ‘Add 1 mL of dichloromethane to the centrifuge tube. Cap the tube and gently mix the layers, being careful to vent the tube frequently. After the146 ‘3. Cleaning Up TECHO0722: Isolating Clove Oil from Cloves Using Steam Distillation initial pressure build-up has subsided, shake the centrifuge tube vigorously to mix the layers efficiently. Swirl the tube. At the same time, gently tap the outside of the centrifuge tube with your index finger to force into the bottom layer any droplets of dichloromethane that are adhering to the sides of the tube. Using a Pasteur pipet, remove the lower dichloromethane layer containing the clove cil into a 10-mL Erlenmeyer flask. Make certain that no water is transferred to the flask. Repeat the extraction process two more times using 1-mL portions of dichloromethane. Combine all three dichloromethane extracts in the same 10-mL Erlenmeyer flask. Anhydrous sodium sulfate (Na,SO,) is iritating and hygroscopic. Do not inhale and ingest this compound. Add approximately 50 mg of anhydrous Na;SO, to the flask containing the dichloromethane extracts. Stopper the flask. Allow the extracts to dry for 5 min. Weigh a clean, dry 25-mL beaker to the nearest 0.001 g and record the mass. Using a Pasteur filter pipet, transfer the dried dichloromethane into the beaker, making certain that no Na;SO, is transferred with the solution. Use two additional 0.5-mL. portions of dichloromethane to rinse the Na;SO, and ensure complete transfer of the clove oil to the beaker. Ina fume hood, place the beaker on the surface of a 40 °C sand bath to ‘evaporate the dichloromethane. Use a gentle stream of air or nitrogen to speed the evaporation. NOTE: When evaporating the dichloromethane, use a gentle siream of air or nitrogen, ‘one you barely feel against your hand. A strong stream of air or ritrogen may blow the solution out of the beaker and product will be lost. When all of the dichloromethane has been evaporated, weigh the beaker to the nearest 0.001 g and record the mass. Subtract the mass of the ‘empty beaker to obtain the mass of the clove oil. ‘Methanol is flammable and toxic. Keep away from flames or other heat sources. Prevent eye, skin, and clothing contact. Use a fume hood. Dissolve the clove oil in 1 mL of methanol. Proceed to the Characterizing the Product Section later in this module. Place your recovered materials in the appropriate labeled collection containers as directed by your laboratory instructor. Clean your glassware with soap or detergent. Wash your hands thoroughly with soap or detergent before leaving the Neaboratorye(© 1998 Cengage Learning TECHO722: Isolating Clove Oil from Cloves Using Steam Distillation “7 MICROSCALE STEAM DISTILLATION Using the Hickman Still Equipment 25-mL beaker marking pen boiling chip microburner 5 mL conical vial, microspatula with screw cap mortar and pestle condenser, with tubing 1-mL pipet? copper metal sponge 4 Pasteur pipets, with latex bulb cotton’ 10-mL round-bottom flask 10-mL Erlenmeyer flask, sand bath* with stopper support ring glass stirring rod support stand 10-mL graduated cylinder 2 utility clamps Hickman still wire gauze “for Pasteur filter pipet ‘tor adjustable micropipet {sand in crystallizing dish on electric hot plate or sand in electric heating well with heat controller Reagents and Properties ‘Molar mass Substance Quantity (ghmol) bp (°C) cloves 05g dichloromethane 4mL 84.93, 40 eugenol” 164.20 254 ‘methanol 1mL 32.04 647 sodium chloride, sat. solution 0.5 mL 58.44 sodium sulfate, anhydrous 50 mg 142.04 ‘product Preview «Grind the cloves with a mortar and pestle * Place the ground cloves and water in the distilling flask + Assemble the steam distillation apparatus + Distill the mixture « Extract the clove oil into dichloromethane « Dry the dichloromethane layer with anhydrous NaSO, + Remove the dichloromethane from the clove oil + Weigh the clove oil‘TECHO722: Isolating Clove Oil fom Cloves Using Steam Disillarion us PROCEDURE A Eee ear ceniainet ea eee ‘Wear departmentally approved safety goggles at all times while in the chemistry laboratory. 1. Conducting Steam Grind 10 whole cloves to a coarse powder, using a small mortar and Distillation pestle. Weigh 0.400-0.500 g of the powder and record the mass to the nearest 0.001 g. ‘Use a microspatula to carefully transfer the ground cloves to a 10-mL. round-bottom flask. Add a boiling chip and 4 mL. of deionized or distilled ‘water. Mix well with a glass stirring rod. Mark the level of the mixture on. the side of the flask. Place a small plug of copper metal sponge in the neck of the Hickman still to help prevent the mixture from foaming over as it is heated. Attach the Hickman still to the round-bottom flask. Assemble the remainder of the apparatus, as shown in Figure 4. Start the flow of water through the condenser. ‘Adjust a microburner flame to lessen the hot central cone. Heat the pot by waving the flame back and forth under the pot. Heat the mixture to ‘maintain a distillation rate of approximately one drop every 5 s. NOTE: Do not heat the mixture too rapidly. The clove mixture tends to foam when ‘rapidly heated. The microbume’ flame can easily be added and withdrawn to control the heating rate Lite ea© 1008 Cengage Leaning “TECHO722: lolating Clove Oil from Cloves Using Steam Distilaion 2. Extracting the Clove Oil 9 When the bottom portion of the Hickman still is full of distillate, remove the flame. Using a bent-tip Pasteur pipet, carefully remove the distillate from the Hickman still. Place the distillate in a 5-mL conical vial. Cap the vial to avoid spillage. eg ae NOTE: A standard Pasteur pipet can be used in a Hickman still model that has a built- in