Fracking has been stopped at the site in Preston New Road following a seismic event measuring 1.55ML (local magnitude) on the Richter scale last night.
The tremor which was detected at 7.46pm on Wednesday night is the largest recorded since fracking began.
Any tremor measuring 0.5 or above means fracking must be temporarily stopped while tests are carried out.
This latest event comes only days after campaigners raised concerns over the number of tremors since Cuadrilla restarted hydraulic fracturing earlier this month.
A spokesman for Cuadrilla, said: “We can confirm that a micro seismic event measuring 1.55ML (local magnitude) on the Richter scale occurred after we had completed the hydraulic fracturing programme for the day at our Preston New Road site.
"Most local people will not have felt it due to its small size. The equivalent ground motion would be similar to a large bag of shopping dropping to the floor.
"Well integrity has been verified and we will now pause operations and continue monitoring for the next 18 hours.
“The Preston New Road exploration site is the most regulated and monitored site in Europe and the systems in place are working as they should.
"Minor movements of this level are to be expected and are way below anything that can cause harm or damage to anyone or their property.
"All the relevant regulators were informed and we have verified that the well integrity is intact”
Jamie Peters, campaigner at Friends of the Earth, said: “It’s obvious that fracking can’t be done without triggering earthquakes.
"This latest quake is a sign that Cuadrilla just can’t stick within the regulations they agreed.
"Even small vibrations at ground level can be the sign of far more damaging impacts deep underground. Earthquake regulations must be maintained for the safety of local communities.
"52 seismic events in just 6 days of fracking and now the biggest earthquake at the site clearly underlines that this is absolutely not the time to start weakening regulations.
A spokesperson for Frack Free Lancashire, said: "Last night, local residents felt the largest earthquake yet from Cuadrilla's operations at Preston New Road.
"Having abandoned their attempts to frack their first well after only being able to fully fracture just two of the planned 42 stages, Cuadrilla are now trying to repeat the process.
"Therefore, it was a certain grim inevitability to last night's events, but what should really give local residents pause for thought, is the fact that these were trailing events and there is nothing that can be done to mitigate them in real-time.
"We deplore Cuadrilla's attempts to downplay and trivialise these quakes by making facile comparisons with milkshakes and bags of shopping being dropped.
"This is a serious issue and we know that seismic activity at similar levels deformed the well bore at Preese Hall, which Cuadrilla failed to report to the relevant authorities for six months.
"This is, in fact, a perfect illustration of why the limits on the seismic traffic light system must not be relaxed. The safety of residents must be prioritised before a private corporation's greedy drive for profits at any cost."