A method for detecting total testosterone in human serum

Abstract

The invention discloses a method for detecting total testosterone in human serum. The method comprises the following steps: adding an internal standard solution containing testosterone internal standard into a test tube containing a human serum sample, and adding methyl tert-butyl ether after mixing Carry out protein precipitation and extract the test substance; after centrifugation, place it at -65~-80°C for 1-2h, then transfer the supernatant to another test tube for drying, and then add a reconstitution solvent to obtain the test sample; The test samples were detected by a high-throughput liquid chromatography tandem quadrupole mass spectrometer; the relative retention time of testosterone and the abundance ratio of the detected quantitative ion pairs were used as qualitative basis, and the internal standard standard curve method was used for quantification. The invention adopts a high-throughput liquid chromatography tandem mass spectrometer to detect the testosterone in human serum, and has short detection time, high flux, good sensitivity and high precision.

Description

technical field [0001] The invention relates to the technical field of analysis and detection, in particular to a method for detecting total testosterone in human serum by high-throughput liquid chromatography tandem mass spectrometry. Background technique [0002] Testosterone is a steroid hormone that is secreted by the testes in men or the ovaries in women, and a small amount is also secreted by the adrenal glands; but in postmenopausal women, testosterone secretion from the ovaries is significantly reduced. In males, it plays an important role in the development of external genitalia and secondary sex characteristics, the biochemical processes of which have not been fully elucidated. In women, its main role is as a precursor of estrogen. At the same time, testosterone may affect many body systems and functions, including: blood production, calcium balance in the body, bone mineralization, lipid metabolism, glucose metabolism and prostate growth. [0003] Most testoster...

AI Technical Summary

Problems solved by technology

[0005] Traditional testosterone detection methods include enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), chemiluminescence immunoassay (CLISA), automated immunoassay system, high performance liquid chromatography, and gas chromatography-mass spectrometry. Some testosterone detection methods mainly have the following problems: First, because testosterone is mainly combined with sex hormone-binding globulin (SHBG) in the human body, and there are a large number of high-affinity binding proteins in the human body, there is serious matrix interference in the detection

However, traditional immunological methods cannot effectively remove matrix interference; secondly, immunological detection methods are based on the recognition of antigen molecules by antibodies. Since there are a large number of compounds similar in structure to testosterone in the human body, the specificity of this detection technology Poor performance, easy to lead to false positives, especially when the concentration of testosterone is low, traditional immunological methods cannot be accurately measured; third, high-performance liquid chromatography and gas chromatography-mass spectrometry, complex pre-treatment, analysis time long and low sensitivity; fourth, the existing high performance liquid chromatography-tandem mass spectrometry and gas chromatography-mass spectrometry have a long detection process and low detection throughput

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