side port. Using a clean Pasteur pipet, add water through the top of the es keep the water level at the mark on the round-bottom flask. ‘Again use the flame to distill the mixture. Continue the distillation until 3 mL of distillate has been collected in the 5-mL conical vial. Dichloromethane is toxic and irritating. Use a fume hood. Clove oil (eugeno!) lnrtating. Prevent eye, skin, and clothing contact. Add 0.5 mL. of saturated NaC! solution to the vial. Using a bent-tip Pasteur pipet and a 0.5-mL portion of dichloromethane, carefully wash down the inside walls of the Hickman still to remove residual clove oil that adhered to the glass. Transfer the dichloromethane to the 5-mL conical vial containing the distillate. Repeat the rinsing with a second 0.5-mL portion of dichloromethane. Transfer the second portion of dichloromethane to the 5-mL conical vial. Cap the vial tightly and gently mix the layers, being careful to vent the vial frequently. After the initial pressure build-up has subsided, shake the vial vigorously to thoroughly mix the layers. Swirl the vial. At the same time, gently tap the outside of the vial with your index finger to force into the bottom layer any droplets of dichloromethane that are adhering to the sides of the vial. Using a Pasteur pipet, transfer the lower dichloromethane layer containing the clove oil into a 10-mL Erlenmeyer flask. Make certain that no water is transferred to the flask. Repeat the exiraction process two more times, using 1-mL portions of dichloromethane. Combine all three dichloromethane extracts in the same 10-mL Erlenmeyer flask. Anhydrous sodium sulfate (Na,SO,) is irritating ZN hygroscopic. Do not Inhale and ingest this compound. esr rea an Premetniy re ovine Add approximately 50 mg of anhydrous Na;SO, to the flask containing the dichloromethane extracts. Allow it to dry for 5 min. Weigh a clean, dry 25-mL beaker to the nearest 0.001 g and record the ‘mass. Using a Pasteur filter pipet, transfer the dried dichloromethane into the beaker, making certain that no Na,SO, is transferred with the solution Use two additional 0.5-mL Portions of dichloromethane to rinse the Na,SO, and ensure complete transfer of the clove oil to the beaker. In a fume hood, place the beaker on the surface of a 40 °C sand bath to evaporate the dichloromethane. Use a gentle stream of air or nitrogen to speed the evaporation.150 3.Cleaning Up Characterizing the Product TECHO722: Isolating Clove Oil ftom Cloves Using Steam Disillaion NOTE: When evaporating the dichloromethane, use a gentle seam of air or nitrogen, ‘one you barely fee! against your hand. A strong stream of air or nitrogen may blow the Solution out of the beaker and product willbe lost. When all of the dichloromethane has been evaporated, weigh the beaker to the nearest 0.001 g and record the mass. Subtract the mass of the empty beaker to obtain the mass of the clove oil. ‘Methanol is flammable and toxic. Keep away from flames or other Sources. Prevent eye, skin, and clothing contact. Use a fume hood. Dissolve the clove oil in 1 mL of methanol. Proceed to the Characterizing. the Product Section below. Place your recovered materials in the appropriate labeled collection containers as directed by your laboratory instructor. Clean your glassware with soap or detergent. Wash your hands thoroughly with soap or detergent before leaving the laboratory. (SIN SS aad Lao a Equipment Bunsen burner 4 Pasteur pipets, with latex bulb developing chamber" pencil 10-mL graduated cylinder 6 test tubes, 13 x 100-mm marking pen 3 x 7-cm TLC plate, silica gel, 2 melting point capillary tubes" with fluorescent indicator metric ruler “oz ar with id or 250 look directly into the UV lamp. lodine (1,) is toxic and corrosive. Prevent eye, skin, and clothing contac inhale and ingest |p. Use a fume hood. id Se Se After developing the plate, immediately mark the eluent front. Dry th Z ase - e plat ina ieee hood. aa the cheater under eect light es in an Ip chamber, as directed by your laboratory instructor, pencil circle the spots on your plate. ae carta ©{© 1998 Cengage Leeming ‘TECHO722; bolting Clove Oil from Cloves Using Steam Distillation 153 3. Cleaning Up Measure the distance from the origin to the eluent front. Measure the distance from the origin to the center of each spot. Record your observations. NOTE: Eugenol acetate should appear as a minor spot at a higher A than that of eugenol. Place your recovered materials in the appropriate labeled collection containers as directed by your laboratory instructor. Clean your glassware with soap or detergent. Wash your hands thoroughly with soap or detergent before leaving the laboratory.“TECHO722: loating Clove Oil from Cloves Using Steam Disillaion 155 Post-Laboratory Questions 1. Calculate the percent yield of clove oil based upon the initial mass of the ground cloves. 2. Give your test results for the reaction of your eugenol product with each of the test reagents. 3, Complete the following reactions, giving the correct structure for each organic product, (@) eugenol + Br. + (©) eugenol acetate + KMnO, > (© eugenol + FeCls + (d) eugenol acetate + FeCls + 4. Complete the following table after performing the TLC analysis on your clove oil sample. Indicate by 1 yes or no answer whether the spots are visible under UV light or I vapors. ‘Compound x uv bk eugenol eugenol acetate other 5. Using your R,s, list the compounds in your clove oil in order of increasing polarity. Briefly explain your answer. ‘© 1008 Cengage Learring© Raoult’s law (d) Dalton’s law (@) essential oil 3, Why is steam distillation preferable to simple distillation for isolating high-boiling natural products?