As many as 20,000 Tesco shopfloor staff could be affected by the claim, which it is estimated could cost the company up to £4bn based on each worker receiving up to £20,000 in back pay over a period of at least six years.
Law firm Leigh Day began legal proceedings against Tesco today, arguing that employees working in the male-dominated distribution centres are paid considerably more than the predominantly female-staffed stores.
Lawyers claim that the company's hourly-paid female store staff earn up to £3 per hour less than their male colleagues, although the value of their work is comparable.
Tesco, which previously said that men on average were paid 14pc more than women in the year through April 2016, said it hadn't received the claims.
Paula Lee, of Leigh Day solicitors, the firm acting for up to 1,000 women who are likely to take test cases, told the BBC it was time for Tesco to tackle the problem of equal pay for work of equal worth.
Thousands of women who work in Tesco stores could receive back pay totalling £20,000 if the legal challenge demanding parity with men who work in the company's warehouses is successful.
The firm has already started submitting claims on behalf of their clients through ACAS, the first stage in the Employment Tribunal process which could potentially last several years.
Tesco is particularly exposed because of its size as well as a recent push to put thousands more staff on its shop floors in a bid to soften its hard-nosed image among United Kingdom consumers.
An equal pay case is being taken against supermarket giant Tesco which lawyers estimate could lead to a bill of £4 billion.
Tesco said that all their staff could progress equally and were paid fairly, whatever their gender or background.
The Tesco case follows similar claims by Leigh Day against two other United Kingdom supermarket operators - Walmart's Asda and Sainsbury.
This comes as research has also revealed that mothers continued to be penalised at work, as they look to find part-time roles.
The Asda case involves almost 20,000 shopworkers and the most recent ruling backed their right to compare their jobs to those of their mostly male colleagues working in distribution centres.
Prime Minister Theresa May is keen to show she is tackling problems faced particularly by younger Britons, who deprived her ruling Conservatives of a majority in a snap election past year by overwhelmingly backing the left-wing Labour Party.