In 1845 he earned his doctorate from the University of Leipzig and shortly after became an associate professor of pharmacology, dietetics, history of medicine and medical literature at the University of Dorpat. In 1849 he was chosen as a full professor of pharmacology. While at Dorpat he created the first pharmacological institute at that school.
In 1867 he became professor of pharmacology and toxicology at the University of Giessen.
Buchheim is remembered for his pioneer work in experimental pharmacology. He was instrumental in turning pharmacology from an empirical study of medicine into an exact science.
He introduced the bioassay to pharmacology, and created a methodology for determining the quantitative and medical aspects of chemical substances. While a student in Leipzig, Buchheim translated Jonathan Pereira"s (1804–1853) handbook of pharmacology from English into German.
He also edited the book, eliminating obsolete and ineffectual medicines and practices, while adding updated information, including a chapter of his own called Art der Wirkung (The Pharmacological Action).
Today at the University of Giessen is the Rudolf Buchheim Institute for Pharmacology.