Application of serum endogenous small molecules in evaluating differentiation degree of laryngeal cancer tumor cells
A tumor cell, endogenous technology, applied in the application field of serum endogenous small molecules in evaluating the differentiation degree of laryngeal cancer tumor cells, can solve the problem of lack of accurate quantitative evaluation methods for the degree of tumor differentiation, and achieve less damage to the body , low cost, enhanced objectivity and accuracy
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[0032] The screening method for biomarkers of laryngeal cancer comprises the following steps:
[0033] 1. Sample preparation:
[0034] Serum samples were thawed in ice-water mixture, took 100 μL and added 400 μL acetonitrile, shaken well, centrifuged at 13000 rpm for 20 min at 4 °C, took the supernatant and dried, added 100 μL 80% acetonitrile-water (v / v), vortexed 2min, centrifuge at 13000rpm for 20min at 4°C, take the supernatant for LC-MS analysis;
[0035] 2. LC-MS analysis test conditions:
[0036] Liquid phase conditions: mobile phase A, water containing 0.1% formic acid; mobile phase B, acetonitrile containing 0.1% formic acid. Gradient elution: 0-2min, 2% B; 2-13min, 2% B-35% B; 13-28min, 35% B-98% B; 28-30min, 98% B; 30-32min, 98% B-2%B; 32-34min, 2%B. Injection volume: 3μL; flow rate, 0.2mL / min; column temperature: 45°C; WatersACQUITYUPLC HSST3 column (2.1mm×100mm, 1.8μm).
[0037] Mass spectrometry conditions: HESI ionization method; spray voltage: positive ele...
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