Solid phase extraction method for various steroid hormones in saliva and method for detecting steroid hormones

A technology of steroid hormones and detection methods, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of poor anti-interference ability of enzyme-linked immunoassay, easy cross-reaction, and large detection interference, etc., to achieve the removal of impurity interference and environment Effects with low impact and short analysis time

Inactive Publication Date: 2019-10-29
深圳健科医学检验实验室
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0005] 1. In addition to extracting steroid hormones, the liquid-liquid extraction method will also extract other substances similar in polarity to the extraction solvent, resulting in many impurities and large detection interference;
[0006] 2. The preparation of nano-h

Method used

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  • Solid phase extraction method for various steroid hormones in saliva and method for detecting steroid hormones
  • Solid phase extraction method for various steroid hormones in saliva and method for detecting steroid hormones
  • Solid phase extraction method for various steroid hormones in saliva and method for detecting steroid hormones

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Embodiment 1

[0076] refer to figure 1 As shown, the method of the present invention is used to carry out pretreatment and tandem mass spectrometry detection on saliva samples and standard music. Such as figure 2 As shown, hydrocorticosterone, testosterone and dehydroepiandrosterone have good peak shape and separation, and the interference of impurities is small, which meets the requirements of quantitative analysis.

Embodiment 2

[0078] Methodology verification linearity experiments were carried out with the method of the present invention. Prepare a mixed standard solution of cortisol, testosterone and dehydroepiandrosterone according to the steps in Table 1, and use the internal standard method for linear regression analysis to calculate the correlation coefficient R2. A linear regression curve such as image 3 , Figure 4 , Figure 5 As shown, the correlation coefficients are all greater than 0.999, indicating that the peak area is proportional to the standard concentration, and the linearity is good.

Embodiment 3

[0080] According to the method of the present invention, samples were prepared for accuracy and precision experiments in the following manner.

[0081] Spike low, medium and high concentrations of standards in saliva samples of known steroid hormone content (cortisolone: ​​0.941ng / mL, testosterone: 5.990ng / mL, dehydroepiandrosterone: 0.218ng / mL) , each concentration prepares 6 parts, and detects its response value, calculates recovery rate and coefficient of variation (CV), the accuracy and the precision of evaluation method, experimental result is shown in Table 5, the result shows that low, medium and high concentration steroid hormone The recovery rate is 90%-104%, and the CV is within 8%, indicating that the method has high accuracy and precision.

[0082] Table 5 Accuracy and precision results

[0083]

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Abstract

The invention discloses a solid phase extraction method for various steroid hormones in saliva. The method comprises activating a solid phase extraction column; adding the solid phase extraction column after a sample is processed; rinsing and eluting the solid phase extraction column, and collecting an eluent; adding an organic solvent and uniformly stirring the mixture, performing a centrifugal treatment at room temperature, and then transferring a product to a 96-orifice plate, sealing a film for a test. The solid phase extraction not only removes impurities, but also greatly improves the detection sensitivity and meets the detection requirement of trace steroid hormones in saliva. Compared with other pretreatment methods, the method is easy to operate, saves the solvent, has little impact on the environment, has a short analysis time and high flux, and can complete the test within 7 minutes. The method uses the isotope internal standard method to quantify the steroid hormones at a low detection error. It is verified by a comprehensive methodology that the method can be used for the detection of hydrogenated corticosterone, testosterone and dehydroepiandrosterone in saliva samples and screening of related diseases. The invention also discloses a method for detecting various steroid hormones after solid phase extraction of saliva.

Description

technical field [0001] The invention relates to the fields of steroid hormone solid-phase extraction and instrumental analysis mass spectrometry detection, in particular to a solid-phase extraction method of various steroid hormones in saliva and a detection method of the steroid hormones. Background technique [0002] With the rapid development of social economy, people's pace of life is accelerated, and the pressure is increased, which affects the secretion of corticosterone, testosterone, dehydroepiandrosterone and other steroid hormones in the body, and anxiety, depression, fear, sexual disorders, Infertility, premenstrual syndrome, sleep disturbance, cardiovascular disease and other diseases. Serum, urine and saliva are commonly used samples for testing human hormones. Since blood drawing itself is a source of acute stress, it may cause an increase in related steroid hormones, and most steroid hormones are combined with serum proteins and have no biological activity. Th...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/062
Inventor 田瑶林张良滔陈婉婷
Owner 深圳健科医学检验实验室
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