LSDby Michael Valentine Smith
Since Hofmann's first trip in 1943, great deal of interest has been generated in the occurrence and properties of various lysergic acid derivatives. Fungi of the genus Claviceps, which grow on rye wheat, rice and other grasses, were the first natural source of these alkaloids to be discovered. In recent years related compounds have been found in the genera Penicillium (the blue-green mold that also produces penicillin), Aspergillus, and Rhizobus (the black bread mold). These compounds are now produced commercially by culturing certain strains of Clavicebs which produce as much as 4 g of ergotamine per Liter of culture medium. Growing pure cultures of fungi is not for amateurs, but those interested will find these references useful: JPS 58,143(1969); App. Microbiol. 18,464 (1969); HCA 47,1052(1964); Lloydia 32,327,401(1969); Can. J. Microbiol. 16,923(1970); CA 61,15314c-f, 67,84858e, 69, 36323w; Biotech. Bioeng. 13,331(1971); CA 76,57736(1972); U.S. Patent 3,483,086; Planta Med. 23,330(1973); J. Pharm. Educ. 36,598(1972); CA 78,41492(1973); French Patent 1,531, 205; German Patents 1,806,984 and 1,909,216; British Patent 1,158,380. For a description of a wild American Claviceps species see Mycologia 66,978(1974).
The occurrence of hallucinogens in the seeds (and to a lesser degree in the leaves and stems) of various members of the family Convolvulaceae (morning glories, etc.) was known to the Aztecs. Seeds of the genera Rivea, Impomoea, and Argyria (Hawaiian baby woodrose) contain lysergic acid derivatives; the woodrose being champion with about one hundred times as much as the other genera (about 7 mg alkaloids/g seeds). In view of the low yield (maximum 10 mg alkaloids/ 100 g seeds) even the famed pearly gates variety of morning glory is not worthwhile extracting, and the trip is commonly a bummer, resembling that produced by scopolamine or ibogaline and unlike that of LSD. However, the lysergic acid amide, etc., can be extracted, hydrolyzed to lysergic acid (as described below for ergot alkaloid hydrolysis), and converted to LSD by any of the methods described. For species variation of alkaloid content see Lloydia 29,35(1966). Crude ergot or woodrose seeds should yield ca. 1 g LSD/kg after conversion of the isolated alkaloids.
Alkaloid Extraction (short method)
Finely grind seeds (preferably woodrose) and add NaHCO3. Extract with ethyl acetate by soaking about one day. Filter and extract the ethyl acetate with tartaric acid solution. Basify the extract with NaHCO3 and extract it with ethyl acetate. Dry and evaporate in vacuum the ethyl acetate to get the alkaloids. Repeat this procedure on the seeds until no more residue is obtained. Alternatively, add 100 ml petroleum ether to 100 g finely ground seeds and let soak about two days. Filter, discard petroleum ether and let seeds dry. Add 100 ml methanol to the seeds and let soak about two days. Filter, repeat extraction with another 100 ml methanol and evaporate in vacuum the combined methanol extracts. The residual yellow oil contains the alkaloids.
For chromatographic purification of ergot alkaloids from seed extracts see Phytochem. 11,1479( 1972). For ergot extraction and separation see also Fr. Patent 2,089,081 (11 Feb 1972) and CA 79,105,457(1973). For a recent review of the ergot alkaloids see R. Manske (Ed.) The Alkaloids, vol. 15: 1-40(1975).
Extraction of Lysergic Acid Amides from Woodrose Seeds or Powdered Ergot
Reduce the seed material to a fine powder in a blender, and spread it out to dry. Grind it again if it is not fine enough after the first time due to dampness. Saturate the powdered seed material with lighter fluid, naphtha or ligroine. When completely saturated, it should have the consistency of soup. Pour it in a chromatography column and let it sit overnight. Remove the fatty oils from the material by dripping the lighter fluid or other solvent through the column slowly and keep testing the liquid that comes through for fats by evaporating a drop on clean glass until it leaves no greasy film. It will take several ounces of solvent for each ounce of seeds. Mix 9 volumes of chloroform with 1 volume of concentrated ammonium hydroxide and shake it in a separatory funnel. When it settles the chloroform layer will be on the bottom. Drain off the chloroform layer. Discard the top layer. Drip the chloroform wash through the column and save the extract. Test continuously by evaporating a drop on clean glass until it ceases to fluoresce under a black light. Evaporate the chloroform extracts and dissolve the residue in the minimum amount of a 3% tartaric acid solution. If all the residue doesn't dissolve, place it into suspension by shaking vigorously. Transfer the solution to a separatory funnel and wash the other vessel with acid in order to get all the alkaloid out. Pour the washings in the funnel also. Basify by adding sodium bicarbonate solution, and add an equal volume of chloroform. Shake this thoroughly, let it settle, remove the bottom layer and set it aside. Once again, add an equal portion of chloroform, shake, let it settle and remove the bottom layer. Combine the chloroform extracts (bottom layers) and evaporate to get the amides.
