Nearly 30 Asda colleagues at the group's head office in Leeds have been axed in a fresh round of redundancies.
One worker said: "It's a nasty shock for all concerned especially as today's redundancies escaped the last cull, thinking they were safe."
The job losses affect the property department at a time when Asda is not planning to open many new stores.
It is understood that some staff who are still on holiday have been informed they have lost their jobs.
In a statement Asda said: “Change is a constant within any retail business and today we have made some changes to the structure of one of our business support functions as part of our ongoing work to ensure Asda is fit for the future.
“Unfortunately these changes mean 28 colleagues from our property team have left us. We’re grateful for their contribution to Asda and wish them the best for the future.”
The move follows the culling of nearly 300 jobs from Asda's head office in Leeds in September as the firm sought to cut costs.
Before the September redundancies, Asda employed 2,500 staff at its Leeds head office.
The firm is battling against stiff competition from discounters Aldi and Lidl, which are around 15 per cent cheaper than Asda, which is the cheapest of the big four grocers.
Just before Christmas Asda told 842 senior shopfloor workers they face a pay cut or redundancy in the new year.
In August the American-owned grocer cut hundreds of jobs in under-performing stores and also asked remaining staff to accept more flexible working arrangements.
The latest round of redundancies comes despite an improvement in Asda's trading.
In November the group reported 1.1 per cent increase in like-for-like sales in the three months to September 30 while net sales rose 3.6 per cent.
At the time Walmart president and CEO Doug McMillon said: “In the UK, Asda delivered positive comp sales again this quarter. The improvements in store experience and price investments are increasing store basket sizes.”
Asda's former CEO Sean Clarke added: “It's great to see that we have now delivered our second consecutive quarter of positive like-for-like growth, and the fifth consecutive quarter of continued sales improvement."
According to Kantar Worldpanel, Asda's sales rose 1.2 per cent in the 12 weeks to December 3.
Fraser McKevitt, head of retail and consumer insight at Kantar Worldpanel, said: "Growth remains in the slow lane at Asda, but they are growing.
"Asda's premium own label is growing at 15 per cent. At the other end Farm Stores is doing well."
The job losses come just a week after Roger Burnley took over as chief executive on January 1.
The first Yorkshireman to take the helm at Asda in over 30 years, Mr Burnley was born in Dewsbury and describes himself as a “born and bred Yorkshireman”.
Mr Burnley first joined Asda in 1997, before being headhunted by Matalan in 2002.
He rejoined Asda last year as chief operating officer with the expectation he would become chief executive over time, but his accession was faster than the market had expected.