In the early 20th century, Carl von Voit and Max Rubner independently measured caloric energy expenditure in different species of animals, applying principles of physics in nutrition. In 1906, Edith G. Willcock and Frederick Hopkins showed that the amino acid tryptophan aids the well-being of mice but it did not assure their growth. In the middle of twelve years of attempts to isolate them, Hopkins said in a 1906 lecture that "unsuspected dietetic factors," other than calories, protein, and minerals, are needed to prevent deficiency diseases. In 1907, Stephen M. Babcock and Edwin B. Hart conducted the single-grain experiment, which took nearly four years to complete.

What was measured in the early 20th century?