The Culture and Extraction of Ergot Alkaloids
Make up a culture medium by combining the following ingredients in about 500 milliliters of distilled water in a 2 liter, small-neck flask:
Add water to make up one liter, adjust to pH 4 with ammonia solution and citric acid. Sterilize by autoclaving. Inoculate the sterilized medium with Claviceps purpurea under sterile conditions, stopper with sterilized cotton and incubate for two weeks periodically testing and maintaining pH 4. After two weeks a surface culture will be seen on the medium. Large-scale production of the fungus can now begin. Obtain several ordinary 1 gallon jugs. Place a two-hole stopper in the necks of the jugs. Fit a short (6 inch) glass tube in one hole, leaving 2 inches above the stopper. Fit a short rubber tube to this. Fill a small (500 milliliter) Erlenmeyer flask with a dilute solution of sodium hypochlorite, and extend a glass tube from the rubber tube so the end is immersed in the hypochlorite. Fit a long, glass tube in the other stopper hole. It must reach near the bottom of the jug and have about two inches showing above the stopper. Attach a rubber tube to the glass tube as short or as long as desired, and fit a short glass tube to the end of the rubber tube. Fill a large, glass tube (1 inch x 6 inches) with sterile cotton and fit 1-hole stoppers in the ends. Fit the small, glass tube in end of the rubber tube into 1 stopper of the large tube. Fit another small glass tube in the other stopper. A rubber tube is connected to this and attached to a small air pump obtained from a tropical fish supply store. You now have a set-up for pumping air from the pump, through the cotton filter, down the long glass tube in the jug, through the solution to the air space in the top of the jug, through the short glass tube, down to the bottom of the Erlenmeyer flask and up through the sodium hypochlorite solution into the atmosphere. With this aeration equipment you can assure a supply of clean air to the Clauiceps purpurea fungus while maintaining a sterile atmosphere inside the solution. Dismantle the aerators. Place all the glass tubes, rubber tubes, stoppers and cotton in a paper bag, seal tight with wire staples and sterilize in an autoclave. Fill the 1-gallon jugs 2/3 to 3/4 full with the culture medium and autoclave. While these things are being sterilized, homogenize in a blender the culture already obtained and use it to inoculate the media in the gallon jugs. The blender must be sterile. Everything must be sterile. Assemble the aerators. Start the pumps. A slow bubbling in each jug will provide enough oxygen to the cultures. A single pump can, of course, be connected to several filters. Let everything sit at room temperature (25øC.) in a fairly dark place (never expose ergot alkaloids to bright light -- they decompose) for a period of ten days. After ten days adjust the cultures to 1% ethanol using 95% ethanol under sterile conditions. Maintain growth for another two weeks. After a total of 24 days growth period the culture should be considered mature. Make the culture acidic with tartaric acid and homogenize in a blender for one hour. Adjust to pH 9 with ammonium hydroxide and extract with benzene or chloroform/iso-butanol mixture. Extract again with alcoholic tartaric acid and evaporate in a vacuum to dryness. The dry material is the salt (i.e., the tartaric acid salt, the tartrate) of the ergot alkaloids, and is stored in this form because the free basic material is too unstable and decomposes readily in the presence of light, heat, moisture and air. To recover the free base for extraction of the amide or synthesis to LSD, make the tartrate basic with ammonia to pH 9, extract with chloroform and evaporate in vacuo. If no source of pure Claviceps purpurea fungus can be found, it may be necessary to make a field trip to obtain the ergot growths from rye or other cereal grasses. Rye grass is by far the best choice. The ergot will appear as a blackish growth on the tops of the rye where the seeds are. They are approximately the same shape as the seeds and are referred to as "heads of ergot." From these heads of ergot sprout the Claviceps purpurea fungi. They have long stems with bulbous heads when seen under a strong glass or microscope. It is these that must be removed from the ergot, free from contamination, and used to inoculate the culture media. The need for absolute sterility cannot be overstressed. Consult any elementary text on bacteriology for the correct equipment and procedures. Avoid prolonged contact with ergot compounds, as they are poisonous and can be fatal.
Since LSD is an indole derivative, it gives a positive reaction (violet color) to the tests given in the indole section. LSD also fluoresces under an ultraviolet light (black light), but so do many other compounds. For infrared spectra of LSD and related compounds, see JACS 78,3087(1956) and J. Forensic Sci.12,538 (1967). For other information on identification see JPS 56,1526 (1967) and JAOAC 50,1362(1967), 51,1318(1968). For a microcrystalloscopic test see J. Pharm. Pharmacol. 22,839(1970). In order to make LSD, lysergic acid is needed. This can sometimes be obtained, but generally one of the lysergic acid containing ergot alkaloids such as ergotamine is more readily available. Ergot is the dried sclerotium of various species of fungi which infect rye (and other grasses), leading to the formation of large purple growths in place of the rye grains. These growths are collected, dried, powdered and the alkaloids extracted. For the extraction procedure see HCA 28,1283(1945), J. Pharm. Pharmacol. 7,1 (1955), JPS 50,201(1961), CA 75,137422(1971). Proc. Indian Acad. Sci. 71B,28,33(1970) gives production from artificially infected rye. Ergot is produced mainly in Europe (especially Switzerland) but some has been grown in the USA (e.g., in Minnesota). This production occurs primarily because of the use of ergotamine and related compounds in medicine (contracting the post-partum uterus, terminating migraineheadaches, etc.). Many of the ergot alkaloids are derivatives (amides) of lysergic acid. Unfortunately, these compounds have little hallucinogenic activity and it is necessary to hydrolyze (split with water) off the amide, producing lysergic acid, and to synthesize a different amide with greater psychedelic activity. This hydrolysis can be done with any of the following compounds or a mixture of them: ergometrine, ergine, ergotamine, ergosine, ergocristine, ergokryptine, ergonovine (ergo- metrine) and methysergide (Sansert). When -ine is added to the name (e.g., ergotaminine) this indicates the isomers which will lead to the production of the inactive iso-LSD. The papers cited here give simple techniques for converting these to the active forms (or see the technique for converting iso-LSD to LSD in method 1 following): HCA 37,820,2039(1954); CA 69,36322(1968); CCCC 34,694 (1969). For a review of the ergot alkaloids see THE ALKALOIDS, Manske and Holmes (Eds.), 8,725(1965), and F. Bove, THE STORY OF ERGOT (1970).
