Business Secretary Vince Cable has repeated his warning that a country the size of an independent Scotland would not be able to support large financial institutions like RBS.
The Liberal Democrat minister reiterated the claim after Lloyds, Clydesdale Bank and RBS confirmed they would relocate south of the border in the event of a vote for separation next week.
During business questions in the Commons, he said: "I think Standard Life in the insurance sector have also made it clear they couldn't remain in Scotland were it to be an independent country.
"This relates to the need for large financial institutions to have a regulator and in some cases lender of last resort.
"A country of the size of an independent Scotland would not be able to support those institutions."
He was replying to shadow business secretary Chuka Umunna who said Scotland's "vibrant" financial services sector was not just important to UK competitiveness but Scotland's own position in the global market place.
The Labour frontbencher went on: "The vote next week is of course for the Scottish people, but does this [the decision of firms to relocate] not illustrate the lorry-load of uncertainty for jobs, competitiveness and growth in Scotland that will come with the break-up of one of the most successful unions the world has ever seen?"
Backbenchers also chose to use the session to flag up their concerns over the prospect of Scottish independence.
Conservative David TC Davies (Monmouth) said: "RBS have today recognised the competitiveness of the UK or at least the southern part of the UK ... the bits that are run by the Coalition Government.
"Would you agree ... that if RBS want to domicile themselves in the southern part of the UK, they should bring their jobs with them and not expect us in the southern part of the UK to underwrite any mistakes they may make in the future?"
In response, Mr Cable said he had met the RBS chief executive a couple of days ago and added that the company was making good progress in sorting out the problems of its balance sheet.
But he went on: "I have been pointing out for quite a long time that the position of RBS in an independent Scotland would be very difficult since its balance sheet I think is 10 times the size of the Scottish economy and it could hardly operate within Scotland as an independent country."
Labour's Andrew Miller (Ellesmere Port and Neston) warned about the impact on Scotland's top universities in the event of a "wrong vote" next week.
Mr Cable replied: "Scottish institutions do benefit disproportionately from UK research.
"They would no longer be guaranteed access to UK funding streams in a similar way, though obviously they will hopefully maintain their excellence."
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