A characteristic peptide segment capable of distinguishing staghorn glue and deerskin glue and its detection method
A technology of characteristic peptide segment and detection method, which is applied in the field of deer-derived characteristic peptide segment and its detection to achieve the effects of strong specificity, simple operation and high sensitivity
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Embodiment 1
[0040] A deer-derived characteristic peptide, the characteristic peptide sequence has 3, as shown in Sequence Table 1:
[0041] Peptide 1:
[0042] Gly-Ser-Asp-Gly-Ser-Val-Gly-Pro-Val-Gly-Pro-Ala-Gly-Pro-Ile-Gly-Ser-Ala-Gly-Pro-Hyp-Gly-Phe-Hyp-Gly- Ala-Hyp-Gly-Pro-Lys;
[0043] Peptide 2:
[0044] Gly-Ser-Asp-Gly-Ser-Val-Gly-Pro-Val-Gly-Pro-Ala-Gly-Pro-Ile-Gly-Ser-Ala-Gly-Pro-Hyp-Gly-Phe-Pro-Gly- Ala-Hyp-Gly-Pro-Lys;
[0045] Peptide 3:
[0046] Gly-Asp-Asp-Gly-Ala-Thr-Gly-Ala-Ala-Gly-Pro-Hyp-Gly-Pro-Thr-Gly-Pro-Ala-Gly-Pro-Hyp-Gly-Phe-Pro-Gly- Ala-Val-Gly-Ala-Lys.
Embodiment 2
[0047] Embodiment 2 buckskin glue A 肽1 / A 肽3 、A 肽2 / A 肽3 Determination of
[0048] Take 10 batches of commercially available buckskin glue samples, 10mg for each batch, add 5ml of phosphate buffer, ultrasonically dissolve the samples completely, centrifuge at 12000rpm for 20min, take 150μl of the supernatant, put it in a 2ml centrifuge tube, and Dilute with 1ml of 50mM PBS, add 1% trypsin (w / v), shake well, enzymolyze at 37°C for 12 hours, add 60μl of 10% TFA solution after enzymolysis to stop the reaction, centrifuge at 12000rpm for 20min, and obtain the enzymolysis of buckhide glue medicinal material solution, stored at -20°C for later use.
[0049] The above-mentioned 10 batches of deerskin gum 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 a 1.7 μm C18 reversed-phase chromatographic column (2.1 μm ×100mm), flow ra...
Embodiment 3
[0056] Embodiment 3 antler glue A 肽1 / A 肽3 、A 肽2 / A 肽3 Determination of
[0057] Take 10 batches of antler samples, follow the method of preparing antler glue in the 2020 edition of "Chinese Pharmacopoeia" to obtain antler glue samples, take about 10 mg of each batch, add 5ml of phosphate buffer, ultrasonically dissolve the samples completely, centrifuge at 12000rpm for 20min, Take 150 μl of the supernatant, put it in a 2ml centrifuge tube, dilute it with 1ml 50mM PBS, add 1% trypsin (w / v), shake well, ultrasonically hydrolyze for 10min, add 60μl of 10% TFA solution after enzymolysis to terminate the reaction, Centrifuge at 12,000 rpm for 20 minutes to obtain an enzymatic hydrolysis solution of antler gum, and store it at -20°C for future use.
[0058] Inject 10 batches of deer antler gum enzymatic hydrolyzate into the LC-MS instrument, the injection volume is 1 μg, and the liquid phase conditions detected by the LC-MS instrument are: the chromatographic column is a 1.7 μm...
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