Steam Distillation of Citral from Lemon Grass Oil A DIS Project in Chemical Separations by Roberto Arbulu (2013-14) Points Pre-Lab 15 Post-Lab Safety and observations 5 Data summary and analysis 125 Spartan 75 Total 40 References J.C Gilbert & S. F, Martin, Experimental Organic ‘Chemistry: A Miniscale and Microscale Approach, ppp. 149-151, Cengage Learning, Boston, MA, 2011. A.B. Padias, Making the Connections®: A How-To Guide.. Fig, 3-14 (p. 144/146) and pp. 153- 155//155-157. Note: If you are performing this experiment prior to performing Experiment 4, refer to adias, pages 130-131, 136// 131-136 and page 8-38 of this manual for details on liquid-liquid extractions. Goals To perform a natural products extraction using the Steam Distilation technique. To use simple distilation/rotary evaporation to remove a solvent + To.use IR spectroscopy to verify the functional ‘groups present in a compound. + Touse Qualitative Organic Characterization tech- riques to confirm the presence of C=C and C=O bonds (Students work in pairs.) Beeuy 666 Experiment 6 Overview Steam distillation is the process of co-cstiling a compound with water It takes advantage Of the insolubilty of most ol-ike organic compounds in water to provide a route fo" StF. rating lower boiling insoluble’ compounds from either higher baling organic comPoner fr physicalplant matter In an immiscible mirture of water and oN) unlike the nora distlation of two misaibe lquids in steam distilation, each component of an immisc, liquid mixture contributes to the total vapor pressure as ifthe other component were 72! present. In ther words the total vapor pressure of the mixture is the sum of the vapor FT Sure of the water plus the vapor pressures of each of the organic components in he 2 ture, Thus, the total vapor pressure ofthe mixture salva’ (wath rare exceptions) Tas ‘than the individual vapor pressures of the liquids in the mixture in accord wath Dalton’ of Partial Pressures: Pr = Pocompone + Pocomgoert2 + Po-compenent3 * Pocnpenttn + Ponater sure, the result ye vapor Presi a at of the Since the boiling point ofthe liquid is inversely proportional to th is at the Boling port ofa mature of niceties rust be towel than ALG Ty lowest boling component. liquid or iquid mixture boils when the total Por FYE noc the liquid is equal to atmospheric pressure. The vapor pressure of water 5760 6S ang ‘The presence of any immiscible organic material contributes to the total poy Prev Tg thus causes the bolting point of water to drop below 100°C. This process allows ccunPe that have very high boting points to be co-distiled at a temperature below 100°C. Coe an example of this. Look up the bp. of Cia for your properties table. Citra} is @ two stereoisomers, geranial and neva. We wil refer tothe rixture as citral for simPHCIY in sunrnry steam dition ivolves a two-phase system with each component TA eae eae vapor pressure One can ure eal gas aw calculations to comput te aaa eee arom the dtilaton We wil use estimated cival concentrations i ne eter thereteal maximum ye of oolted cial The tower ast lemon as os toposes some ofthe more heat and a-sentive funcional dao ea conbl aldehydes and paiyenes rom ondaton. Using tis process the Fe re cls called ctroneal can ready be foated from lemon grass ol “The website http/lwwrw.chemicalland2t.convspecialtychem/perchem/CITRONELLALAt Terr that Citronella s the common name fora grass. C/mbopogon nardus, native to India ret Southeast Asia, Citronelaolis a yellowish essential ol distilled from the leaves of either Sf two grasses, Cymbopogon nardus or C. wanteranus. This aromatic oils inexpensive and. widely used in perfumes and soaps. [tis also best known as an insect repellent. Citronella, the main component of eitonelia oil is a clear to yelow oily liquid tis soluble in alcohol ‘and fixed ofls: and iis Insoluble in glycerin and water. Its a component of balm mint. It is Used in perfumes and flavoring sweet, flora. rose, waxy, citrus, green Literature bp, values range from 20! to 207°C. The approximate weight