Hinshelwood, a Londoner, was educated at Oxford University, where he was elected to a fellowship in 1920 and obtained his doctorate in 1924. In 1937 he became Dr. Lee's Professor of Chemistry at Oxford. He retired in 1964 when he moved to Imperial College, London, as a senior research fellow.

Hinshelwood worked mainly in the field of chemical reaction kinetics. He produced a major text on the subject, The Kinetics of Chemical Change in Gaseous Systems (1926) and, in 1956, shared the Nobel Prize for chemistry with Nicolay Semenov for his work. He later applied his work to a relatively new field in his book, The Chemical Kinetics of the Bacterial Cell (1954).

In some papers published earlier, in 1950, Hinshelwood came very close to the true meaning of DNA, established by Jim Watson and Francis Crick three years later. He declared that in the synthesis of protein the nucleic acid guides the order in which the various amino acids are laid down. Little attention was paid to Hinshelwood's proposal at the time although Crick later declared it to be the first serious suggestion of how DNA might work.

Hinshelwood was a linguist and classical scholar as well as a scientist; he had the unique distinction of serving as president of both the Royal Society (1955–60) and the Classical Association. He was knighted in 1948.