The campaign, created by Rainey Kelly Campbell Roalfe/Y&R, boasts that a Virgin train journey emits three-quarters less carbon dioxide than a comparable trip by air.
The move is another sign of how the environment is increasingly becoming a battleground for travel companies in their marketing campaigns.
Only last month, the ASA ruled that EasyJet had "inaccurately portrayed" the environmental benefits of its planes.
Last year, Ian Pearson, the environment minister, labelled Ryanair "the irresponsible face of capitalism" over its refusal to join the EU's emissions trading scheme.
Virgin Trains has already submitted evidence to the ASA including data from the Edinburgh Centre of Carbon Management, whose figures form the basis of the claims in the campaign.
EasyJet has also alleged to the ASA that Virgin Trains has falsely inflated its passenger numbers to manipulate its green credentials and failed to disclose whether the company uses electricity from nuclear power stations.
In a separate complaint, the ASA will investigate a Shell ad following a complaint from Friends of the Earth.
The pressure group has drawn attention to an ad featuring flowers spouting from a Shell refinery as opposed to smoke. The ad claims Shell recycles waste carbon dioxide.