The on-chip measurement of absorbing species has proven to be challenging, particularly with respect to the sample pathlengths available in a miniaturised system. This paper demonstrates how the principles of total internal reflection can be utilised to form a liquid-core waveguide along a single microfluidic channel, increasing the sampling pathlength to 5 mm while maintaining a detection volume of ≤1 µL. This was achieved using the Teflon fluoropolymers PTFE, FEP and AF as cladding for the liquid-core waveguide. In conjunction with a 3D chip architecture, the use of the liquid-core waveguide enables more efficient use of the probing light beam along with easy and effective coupling of the source, microfluidic chip and the detection system. The confirmation that waveguiding was occurring was successfully demonstrated and the subsequent spectrophotometric analysis of crystal violet provided a linear calibration with reproducibility (<2.4% RSD) and limits of detection (<1.3 µM), comparable to absorbance measurements made with a standard UV-Vis spectrophotometer.