The four-piece group, who performed 'Heal The World' in a Michael Jackson-themed show, found themselves in the bottom two alongside Spanish contestant Ruth Lorenzo, a member of Dannii Minogue's 'Over 25s' category.
Both acts went on to perform songs of their choice in the sing-off. While Girlband picked Aerosmith's 'I Don't Wanna Miss A Thing', Lorenzo belted out a rendition of Prince's 'Purple Rain'.
As usual, it was then left to the four judges to decide the contestants' fates. However, for the second week running, the vote went to deadlock after Girlband were backed by Louis Walsh and Simon Cowell, while Lorenzo was supported by Minogue and Cheryl Cole.
Show host Dermot O'Leary confirmed that producers would revert to the earlier viewer vote before announcing that Girlband were the least popular act and would be going home.
Walsh, the girls' mentor, said: "They sang their hearts out, they did their best - I'm absolutely glad I picked them. If I was doing it all over again I would pick these girls."
Band member Phoebe Jay Lau, 17, added: "It's been the most amazing journey ever - I'm just so happy we even got this far."
The result means that Walsh now has just one act, boyband JLS, left in his 'Groups' category after two weeks of live shows.
X Factor continues next Saturday at 7.45pm on ITV1 with a Big Band theme night.