Ergot Alkaloid Hydrolysis
JBC 104,549(1934); HCA 47,1929(1964). Perhaps the best method is Hofmann's modern hydrazine hydrolysis given later, since this disposes of the necessity for isolating the lysergic acid (I); otherwise the following alkaline hydrolysis can be used: Dissolve 20 g of the alkaloid (e.g., ergotamine) in 200 ml 1M KOH in methanol (i.e., dissolve 56 g KOH pellets in 1L 100% methanol) in a 1 L heavy walled vacuum flask and evaporate in vacuum the methanol at room temperature. To prevent the solution from cooling, and thus greatly prolonging the evaporation time, put the flask in a pan of water kept at room temperature by gentle heating or by running warm water through it. Add 400 ml 8% KOH in water to the residue and boil for one hour (under N2 if possible, this can be done by filling the flask with a N2 stream and loosely stoppering or by allowing a gentle stream of N2 to flow through during heating). Cool, acidify with dilute sulfuric acid and shake in separatory funnel with 1 L ether. Discard the upper ether layer and filter with vacuum the aqueous suspension of lysergic acid (I). Wash precipitate with 20 ml dilute sulfuric acid. To recover the small amount of (I) remaining in solution, basify with Na carbonate and bubble C02 through it. Filter and add precipitate to first batch. Some isolysergic acid will remain in solution and can be precipitated by adding 10% HNO3. It can be converted to (I) by adding 3 ml 10% KOH for each 0.1 g acid, boiling on steam bath for one hour under N2 (if possible) and precipitating by acidifying with glacial acetic acid. Maximum yield is about 9 g (I) for 20 g ergotamine. A shorter method of hydrolysis which may work as well follows: dissolve 20 g alkaloid in 300 ml methanol and 300 ml 40% KOH and reflux two hours under N2 (if possible). Cool, saturate with CO2 and evaporate in vacuum. Extract the residue with hot ethanol three times and dry, evaporate in vacuum the combined ethanol extracts to get (I). Under ordinary conditions, about 20% of (I) will be converted by the action of hot water, etc., to the inactive isolysergic acid. Most of this remains in solution and can be isomerized to (I) as described above, or it can be converted to iso-LSD by any of the methods described later and isomerized to LSD (see method 1). It is unnecessary to purify (I), but this can be done as follows: dissolve 9 g (I) in 20 ml NH4OH, filter and concentrate in vacuum at room temperature to precipitate (I). After filtering, the grey crystals can be further purified by dissolving in boiling water and cooling in ice bath to precipitate (I). Melting point should be about 240o (decomposes). Alternatively, the dark-colored (I) resulting from hydrolysis can be shaken with 2x400 ml 2 M NH4OH in ethanol, and the combined extracts evaporated in vacuum to give (I). Dissolve the remaining residue in 500 ml hot methanol, cool to 0ø and filter out the (I) (recrystallize-water). Can remove colored impurities by shaking solution with decolorizing carbon and filtering. Recently a method for increasing the yield of (I) about 10% using 2.5% hydrazine hydrate was described (CA 69,36323(1968)). Dissolve 7 g alkaloid in 200 ml 6 N KOH in methanol and 200 ml ethanol, add 10 ml hydrazine hydrate and boil four hours under N2 (if possible) and proceed as above. Finally, the (I) must be thoroughly dried by heating at about 110ø/1 mm for two hours or 150ø if ordinary lab vacuum of 15 mm is used. A forced water vacuum (about 25 mm) can be used here as elsewhere. An oil bath (e.g., mineral oil) will allow temperature regulation.
There are certain aspects of LSD production which are common to all synthetic methods. The first is a certain degree of danger; each uses dangerous reagents and solvents. Hydrazine and hydrazine hydrate are both violent poisons, and each can cause severe skin burns and eye damage. The vapor of each is irritating, and can cause severe eye irritation as well as liver and blood damage, but the symptoms don't always manifest right away, sometimes appearing three or four days after exposure, so it is easy for exposure to be much more dangerous than is immediately realized. In addition, anhydrous hydrazine is a sensitive and violent explosive, the explosion of which can be set off by certain types of stainless steel and such common things as wood and rust. Both trifluoroacetic acid and sulfur trioxide will cause very severe skin burns, and their vapors are extremely irritating. Sulfur trioxide is such a strong dehydrating agent that it chars organic material, and its heat of dehydration is so high that it will start a fire if spilled on wood, which could prove fatal were flammable solvents in use at the time or stored nearby. Phosgene is very poisonous; so insidious that it was used as a war gas in World War I. One deep breath can cause immediate collapse and death, and as it is not irritating there is no gag reflex to prevent one from taking that deep breath. Doses which are not high enough to be immediately lethal may not be noticed at all at the time of exposure, yet lead to death within 24 hours. Sub-lethal doses cause pulmonary edema and serious respiratory disability; again, the symptoms can appear well after an exposure which was hardly noticed. Diethylamine, used in every LSD synthesis, has a very low flash point, and its vapor is irritating. The vapor of DMF is also irritating, and prolonged exposure can cause liver damage. In fact, most of the solvents used in LSD production are either flammable or toxic or both. In addition to all the above, the starting material, the ergot alkaloids, is as a class quite toxic, and clean working conditions are necessary when working with it. Ergot alkaloid poisoning, known in the Middle Ages as Saint Antony's fire, can actually cause one's limbs to blacken, shrivel, and fall off! Any woman working with these compounds should also be aware that many of them are oxytoxics, that is, they cause uterine contractions, and are so used to induce labor, etc.
