B vitamins (including folate, vitamin B2, vitamin B6 and vitamin B12) and methionine are essential for methylation reactions, nucleotide synthesis, DNA stability and DNA repair. However, epidemiological evidence among Chinese population is limited. The objective of this study was to evaluate B vitamins and methionine in relation to colorectal cancer risk in a Chinese population. A case-control study was conducted from July 2010 to April 2019. A total of 2502 patients with colorectal cancer were recruited along with 2538 age (5-year interval) and sex matched controls. Dietary data was collected by using a validated food frequency questionnaire. Multivariable logistic regression was used to assess odds ratios (ORs) and 95% confidence interval (CI). The intake of folate, vitamin B2, vitamin B6 and vitamin B12 was inversely associated with colorectal cancer risk. The multivariable ORs for the highest quartile versus the lowest quartile were 0.62 (95%CI 0.51, 0.74; Ptrend < 0.001) for folate, 0.46 (95%CI 0.38, 0.55; Ptrend < 0.001) for vitamin B2, 0.55 (95%CI 0.46, 0.76; Ptrend < 0.001) for vitamin B6 and 0.72 (95%CI 0.60, 0.86; Ptrend < 0.001) for vitamin B12, respectively. No statistically significant association was found between methionine intake and colorectal cancer risk. Stratified analysis by sex showed that the inverse associations between vitamin B12 and methionine intake and colorectal cancer risk were found only among women. This study indicated that higher intake of folate, vitamin B2, vitamin B6 and vitamin B12 was associated with decreased risk of colorectal cancer in Chinese population.