The learning and teaching of different singing styles, such as operatic and Chinese folk singing, was often found to be very challenging in professional music education because of the complexity of varied musical properties and vocalizations. By studying the acoustical and musical parameters of the singing voice, this study identified distinctive tuning characteristics of a particular folk music in China-Hua'er music-to inform the ineffective folk singing practices, which were hampered by the neglect of inherent tuning issues in music. Thirteen unaccompanied folk song examples from four folk singers were digitally audio recorded in a sound studio. Using an analyzing toolkit consisting of Praat, PeakFit, and MS Excel, the fundamental frequencies (F0) of these song examples were extracted into sets of "anchor pitches" mostly used, which were further divided into 253 F0 clusters. The interval structures of anchor pitches within each song were analyzed and then compared across 13 examples providing parameters that indicate the tuning preference of this particular singing style. The data analyses demonstrated that all singers used a tuning pattern consisting of five major anchor pitches suggesting a nonequal-tempered bias in singing. This partly verified the pentatonic scale proposed in previous empirical research but also argued a potential misunderstanding of the studied folk music scale that failed to take intrinsic tuning issues into consideration. This study suggests that, in professional music training, any tuning strategy should be considered in terms of the reference pitch and likely tuning systems. Any accompanying instruments would need to be tuned to match the underlying tuning bias.