Abstract: Different medicinal herbs play an important role in shaping the characteristic flavor of traditional herbal chicken soup. In this study, chicken soup prepared with Radix Astragali (HQ), Radix Codonopsis (DS), Fructus Lycii (GQ), and Fructus Jujubae (HZ) was used as the control (CK). Four model soups were established by individually omitting one of these herbs: -HQ, -DS, -GQ, and -HZ. The characteristic flavors and their sources were compared using intelligent sensory techniques, gas chromatography–mass spectrometry (GC–MS), free amino acid analysis, flavor nucleotide analysis, and orthogonal partial least squares-discriminant analysis (OPLS-DA). The results showed that the electronic nose and electronic tongue could effectively detect and distinguish differences in odor and taste among the five groups. The omission of herbs mainly led to a decrease in the response values of the hydrogen sulfide-sensitive sensor (W1W) and the umami sensor (NMS). Further analysis revealed that the omission of herbs led to a reduction in the total content of volatile flavor compounds. OPLS-DA and variable importance in projection (VIP) analysis identified ten volatile flavor compounds as the main contributors to the overall flavor profile, including 2,3-butanedione, 1-hexanol, dipentene, hexanal, 2-methylbutanal, and phytol. Compared to the other omission groups, the -HQ group showed the greatest distance from both these key compounds and the electronic nose sensor signals, indicating that Radix Astragali has the most significant impact on the overall flavor of the chicken soup. Additionally, the levels of key umami compounds (Glu and 5’-IMP) were reduced in the herb-omitted groups. In conclusion, the absence of any herb affects the characteristic flavor of the soup, with Radix Astragali contributing the most notably to its distinctive flavor profile.