Camel milk characteristic peptide fragment combination and identification method
A technology of camel milk and peptides, applied in the field of proteomics, can solve problems such as difficulty in guaranteeing product quality and shortage, and achieve the effects of good accuracy, strong applicability, stability and accuracy
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Embodiment 1
[0104] Example 1 Screening of Camel Milk Characteristic Polypeptides.
[0105] Step 1. Pretreatment of camel milk standard samples
[0106] (1) Determination of total protein content of pure camel milk powder solution
[0107] Take the pure milk powder of dromedary camel and pure milk powder of bactrian camel respectively as standard samples of camel milk, mix pure camel milk powder with pure water at a concentration of 0.01g / mL, vortex and mix well, and shake at 37°C for 30 minutes to make it After fully dissolving, dilute 20 times to prepare a working solution, and measure the total protein content; use the Coomassie brilliant blue method (Bradford protein concentration determination kit) to determine the total protein concentration, with bovine serum albumin (BSA, 5mg / mL) as the standard For protein, prepare standard solutions of 50 μg / mL, 100 μg / mL, 200 μg / mL, 400 μg / mL, 500 μg / mL, 800 μg / mL, 1000 μg / mL with PBS diluent, and then detect the total protein content of the sa...
Embodiment 2
[0152] Example 2 Screening of parameters for quantitative detection of characteristic peptides and detection of actual samples and simulated adulterated milk powder
[0153] 1. Optimize the quantitative parameters of characteristic peptides
[0154] The quantitative characteristic peptides screened in Example 1 were tuned with the MRM mode of liquid chromatography-tandem mass spectrometry described in Step 3 (5) of Example 1 to optimize important parameters such as reaction ion pairs, collision energy, declustering voltage, and retention time , all quantitative characteristic peptide mass spectrometry parameters are shown in Table 3.
[0155] Specifically, Shimadzu’s EXION AC-LC system was used for chromatographic separation, and the chromatographic column was ACQUITY UPLC BEHC8 (2.1×100mm, 1.7μm); the column temperature was: 60°C; the flow rate was 0.4mL / min; the injection volume was 5μL ; Mobile phase A selects 0.2% formic acid water for use, and mobile phase B selects 0.2%...
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