Antler characteristic peptide fragment and detection method thereof
A technology of characteristic peptides and detection methods, which is applied in the field of characteristic peptides that can distinguish deerskin glue from deer antler glue and its detection, can solve the problem of high price, and achieve the effects of strong specificity, good technological progress, and simple operation
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0032] A deer-derived characteristic peptide, said characteristic peptide sequence has 2, as shown in Sequence Table 1:
[0033] Pep-G:
[0034] Gly-Phe-Hyp-Gly-Thr-Hyp-Gly-Leu-Hyp-Gly-Phe-Lys
[0035] Reference peptide (Pep-R):
[0036] Gly-Glu-Hyp-Gly-Glu-Hyp-Gly-Gln-Thr-Gly-Pro-Ala-Gly-Ala-Arg.
Embodiment 2
[0037] Embodiment 2 Determination of Pep-G and Pep-R
[0038] Take 5 batches of deerskin glue samples and 5 batches of deerskin glue samples, take 10 mg for each batch, add 5 ml of phosphate buffer solution, ultrasonically dissolve the samples completely, centrifuge at 12000 rpm for 20 min, and take 150 μl of supernatant , placed in a 2 ml centrifuge tube, diluted with 1 ml of 50 mM PBS, added 1% trypsin (w / v), shaken well, digested at 37°C for 12 hours, and then added 60 μl of 10% TFA solution to stop the reaction , Seppak C 18 Desalting treatment, centrifugal concentration and drying, reconstitution with pure water, determination of peptide concentration, centrifugal concentration and drying again, and storage at -20°C for future use.
[0039] Dion U3000 NanoRSLC nanoliter liquid phase system, the chromatographic column is 5 μm Reprosil C18AQ (75 μm × 150 mm), and the LC-MS / MS mass spectrometry system analyzes the sample. Reconstitute deerskin glue and deer antler glue enz...
Embodiment 3
[0043] Example 3 Deerskin glue and antler glue A G / A R Determination of
[0044] Take 10 batches of deerskin glue samples and 10 batches of deer horn glue samples, take 10 mg for each batch, add 5 ml of phosphate buffer, ultrasonically dissolve the samples completely, centrifuge at 12000 rpm for 20 min, and take 150 μl of supernatant , placed in a 2 ml centrifuge tube, diluted with 1 ml of 50 mM PBS, added 1% trypsin (w / v), shaken well, digested at 37°C for 12 hours, and then added 60 μl of 10% TFA solution to terminate the digestion React and centrifuge at 12,000 rpm for 20 min to obtain deerskin glue and deer antler glue enzymatic hydrolysis solution, which are stored at -20°C for later use.
[0045] The above 10 batches of deerskin glue and deer antler glue enzymatic hydrolysis solution were injected into the LC-MS instrument, the injection volume was 1 μg, and the liquid phase conditions detected by the LC-MS instrument were: the chromatographic column was 1.7 μm C18 re...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More