Ms Hartland will be the Australian Secret Intelligence Service's 13th director-general and the first woman to lead the department.
She was the first female deputy director-general of domestic intelligence agency ASIO.
She also previously served as the Employment Department secretary from 2017 to 2020 and was the department's deputy from 2006 to 2011.
Ms Hartland said she was honoured by the appointment to the new position.
"With an increasingly complex geo-strategic landscape, intelligence will continue to be critical to securing Australia's safety, prosperity and sovereignty," she said.
"To lead an organisation that so significantly contributes to Australia's national security is a true privilege."
Ms Hartland's appointment comes after the retirement of Paul Symon, who led ASIS for five years.
In a joint statement, Prime Minister Anthony Albanese and Foreign Minister Penny Wong thanked Mr Symon for his service.
"Mr Symon's career has been one of service to Australia. He has contributed with distinction in senior Defence and intelligence roles both domestically and internationally," the statement said.
"We have greatly valued his experience, integrity and counsel and wish him and his family well."
Ms Hartland will begin her new role as the head of ASIS on February 20.
Mr Albanese said the position would be a crucial one at a time of growing international tensions.
"At a time when Australia's external environment is being reshaped, the work of ASIS is critical to defending our interests and keeping Australians safe," the statement said.
"Ms Hartland will bring excellent strategic, operational and people leadership to her new role."
Opposition Leader Peter Dutton said the coalition welcomed the appointment of Ms Hartland.
"With an eminent and distinguished career in the Australian public service and the private sector, Ms Hartland is well-placed to deliver the national security leadership that this role requires," he said.
"As the international environment becomes more fractious and challenging, the work of ASIS remains pivotal to Australia's defence and national security."