The concentrations of PYY, GLP-1, and NTS in the Caco-2 cell culture medium. IH significantly increased PYY (102 ± 0.368%), GLP-1 (113 ± 2.36%), and NTS (203 ± 11.5%) concentrations in the culture medium. The living cell numbers of normoxia treated cells and IH treated cells are similar levels because the mRNA levels of house keeping genes in IH treated cells were similar to those in normoxia treated cells (91.12 ± 4.196%, p = 0.1484). The measurable ranges were 0.15–10 ng/mL for PYY, 3.91–250 pg/mL for GLP-1, and 0.1–1000 ng/mL for NTS. We measured the concentrations of the peptides directly without any concentrations and dilutions. As the values were within the measurable ranges, the concentrations in the medium are comparable with those in human plasma. Data are expressed as mean±SE for each group (n = 6). The statistical analyses were performed using Student’s t-test.