We present ultrafast z-scan measurements of the two-photon absorption (TPA) spectra of a pair of two-dimensionally conjugated quadrupolar donor/acceptor (D/A) chromophores. The all-donor substituted species displays a peak TPA cross section $[{\sigma }^{\left(2\right)}=520\pm 30\mathrm{GM}]$ that is more than twice that of the D–A species $[{\sigma }^{\left(2\right)}=240\pm 20\mathrm{GM}]$ . Unlike previous structure–property studies that have evaluated TPA behavior for D/A molecules through the comparison of dipolar and quadrupolar compounds, both molecules investigated herein are quadrupolar, ultimately providing a more consistent evaluation of the effects of donor and/or acceptor substitution on the TPA of conjugated chromophores.