of ciral from lemon grass is quoted at {60% to 85% depending on the sources. For our sample, use 75% by weight to compute the theoretical yeld (use the volumes, not the masses o do the calculations) eit ten crs tC (MARA A A AAARAAMBMAHKALAAAARARARARAARAMSCEHALEORESSteam Distillation of Citral from Lemon Grass Oil ‘The logistics of steam distitation ae fain simple. Since the desired oil component very Sparnaly soluble water i's going to take a substantial amount of the ol/water emulsion trelatively speaking: 20-25% the cil volume) to “carry over" even small amounts ofthe de sited compound. We compensate fo this propertion ‘by using a somewhat larger round bottom flask (100 mL relative to 25 mofo to hale the larger Volume of water that s necessary. We equip our distr lation flask with a Claigen adapter to permit us to at- tach a separatory funnel wth additional water, if Needed. See Padias, Figure 3-22, page 155/157 and the figure at right ‘The lag-time to bring the oilfwater mixture to its boil: ‘ng point may be reduced by pre-heating the heating ‘mantle to about 60°C. Use the short glass or metal thermometer to verty this You should set your Variac to about 30, Pre-heating the rare ‘means that you have to Randle it carefullydo not touch the inside of the manti, The heating mantle should be ints final location/postion when you beatn the heating Proce Be careful when you put the round bottom flak in place and add the olwater mixture fo using a short stem plastic funnel Qualitative Functional Groups Tests of Citral, C=C and C=O Groups ‘Ctrl is a simple molecule containing a hydrocarbon backbone and two functional grOUPS. We wail use simple chemical tests to verify the presence of the functional groups. “To test for the presence of a C=C group, we will take advantage of the propensity of these {groups to add bromine across the double bond. Review the halogenation of alkenes section fof Carey's Organic Chemistry text, Citra also has an aldehyde group. Aldehyde functional ‘groups are generally characterized by an oxidative process, using either chromic acid solu- tion (Jones reagent), or the Tollens' tet, which uses siver redox chemistry. We will use the Tollens'test, since it generates a ess problematic waste “The Tollens’ test gives a postive result for aldehydes. n the presence of AgNO, [silver ni- trate) and hydroxide (HO), the aldehyde gets oxidized. At the same time, the silver is re- duced from Ag’ to Ag”. The Ag? gets deposited on the surface of the glassware to form @ silver mirror Thus, the appearance of the silver mirror is proof that the test compound was an aldehyde. Pee gee Ned aoe, a “The detailed equations for this process: 4. AgNO, + NOH AgiNO,),OH [add NH,| > (AgINH,),}OH" [not balanced 2, ZAGINH,)’ + RCHO+30H" 2g" +RCOO"+4NH,+2H,O balanced) 6768 Experiment 6 Safety and Waste Disposal ‘Wear gloves when handling all reagents/chernicals. Exercise care in handling a hot heating mantle. perature sr room tem Do not handle hot glassware: allow the glassware to c001 #0" 00! before changing or disassembling. yrtner when 2 Fe Poin the opening of a separatory funnel aay rom yousyou venting. ine in DCM) + Bxereae particular care when handing bromine stutons (rern"= + Exercise particular care when handling Tollens'test reagen'® ed waste containet + Dispose of halogenated reagents/mixtures in the halogenat 1 hood < e reagent ho + Dispose of silver-coated vials in the "USED VIALS" container in the reaget only! may be rinsed stilling flask. ion inthe distilling Hash may OS ign + The water/oil mixture that remains after distillati Pahari a the sink (lemon g7@ss a ‘out with acetone and water and flushed down natural product), Experimental Come poston of ‘Incorrect poston of fhomemstoreub thermometer bu Be certain you assemble the glass thermometer/rubber adapter/glass adapter pieces prior to adding to the top ofthe stil-head three-way connector. Use ‘3 dab of grease to lubricate the thermometer so that it will slide smoothly through the rubber adapter. Ths will minimize the chance