Isolation of (+)-Limonene from Citrus

Read Steam Distillation in Zubrick and Stereochemistry, Ch. 5, in Wade. 6th ed.
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Limonene, the chief component of orange oil, is widely used as a fragrance and flavoring, as well as a cleaning solvent. Limonene is an example of a terpene, a class of natural products biosynthesized by the assembly of isoprene units into various structures. Many terpenes are responsible for the odors of plants like eucalyptus, pine, mint, lavender, rose, and others. Organic chemists use terpenes and other natural products as chiral starting materials for complex chemical syntheses or as inspirations for pharmaceuticals. Some natural products are attractive synthetic targets because of interesting or unusual structural features or medicinal applications. Isolation of natural products typically involves multiple extractions and chromatographic steps, but certain organic oils can be freed of contamination by subjecting them to a process known as steam distillation. In todays lab, you will perform a steam distillation to isolate limonene from the peelings of citrus fruit. You will then use IR spectroscopy and polarimetry to analyze your isolated limonene.
isoprene skeleton

Steam Distillation Normally, a liquid boils when its vapor pressure is equal to the surrounding pressure. Under ordinary circumstances, surrounding pressure is equal to atmospheric pressure, which in the Phoenix area is approximately 1 atmosphere (760 mm of mercury). A solution (a homogeneous mixture of two or more miscible liquids) will boil when the combined vapor pressures of its dissolved components is equal to the surrounding pressure. The pressure of each component in a solution is related to its concentration in the mixture, and so the boiling point of a solution, or homogeneous mixture, is normally between the boiling points of the individual components. On the other hand, a heterogeneous mixture will also boil when the combined vapor pressures of its components is equal to the surrounding pressure. However, because the liquids in a heterogeneous mixture are immiscible, the vapor pressures of the individual components are independent of one another and not related to their concentrations. Since the two liquids are immiscible, they independently exert vapor pressures against the

external pressure, and when the sum of the two partial pressures is equal to the external pressure, boiling occurs. Thus, the total vapor pressure of a heterogeneous mixture is given by the following equation: Ptotal = PA + PB (Eq. 1)

where Ptotal is the total pressure of a system at a given temperature, and PA and PB are the individual pressures of components A and B at the given temperature. Generally, a heterogeneous mixture will boil at some temperature below the boiling point of either component, the result of the combined effects of both compounds. We can take advantage of the fact that many water-insoluble liquids and solids behave in the manner described above for heterogeneous mixtures, volatilizing at temperatures below their boiling points. The effect described above is exploited in a technique called steam distillation, where an organic compound of moderate volatility and vapor pressure is distilled as part of a heterogeneous mixture with water. The boiling point of the mixture is slightly below 100 C, the boiling point of water. At this temperature, a fraction of the distillate will be the compound of interest. The greater the vapor pressure of the organic compound, the larger the fraction that will co-distill with the water. This technique is considerably more gentle than regular distillation, since some organic compounds can decompose at temperatures approaching their higher boiling points. Experimental Procedure Work with a partner for this experiment. Peel about 10 medium-sized citrus with a sharp knife or peeler. Remove just the exterior, brightly colored portion of the peel, called the zest, as the white material underneath (the pith) contains little or no limonene. Determine the mass of the peelings, then place them in a blender. For best results, try to have about 100 g of peelings. Add about 150 mL deionized water, put the lid on the blender, and blend the mixture until a smooth puree is obtained. Pour the mixture into a 500 mL round bottom flask equipped with several boiling stones. Attach the flask to the steam distillation system. There will be an example setup in the lab for you to inspect before assembling your own. Note that instead of the thermometer adaptor, you will be placing a separatory funnel containing water at the top of the Y-adaptor. This allows you to add water to the distillation pot as the distillation proceeds. Make sure that the pieces of your distillation apparatus are properly clamped together and securely clamped to a ring stand. Heat the contents of the distillation flask to boiling using a heating mantle placed on a ring clamped to the ring stand. Heat uniformly and slowly to avoid scorching the mixture. Heating too rapidly could cause the mixture to bump (splash violently) into the condenser and contaminate the condensate. If this happens, you will need to stop the distillation, let the apparatus cool, clean the system and begin the distillation process again. Periodically add portions of distilled water from the separatory funnel to the distillation flask to replace the water that has been distilled over. Try to maintain a fairly constant liquid level in the distillation flask. Continue collecting distillate until the receiving flask is about half full. If the steam distillation was successful, you should

notice an upper layer of limonene in the receiving flask. If necessary, replace the receiving flask and collect more distillate to improve your yield of limonene. When you are finished distilling, remove the heat source. Transfer the distillate to a separatory funnel. Allow the layers to separate and discard the lower, aqueous layer. Dry the limonene over sodium sulfate (see Drying Agents in the Zubrick text). Filter it through a Pasteur pipet equipped with a cotton plug into a clean, tared vial and find the mass of the isolated limonene. Use this mass to calculate percent yield. Obtain an IR spectrum of your limonene. After obtaining the IR spectrum, find the new mass of your limonene and pool your sample with several other students. Measure the optical rotation of the sample using the polarimeter. Your instructor or the stockroom coordinator will show you how to operate the polarimeter. Waste Disposal and Cleanup Place excess pulp into the containers marked Orange Pulp. Place peeled fruit in the trash cans. Do not pour pulp into the sink. Do not clean the plastic blenders with acetone; use water only. Dispose of the aqueous layer from the distillation and any excess limonene and ethanol in the waste container in the main fume hood. Discussion and Conclusions 1) 2) Calculate your percent yield of limonene based on the mass of peel you used. Calculate the specific rotation of the limonene sample using the equation

cl

where t is the specific rotation at temperature t and wavelength , is the observed rotation, c is the concentration of the sample in g/mL, and l is the light path in dm. How does your experimental value compare with the literature value of 125.6 at 20 C using the D line of the sodium lamp?

3) Identify diagnostic peaks in the IR spectrum that support the identity of your product as limonene. 4) Assign the absolute configuration of the chirality center in limonene as R or S.