DGCA took the action after a spot audit on Air India Limited in January to assess adherence to regulations.
Fleet-wise random reports were scrutinized by the DGCA during the audit process.
According to the aviation regulator, evidence indicates that the airline operated "flight(s) with both flight crew aged above 60 years flying together in a few instances."
This led to the breach of Sub Rule (2) of Rule 28 A of the Aircraft Rules, 1937.
"The operator was also found deficient in providing adequate weekly rest, adequate rest before & after ultra-long range flights and adequate rest on layover to the flight crew, which violates the extant provisions of the Civil Aviation Requirements pertaining to FDTL," the regulator said in a release.
On March 1, the regulatory body sent a show cause notice to Air India regarding the matter.
The aviation watchdog stated that the fine was imposed because the response provided by the airline to the show cause notice was deemed unsatisfactory.
Following the unsatisfactory response given by the operator, a penalty of Rs 80,00,000 has been imposed on the operator, the release said.
The airline was also found to have overlapping duties and inaccurately marked training records among the issues identified.