The former world indoor champion could only manage 10.29 secs - well outside the 10.18 sec mark he needs to achieve to be considered for his controversial spot on Team GB.
The top two sprinters at this month's Aviva UK Championships in Birmingham will secure their London 2012 places, provided they have achieved the qualifying standard, and another spot will be awarded at the selectors' discretion.
So far only Adam Gemili and James Dasaolu are the right side of the required line, with Chambers currently ranked seventh in the UK - he doesn't even have the B standard yet.
After winning his battle to earn a prospective place at London 2012, due to his previous two-year doping conviction, it would be a supreme irony if the 34-year old now failed to get selected.
This time last year Chambers had already clocked 10.01 secs in Brazil and gone below the current A standard in five races.
But his best time of the season remains the 10.28 secs he ran when finishing behind triple Olympic champion Usain Bolt in Ostrava.