A method for establishing a protein fingerprint of fresh Cordyceps sinensis and its application
A technology of Cordyceps sinensis and fingerprints, which is applied in the direction of instruments, measuring devices, scientific instruments, etc., can solve the problems of cumbersome preparation and gel making process, poor repeatability of gel spectra, expensive instruments, etc., and achieve simple sample pretreatment and processing methods Effects of Gentle, Sample-Loss High-Throughput Analysis
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Embodiment 1
[0060] The preparation method of embodiment one need testing sample
[0061] (1) Method 1 for preparing the test solution
[0062] Weigh 1.00 g of Cordyceps sinensis sample, add 7 mL of ultrapure water according to the weight to volume ratio, and homogenize. The homogenization speed is 3000 rpm, the homogenization time is 10 min, and the temperature is controlled at 5 ° C. The samples that were homogenized until there were no obvious particles were taken out and extracted by ultrasonic. The extraction time was 40min, the ultrasonic power was 400W, and the temperature was controlled at 15°C. After that, the samples were placed in a -20°C refrigerator to freeze. The samples were then removed, thawed, and the homogenization and sonication steps repeated. The samples after ultrasonic extraction were centrifuged, and the centrifugation temperature was controlled at 4° C. to obtain a supernatant, which was freeze-dried. Add 1.1mL ultrapure water to the lyophilized sample to rediss...
Embodiment 2
[0071] Example 2 The establishment method of the protein fingerprint of Cordyceps sinensis
[0072] (1) The preparation method of need testing solution
[0073] Prepare 12 batches (S1-S12) of test solution samples according to preparation method 3 in Example 1.
[0074] (2) Chromatographic analysis conditions: high performance liquid chromatography column Welch Xtimate C4 (4.6 × 250mm, 3μm), the filler of the chromatographic column is tetraalkylsilane bonded silica gel, gradient elution is adopted, and the mobile phase A is 0.1% formic acid - aqueous solution, the mobile phase B is 0.1% formic acid-acetonitrile solution, the flow rate is 0.6 mL / min, the ultraviolet detection wavelength is 214 nm, the column temperature is 30° C., and the injection volume is 20 μL.
[0075] The gradient elution conditions are:
[0076] 0min→10min: 18%B→21%B;
[0077] 10min→40min: 21%B→30%B;
[0078] 40min→50min: 30%B→31.5%B;
[0079] 50min→70min: 31.5%B→33%B;
[0080] 70min→100min: 33%B→3...
Embodiment 3
[0089] Example 3 Application of protein fingerprints in identifying Chinese medicinal materials of Cordyceps
[0090] (1) Preparation of the test solution: prepare 13 batches of S11-S25 test solution according to the preparation method 3 in Example 1.
[0091] (2) Chromatographic analysis conditions: the same as in Example two.
[0092] (3) Mass spectrometry analysis conditions: the same as in Example 2.
[0093] (4) Determination: Draw 20 μL of the test solution and inject it into the instrument, and perform the determination according to the above method to obtain the protein fingerprint.
[0094] (5) Results: if image 3The above are the protein fingerprints of 13 batches (S13-S25) of counterfeit Cordyceps sinensis and Cordyceps fermented products. According to the same method, the protein fingerprints of 13 batches of counterfeit Cordyceps sinensis and fermented products were evaluated for similarity, and the results were imported into the "Similarity Evaluation System ...
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