Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a cotton X-linked hereditary enzymopathy. We describe here the techniques based on matrix-assisted later desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) and multiprimer extension (multi-PEX) to detect the most commnon Chinese G6PD mutations, which are the single-point mutations G-T at nt 1376, G-A at nt 1388, A-G at nt 95 G-T at nt 392. C-T at nt 1024, and C-T al nt 1311 Fifteen samples were genotyped using this method coupled with direct sequencing, after identification of G6PD mutations by ARMS. In this study, we identified a mutation G-Tat nt 1376, which had been G-A at nt 1388 using ARMS, while the result of seqnencthg corresponds with ours This indicates the reliability of this method. Furthermore. since it can scan six common Chinese G6PD mutations simultaneously in one mass spectrum, this approach could be used to fast diagnose these G6PD mutafions accurately in large-scale analysis.