There are certain procedures common to all syntheses of LSD which are based upon the sensitive nature of ergot compounds in general. Natural ergot alkaloids, lysergic acid, LSD, and the intermediate products associated with the various syntheses are all to a varying degree unstable. Even the most stable of these compounds will readily decompose under any but moderate conditions. Thus precautions must be taken against light, moisture, oxygen, and heat. Light of the ultraviolet region promotes addition of water at the delta-9-10 double bond to form the lumi-compounds. Thus reactions are best carried out in the light of red or yellow photographic darkroom bulbs, and storage should be in opaque or amber bottles. Most of the reactions involved in LSD synthesis require anhydrous conditions for good yield, and so protection must be made against moisture during the actual production. Furthermore, the final product must be thoroughly dried to prevent possible formation during storage of the lumi-compounds as mentioned above. Oxidizing agents, including atmospheric oxygen, will decompose ergot compounds. For this reason, all reactions are carried out in an atmosphere of an inert gas such as nitrogen. The danger of oxidation increases with temperature, so this precaution is of course most important with those reactions proceeding at elevated temperature. Various methods have been devised to prevent oxidation during storage. The most obvious is to store the LSD in nitrogen filled containers, but the excellent protection thus afforded is of course lost when the bottle or ampule is opened. Another method is to use an antioxidant; Brown and Smith recommend ascorbic acid. A more sophisticated method, recommended on the highest authority, is to make LSD maleate rather than the tartrate. Both maleic and tartaric acids are dicarboxylic, but the pK2 of maleic acid is too low to form a salt with LSD. Thus there is a free carboxyl group in LSD maleate, which group will serve to prevent oxidative decomposition. Excessive heat will cause decomposition of LSD and its precursors, and will also increase the possibility of racemization. Thus reactions at elevated temperature are not unnecessarily prolonged, nor are temperatures unnecessarily raised. All drying is done in vacuo in an inert atmosphere, and long term storage should be under refrigeration.
I want to emphasize that "legal acid" can be obtained if other amines are substituted for diethylamine in LSD synthesis. These other lysergamides should give identical trips, but most of them are less potent than LSD. Precise potency data do not exist, so it remains for an enterprising chemist to gain immortality by adding each of the following amines (and any others that come to mind) to separate aliquots of the final step of LSD synthesis (they could easily be done simultaneously), isolating the tartrates and assaying them for potency: piperidine, diisopropylamine, ethylisopropylamine, ethylpropylamine, methylethylamine, methylisopropylamine, tetrahydrooxythiazine, tetrahydroisoxazine, dioxazole, 2-methylmorpholine, 2,5-dimethyl (or dimethoxy) pyrrolidine, cyclo-butyl-amine, cyclopentylamine, etc. Published potency data expressed as a fraction of LSD activity follow: pyrrolidide (1/20), dimethylamide (1/20), morpholide (1/10 or 1/3), ethylpropyl (1/3), dipropyl (1/10), methylethyl (less than 1/10), methylpropyl (less than 1/10).
LSD via the Hydrazide HCA 38,429(1955), HCA 26,953(1943)
|Extract||Quantity||Total Solvent Used|
Continue until clear lower layer appears.
The extracts from the reaction are combined and shaken up with a little anhydrous magnesium sulfate (120 grams) and filtered. The filtrate is evaporated to dryness in the rotary vacuum evaporator, care being taken not to heat the extracts or the residual syrup above 55øC. A good mechanical vacuum pump and effective cold traps in the line are necessary to remove the residual dimethylformamide from the residue. A brown to black bubbly residue should remain when evaporation is complete. This residue contains the amide product and considerable impurities. A general method of purifying the amide follows.
The material to be purified (the above residue or other material containing N,N-diethyl lysergamide) is taken up in 1200 ml of methylene chloride containing 20% benzene and applied to a chromatographic column containing two pounds of basic alumina 100-200 mesh Brockmann activity two or three. The column is eluted with nine liters of methylene chloride containing 20% benzene. At this point, the column when viewed with visible light should display three distinct color bands. The uppermost band will be a dark brown or greyish color, the next band will be a reddish brown color, and the lowest band will be a light brown or tan color. The eluant should be amber colored. The column may now be eluted with about one liter methylene chloride containing 0.5% methanol. This will bring the reddish band nearly to the bottom of the column. At no time should any portion of the reddish band be eluted from the column. If any of the reddish band reaches the bottom of the column, elution should be stopped. Next, the total eluant is shaken up with 30 grams of activated charcoal (Norit A) mixed with 30 grams of alumina and filtered. The filter is washed with 600 ml of methylene chloride and the total filtrate taken to dryness on the rotary vacuum evaporator. Care is taken not to heat the residue or solution above 55ø C. The residue is taken up in one liter of benzene and immediately taken to near dryness when another liter of benzene is added to dissolve the residue and the solution is again taken to near dryness. This procedure is repeated until four to five liters of benzene have been added and evaporated. The residue is finally taken to complete dryness at about 45ø C. If sufficient benzene has been added and evaporated, a light tan bubbly, crystalline material will fill the interior of the evaporator flask. It is important that this residue be completely dry before proceeding. The evaporation of benzene from the residue aids removal of solvents and other volatile materials (as azeotropes) which promotes formation of the bubbly crystalline structure in the residue. 700 ml of petroleum ether is next added to the evaporator flask which is then removed from the evaporator and tightly stoppered. The flask is shaken vigorously to loosen the residue from the sides of the flask. Usually all the material comes loose from the flask and forms a slurry in the petroleum ether. If necessary, a bent wire rod may be used to scrape material from the flask. The slurry is now decanted into a buchner funnel and filtered. The filtrate is used to further wash material from the flask into the filter funnel. The filter cake is sucked as dry as possible and then dried to a constant weight under high vacuum at 45øC in a dessicator.
Yield: approximately 130 grams N,N-diethyllysergamide MW 323.42.
