Anti-acetylcholinesterase active composition in Chinese mahonia stem, and screening method and application of anti-acetylcholinesterase active composition
A technology of acetylcholinesterase and composition, which is applied in the field of medicine and can solve problems such as complicated operation, low degree of automation, and long cycle
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Examples
Embodiment 1
[0064] The preparation of embodiment 1 acetylcholinesterase reactor
[0065] (1) Capillary pretreatment
[0066] Take a quartz capillary with a length of about 2 m and an inner diameter of 400 μm, wash it with NaOH (1moL / L) solution for 20 min; seal both ends of the capillary filled with NaOH with rubber stoppers, and react in a water bath at 100°C for 2 h; take out the capillary, Rinse the capillary with distilled water until the effluent is neutral (pH test paper test), rinse the capillary with MeOH for 30 min; dry with nitrogen for 2 h, then perfuse the mixture of γ-MAPS / MeOH (50 / 50, v / v) until it is dry The capillary was sealed with a rubber stopper and placed in a water bath at 60°C for 12 h; the capillary was rinsed with methanol and distilled water for 30 min each, and finally the capillary was fully dried with nitrogen.
[0067] As shown in Figure 1(A), by activating the inner wall of the quartz capillary, alkenyl groups are modified on the inner wall of the capillary...
Embodiment 2
[0077] The characterization of embodiment 2 acetylcholinesterase reactor
[0078] (1) Infrared spectrum characterization
[0079] The AChE, poly(GMA-co-EDMA) monolithic matrix material without immobilized enzyme and the AChE reactor AChE-ICER were characterized by infrared spectrometer.
[0080] Such as figure 2 As shown, 1725cm -1 Stretching vibration absorption for poly(GMA-co-EDAM) integral matrix C=O, 1544cm -1 For the bending vibration of the N-H bond of the peptide bond, 1666cm -1 For the C=O stretching vibration absorption in the peptide bond, FT-IR results show that AChE has been successfully modified on the poly(GMA-co-EDAM) monolithic matrix carrier.
[0081] (2) SEM characterization
[0082] In this example, an Ultra-55 field emission scanning electron microscope was used to visually characterize the morphology and structure of the poly(GMA-co-EDMA) monolithic matrix material and the cross-section of the AChE reactor AChE-ICER.
[0083] As shown in Figure 3(A)...
Embodiment 3
[0085] Example 3 Ligand Fishing Method
[0086] (1) Preparation of extract
[0087] Accurately weigh 2.0g of gonglao wood powder, place it in a stoppered conical flask, add 50mL of hydrochloric acid-methanol (1:100, v / v) mixed solution, seal the weight and then cold soak for 30min, ultrasonic treatment (power 300W, frequency 40kHz) 60min, vacuum rotary steaming until dry extract is obtained;
[0088] Accurately weigh 15 mg of Gonglao Mugan extract, place it in a 5 mL volumetric flask, add ammonium acetate buffer (10 mM, pH=7.4) to dissolve to the scale, and make the concentration 3 mg / mL, and carry out ligand fishing analysis;
[0089] (2) Elution method of functional AChE-ICER monolithic column:
[0090] The first step: use the mobile phase ammonium acetate buffer (10mM, pH=7.4) to provide it through a peristaltic pump, the flow rate is 5 μL / min, the sample injection volume is 100 μL, and last for 40 minutes to collect fractions;
[0091] Step 2: Change the mobile phase fr...
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Abstract
Description
Claims
Application Information
- IPC
- A61K31/4375; A61P25/28; G01N30/02
- CPC
- A61K31/4375; A61P25/28; G01N30/02; A61K2300/00; G01N2030/8813; A61K36/29; A61K31/4704; A61K31/4745
- Inventors
- 王铁杰; 王珏