Robert Henry Pulleine was born on 7 June 1869 in Picton, New Zealand, but when he was a young child he moved with his family to Fiji where they lived for several years. In 1880 the family moved to Tasmania and then settled in Adelaide the following year when his father, FA Pulleine, was appointed the first Registrar of the South Australian School of Mines. Robert joined the Public Library in 1885 as a cadet, and later became a teacher at the Adelaide Collegiate School at North Adelaide. He commenced a medical degree at the University of Adelaide in 1892. After completing his medical studies at the Sydney Medical School and the Royal Prince Alfred Hospital in 1898 he took up a medical practice in Queensland. In 1899 he married Ethel Williams and moved back to Adelaide with his wife. In 1905 he and his wife and their two children travelled to Gottingen in Germany where he undertook specialist medical studies. Pulleine returned to Adelaide in 1907 with his family. He established a practice as an eye, ear, nose and throat specialist and soon became a leading authority in that field. He became a Fellow of the Royal Australasian College of Surgeons and served as President of the South Australian Branch of the British Medical Association from 1927-1928.

Pulleine had developed an interest in natural history in his youth, particularly ornithology, marine biology and entomology, and also became interested in botany, particularly the study of cacti and other succulent species. He joined the Royal Society of South Australia in 1907 and later became a life member. He served several terms as Vice-President in the 1910s and 1920s and served as President from 1922-1924. He joined the Royal Geographical Society of Australasia (South Australian Branch) in 1914 and later served for many years on the council of that Society, and was elected President from 1932-1933. He was a member of the Linnean Society of NSW and the South Australian Ornithological Association, as well as the Australian Association for the Advancement of Science and the South Australian Branch of the British Science Guild.

He developed an interest in anthropology and archaeology, and made a number of visits to Tasmania to collect Aboriginal artefacts. He was a founding member and first President of the Anthropological Society of South Australia and an inaugural member of the Board for Anthropological Research (BAR). He participated in BAR expeditions to Wilgena in 1925, Koonibba in 1928, MacDonald Downs in 1930, Cockatoo Creek in 1931 and Ernabella in 1933. He published several papers on Aboriginal archaeology, and others on arachnology, botany and a range of medical subjects.

Pulleine died in Adelaide on 13 June 1935 after a short illness. He was survived by his widow, a son and four daughters. During his life, Pulleine accumulated a large collection of Aboriginal ethnographic items and an extensive library. The South Australian Museum acquired some of his ethnographic material after his death.

Pulleine published the following papers on Aboriginal anthropology and archaeology:

RH Pulleine, 'Old native camps at Commodore Point, Encounter Bay', Transactions of the Royal Society of South Australia, 45, 1921, pp. 278-80.

RH Pulleine, 'Cylindro-conical and cornute stones from the Darling River and Cooper Creek', Transactions of the Royal Society of South Australia, 46, 1922, pp. 304-8.

RH Pulleine, 'On the discovery of supposed Aboriginal remains near Cornwall, Tasmania', Transactions of the Royal Society of South Australia, 48, 1924, pp. 83-6.

RH Pulleine 'Cylindro-conical stones from Arcoona, Pimba, South Australia', Transactions of the Royal Society of South Australia, 50, 1926, p. 179.

RH Pulleine, 'Rock carvings (petroglyphs) and cave paintings at Mootwingee, New South Wales', Transactions of the Royal Society of South Australia, 50, 1926, p. 180-2.

RH Pulleine, 'The Tasmanians and their stone culture' (Presidential address), Report of the Nineteenth Meeting of the Australian Association for the Advancement of Science (1928 Hobart), 19, 1929, pp. 294-314.

RH Pulleine & H Woollard, 'Physiology and mental observations on the Australian Aborigines', Transactions of the Royal Society of South Australia, 54, 1930, pp. 62-75.

HK Fry & RH Pulleine, 'The mentality of the Australian Aborigines', Australian Journal of Experimental Biology and Medical Science, 8 (3), 1931, pp. 153-67.

RH Pulleine, 'Botanical colonisation of the Adelaide Plains', Journal of the Royal Geographical Society of Australasia (SA Branch), 35, 1934, pp. 31-65.