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CAS:2492-87-7
Appearance:White to yellow solid
Storage:Powder : -20℃, 2 years;In solvent(mother liquid): -20℃, 1 month; -80℃, 6 months
Purity:HPLC≥98%
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Examples of using this product（for reference only）
In Vitro：
α-Glucosidase inhibitory assay（5 mM PNPG，37℃， 30 min, 405nm）
10 μL of α-glucosidase, 10 μL of 0.1 mg/mL Up, Up-3, Up-4, and Up-5, 80 μL of PBS buffer (0.02 mol/L, pH 6.8) and 10 μL of 5 mM PNPG were mixed in a 96-well plate, and then the mixture was incubated at 37℃ for 30 min. The presence of the product, p-nitrophenol (PNP) was monitored by measuring absorbance at 405 nm in a plate reader. Acarbose was used as a positive control. Reactions without inhibitor were also performed in parallel serving as controls. The PNP concentration was determined from a standard curve. The relative activity was expressed as a percentage of the PNP formation in the presence of acarbose or polysaccharides compared with that of the control reactions. Subsequently, the α-glucosidase inhibitory activity of acarbose, Up-3 and Up-4 at different concentrations (0.01, 0.03, 0.05, 0.08, 0.1, 0.3, 0.5, 1.0, and 2.0 mg/mL) were determined according to the method described above to calculate the IC50 value.
Reference：
Zhong QW, Zhou TS, Qiu WH, Wang YK, Xu QL, Ke SZ, Wang SJ, Jin WH, Chen JW, Zhang HW, Wei B, Wang H. Characterization and hypoglycemic effects of sulfated polysaccharides derived from brown seaweed Undaria pinnatifida. Food Chem. 2021 Mar 30;341(Pt 1):128148. doi: 10.1016/j.foodchem.2020.128148. Epub 2020 Sep 22. PMID: 33038776.
Determination of b-Glucosidase activity（2.0 mg/mL PNPG, 60s, 405nm）:
Firstly, 30.0 mL of standard b-Glucosidase solutions (1.5-21.0 U/mL) were diluted by 50.0 mL of NaH2PO4-H3PO4 (10.0 mM, pH=5.0), respectively. Then, 10.0 mL of pNPG (2.0 mg/mL) was added to initiate the enzymatic reaction. After incubation for 60 s, the absorbance of product pNA was measured at 405 nm. Finally, the calibration curve was obtained. Subsequently, 30.0 mL of crude almond extract solution was diluted by 50.0 mL of NaH2PO4-H3PO4 (10.0 mM,pH ? 5.0). Then, 10.0 mL of pNPG (2.0 mg/mL) was added to initiate the enzymatic reaction. The absorbance of product pNA was measured at 405 nm after incubation for 60 s. The solution without pNPG was used as background solution. In this experiment, all assays were performed in triplicate.
Reference：
Chen GY, Zhang H, Yang FQ. A simple and portable method for β-Glucosidase activity assay and its inhibitor screening based on a personal glucose meter. Anal Chim Acta. 2021 Jan 15;1142:19-27. doi: 10.1016/j.aca.2020.10.047. Epub 2020 Oct 27. PMID: 33280697.
