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Diacylglycerol (DAG) supplementation has been shown to be associated with the reduction of fasting serum triacylglycerol (TAG) concentration, although the extent of the association is uncertain. We quantitatively examined the effect of dietary DAG on fasting serum TAG concentration by conducting a meta-analysis of randomized controlled trials. Potential papers were searched from electronic databases of Medline, Embase and Cochrane Library. Information was extracted and the net change of fasting serum TAG concentration was used as the primary outcome to examine the effect of DAG in Review Manager 4.2. Six papers with seven independent studies (298 subjects) were included into the statistic pooling. Meta-analysis with random effect model showed that DAG did not reduce the fasting serum TAG concentration (WMD: -0.07 mmol/L; 95% CI: -0.21 to 0.08 mmol/L; P = 0.37). Sensitivity analysis indicated the robustness of overall results. Fail-safe number analysis indicated that 18 studies with positive effect were necessary to reverse the reported non-significant efficacy of DAG. Weight estimation analysis indicated that the effect of DAG was influenced to some extent by the initial fasting serum TAG concentration. In conclusion, DAG supplementation did not reduce the fasting serum TAG concentration significantly compared with TAG, but some effects were suggested in diabetic patients with hypertriglyceridemia.