HRT applied for an exemption from the 107 pc qualifying rule in Australia but it was thrown out by FIA stewards.
The rules are the rules. Now we're focused on the next race
"Our times were not good enough, but given the number of laps we've done this weekend, we probably are not so bad," said the team's Indian driver Narain Karthikeyan.
But the stewards, including former grand prix winner Johnny Herbert, did not agree that "exceptional circumstances" explained the Spanish team's lack of pace.
The grid will therefore feature 22, and not 24, cars for Sunday's Melbourne race.
"The car is capable of running a lot quicker than today," insisted team boss Colin Kolles. "We'll be back stronger than ever in Malaysia."
Added Italian driver Vitantonio Liuzzi: "The rules are the rules. Now we're focused on the next race."