New purification method and testing technology of ochratoxin A in traditional Chinese medicines
An ochratoxin and detection technology technology, which is applied in the field of preparation of aptamer affinity columns, can solve the problems of improper processing and storage, difficult detection, complex matrix and the like, and achieves simple and easy preparation process, low detection cost and production cost. low effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0026] Embodiment 1: the preparation of AAC
[0027] (1) Preparation steps:
[0028] a. Aptamer refolding: Dissolve 2.7nmol of 5'amino-modified aptamer in 400μL buffer (200mM Na 2 HPO 4 5mM MgCl 2 , pH8.0), renaturation at 75°C for 5 minutes, and left at room temperature for 30 minutes;
[0029] b. Washing: Take about 200 μL of NHS-activated sephaorse4FF gel and wash with 1 mL HCl (1 mM, pH 3.0) rapidly and repeatedly for 5 times;
[0030] c. Coupling: Remove excess hydrochloric acid, quickly add refolded aptamer buffer, adjust pH to 8.0, and shake for 3 hours at room temperature;
[0031] d. Blocking: After the reaction is completed, use 3mL Na 2 HPO 4 (200mM, pH8.0) to wash, add 1mL Tris-HCl buffer (0.1M, pH8.0) to react at room temperature for 2h to block the remaining active sites;
[0032] e. Washing: alternately wash 5 times with 2mL Tris-HCl buffer (0.1M, pH8.0, containing 0.5M NaCl) and 2mL acetate buffer (0.1M, pH4.0, containing 0.5M NaCl);
[0033] f. Column ...
Embodiment 2
[0036] Embodiment 2: AAC purification UPLC-FLR detects OTA in ginger powder
[0037]1. Sample extraction: Accurately weigh 20.0g of ginger powder (accurate to 0.1g), add 60mL of acetonitrile-water (60:40, v / v) solution, ultrasonically extract for 15min, coarse filter with quantitative filter paper, take 5mL of filtrate and add 45mL BBS 2 (pH8.0), mix well, filter with glass fiber filter paper, and the filtrate is set aside.
[0038] 2. Purification: take AAC column and use 3mL BBS 1 For activation, pipette 3 mL (equivalent to 0.1 g sample) of the above-mentioned diluted filtrate to pass through the column, and adjust the flow rate to 1 drops / s. with 1mL BBS 1 Rinse the column until the air has completely passed through the column. Add 1 mL of pure methanol, elute at a flow rate of 1 drops / s, and collect the eluate. The eluate was blown dry with nitrogen at 45°C, and finally the volume was adjusted to 0.5 mL with methanol-water (50:50, v / v), shaken and mixed, centrifuged a...
Embodiment 3
[0074] Example 3: AAC purified UFLC-MS / MS detection of OTA in traditional Chinese medicine
[0075] 1. Sample extraction: Accurately weigh 2.0g of medicinal material powder (accurate to 0.1g), add 8mL of acetonitrile-water (60:40, v / v) solution, vortex at 2400rpm for 2min, let stand for 30min, and centrifuge at 4000rpm 10min, take 3mL filtrate and use BBS 2 Dilute to 30mL, mix well, and adjust the pH to 7.5 with hydrochloric acid, filter with glass fiber filter paper, and use the filtrate for later use.
[0076] 2. Purification: take AAC column and use 3mL BBS 1 For activation, pipette 2 mL (equivalent to 50 mg sample) of the above-mentioned diluted filtrate to pass through the column, and adjust the flow rate to 1 drops / s. with 1mL BBS 1 Rinse the column until the air has completely passed through the column. Add 1 mL of pure methanol, elute at a flow rate of 1 drops / s, and collect the eluate. Dry the eluent with nitrogen at 45°C, add 5 μL of internal standard working solu...
PUM
Property | Measurement | Unit |
---|---|---|
adsorption capacity | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com