The material remaining on the column may be removed with methanol, evaporated in a vacuum and recycled through the isomerization and subsequent procedures by itself or combined with fresh material. Also, all leftover solutions and residues may be neutralized with sodium bicarbonate, evaporated in vacuo, extracted with ammoniacal chloroform, the extract evaporated to dryness, and the residue re-used.
130 grams of N,N-diethyllysergamide is dissolved in 400 ml methanol and filtered. The filter is washed with 30 ml methanol and the filtrate and washing is poured into a one liter beaker. 30 ml more methanol is used to further wash the filter and filter flask and the wash is also poured into the beaker. 130 ml of diethyl ether is now added to the contents of the beaker. The beaker is gently warmed on a hot plate and 32.0 grams of tartaric acid are then added with constant stirring and warming until they are completely dissolved. The beaker is then allowed to cool. Crystallization of the tartrate usually begins as soon as the tartaric acid dissolves completely. The beaker and contents are refrigerated for at least four hours. Occasional stirring of the crystallizing solution will produce smaller crystals, whereas if the solution is left unstirred during the crystallization, larger crystals will grow. Either is satisfactory. After the beaker has been allowed to stand in the cold four hours or more, the contents are filtered off on a 110 mm buchner funnel with suction. The crystals are washed on the funnel with first 200 ml of a two part methanol:one part ether mixture, and then with 250 ml of a two part ether:one part methanol mixture. Next the crystals are washed with 600 ml of ether and sucked dry. The filter cake is broken up and allowed to air dry in a warm, dark place.
First crop yield: Approximately 80 grams pale yellow to white needles.
The mother liquors and the two washes containing methanol are collected and combined. A one normal solution of potassium hydroxide in methanol is added in approximately equal volume to the combined washes and mother liquors. The solution is then filtered and the filter washed with a few ml of methanol. The filtrates are allowed to stand at room temperature for two to three hours to re-equilibrate the iso-lysergic acid amides from the mother liquors. About 500 ml of water is then added and the mixture extracted with 2.5 liters of methylene chloride in divided portions in a separatory funnel. The combined extracts are shaken with 25 grams of anhydrous magnesium sulfate and filtered. The filtrate is taken to dryness on the rotary vacuum evaporator, care taken not to heat above 55øC. The material is purified in the same manner as that from the original reaction mixture using approximately one fourth the quantities of solvents and alumina as for the original.
Second crop yield: Approximately 20 grams white needles.
The mother liquors may again be worked up as before, or alternatively, they may be saved and included in subsequent batches.
Third crop yield: Approximately 5 grams white needles.
Total yield: Approximately 105 grams N,N-diethyllysergamide tartrate MW = 430.51 (includes one mole methanol per mole of amide).
The residue from the previous step is taken up in two liters of chloroform and filtered with suction through a column 50 millimeters in diameter packed with 400 grams of basic alumina, Brockman activity 1. The filtrate is then refiltered through the same column in the same manner four or five times until the filtrate appears light amber and further repetition of this process fails to remove significant color from the filtrate. The column is now eluted by adding several liters of fresh chloroform to the top and sucking it through into the previous filtrate. Sufficient chloroform should be added to remove all blue fluorescent material from the column but not greenish or yellow (use a blacklight in a darkened room). A band of greenish yellow material should remain in the upper 2/3 of the column when viewed in ultraviolet light (blacklight). The total filtrate is taken to dryness in vacuo in a three liter round bottom flask on a rotary evaporator over a 60ø hot water bath. The residue is taken up in 500 ml of benzene and again taken to dryness in the same manner. 500 ml of benzene is again added and taken to dryness. The flask is left on the evaporator under full vacuum for a considerable length of time after the residue appears dry to remove any traces of dimethylformamide that may still remain. A bubbly, crystalline residue should fill the interior of the flask at the end of this step. If any tarry, gummy appearing material appears to remain on the sides of the flask, repeat the addition of 500 ml of benzene and evaporate to dryness again to get a glassy, crystalline appearance. When the material in the flask is totally dry, remove the flask from the evaporator and add sufficient petroleum ether (a commercial mixture of hexanes is excellent for this purpose) to the flask to be able to swirl the crysþalline material around and loosen it from the sides of the flask. Filter this slurry on the buchner funnel with a fritted glass disk and use the filtrate to further wash the remaining material from the evaporator flask into the buchner funnel. Suck the material dry on the funnel and then place in a vacuum dessicator and dry to a constant weight. Record the dry weight of this material N. Calculate the weight of one equivalent of tartaric acid as follows:
Weight of tartaric acid =.232N
Add methanol to a small beaker in a quantity equal to four times N in milliliters. Dissolve the dried material of weight N in this. Dissolve one equivalent of tartaric acid in the same solution, warming the solution gently and stirring. Slowly with stirring, add ether to the solution in the quantity of no greater than.5 N ml. Addition of ether causes a precipitate which dissolves quickly. Ether should be added dropwise with stirring between drops to dissolve any precipitate before addition of the next drop. Crystallization of LSD tartrate should begin shortly after or during addition of the ether. This precipitate does not dissolve and should not be confused with the precipitate caused by the addition of ether. The mixture should be stirred until the solution becomes thickened by formation of crystals. Once crystallization of LSD tartrate is begun it is unnecessary to continue addition of ether. The beaker should be refrigerated several hours and the contents then filtered on a buchner funnel with a fritted glass disk. The crystals are sucked dry and washed with 2.0 N milliters of methanol previously chilled below -5øC and then with 4 N milliliters of a 1:1 mixture of cold ether and methanol. The crystals are sucked completely dry, washed with 8 N ml of ether, sucked dry, and placed in a vacuum dessicator to remove last traces of solvent.
