Chitosan oligosaccharide complex and preparation method thereof
A technology of chitosan oligosaccharides and complexes, which is applied in the field of chitosan oligosaccharides complexes and their preparation, can solve the problems that the application of preparation methods has not been reported, and achieve shortening of the incubation period and logarithmic growth phase, lower blood sugar, good effect effect
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Embodiment 1
[0036] In this embodiment, a chitosan oligosaccharide complex, wherein the deacetylation degree of the chitosan oligosaccharide is ≧95%, the molecular weight range is 1000-3000Da, and the content of manganese is 5%-8%. Oligochitosan and Mn +4 Coordination forms chitooligosaccharides to chelate manganese. The preferred preparation process is as follows:
[0037] 1. Preparation of specific molecular weight chitosan oligosaccharides:
[0038] ① Weigh 100 grams of chitosan (deacetylation degree ≧ 95%) sample and dissolve it in 1 L of glacial acetic acid solution with a mass percentage of 2%.
[0039] ② Add 2 grams of chitosanase (enzyme activity unit 180U / g), and stir at 45°C for 3 hours.
[0040] ③ Add 42 grams of sodium hydroxide, let stand at room temperature for 30 minutes, centrifuge at 8000 rpm / min for 10 minutes, and discard the precipitate.
[0041] ④ After centrifugation, the supernatant was ultrafiltered with a hollow cellulose membrane with a molecular weight cut-of...
Embodiment 2
[0049] Experiment on the effect of chitosan oligosaccharide chelated manganese on pancreatic beta cell line NIT-1:
[0050] The islet β cell line NIT-1 was cultured in DMEM medium containing 100 mL / L fetal bovine serum, and after growing to a monolayer in the culture flask, it was digested with 2.5 g / L trypsin to make a single-cell suspension to The concentration of 5.0×107 / L was inoculated on a 96-well plate. In the experimental group, different concentrations of chitosan oligosaccharide chelated manganese were added, and cultured in a constant temperature incubator with 50 mL / L CO2, 37°C and saturated humidity. The culture solution of the sample was used as the control group, which was taken out at 24, 48, 72, 120, 144 and 168 hours after adding the sample, and the absorbance value (A value) was measured at 492 nm on a microplate reader by MTT method.
[0051] figure 1 The effects of different concentrations of chitosan oligosaccharide chelated manganese on the proliferatio...
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