They claimed that if the allegations against him, that he had an affair with a pupil while a teacher at her school, were true then he should be sacked.
The latest row follows revelations in The Independent earlier this year that Mr Woodhead had told a meeting of student teachers that relationships between pupils and teachers could be "experiential and educative". He later apologised for the remark amid a barrage of criticism, although he retained the support of ministers.
Under the Sexual Offences (Amendment) Bill, currently passing through the Commons, teachers face up to two years in jail if they have sex or "any sexual activity" with pupils aged under 18 at their school.
Last night, heads called on Tony Blair to investigate the latest claims, which were made yesterday in a Sunday newspaper by Mr Woodhead's former wife, Cathy.
She claimed that Mr Woodhead had admitted to an affair with Amanda Johnston, a pupil at Gordano School near Bristol. Mr Woodhead has always maintained that the relationship did not start until they had both left the school.
Ms Johnston has backed his version of events, saying in a statement there was nothing improper about their relationship while they were at school.
But in a newspaper article yesterday Mrs Woodhead wrote: "I have become increasingly aware that my silence has condoned his lying and, despite further pressure, I am no longer willing to stand back and see this continue."
Yesterday, David Hart, general secretary of the National Association of Head Teachers, said: "If the allegations are proved to be true then I think in the interests of all concerned a parting of the ways is inevitable.
"If it's clear beyond doubt that he has been lying then there is no alternative to his going. The Government cannot just sit on its thumbs. It is no longer a question of whether the Chief Inspector made indiscreet comments to a group of university students. It is a question of whether the Chief Inspector lied on television and elsewhere."
John Dunford, general secretary of the Secondary Heads Association, said Mr Woodhead's "position becomes increasingly untenable" if the allegations prove to be true.
"Ofsted [Office for Standards in Education] has such an influence on the lives of so many teachers and children it is essential to have someone of complete integrity at the head of it. It is necessary for this to be investigated. It behoves the Prime Minister to investigate this, otherwise the credibility of Chris Woodhead is damaged.
"Mr Woodhead seems to claim that he is responsible to the Prime Minister and if that is the case the Prime Minister will want to know the answer to some questions," he said.
Peter Smith, general secretary of the Association of Teachers and Lecturers, added: "The fundamental question is: has he, as his wife alleges, consistently lied about this relationship? If that is the case, he must ask himself whether he has compromised his moral authority to continue as Chief Inspector of Schools."
Senior government sources declined to comment on the claims.
Mr Woodhead said last night: "I have got nothing to add to what I am already on record as saying, nothing else. My position is perfectly clear and it has been clear for four years."Reuse content