Lots of experiments led up to the discover of hormones but the most defining experiment was by Bayliss and Starling in 1902 who showed that when the acid secreted in your stomach in response to food the acid would stimulate the release of a hormone, they called secretin that would stimulate the release of pancreatic juice. All this happened in the absence of nerves showing that internal secretions, now know as hormones, did not require nerves and did exist.
It’s also quite interesting that although Bayliss and Starling came up with the term “hormone” in 1902, there were certain diseases discovered and named before this even though the cause was down to hormones. For example, Grave’s Disease acquired its name in 1835; Addison’s Disease was named in 1849.
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