Abstract:To optimize the extraction process of Congrong Zaoren Wumei Beverage (CZWB, composed of Cistanche Herba, Ziziphi Spinosae Semen, and Mume Fructus) and evaluate its sleep-improving effects, an HPLC method for the simultaneous determination of echinacoside, verbascoside, spinosin, and citric acid was established. Using the content of these four compounds and polysaccharides as evaluation criteria, the CZWB extraction process was optimized through orthogonal experiments, followed by validation and scale-up trials. The sleep-improving effects of the extract were investigated in mice using direct sleep tests, open-field tests, subthreshold-dose sodium barbital sleep tests, and sodium barbital sleep latency tests. The results demonstrated that the optimal extraction conditions for the CZWB extract were an extraction temperature of 100 ℃, a solid-to-liquid ratio of 1:12 (g•mL-1), and an extraction time of 2 h. Continuous administration of the extract for 30 d did not induce direct sleep effects in mice but significantly alleviated anxiety in the open-field test, indicating a certain sedative effect. The extract enhanced the sleep-inducing effects of subthreshold-dose sodium barbital (sleep rates for low, medium, and high doses were 33.33%, 41.67%, and 50.00%, respectively) and significantly shortened the sleep latency induced by sodium barbital (sleep latencies for low, medium, and high doses were 22.77, 22.18, and 14.93 min, respectively). This study optimized the extraction process for CZWB, and the results demonstrate its sleep-improving effects. As such, they provide a scientific foundation for developing sleep-enhancing products derived from Cistanche Herba.