This work presents the development and validation of a capillary zone electrophoresis method for calcium ions release quantitation from composite biomaterials with potential application in bone tissue engineering. Calcium ions quantitation was made using a fused silica capillary (40 cm, 75 m ID) and background electrolyte containing 5 mM imidazole, 6 mM α-hydroxyisobutyric acid, 1 mM 1,4,7,10,13,16-hexaoxacyclo-octadecane and 20 % w/v methanol, at pH 4.5. Indirect UV-detection mode at 214 nm was performed. The separations were achieved using normal polarity at 6 kV, a cartridge temperature of 25 °C and 0.5 psi for 5 s for sample introduction. Parameters of validation such as specificity, linearity, limit of detection and quantitation, accuracy, precision and robustness were evaluated according to the International Conference on Harmonization guidelines. The new developed method resulted to be suitable for calcium determination from composite biomaterials for potential application in bone tissue engineering.