Isolation of Trimyristin and Cholesterol

Two Microscale Extractions for One Laboratory Period

Martha M. Vestling State University of New York College at Brockport, Brockport, NY 14420 The isolation of trimyristin from nutmeg and cholesterol from gallstones are two classic beginning organic laboratory experiments. One or the other appears in most laboratory textbook^.'.^ They are chosen t o illustrate the extraction of a natural oroduct from a solid while providing practice in filtering a n d recrystallizing. Each experiment Gkes about one 3-4-h laboratory period. The major expenses and major safety problems associated with the classic experiments are due to the quantities of organic solvents used per student. When these two experiments are converted to microscale, less than 10 mL of organic solvent per student is required for both exoeriments (<5 mL Der student oer isolation). -~~ Eachstndent starts with 200 mg oiground nutmeg and 100 mg of crushed gallstones. The first step in each isolation has the students shaking the solid with 2 mL of diethyl ether in a closed 1-dram vial. The vial should have an inert cap liner. After 15 min the solids are removed by filtration through an appropriately packed Pasteur pipet. Evaporation of the ether, followed by recrystallization, produces clean product. Each student needs only four 1-dram vials, 4 Pasteur nioets. and a small s ~ a t u l aThe organic solvents are . dispeniekeh'in hood using-a calibrated kasteur pipet [a Pasteur pipet connected viasmall piece of rubber tubing to a 1-mL or 2.5-mL plastic disposable syringe without a needle (see footnote 26 for a ~ i c t u r e ) lA packed Pasteur pipet can .. . be clamped to a ring stand using a small, thre&pronged clamp or with a standard wooden clothes pin and a binder clip. Solvent should be evaporated in a hood using either a heated sand bath and boiling chips or by gently blowing nwav the solvent with comoressed air. The solids are collected o small Hinch funnels (vacuum filtration). ; At Brockport all students isolated visible amounts of both products. Typically 216 mg of nutmeg will yield 34 mg of trimyristin while 108 mg of gallstones will give 75 mg of cholesterol. During the recrystallization of trimyristin from acetone, care must be taken to keep the solutions cold, and only very small amounts of cold acetone should be used to wash the crystals in the Hirsch funnel. During the decolorizing of the gallstone extract, cholesterol forms on the Pasteur pipet . . tip and should be washed with ether into the collection vial.

clamp here
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cotton

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(a)The packed Pasteur p i p l tor finering and drying tha.nutmeg ether extract: (blthe Packed Pasteur pipet for filtering, decolarizing,and drying the gallstone ether extract.

Experimental

Presented at the 10th Biennial Conference on Chemical Educations, Purdue University, West Lafayette, IN, August 2, 1988. 'Trimyristln: (a) Roberts. R. M.; Gilbert, J. C.; Rodewald, L. B.; Wingrove, A. S. Modern Experimental Organic Chemistry, 4th ed.; Saunders: New York, 1985; (b) Fessenden, R. J.; Fessenden, J. S. Techniques and Experiments for Organic Chemistry: Grant: Boston. MA. 1983; (c) Marmor. S. Laboratory Methods in Organic Chemisby; Burgess: Minneapolis, MN, 1981: (d) Doyle, M P.: Mungall, W. S. . Experimental Organic Chemishy; Wiley: New York, 1980. Cholesterol:(a)Pavia, D. L.; Lampman, G. M.; Krlz, G. S. introduction to Organic Laboratory Techniques, 3rd ed.; Saunders: New York. 1988; (b) Fieser, L.; Williamson, K L. Organic Experiments, 6th ed.; . Heath: Lexington. MA. 1987; (c)Williamson, K. L. Microscale Organic Experjments: Heath: Lexington, MA 1987; (d) Todd, D. Experimental Organic Chemistry: Prentice-Hall. Englewood Cliffs. NJ. 1979.

Procedure for Trirnyristin 1. Weigh a 1-dram vial that has a Teflon-lined cap. Place approximately 200 mg of nutmeg in the vial, and weigh again. A weighed sample in a vial will he on display so that you can see about how much nutmeg to add. 2. Use the calibrated Pasteur oinet in the hood next to the diethvl ether container to add 2.00 mL hfdiethyl ether to the vial. 3. Cap the vial tightly, and shakr the mixture for 15 min. Your hand will heat the wal t u nrar the hoilrng point of diethyl ether. 4. Cool the closed vial in cold running tap water. 5. Pack a Pasteur pipet with a lowe wad of cotton, 2 rnm of sand, and 1 cm of anhydrous sodium sulfate. See part a of the figure. Clamp the packed pipet to a ring stand. 6. In a hood, cautiously open the vial. With a clean Pasteur pipet, transfer the contents of the vial to the packed Pasteur pipet, and let the solution drip through the packing into a clean vial. Rinse the packing with 0.5 mL of clean diethyl ether. 7. In a hood, attach a piece of rubber tubing with a Pasteur pipet at one end to the air jet. Gently blow away the solvent. This process should take about 5 min. If the air is flowing too fast, you will blow the whole mixture into the hood, so be careful. 8. Add 0.5 mL of acetone to the residue in the vial. In a hot sand bath. swirl the acetone mixture until the residue has iust dissolved. l&&e the vial from the heat and rap it. Let the vial stand for 10 mi" at n,om temperature and then in ice for 5 mm. 9. Collect your now-whitesolid in a Hirsch funnel. Then transfer it to a clean weighed vial. 10. Weigh. Determine the melting point. Procedure far Cholesterol 1. Place 100 mg of crushed gallstones in a clean, weighed 1-dram vial equipped with a Teflon cap liner. 2. In a hood, transfer 2.0 mL of diethyl ether to the vial with a calibrated Pastenr pipet. 3. Cao the vial. and shake for 15 min , 4. I'ncka ~ a & r pipet withasmall plug ofcotton. 2mmof sand. 2 rnm oianhydrous sodium ~ulfatr, 1 cm of eharcdanhydrous and sodium sulfate mix. Src part h of the f~gure. Clamp thr packed pipet to a ring stand. 5. In a hood, using a clean Pasteur pipet, transfer the contents of

274

Journal of Chemical Education

the vial to the packed Pasteur pipet, and let the solution drip through the packing into a clean vial. Rinse the original vial and the nacked Pnateur ~ i ~witht a little ether.. esoeciallv if a white crust e r ..-.. . forms on the tip of the Pasteur pipet. 6. In a hood, gently hlaw away the solvent (use a piece of rubber tubing with a Pasteur pipet a t one end and the air jet a t the other). I. Add 1.0 mL of acetone to the residue in the vial. Swirl the
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mixture in a hot sand bath until the solid is just dissolved in the acetone. Remove the vial from the sand bath and cap. Let the solution stand 10 min st room temperature and then 5 min in an ice bath. 8. Collect your cholesterol, the white solid, in a Hirsch funnel. Then transfer it t o a clean weighed vial. 9. Weigh. Determine the melting point.

Volume 67

Number 3

March 1990

275