The study aimed to investigate RRR-α-Tocopherol and synthetic α-Tocopherol stereoisomers in maternal plasma, cord plasma and breast milk from different regions of China, providing a reference for further guidance on maternal diet and the potential need to supplement mothers with vitamin E. Two hundred and twenty-one sample sets from maternal plasma, cord plasma and three stages of milk (colostrum: 1–5 d after delivery; transitional milk: 10–15 d; mature milk: 40–45 d) were collected longitudinally in six regions of Shanghai, Guangzhou, Tianjin, Chengdu, Lanzhou and Changchun. α-Tocopherol and the stereoisomers were determined by HPLC with a fluorescence detector. The RRR configuration accounted for more than 80 % of α-Tocopherol in maternal plasma, cord plasma and breast milk. Overall, there were regional differences both in α-Tocopherol and RRR. There were significant correlations between α-Tocopherol and RRR in maternal plasma, cord plasma and milk (P < 0.001). As well as negative correlations among α-Tocopherol, RRR and weight-related indicators, which refer to pre-gestation weight and BMI, pre-delivery weight and BMI and pregnancy weight gain, in almost all of samples. This study suggested that RRR-α-Tocopherol was the dominant configuration of α-Tocopherol and the main active form of vitamin E in the early life, guiding the rational supplement of pregnant women and the addition of α-Tocopherol in infant formula milk powder.