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Although environmental exposure to multiple metals is common, epidemiological studies on the associations of exposure to 23 metals with kidney function have not been analyzed. We aimed to investigate the associations of 23 metals levels with renal function. We conducted a cross-sectional study in four rural regions of Hunan province. Plasma and urine metals levels were determined by inductively coupled plasma mass spectrometer (ICP-MS). Two-level logistic regression was used to investigate the associations of metals levels with estimated glomerular filtration rate (eGFR) with adjustment for confounding factors. We conducted a sensitivity analysis of the results using the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation. A total of 3553 participants completed the investigation. Five metals (plasma arsenic and molybdenum; urine copper, rubidium, and strontium) were identified to be significantly associated with renal function. Participants in the highest quartile of plasma arsenic and molybdenum were at 17.95 (95% CI: 6.35–50.76) and 24.23 (95% CI: 7.42–79.19) fold risk of abnormal eGFR, respectively, compared with the lowest quartile. The highest quartiles of urine copper, rubidium, and strontium were associated with 3.70 (95% CI:1.92–7.14), 0.16 (95% CI:0.07–0.37) and 0.08 (95% CI: 0.03–0.21) fold risk of abnormal eGFR. The sensitivity analysis revealed that plasma arsenic, molybdenum and urine copper, rubidium and strontium levels retained similar associations with abnormal eGFR. Plasma arsenic and molybdenum, and urine copper are risk factors for abnormal renal function, while urine rubidium and strontium are protective factors for renal function. • A cross-sectional study was conducted in four rural regions of Hunan province. • The associations of multiple metal levels with renal function was investigated. • Multiple metals in plasma and urine are associated with abnormal eGFR. [ABSTRACT FROM AUTHOR]