This study aimed to investigate the glycemic and insulinemic effects of lunch timing based on a fixed feeding window, and the effects of apple preload on postprandial glucose and insulin responses after nutrient-balanced lunch and the subsequent high-fat dinner in healthy participants. Twenty-six participants completed four randomized, crossover experimental trials: (1) early standardized lunch at 12:00 (12S); (2) apple preload to 12S (12A+S); (3) late standardized lunch at 14:00 (14S); and (4) apple preload to 14S (14A+S); wherein twenty participants' blood samples were collected for insulin analysis following the lunch trails. In each experimental trial, each participant equipped with a continuous glucose monitor (CGM) was provided with a standardized breakfast and a high-fat dinner to be consumed at 8:00 and 18:00, respectively. The late lunch (14S) resulted in significantly elevated glucose peak, delayed insulin peak time, decreased insulin sensitivity, and increased insulin resistance following the lunch; also decreased glycemic response following the subsequent dinner and larger blood glucose fluctuation over the 24-h period compared with the 12S. The 14A+S significantly reduced the glucose peak, the insulin peak time and the glycemic variability following the lunch, also the 24-h glycemic variability compared with the 14S. The insulin sensitivity was significantly improved in the 12A+S, compared with that of the 12S. In conclusion, the present study found that an extra 2-h inter-meal fasting before and after lunch resulted in elevated glycemic response in both macronutrient-balanced meal and high-fat meal in healthy subjects. The negative impact of a late lunch could be partly reversed by the apple preload, without a trade-off of insulin secretion.