The total filtrates from the crystals (mother liquors plus washings) are made basic by addition of 2 molar ammoniacal ethanol in approximately equal volume and allowed to stand several days at room temperature when the mixture is filtered and taken to dryness and treated in the same manner as the residue from step 2 for a second crop of crystals.
Ergotamine tartrate (10g) is added to a stirred de-aerated (nitrogen stream) solution of 38 g potassium hydroxide in 100 ml of methanol and 200 ml of water. The solution turns pink to red. The solution is heated to reflux and the methanol is slowly removed using a partial takeoff. Methanol is allowed to distill until the pot temperature reaches 90-95ø C. The mixture is then maintained at total reflux until the evolution of ammonia ceases (hold pH paper in outlet of reflux condenser to test for ammonia). Nitrogen should be bubbled through the mixture to entrain the ammonia. The hot dark solution is then allowed to cool somewhat and then cautiously acidified with a mixture of 60 ml acetic acid and 60 ml water. The resulting hot solution is quickly treated with Norite "A" decolorizing carbon and filtered hot. The clear purple-hued filtrate is allowed to cool to room temperature (crystallization begins) and then in an ice bath or refrigerator. The crystalline precipitate of lysergic acid (grey to purplish white) is collected, washed with a small amount of cold water (5 ml), followed by cold methanol (5 ml) and ether. Yield 3.2 to 3.8 g. Digestion of the crude acid with about 50 ml of methanol (to remove some of the colored impurities) gave after cooling to 0-10øC and filtering an almost quantitative recovery of lighter colored acid. This material is suitable for conversion into LSD.
Lysergic Acid Diethylamide
1. Sulfur Trioxide-Dimethylformamide Comþlex
Into a carefully dried two liter three necked round bottomed flask fitted with a mechanical stirrer, thermometer and a pressure equalizing dropping funnel protected from the atmosphere with a CaCl2 drying tube, was placed approximately one (I) liter of freshly distilled dimethylformamide (DMF) (a one to three degree fraction BP ca. 62-63øC/20 mm). The DMF was cooled to 0-5ø C by means of an external ice-salt-water bath. Sulfur trioxide (Sulfan B) (ca. 100 g) was then placed in the dropping funnel and added dropwise over a period of 30 to 40 minutes to the stirred DMF. The temperature is carefully maintained between 0 and 5ø C throughout the course of the addition. Stirring is continued thereafter until all of the crystalline material is brought into solution. The resulting reagent solution is then transferred into a suitable reservoir fitted with an automatic burette (protected from the atmosphere with a Drierite tube) and refrigerated. If kept dry, the reagent will be good for a month or two even though it will turn yellow and then orange in color. The molarity of the reagent is then determined by titration against standard base. An aliquot (1 or 5 ml) is first diluted with water (20 or 100 ml) to convert the sulfur trioxide-DMF complex into sulfuric acid. The resulting solution is titrated to phenolphtalein end-point with standard 0.1 or 0.01 N aqueous alkali (NaOH or KOH) to determine the molarity (1/2 of the Normality). It should be in the range of 0.9 to 1.2 depending on the amounts of 503 and DMF used.
2. Lysergic Acid Diethylamide (LSD)
For best results all lysergic acid and LSD solutions should be protected from direct light (yellow light is non-damaging) and the working temperatures should never exceed 25ø C. Lysergic acid monohydrate (7.15 g, 25.0 mmol on a 100% basis) and lithium hydroxide monohydrate (1.06 g, 25.0 mmol) were added to 200 ml of anhydrous methanol and stirred until complete solution occurs. Use magnetic stirrer and keep solution under dry nitrogen in the dark. The solvent methanol is then removed by evaporation under reduced pressure to leave a frothy glass-like residue of lithium lysergate. A solution of the calculated amount of tartaric acid is prepared in methanol (ca. 8 ml/g). Approximately 1/2 of the methanol to be used and 20% of the tartaric acid solution is added to the flask containing the LSD base. The flask is swirled and/or shaken until the solid material has dissolved (5-10 minutes) and the solution is then transferred into an Erlenmeyer flask. The balance of the methanol, in two portions, is used to complete the transfer. At this point the rest of the tartaric acid solution is added. lt may be helpful to titrate the solution to an end-point pH of 5.3, since adding excess tartaric acid solution inhibits crystallization somewhat. However this is optional. If seed crystals are available, they should be added at this point. Crystallization should begin within a 1/2 hour: The flask should then be refrigerated for 12-24 hours at 5-10ø C and then for another 12 hours at -10 to -20øC. For 5 g of LSD base 1 g of tartaric acid in 7-8 ml methanol and an additional 17-18 ml of methanol are used. The crystalline mass of needles is broken up and the cold solution filtered (suction). The filter cake is sucked dry and then washed with anhydrous ether. lf necessary the product may be recrystallized from methanol using 5 ml for each gram. The snow white product melts at 198-200øC.
3. Recrystallization Procedure
The crude tartrate (10 g) is placed in a 125 ml Erlenmeyer flask and boiling methanol (50 ml) is added and the mixture stirred and heated for a minute or two (no longer) until solution is complete. The hot solution is quickly filtered through a previously warmed buchner funnel and the filtrate cooled immediately by swirling in a cold water bath until the temperature drops to 25ø C. Crystallization should be well on the way by this time. The mixture is further cooled to 5 to 10øC and then to -10 to -20øC as previously described, to complete the crystallization. Recovery is between 50 and 70%.