that you will break the thermometer and push it through the palm of your hand! (See Figure 3, p.B-17) Refer to the figure to be sure you have proper placement of the thermometer bulb.YUU EU eb bee eee uUuuueewn~~— Wi UwvaiTs Steam Distillation of Citral from Lemon Grass Oil 4 Figure 6-1 Keck cip—Use at Every ‘Figure 6-2. Round Bottom Fask wit Ground Glass Jomnt Distilled ital Samele Determine the weight of25 mLof lemon grass i and then place it 3-4 tetion baling iPS ad GD of wate fos Dn tout as aut ask fr star stiston except replace the steam net ube wih aglassoTefion stopper. You may need 0 add hots ‘water abit later: you wil substitute a separatory funnel in place ofthe stopper to-do So Make sure to grease the glassware's ground glass connectors and place all Keck clips ‘before finalizing the apparatus to ensure a tight fit. Fit the curved adapter ‘with a 50 mL round bottom, which you have pre-marked to show 30 ml of volume. ‘Use your graduated. cylinder ‘to measure out this volume, The 50 mL flask need not be dried for collection purposes. eat the quid to boing setting your Varia to atleast 70/100), and cover the Clase stihead and the neck (only of the rb fas. so tat te dstilation proceeds as rapily 2$ possible. Note the temperature ofthe vapors in your lab notebook. Avotd applying excess Feat to the stil pot especially when the volume of water inthe ask has been reduced Pe- Tow about 30 mL: add more water to maintain a volure greater than 30 ml. Continue the dititation until ol droplets no longer appear in the distillate; this should occur after about 50 mi of dstlate has been collected. If droplets continue to form after 30 ml. of cistiate collect about 5 mL more and then stop. Note that it maybe dificult to see the oil droplets the condenser after several minutes of vapor flow. Do the best you can; Keep an eye on the cil layer inthe collection fask to estimate the progress ofthe oils distillation. ‘Allow the distillate to cool to room temperature or below, using an ice-water bath if neces- tary. Transfer the cool stilt toa separatory funnel and extract tvice, using two 15 mL portions of DCM each te. The funnel shouldbe vented frequently to avoid the buildup of pressure. Transfer the organic layer from a separatory funnel to a SO ml Erlenmeyer flask, nd add about 05-109 of anhydrous calcium chloride. Allow the solution to remain in ‘contact with the drying agent until the organic layer is dry, when it tums completly clea. Be sure there is enough calcium chloride to handle the level of water that may be present. ‘The drying agent should move loosely when swirled i sufcient quantity is used, Pour the dried organic solution into a tared 100 mi round bottom flask. Typically, we might equip the flask for simple distilation and evaporate the solvent using simple distilation. However, it may be simpler to remove the DCM via rotary evaporation: this facilitates the collection. of the DCM, After the methylene chloride is completely removed, the gold to dark amber residue is crude cial. Not ts smell. Take an IR spectrum and compare it tothe reference standard shown below. 6970 Experiment 6 Qualitative Tests (Your TA will announce if these will be done—be prepared to do them.) First, we will test for Unsaturation: Using a 10 mL screw-cap vial, place approximately 1 ml. of bromine in DCM solution. Note its deep, red-brown color. Now add, via a 1 mL syringe, 3 to 5 drops of citral. Note any color change. f there is not a significant lightening of the color of the solution, add another 3 to 5 drops of citra. ‘The Tollens’ Test for the aldehyde group: In a 10 ml. vial, dissolve 2 drops of the test com- Pound in 0.5 mL of dilethyleneglycol}aiethylether. To another 10 mL vial add 0.5 ml AgNO, solution and 0.5 mL NaOH solution; a precipitate should form. Add the NH, (arimonial so" lution dropwise until the precipitate fully dissolves. Transfer both solutions to a fresh 20 mL. vial, cap the vial and shake vigorously. Citral should give an Ag precipitate; it should coat the insides of the test tube as a "silver mirror” Please return vials with (+) test results to the Used Vials box for cleaning, 21 50] eT moe eC eee cf a ympare this tothe Rf your sample rhe igure 6-3 I specrum of commercial tal Compare ~ class‘ samples; note the differences.ee ‘periment 6: Steam Distillation of Citral from Lemon Grass Oil nm Name Date Section # 1. We used the Bromine in DOM method to test for Unsaturation. We also took 2 IR spectrum of the compound. However, neither method coud really give direct answer for the number of C=C bonds present. What methods could we use fo discern ths information? 