The aim of the study was to see the diurnal variation of nutrients metabolism and their regulation under the management of large-scaled production. The hepatic transcriptional and serum metabolic studies on circulating nutrient metabolism were investigated in diurnal laying hens. Liver and blood were collected from 36 hens that were slaughtered at 3:30, 7:30, 11:30, 15:30, 19:30, and 23:30 (n = 6), respectively. The serum amino acid, fatty acid and glucose levels, as well as the hepatic transcriptome were analyzed. The results revealed that the circadian clock genes such as Bmal1, Clock, Per1, and Cry2 displayed circadian rhythms in hen livers. The genes related to circulating nutrient transportation, lipogenesis, lipid catabolism, sterol metabolism, and oxidative/anti-oxidative systems also oscillated. However, the nadir of glucose was observed at 7:30 and peaked at 11:30 in the day. Amino acid levels peaked mainly at night, and most amino acids exhibited circadian rhythms based on CircWave analysis. With the exception of undecanoic acid (C11:0), myristoleic acid (C14:1), cis-11, 14-eicosenoic acid (C20:2), and (cis-4, 7, 10, 13, 16, 19-docosahexaenoic acid) C20:3N6 fatty acids, others peaked at 7:30 and 15:30. The results indicated that the hens required more glucose in the early morning. More proteins should be ingested late in the day, since protein catabolism occurred mostly at night. To remove the redundant fats and lipids, fewer should be ingested, especially during the night. All these results would help to design a more accurate nutrition schedule for improving the performance of laying hens in the future.