Liquid chromatogram tandem mass spectrum measuring method for pregnanediol-3-glucuronic acid content in serum
A glucuronic acid and liquid chromatography technology, applied in the field of blood physical and chemical testing, to achieve good repeatability, improve the chromatographic separation process, and improve the effect of specificity and sensitivity
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example 1
[0027] 1. Instruments and reagents: Agilent 1200 liquid chromatography (Agilent, USA), equipped with G1329A autosampler, G1311A quaternary mixing pump and G1316A, column oven; API 4000 triple quadrupole tandem mass spectrometer (Applied Biosystems, USA) ), equipped with electrospray ion source (ESI) and analyst 1.5 software data processing system.
[0028] The PDG standard was purchased from Sigma-Aldrich Company, with a purity greater than 96%; both formic acid and acetonitrile were chromatographically pure, and the water used in the experiment was deionized water.
[0029] 2. Sample processing: Serum was thawed at 4°C, centrifuged, then pipetted 100 µL of serum into a 1.5 mL centrifuge tube, added 300 µL of methanol, vortexed for 30 s, and centrifuged at 4°C for 15 min (15000× g ), pipette 200 µL of the supernatant into a chromatographic vial with an inner insert, and wait for LC-MS / MS analysis.
[0030] 3. Determination method: draw the standard solution (concentrations...
example 2
[0031] Example 2: According to the assay method described in Example 1, 8 serum samples were selected, and the PDG content was measured as shown in Table 2.
[0032] Table 2 Content levels of PDG in samples (ng / mL)
[0033] serial number
[0034] Among them, samples 1-4 are serum samples of healthy female volunteers, and samples 5-8 are serum samples of patients with thyroid cancer. Serum PDG levels in patients with thyroid cancer were lower than those in healthy volunteers. PDG may serve as a potential biomarker for thyroid cancer. In the next step, the sample size will be expanded, and the method will be used for detection, and the level range of PDG in the serum of healthy people and thyroid cancer patients will be established respectively.
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