Attributing to the rapid demand expansion for the edible medicinal materials in the market, the limited throughput of high-performance liquid chromatography–multiple reaction monitoring (HPLC–MRM) cannot fully address the measurement workload for a huge number of testing samples. Hence, it is urgent to pursue more efficient approaches for the quality evaluation. Because of the greater selectivity of MRM cubed (MRM) over MRM, there might be a chance to omit the time-intensive LC separation. In current study, we attempted to develop a direct infusion (DI)–MRM program, and the applicability was thereafter assessed through simultaneous determination of four ganoderic acids (GAs) in one of the most famous tonic herbal medicines namely Ganoderma (Chinese name: Lingzhi). Primary parameters such as Q1 > Q3 > Q ion transitions, collision energy (CE), and excitation energy were optimized by programming online energy-resolved mass spectrometry with authentic compounds. A single DI–MRM measurement merely costed four minutes, and in spite of the wide occurrences of isomers, satisfactory selectivity was achieved. Method validation assays demonstrated the method to be sensitive, precise, accurate, and reproducible. The quantitative results from DI–MRM were also justified by conducting LC–MRM measurements in parallel. Significant differences occurred for the content patterns between the two original sources namely Ganoderma lucidum and G. sinense, and, moreover, either cultivar or harvest time showed dramatical influence on the quantitative features of the four targeted GAs. More importantly, DI–MRM is a meaningful analytical option for rapid quantitative analysis of herbal medicines, because of the comparable reliability, nonetheless, less consumptions of both measurement time and solvent, compared with LC–MRM.