4. Additional Crops of Crystals
The mother liquors from initial crystallizations and from re-crystallizations of LSD can be concentrated by evaporation under reduced pressure to produce additional crops of crystals. The second and third crops of crystals are usually dirty enough to require re-crystallization. After three crops, the mother liquors usually become very syrup-like. They then contain mostly iso-LSD (as the tartrate salt). The iso-LSD salt can be converted back into the base by the addition of methanolic KOH or potassium methoxide to the mother liquor. The resulting mixture should be added to a separatory funnel containing salt solution and ethylene dichloride. The LSD base is extracted into the ethylene dichloride layer (the lower layer). The lower layer is removed and fresh ethylene dichloride used to extract the last traces of LSD base from the salt water-base mixture. The ethylene dichloride extracts are combined, dried with MgSO4, decolorized and filtered through diatomaceous earth as earlier. The resulting ethylene dichloride solution may be combined with the chloroform solutions of iso-LSD which eluted from the chromatographic column. The combined solution may be evaporated to dryness under reduced pressure.
The dry iso-LSD base can then be dissolved in methanol and potassium methoxide added. The resulting mixture is stirred for about 30 minutes. During this time isomerization takes place; about 70% of the iso-LSD is converted into the desired "normal" form of LSD. The methanolic solution is poured into a separatory funnel containing salt water solution and ethylene dichloride. The salt water layer is repeatedly extracted with ethylene dichloride to separate the LSD base from the water-base mixture. The ethylene dichloride extracts are combined, dried with MgSO4, decolorized and filtered. The ethylene dichloride solution is then evaporated to dryness under reduced pressure. The resulting dry LSD base is chromatographed on basic alumina (activity grade 1) as previously described. The blue band is collected as before, evaporated and converted into the tartrate salt. The iso-LSD band may be collected and saved for further re-cycling. NOTE: If you only have mother liquors to isomerize, the second mixing with potassium methoxide is unnecessary. Simply prolong the initial mixing to about 1/2 hour.
Of the many attempts at the total synthesis of lysergic acid from simple starting materials, only two have been successful (JACS 78,3087(1956), which is very complicated, and CA 74,3762 (1970), which follows). However, it is very likely that some of the intermediates in each attempt are psychedelic. ln fact, this is one of the most promising and least investigated areas of psychedelic chemistry. Following are some references to the synthesis of intermediates: CCCC 33,1576(1968); HCA 33,67,375,1796, 2254,2257(1950), 34,382(1951), 35,1249,2095(1952), 36,839 1125,1137(1953), 37,1826(1954), 38,463,468(1955), 44,1531 (1961); JCS 3399,3403(1954); BSC 861(1965); JOC 10,76 (1945), 26,4441(1961), 29,843(1964); Chem. and Ind. 1151 (1953); CJC 41,2585(1963); CPB 12,1405,1493(1964),13,420 (1965), 14,1227(1966); JACS 61,2891(1939), 67,76(1945), 71,761(1949), 73,2402(1951), 78,3087(1956), 79,102(1957), 82,1200(1960), 88,3941 (1966); BER 86,25,404(1953), 87,882 (1954), 88,370,550(1955), 89,270,2783(1956), 90,1980,1984 (1957), 93,2024,2029 (1960), 96,1618(1963),100,2427(1967), 101,2605(1968); JMC 8,200(1965); C.R. Acad. Sci. Paris 264 (C), I 18(1967), 265 (C), 110( 1967); BSC 1071 (1968); CJC 41,2585(1963); CPB 12,1405,1493(1964), 13,420(1965), 14, 1227(1966); JCS (P.T.1.) 1121(1972), 760(1973), 438(1973); CA 78,71830,72200(1973).
Various analogs containing part of the LSD structure have been synthesized, but few have any activity. See JPS 60,809(1971) for a review of these compounds. For other LSD analogs see JMC 16,804,1015(1973); BSC 2046(1973); CA 48,4489(1954); JPS 62,1881(1973). Some other useful references on LSD chemistry: U.S. Patents 3,856,821 and 3,856,822; Swiss Patent 517,680(1970); Belgian Patent 738,926; French Patent 1,368,420 and addition 91,948 (1968). Total Synthesis of LSD German Patent 1,965,896 (1 Oct 1970). German Patent 1,947,063 is the same as 1,965,896. For synthesis of 5-Br-isatin from isatin see CA 33,2516(1939) or BER 40,2492(1907). For the use of cycloaliphatic or aromatic esters in place of methyl- 6-methylnicotinate or of isatin or 4 or 5 chloroisatin or 4-bromoisatin in place of 5-bromoisatin see French Patent Addition #2,052,237 (14 May 1971).
Total yield of LSD from starting materials is probably about 1%
Mix 32.8 g (0.217M) methyl-6-methylnicotinate (other alkyl groups can replace either methyl group) with 45.2 g (0.2M) 5-bromoisatin (apparently 4-Br or 4 or 5 Cl isatin will also work) in a 250 ml flask at 100ø in an oil bath and raise the temperature of the bath to 180ø over 15 minutes. Lower temperature to 170ø and let react for 70 minutes. Cool and then grind the solid as fine as possible in a mortar. Recrystallize from 150 ml dimethylformamide and wash with ether to get 40 g (57%) methyl-a(5-bromo-3-isatylidene)-6-methylnicotinate (I). Suspend 10 g (I) in 250 ml glacial acetic acid and heat to boiling. Add in small portions over 30 minutes excess powdered zinc. Reflux 1 hour, filter and evaporate in vacuum and recrystallize the residue from dioxane to get 9.7 g (95%) methyl-a-(5-bromo-2-oxindol-3-yl)-6-methylnicotinate (II). To a suspension of 18 g dry NaBH4 in 300 ml dry tetrahydrofuran add with stirring at 0ø over 30 minutes about 75 g BF3 etherate. Stir 3 hours at 0ø, add 18 g (II) and heat exactly 20 minutes at precisely 22-24ø. Add carefully 150 ml concentrated HCl while cooling in an ice bath. Add 200 ml water and stir 12 hours. Basify, extract the product with ethyl acetate and dry, evaporate in vacuum to get 11 g of residue which recrystallizes from methanol to give methyl-a-(2,3-dihydro-5-bromo-3-indolyl)-6-methylnictotinate (111).
