The mode-converted (Longitudinal to Transverse, L-T) ultrasonic scattering was utilized to characterize the microstructural anisotropy on three surfaces of samples cut from the low-scattering and high-scattering regions of a raw titanium alloy Ti-6Al-4V billet, respectively. The L-T ultrasonic measurements were performed in two perpendicular directions using two focused transducers with a 15 MHz center frequency in a pitch-catch configuration. The root mean square (RMS) of ultrasonic scattering was calculated for each L-T measurement and a Gaussian function was used to fit each RMS to determine the RMS amplitude. The ratio of RMS amplitudes for L-T measurements performed in two perpendicular directions was calculated to characterize the microstructural anisotropy on the measured surface of a sample. The results show that the amplitude of L-T ultrasonic scattering is highly dependent on the microstructural anisotropy. The microstructural isotropy was considered on the x-y planes of all samples, while the high anisotropy was seen on the x-z and y-z planes of all low-scattering and high-scattering samples. In addition, the microstructural anisotropy measured on the x-z planes of the low-scattering and high-scattering samples gradually increases and decreases, respectively, from the outside diameter (OD) to the centerline (CL) of the billet. The anisotropy measured on the y-z planes of the low-scattering samples slightly decreases and then increases towards the center, while the anisotropy measured on the y-z planes of the high-scattering samples continuously increases towards the center. The variation of microstructural anisotropy in the titanium alloy Ti-6Al-4V billet with duplex microstructure was quantified with the L-T ultrasonic method and the results agree well with micrographs shown in Ref. [18]. The mode-converted ultrasonic scattering method provides a NDE method to characterize microstructural anisotropy, which can be used as an NDE tool for quality control.