2, Why do we not perform a simple dstlation on lemon grass oll at atmospheric the advantage of steam distila~ pressure to separate the components? What's tion? Can you imagine another distillation method that might serve the same purpose? (See Padias, page 150//152) 3. Lookup the structures of Carvone, Cuminaldehyde and Eugenol. Each of these natural products has similar structures to each other and Citra, Make a small {able with the molecule names at the top of the colurnns and the two functional {group tests as the rows. Indicate wich qualitative test would give (+) results. fs {t possible, without spectroscopic methods to unequivocally cfferentiate these four molecules using wet methods? 4, Research the *H NMR of a mixture of neral and geranial (citral) from the ‘SigmaAlrich com website. Be prepared to answer questions about those spec tra on quizzes and exams. Can you explain why there are differences, especially e inthe chemical shifts ofthe vinyl hydrogens? Compare the commercial NMR of citral to that of a student spectrum below. 'e ° Questions72 ” Experiment 6—Spartan Exercises: Steam Distillation of Citral from Lemon Grass Oi rass Oil 2. Based on the relative energies, doe: 3. Consider the dipole mome! Experiment 6 2 4 © © © © Spartan Exercises Procedure Build the following C,,},,0 structures in Spartan: ol) Note that Spartan does not care if you a build a structure wath a lot of bond ea angle distortion, Click on the Minimize button to clean up ualy structures. Be sure to build the correct geometric {isomer of citral (geranial net neral pane me Click Setup->Caleulations. : EN ore eae Set up an equilibrium geometry optimization using HF wath the SG bass set on your Sct and sub hej, Ones Me click 4 spreadsheet and add relatve energy {in kealmol) as well as dipole moment to fe spreadsheet. Use the calculated data to answer the folowing questions Questions 1. To build camphor, open a new file and start with cyclopentane (the solid lines in re figure here) from the Rings pull-down menu in the Model Kit. Add sp® carbons whers porropnate to provide the methyl and methylene groups to make the camphor and myrtenol molecules (the image below should help). a 1s the most stable structure make sense? In the Spartan box next to the citral molecule number (M0001. for sPrrmple) so that when you set up the relative energies the program wil st citral as Sear reference structure and give ta 0.00 kcal/mol reatve energy. Remem et that rela- yeu ergy ean be used to look at different conformations of the same Siruchtt and te Sie bg used for comparing structural isomers with the same molecular formula spreadsheet, you should check the the relative stability of the compounds, consider the ims. structural strain from sp® hybridized atoms, and jings/ring sizes that might be present. Hint: When comparing hybridization of the ator number of ri int of the molecules above. Do the dipole moment direction cor magnitude seem appropriate forthe main functional groups? Why? 4. Make a copy of the geranial structure, lick on Build New Structure (Fle ment, go into it to neral. You can do this by clicking the builder menu to paste it and then isomerize it ee pedsinle bond next to the C=O, now holding the alloption key right click and drag, tDrotate the fragment 180 degrees, Run a 3-21G geometry optimization and compare the total energy of neral to geranial. What is the relative energy in kcal/mol between the two isomers?