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

Inactive Publication Date: 2018-09-14
CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The method of directly deproteinizing the sample and measuring the PDG content in serum by LC-MS / MS has not been reported yet

Method used

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  • Liquid chromatogram tandem mass spectrum measuring method for pregnanediol-3-glucuronic acid content in serum
  • Liquid chromatogram tandem mass spectrum measuring method for pregnanediol-3-glucuronic acid content in serum
  • Liquid chromatogram tandem mass spectrum measuring method for pregnanediol-3-glucuronic acid content in serum

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a liquid chromatogram tandem mass spectrum measuring method for pregnanediol-3-glucuronic acid content in serum. The method is characterized in that after deproteinization is performed on a serum sample, liquid chromatogram introduction-electrospray ionization-tandom mass spectrometer measurement are carried out, an external standard method is adopted to quantify analyte, and the pregnanediol-3-glucuronic acid content level in the serum can be quickly and accurately detected. The novel measuring method for the pregnanediol-3-glucuronic acid content in the serum is provided, the tandem mass spectrometry improves the selectivity and accuracy of the novel method, the quantitation limit of the pregnanediol-3-glucuronic acid is 0.05 ng / mL, the recovery rate of the methodis 95.5-99.4.0%, and RSD is less than 2.1%. Through the chromatographic column and selectivity and optimization of elution requirements, the chromatograph process is better improved, the chromatographic analysis time is shortened, and only 15 minutes are needed. The method has the advantages that the measuring result is accurate, the sensitivity is high, and measuring interference is little.

Description

Technical field: [0001] The invention belongs to the technical field of physical and chemical testing of blood, and specifically relates to a method for measuring the content of pregnadiol-3-glucuronic acid in serum by using liquid chromatography-electrospray ionization-tandem mass spectrometer. Background technique: [0002] Progesterone, secreted mainly by the corpus luteum and placenta, is the main naturally occurring human progesterone. Abnormal levels can affect reproduction, neurodevelopment and the immune system. Progesterone acts through the progesterone receptor, which belongs to the nuclear hormone receptor superfamily and is abnormally expressed in thyroid and breast cancers. Previous studies have shown that progesterone receptor is rarely expressed in medullary thyroid carcinoma, and immunohistochemical assessment of progesterone receptor expression can help predict the prognosis of patients with papillary thyroid carcinoma. Thyroid cancer (TCC) is a common end...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 胡清源陈建侯宏卫陈欢罗彦波何昀潞
Owner CHINA NAT TOBACCO QUALITY SUPERVISION & TEST CENT
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