AIM: To find specific metabolic markers for women entering peri-menopausal period and patients with menopausal syndrome based on 1H-NMR metabolism serum metabolite analysis combined with pattern recognition. METHODS: Eighty patients with menopausal syndrome, 40 peri-menopausal women of the same age and 40 healthy women of childbearing age as controls were selected. Then the serum samples were detected by nuclear magnetic resonance spectroscopy (1H-NMR), and the metabolites were compared by partial least square discriminant analysis (PLS-DA) and orthogonal partial least square discriminant analysis (OPLS-DA). RESULTS: There were nine serum metabolites with significant differences among the three groups of menopausal syndrome patients and the healthy women during the perimenopausal and childbearing period, which include lactic acid, acetic acid, glutamic acid, pyruvate, creatine, TMAO, N-acetylglycoprotein, alanine and choline. Compared with the healthy women of childbearing age, there were six serum metabolites of menopausal syndrome decreased significantly, but they had no diagnostic value yet. Compared with the healthy peri-menopausal women, the contents of alanine and TMAO were significantly increased, while the contents of glutamate, pyruvate, N-acetylglycoprotein and choline were significantly decreased in patients with menopausal syndrome. And the changes of glutamic acid, pyruvic acid, N- acetylglycoprotein and choline had certain diagnostic value. CONCLUSION: Peri-menopausal period is a special physiological stage of women's transition from reproductive period to old age. Though healthy women have some changes compared with childbearing age, they do not have diagnostic value. However, the differential metabolic markers in patients with menopausal syndrome have a certain diagnostic value, which can provide a certain objective basis for the clinical diagnosis of menopausal syndrome.