H835 50

The GOPs sample was provided by Jilin Taigu Biological Engineering Co., Ltd. (Jilin, China). It was extracted from the roots of Panax ginseng C.A. Meyer by enzymatic hydrolysis, which was planted in Jilin province, China. In brief, ginseng roots were cleansed, minced, homogenized in distilled water, and treated by complex protease (3000 U/g protein) at 40 °C for 3 h after adjusting the pH to 8.0 by sodium hydroxide. Next, nanofiltration, cryoconcentration, decolorization, purification, and spray drying were performed to obtain GOPs powders [19 (link),21 (link),22 (link)].
After purification by high-performance liquid chromatography (HPLC, Waters Corporation, Milford, MA, USA) using a Phenomenex C18 column (10 mm × 250 mm), the sample was measured by LDI-1700 matrixassisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS, Liner Scientific Inc., Reno, NV, USA). The identification results showed that 95.42% of GOPs had a molecular weight between 180 and 1000 Dalton. Amino acid accounted for 3.94%, which was further analyzed by an automatic amino acid analyzer (H835-50, Hitachi, Tokyo, Japan). The amino acid composition is shown in Table 1 [19 (link),24 (link)].

The GOP sample, which was derived from the roots of Panax ginseng C. A. Meyer planted in Jilin province, China, was provided by Jilin Taigu Biological Engineering Co., Ltd. (Jilin, China). In brief, ginseng roots were firstly boiled at 120–128 °C, 0.12–0.16 MPa for 0.5–1.5 h and then the concentrated solution was centrifuged. The supernatant was treated by enzymolysis with 5% special enzyme of ginseng for 3–5 h after adjusting the pH to 7.0 with sodium hydroxide. Then it was adsorbed with 3%–5% of active carbon, and the carbon was removed by filtration. After the filtrate was concentrated by vacuum concentration, the condensed liquid was dried by spray drying to obtain GOP powders.
The GOP sample was purified by HPLC (Waters Corporation, Milford, MA, USA) using a Phenomenex C18 column (10 mm × 250 mm), and the molecular weight distribution of the GOP sample was measured by LDI-1700 matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS, Liner Scientific Inc., Reno, NV, USA). In addition, amino acid composition was further analyzed by an automatic amino acid analyzer (H835-50, Hitachi, Tokyo, Japan). The result showed that 95.42% of the GOP sample had a molecular weight between 180 and 1000 Da. Amino acids accounted for 3.94%, and the amino acid composition is shown in Table 1.

BCOP were derived from the bone of bovine by enzymatic hydrolysis and donated by Hua Peptide Bioengineering Co., Ltd. (Beijing, China). Oligopeptide samples were purified by high performance liquid chromatography (HPLC, Water Corp., Milford, M222KOA, USA) using a Phenomenex C18 column (10 mm × 250 mm), and the molecular weight distribution of the oligopeptide sample was determined by LDI-1700 matrix-assisted laser desorption ionisation time of-flight mass spectrometry (MALDI-TOF-MS) (Linear Scientific Inc., Reno, NV, USA). Then the amino acid composition was analysed with an automatic amino acid analyser (H835-50, Hitachi, Tokyo, Japan), and the amount of free amino acids was measured by HPLC. After HPLC purity and MALDI-TOF-MS analysis, we found that the content with a relative molecular mass greater than 2000 was 1.96%, between 1000 and 2000 was 5.27%, and less than 1000 (small molecule oligopeptides) in BCOP was 92.77%; the content of free amino acids amounted to 2.73%. The amino acid composition is shown in Table 5.
AIN-93G rodent diet were purchased from HFK Bioscience Co. Ltd. (Beijing, China). The dietary ingredients were thoroughly mixed together into granules and then air-dried at room temperature.