Former US Open winner Michael Campbell finished just outside the qualifying places for next month's Open Championship at Sandwich.
The New Zealander was among 288 players competing for 12 qualifying places over four courses in south-east England.
He carded rounds of 68 and 72 for a four-under-par total at Prince's, one shot outside the qualifying mark.
Another to miss out was former Ryder Cup player Paul Broadhurst, who shot a 67 but then a 75 at Royal Cinque Ports.
Broadhurst, whose 63 at St Andrews in 1990 still stands as the joint best round in a major, missed out by three at the Kent course.
Oxfordshire amateur Craig Hinton, who in February, was in intensive care for a week after being diagnosed with myopericarditis, an inflammation of the heart, won there with a five-under score, leaving four players on four under in a play-off on Wednesday.
Andrew Johnston from North Middlesex and Wellingborough's Simon Lilly were the two players to progress.
Campbell, now 728 in the world rankings, made 12 consecutive Open appearances before missing out at the 150th Championship last year.
"This is a very humbling experience for me," said Campbell, who pushed Tiger Woods into second place at Pinehurst in 2005.
"Having won a major and enjoying the exemptions that go with it for the past few years, there's a danger of taking all that for granted.
"But this experience certainly focuses your attention and gets your mind back on the job."
He can still qualify for the tournament if he finishes in the top five at the French Open, starting on Thursday, or in next week's Scottish Open.
Three players from each of the four courses; Littlestone, Prince's, Royal Cinque Ports and Rye, go through to the Open at Sandwich starting on 14 July.
Lake District professional Simon Edwards won at Prince's, the only course not to go into a second day on Wednesday, on seven under. Thomas Shadbolt and Francis McGuirk, on his home course, also qualified.
Andy Smith from Tudor Park won by one shot with a four under total at Littlestone and said: "I only entered qualifying this year because the events were in Kent. I haven't got much money at the moment and I have been doing all kinds of part-time jobs to keep myself going — labouring, landscape gardening for one of my friends, and a bit of work as a carpenter's mate.
Austria's Markus Brier, twice a winner on the European Tour, and Burnham and Berrow's Lee Corfield also went through in a play-off after five players finished three under.
Amateur Tom Lewis from Welwyn Garden City, who captured the Boys Amateur Championship at Sandwich two years ago, won by three shots at Rye after rounds of 63 and 65 left him at eight under.
Oxford's Adam Wootton was five under, while Mark Laskey, from The Shire, London edged out Inder Van Weerelt of the Netherlands in a play-off after both players had finished three under.