Ms Totterdell is currently group chief executive officer at St George’s, Epsom and St Helier University Hospitals and Health Group.
She will be taking over from Judith Paget, who is moving to become director general for strategy within Welsh Government.
Health secretary for Wales Jeremy Miles said: “I would like to thank Judith for her four years as chief executive of NHS Wales and for the leadership she has provided to NHS Wales and the Welsh Government’s health, social care and early years department through the pandemic and its aftermath.
“Judith has been at the helm during an extraordinary period for the NHS, and I wish her all the best for her new role.
“I am very pleased that Jacqueline Totterdell will be joining us, bringing with her a wealth of experience in the healthcare sector and an impressive track record at St George’s.
“I look forward to working with Jacqueline as we focus on delivery and improvement and help the NHS adapt for the future.
“This includes building on the progress we’re making in Wales on waiting times, and to go further, to deliver the faster access to treatment that people in Wales want.”
Ms Totterdell said: “After a long career in the NHS, having the opportunity to lead health and social care services in Wales is something very special.
“Joined-up services that achieve better outcomes for the citizens and communities we serve is a vision that most of us who work in the public sector aspire to.
“Being able to improve services on a whole-country level is what attracted me to this role and I’m really looking forward to working with everyone to make a real difference for people in Wales.”