A method for identifying specific peptides of protein-containing traditional Chinese medicine
An exclusive, peptide technology, applied in the field of its processed products, can solve the problem of insufficient sample size, false positives, unable to find exclusive peptides, etc., to achieve the effect of strong scientificity and convenient operation
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Embodiment 1
[0046] 1. Identification of specific peptides in deerskin glue
[0047] (1) Sample preparation
[0048] Genuine product: different batches of buckskin glue;
[0049] Counterfeit products: different batches of donkey-hide gelatin, cowhide glue, pigskin glue, and horsehide glue.
[0050] Take 1.0mg of various gum powders, put them in a 1.5ml EP tube, add 50mM Tris buffer, pH=8.5, ultrasonically extract for 30 minutes, centrifuge at 16000g for 15 minutes, take the supernatant, add 10μg of trypsin, Enzymatic hydrolysis at 37°C for 12 hours, adding 10% TFA to terminate the enzymatic hydrolysis, centrifuging at 16000g for 15 minutes, and obtaining the precipitated sample after enzymatic hydrolysis;
[0051] (2) Then use Seppak C18 to desalt
[0052] The specific method is: first use acetonitrile (ACN) to activate Seppak C18 three times, each 1ml, 0.1% trifluoroacetic acid (TFA) solution balance 3 times, each 1ml, the sample after the enzymolysis, 0.1% TFA wash 3 times, 1ml each ...
Embodiment 2
[0062] Example 2 Identification of Specific Peptides in Angular Animal Medicines
[0063] (1) Sample preparation
[0064] Genuine product: different batches of buffalo horns;
[0065] Counterfeits: Different batches of goat horns, horse antlers, and pig's hooves.
[0066] Take 1.0 mg of various horn-like medicinal material powders, put them in a 1.5 ml EP tube, add 1 ml of 4% SDS Tris buffer to dissolve, ultrasonically extract for 30 minutes, centrifuge at 16,000 g for 15 minutes, add acetone for precipitation (final concentration of acetone is 80%) 4 After 1 hour, centrifuge at 16000g for 15 minutes, discard the supernatant, wash twice with acetone, centrifuge, evaporate the acetone to dryness, add Tris buffer solution containing 8M urea to dissolve, and then use Tris buffer solution to dilute the urea concentration in the solution to below 1M , add 10 μg of trypsin, hydrolyze at 37°C for 12 hours, add 10% TFA to stop the hydrolysis, centrifuge at 16,000 g for 15 minutes, a...
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Abstract
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