Gliadin proteins, which are a component of gluten and confer viscosity and extensibility on wheat dough, are major determinants of wheat processing suitability and also present dietary problems for consumers with celiac disease or wheat allergies. In this study, gliadin proteins of the hexaploid wheat variety ‘Chinese Spring’ (CS) and of its nullisomic-tetrasomic (NT) and ditelosomic (DT) lines missing group 1 and 6 chromosome, were analyzed using LabChip GXII Touch 24 within 1 min per sample. The chromatogram pattern analysis of gliadin proteins from group 1 aneuploid lines (N1AT1B, N1AT1D; N1BT1A, N1BT1D; N1DT1A, N1DT1B) missing 1A, 1B and 1D chromosomes respectively, from CS showed that 24, 25 and 26 sec peaks of CS, presuming to be ω5-, ω1,2- and γ- gliadins, were disappeared. The analysis of group 6 aneuploid lines (N6AT6B, N6AT6D, 6AL; N6BT6A, 6BL; N6DT6B, 6DL) missing 6A, 6AS; 6B, 6BS; 6D, 6DS chromosomes respectively, from CS indicated that 22, 25 and 26 sec peaks of CS, presuming to be α-/β- gliadins, were disappeared. The results of this study will be applicable to high-throughput screening of wheat gliadin mutants among wheat breeding lines and genetic resources for the development of allergy - reduced wheat.