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Hofmann became an employee of the pharmaceutical / chemical department of Sandoz Laboratories (now a subsidiary of Novartis), located in Basel as a coworker with professor Arthur Stoll, founder and director of the pharmaceutical department. He began studying the medicinal plant Drimia maritima (squill) and the fungus ergot, as part of a program to purify and synthesize active constituents for use as pharmaceuticals. His main contribution was to elucidate the chemical structure of the common nucleus of the Scilla glycosides (an active principal of Mediterranean squill) .Wile researching lysergic acid derivatives, Hofmann first synthesized LSD on 16 November 1938.

The main intention of the synthesis was to obtain a respiratory and circulatory stimulant (analeptic) with no effects on the uterus in analogy to nikethamide (which is also a diethylamide) by introducing this functional group to lysergic acid. It was set aside for five years, until 16 April 1943, when Hofmann decided to reexamine it. While re-synthesizing LSD, he accidentally absorbed a small amount of the drug through his fingertips and discovered its powerful effects. He described what he felt as being:

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