Abstract  [Aim]  To evaluate the effect of chicken manure supplementation on the growth of Hermetia illucens larvae (HiL), and determine the optimal proportion of chicken manure (PCM) to add to feed. [Methods]  Different ratios of chicken manure were added to soybean meal and fed to HiL. The growth status of HiL was evaluated by measuring changes in larvae weight and dry matter. Fatty acids and water-soluble components were extracted from larvae using petroleum ether and distilled water, respectively, combined with ultrasonic technology. To investigate the accumulation of key nutrients in the larvae extract, the bicinchoninic acid and phenol-sulfuric acid methods were used to measure the water-soluble protein and sugar content, respectively. To determine the effect of chicken manure feed supplementation on larval growth, Gas Chromatography-Mass Spectrometry (GC-MS) and a High-Performance Gel Permeation Chromatography-Evaporative Light Scattering Detector (HPGPC-ELSD) were used to analyze the chemical composition of fatty acids and the molecular weight distribution of water-soluble macromolecules in larvae, respectively. [Results]  Growth measurements from HiL indicated a significant decrease in larval weight gain when the proportion of chicken manure in the feed exceeded 30%. Additionally, when the proportion exceeded 50%, the dry weight ratio significantly decreased (P<0.05). Nutrient accumulation analysis revealed that when the proportion of chicken manure added to feed was ≤30%, the fatty acid content of the larvae significantly increased, but decreased as the proportion increased. However, the content of soluble protein and sugar components remained stable (P<0.05), indicating that moderate chicken manure supplementation promotes fatty acid accumulation. GC-MS analysis did not find any significant changes in the chemical composition of lipids, with lauric acid, oleic acid, and palmitic acid identified as common primary components. The HPGPC-ELSD results demonstrated obvious chromatographic peaks at retention times of 16.5 min and 20 min when the feed incuded ≤30% chicken manure. However, the first main chromatographic peak appeared to split when the proportion was increased to 40% and 50%. At 80%, the second main peak disappeared, indicating that a high proportion of chicken manure may cause macromolecular fracture or inhibit low molecular weight polymerization or production. [Conclusion]  The quantity of chicken manure added to soybean meal influences the HiL growth and nutrient accumulation. A high proportion of chicken manure can inhibit larval growth. However, when the proportion of chicken manure does not exceed 30%, its effect on larval growth and nutrient accumulation is not significant. Therefore, based on the findings of this study, a maximum of 30% chicken manure is considered the optimal proportion for supplementing HiL feed.