Food fortification is one strategy for addressing micronutrient deficiencies among the population groups at risk. Non-compliance with fortification standards hinders the success of fortification programs. This is due to a lack of techniques to rapidly check the amounts of the added fortificants. Fourier transform - near-infrared (FT-NIR) spectroscopy is a fast and reliable technique that would be used to ensure adherence to requirements. This study aimed to investigate the potential of using FT-NIR spectroscopy to predict the amount of retinol in fortified maize flour. 150 fortified maize flour samples were used in this study. Partial least squares regression (PLS-R) was used to build calibration models based on the retinol reference values obtained by high-performance liquid chromatography (HPLC), and fortified maize flour NIR spectra acquired from the FT-NIR spectrophotometer. Two calibration models were developed to predict retinol above and below 1.0 mg/kg. The performance metrics of model one developed to predict retinol 1.0 mg/kg were: R2c = 0.81, RMSEE = 0.08, RPD = 2.29 and R2v = 0.82, RMSEP = 0.09, RPD = 2.07 for the calibration and validation, respectively. The second model developed to predict retinol ≥ 1.0 mg/kg had the following performance metrics: R2c = 0.93, RMSEE = 0.16, RPD = 3.58 and R2v = 0.81, RMSEP = 0.22, RPD = 2.43 for the calibration and validation, respectively. Overall, the findings demonstrated that FT-NIR spectroscopy can be utilised to reliably predict retinol levels in fortified maize flour samples. FT-NIR spectroscopy, by replacing time-consuming and laborious wet chemistry laboratory procedures, has the potential to be used for rapid regulatory monitoring of fortification compliance for a large number of samples.