A saliva or urine metabolic spectrum molecular weight calibrator kit and preparation method and use method thereof
A technology of molecular weight and calibrator, applied in the field of mass spectrometry, can solve the problems of uneven height, limited scope of application, complex composition of metabolic molecules, etc., and achieve the effects of convenient preparation, low cost, and convenient use
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Embodiment 1
[0068] Example 1 Extraction and Preparation of Metabolic Molecular Calibrators from Saliva Samples
[0069] Step 1, obtain a saliva sample, and remove its mucin, cell debris and bacteria through filtration, centrifugation and other steps to obtain a clear and transparent liquid;
[0070] Step 2, take 10-50 μL of clear and transparent saliva sample, add 1-5 times the volume of extraction solvent, undergo shaking incubation and centrifugation steps to precipitate proteins, and the upper layer is a solution containing metabolites;
[0071] In step 3, a solution containing metabolites is obtained from the sample in step 2, and the solvent thereof is evaporated to dryness to obtain a calibrator of metabolites.
[0072] Step 4, redissolving the metabolite calibrator in a dissolving solution to prepare a metabolite calibrator solution. As a preference, the dissolving solution of the metabolite calibrator extracted from the saliva sample is a mixed solvent of methanol, acetonitrile a...
Embodiment 2
[0073] Example 2 Extraction and Preparation of Metabolic Molecular Calibrators from Urine Samples
[0074] Step 1, obtain a urine sample, remove cell debris, impurities and high-concentration salts through filtration, dialysis, centrifugation, dilution and other steps to obtain a clear and transparent liquid;
[0075] Step 2, take 10-50 μL of clear and transparent urine sample, add 1-5 times the volume of extraction solvent, undergo shaking incubation and centrifugation steps to precipitate proteins, and the upper layer is a solution containing metabolites;
[0076] In step 3, a solution containing metabolites is obtained from the sample in step 2, and the solvent thereof is evaporated to dryness to obtain a calibrator of metabolites.
[0077] Step 4, redissolving the metabolite calibrator in a dissolving solution to prepare a metabolite calibrator solution. As a preference, the dissolving solution of the metabolite calibrator extracted from the urine sample is a mixed solven...
Embodiment 3
[0079] Example 3 Molecular Weight Determination of Metabolic Molecular Weight Calibrators and Acquisition of Molecular Weight Calibration List
[0080] Step 1: Add 100-1000 μL of dissolving solution to the metabolite calibrator, shake and redissolve to prepare a calibrator solution, dispense 10 μL / tube into a single tube, and store at -80°C when not in use;
[0081] Step 2, adding pure mixed metabolites composed of lactic acid, taurine, uric acid, palmitic acid and sodium tetraphenyl boron into the calibrator solution to form a molecular weight calibrator containing an internal standard;
[0082] Step 4: Spot the metabolite calibrator solution containing the internal standard on the nano-mass spectrometer chip, 0.5-2 μL / well, 10-20 sites of the molecular weight calibrator under the two detection modes, and dry naturally. For the specific structure of the nano mass spectrometer chip, please refer to the applicant's CN110530965A patent;
[0083] Step 5: Put the sample stage int...
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