Melamine Monitoring in Milk by Reversed-Phase High-Performance Liquid Chromatography Technique

• Publish Date: 16 - July - 2025

Research Abstract:

A rapid, selective, and sensitive reversed-phase high-performance liquid chromatography (HPLC) method was developed to determine melamine in milk. The mobile phase was methanol:phosphate buffer at pH = 3 (20:80 v/v). An SGE ProteCol-GP125-C18 (4.6 x 250 mm, 5 µm) column was used to perform the separation, and the detector was a photodiode array at a wavelength of 236 nm.. The parameters such as mobile phase ratios, pH values, and buffer concentrations were tested. The method demonstrated the validation and ability to separate melamine from with high accuracy in less than 5 min, the retention time of melamine was 4.1 min. The method exhibited an acceptable linearity within the concentration range of (100-1000 ng/mL) with a correlation coefficient r2 = 0.999. Under optimum conditions, the limit of detection (LOD) and limit of quantification (LOQ) were 22.65ng/mL and 68.64ng/mL, respectively, with good precisions. The developed method was successfully applied to liquid and powdered milk and infant formula samples, the melamine peak was completely separate from the milk components, and the recovery of melamine was within 87.26- 100.40% in spiked milk containing 250 and 500 ng/mL, with RSD% of 1.34-3.71%.

Publishers:

Mudar Sadiq