The following step may be unnecessary but it gives stability to (III). The acetyl group can be split off at the end of synthesis, but this is unnecessary since the 1-acetyl-LSD is as active as LSD.
Treat 12 g (Ill) at room temperature for 24 hours with acetic anhydride then hydrolyze and extract to get 11.5 g residue which is ground in petroleum ether and recrystallized from cyclohexane (can chromatograph on alumina and elute with petroleum ether to wash out an oil, then with benzene containing 5% ethyl acetate to elute the produce) to give methyl-a-(1-acetyl-5-bromo-2,3-dihydro-3-indolyl)-6-methylnicotinate (IV). Heat 5 g (IV),12.5 ml acetone,12.5 ml methanol and 1.8 ml methyl iodide for 18 hours in a Carius tube at 70-80ø. Cool, filter, wash with acetone and recrystallize from methanol to get methyl-a(1-acetyl-2,3-dihydro-5-bromo-3- indolyl)-1,6-dimethylnicotinate iodide (V). To 9.4 g (V) in 250 ml water and 250 ml methanol at 35ø add over 5 minutes 2.9 g KBHþ and stir 10 minutes. Add 2.9 g more KBH4 and stir 30 minutes. Evaporate in vacuum and extract the residue with methylene chloride to get 6.2 g oily mixture containing about 2 g of the d isomer (can separate by chromatography if desired) of methyl-a(1-acetyl-2,3-dihydro-5-bromo-3-indolyl)-6-methyl- 1,2,5,6-tetrahydronicotinate (VI). To a suspension of finely powdered NaNH2 (6.1 g) in 2 liters dry ammonia, add with stirring 8 g (VI) in 50 ml dry tetrahydrofuran. Stir 1 hour, add NH4Cl and evaporate the ammonia as fast as possible in a nitrogen stream. Extract at pH 8 with methylene chloride to get 6 g (can chromatograph on 300 g silica gel and 250 g Celite and elute with 98% benzene-2% absolute ethanol and evaporate in vacuum) of methyl-1-acetyl-2,3-dihydro-lysergate (VII). (VII) can be converted to 2,3-dihydro-LSD (not to be confused with 9,10-dihydro-LSD, which is inactive), which is about ten times less active than LSD. (VII) can be converted to lysergic acid prior to conversion to LSD, which will triple the yield in terms of LSD activity (considering 30% yield). The process (which follows) is somewhat complicated and an easier dehydrogenation process may work. 2,3-dihydro-LSD can be converted directly to 12-hydroxy-LSD, which has about the same activity as LSD and this process is also given below.
Dissolve 4 g (VII) in 78 ml 1.5% KOH and reflux five minutes (under N2 if possible). Add 8.5 g Na arsenate hydrate and 16 g Raney-Ni (wet) (deactivated by boiling in xylene suspension - see JOC 13,455(1948)) and reflux twenty hours (under N2 if possible). Filter, precipitate lysergic acid by taking pH to 5.6 with HCl; filter and wash precipitate with water to get 1 g lysergic acid. Evaporate in vacuum the filtrate to get more product.
Warm to dissolve 1.5 g 2,3-dihydro-LSD in 5 ml acetone, 40 ml water and 40 ml saturated NaHCO3. Cool to 20ø and add all at once with vigorous stirring 2.46 g potassium nitro-sodisulfonate dissolved in 90 ml water and 10 mI saturated NaHCO3. After 1 minute, extract 7 times with ethylacetate, wash the combined extracts with water, dry and carefully remove solvent to get a mixture of 12-OH-LSD, LSD and starting material which can be chromatographed to give about 0.2 g 12-OH-LSD. The following method of converting (IV) to the diethylamide (which can probably be used in place of (IV) to give the diethylamide of (V), (VI), and (VII)) will probably also work admirably for (VII) or lysergic acid. Reflux 0.5 g (IV) with 0.5 g KOH in 30 ml methanol for 4 hours. Evaporate in vacuum and add water to the residue. Adjust the pH to between 5 and 6 and filter or centrifuge to get 0.3 g of the free acid. Suspend 1.8 g of the acid in 125 ml chloroform, cool to -5ø and add 0.5 g triethylamine, then 0.6 g ethylchloroformate and stir 45 minutes. Add 2 ml diethylamine and stir 3 hours at room temperature to get, after the usual workup, 1 g of the diethylamide (recrystallize from benzene).
* Starred chemicals are carefully watched.
|**** diethylamine||500 g (725 ml)|
|methylethylamine||50 g (75 ml)|
|ethylisopropylamine||50 g (75 ml)|
|diethyl ether||5 lbs|
|potassium hydroxide pellets||2 Ibs|
|activated charcoal powder||100 g|
|tartaric or maleic acid powder||200 g|
|small cylinder N2 or N2O or freon|
|HCL or sulphuric acid concentrated||500 ml|
|chloroform||5 L (optional but desirable)|
|ethanol||9 L (optional but desirable)|
|NH4OH concentrated||500 ml (optional but desirable)|
IN ADDITION AT LEAST ONE OF THE FOLLOWING SETS OF REAGENTS
|methylene chloride||10 L|
|N,N carbonyldiimidazole||200 g|
|** anhydrous hydrazine||4 L|
|HCl conc.||1 L|
|sodium nitrite||1 kg|
|** phosphorous oxychloride||200 ml|
|acetonitrile or DMF||15 L|
|** trifluoroacetic anhydride||500 g|