Solid-phase extraction method of steroid hormones in serum or plasma

A steroid hormone and extraction technology, which is used in measuring devices, instruments, scientific instruments, etc., can solve the problems that steroid hormones cannot completely contain the main hormones, the content of steroid hormones is low, and the content span is large, so as to reduce errors and reduce the use of reagents. The effect of reducing the amount and improving the detection sensitivity

Active Publication Date: 2020-12-08
北京金域医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The content of steroid hormones in the human body is low, the content spans a wide range, and is distributed from ng level to pg level, and there are many structural similarities, and even isomers exist; in addition, the matrix of serum or plasma is complex, so it is very difficult to It is difficult to efficiently and accurately extract steroid hormones from serum or plasma at the same time
At present, there are few reports on the extraction of steroid hormones from serum or plasma by solid phase extraction; and the extraction methods of steroid hormones reported can not completely include the main hormones of the entire steroid hormone metabolic pathway

Method used

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  • Solid-phase extraction method of steroid hormones in serum or plasma
  • Solid-phase extraction method of steroid hormones in serum or plasma
  • Solid-phase extraction method of steroid hormones in serum or plasma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Step 01: Take 100 μL serum sample and put it in the EP tube; add 10 μL internal standard working solution to the serum sample; add 100 μL pure methanol and mix to precipitate protein; then add water and mix for 1 min, adjust the volume ratio of the organic phase to the overall aqueous phase to 1 : 2; 4°C, 12000rpm, centrifuged for 5min to collect the supernatant to obtain the pretreated sample;

[0055] When preparing the internal standard working solution, take 20 μL of the internal standard, add 180 μL of water, vortex and mix for later use.

[0056] Step 02: Activate the wells of the solid phase extraction plate with 200 μL methanol and 200 μL ultrapure water in sequence;

[0057] Step 03: Add 250 μL of pretreated samples to the wells of the activated SPE plate;

[0058] Step 04: Add 200 μL of 10% acetonitrile aqueous solution (the first eluent) and 200 μL of n-hexane (the second eluent) to the wells of the solid phase extraction plate successively for rinsing to ma...

Embodiment 2

[0062] Step 01: Take 100 μL of plasma sample and put it in the EP tube; add 5 μL of internal standard working solution to the plasma sample; add 80 μL of pure methanol and mix to precipitate protein; then add water and mix for 1 min, adjust the volume ratio of the organic phase to the overall aqueous phase to 1 : 2; 4°C, 13000rpm, centrifuged for 5min to collect the supernatant to obtain the pretreated sample;

[0063] When preparing the internal standard working solution, take 20 μL of the internal standard, add 180 μL of water, vortex and mix for later use.

[0064] Step 02: Activate the wells of the solid phase extraction plate with 200 μL methanol and 200 μL ultrapure water in sequence;

[0065] Step 03: Add 250 μL of pretreated samples to the wells of the activated SPE plate;

[0066] Step 04: Add 200 μL of 10% acetonitrile aqueous solution (the first eluent) and 200 μL of n-hexane (the second eluent) to the wells of the solid phase extraction plate successively for rins...

Embodiment 3

[0070] Step 01: Take 100 μL serum sample and put it in the EP tube; add 15 μL internal standard working solution to the serum sample; add 120 μL pure methanol and mix to precipitate protein; then add water and mix for 1 min, adjust the volume ratio of the organic phase to the overall aqueous phase to 1 : 2; 4°C, 12000rpm, centrifuged for 5min to collect the supernatant to obtain the pretreated sample;

[0071] When preparing the internal standard working solution, take 20 μL of the internal standard, add 180 μL of water, vortex and mix for later use.

[0072] Step 02: Activate the wells of the solid phase extraction plate with 200 μL methanol and 200 μL ultrapure water in sequence;

[0073] Step 03: Add 250 μL of pretreated samples to the wells of the activated SPE plate;

[0074] Step 04: Add 200 μL of 10% acetonitrile aqueous solution (the first eluent) and 200 μL of n-hexane (the second eluent) to the wells of the solid phase extraction plate successively for rinsing to ma...

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Abstract

The invention discloses a solid-phase extraction method of steroid hormones in serum or plasma, which comprises the following steps: (1) adding an internal standard working solution into a serum or plasma sample, uniformly performing mixing and standing, adding methanol to precipitate proteins, adding water to regulate the volume ratio of an organic phase to a water phase to 1: 2, and performing centrifuging to obtain a pretreated sample; and (2) activating the solid-phase extraction plate holes, adding the sample pretreated in the step (1) into the activated solid-phase extraction plate holes, sequentially performing treating by using a leacheate and an eluent, collecting fractions treated by the eluent by using a collecting plate, adding water into the collecting plate holes, carrying out vortex uniform mixing, and sealing a film. By adopting the extraction method provided by the invention, 19 steroid hormones containing the whole steroid hormone metabolic pathway can be obtained atthe same time through one-time solid-phase extraction treatment.

Description

technical field [0001] The invention relates to the technical field of solid phase extraction of steroid hormones, in particular to a solid phase extraction method of steroid hormones in serum or plasma. Background technique [0002] Steroid hormones in the human body can be divided into adrenal cortex hormones and sex hormones. Steroid hormones are fat-soluble hormones, which are produced from cholesterol through a series of enzyme-catalyzed metabolism. They are all cyclopentane polyhydrophenanthrene derivatives in structure. Steroid hormones are divided into adrenocortical hormones and sex hormones. Adrenocorticoids include mineralocorticoids and glucocorticoids; sex hormones include progesterone, androgens, and estrogens. Steroid hormones regulate human metabolism through the endocrine system, and play an important role in the growth, development, reproduction and maintenance of internal environment stability. [0003] Steroid hormone detection can be used for screening...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/88
CPCG01N30/06G01N30/88G01N2030/045G01N2030/062G01N2030/8822
Inventor 孔许净胡滨陈宝荣任朋樊晓敏于蕾侯生根彭宝生
Owner 北京金域医学检验实验室有限公司
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