Screening and follow-up of neonatal hyperbilirubinemia has been done with the use of bilirubin serum levels. This method is invasive and exposes patients to anaemia. A transcutaneous measurement, the Bilispect®, offers an attractive alternative. The concordance between serum and transcutaneous bilirubin levels using the Bilispect®, and the correlation of these values according to prematurity and phototherapy in Latin American neonates were estimated. Simultaneous measurement of serum and transcutaneous bilirubin levels were taken in 239 Colombian new-borns. Concordance was estimated with the Bland-Altman method and Lin concordance index. The Lin concordance index was 0.747 (95% CI 0.691, 0.804). According to Bland-Altman, mean difference was –0.151 (SD 2.510; 95% limits of agreement –5.070, 4.769). Pearson's correlation was 0.79 without phototherapy, 0.72 with phototherapy, 0.76 in preterm and 0.73 in term new-borns. This device has a high potential due to its easy handling. Results are useful to improve the software performance.