Lysergic Acid from Claviceps paspali Cultures

Isolation of Lysergic Acid from Claviceps paspali Cultures by Werner Schlientz and Benno Sutter Switzerland Januanry 11,1965 

The process which consists of adsorbing the acids on activated carbon in presence of a reducer in an inert gas atmosphere then eluted with aqueous-alcoholic solutions of very volatile bases. The adsorption of the acids on the activated carbon can be carried out in a wide field of pH, for example between pH 1 and pH 13. With the values extremes of the pH, for example at pH 1 and for pH 13, it is simply necessary to use more activated carbon to obtain an adsorption complete. 

It is perfectly possible also to carry out adsorption and filtration in an inert atmosphere of gas, for example under nitrogen or carbon dioxide, the activated carbon used having to be heated with approximately 300° under an inert gas before being used, then, adsorption and filtration carried out, the activated carbon charged with the adsorbed product is eluated, for example by means of an aqueous-methanolic ammoniacal solution. 

Example 1-100 Liters of a culture filtrate of the new stock of Claviceps paspali Stevens and Hall [Helvetica ChimicaActa 47,4,1052 (1964)] containing according to the colorimetric analysis 870 mg/liter of alkaloids (relatively has a molecular weight of 268) one adds 800 grams of potassium metabisulfite then 2.5 kg of activated carbon, for example of charcoal or of coal of blood, one agitates the whole during approximately fifteen minutes and one separates by filtration the activated carbon. 

One washes this carbon with 20 liters of distilled water and then eluates with 120 liters of methanol containing 10% of concentrated ammonia. The eluate is concentrated with 2 liters under 20 mm of mercury and at 30° and the carboxylic acid, which crystallises at the end of twenty-four hours, is dried at 80° under 1 mm of mercury. 

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Cell-free extracts have been obtained from Claviceps paspali cultures which catalyse the formation of (115) and (120) from tryptophan and isopentenyl pyrophosphate. Both compounds were incorporated into lysergic acid amide. Another unidentified compound has been isolated as a product of the incubation, derived from (120) and convertible into lysergic acid amide " cf. ref. 9. 

Lehtola, T, Huhtikangas, A., Lundell, J. and Vallanen, T. (1982) Radioimmunoassay of lysergic acid derivatives from Claviceps paspali fermentation cultures. Planta Med., 45,161. 

Lysergic acid methylcarbinolamide (XXXVI) crystallizes from chloroform in long prisms, mp 135° (dec.), [a]f,° + 29° (c = 1.0 in dimethyl-formamide). The alkaloid, which was isolated together with ergine and erginine from saprophytic cultures of Claviceps paspali (69), easily decomposes in a weak acid solution to form ergine and acetaldehyde. 

Example 6-100 Liters of a very acid filtrate of culture (pH from approximately 2) of Claviceps paspali Stevens and Hall, whose content, determined by colorimetry, is 1.2 grams per liter (relative with a molecular weight of 268), one adds, while agitating, 500 grams of sodium dithionite and 2.0 kg of activated carbon (Carboraffin). One separates coal by drying, one washes it with 20 liters of water and one subjects it to an elution with 120 liters of methanol added with 10% concentrated ammonia. One concentrates the eluate to 8 liters. One separates by filtration the lysergic acid which crystallized, one washes... 

The synthesis of dimethylallyltryptophan (132) by a crude extract of Claviceps purpurea from tryptophan and dimethylallyl pyrophosphate recorded earlier has been reported again recently. In addition to (132), the formation of (133) was observed. (The latter compound, with unspecified stereochemistry around the double bond, has also been isolated from a C. purpurea culture ). It was found further that both (132) and (133) could act as precursors for lysergic acid amides in C. paspali cultures. Both (133) and its (Z)-isomer have been found to act as precursors for elymoclavine (137) but not chanoclavine-I (138) or agro-clavine (136), which are considered to be normal intermediates in elymoclavine biosynthesis.It may be concluded, however, from the combined evidence, that elymoclavine, lysergic acid, and related compounds may normally be formed along an alternative pathway via these allylic hydroxy-compounds. 

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