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Method for simultaneously detecting 11 steroid hormones in serum

A steroid hormone and serum technology, applied in the field of blood analysis, can solve the problems of low concentration of target compounds, large number of clinical samples, complex substrates, etc., and achieve the effect of improving sensitivity and reproducibility, good accuracy and short analysis time.

Pending Publication Date: 2020-09-29
浙江湖州三体生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In view of this, the present invention provides a method for simultaneously detecting 11 steroid hormones in serum. The detection method has good accuracy, high sensitivity, and good reproducibility, and can limit the number of clinical samples, complex substrates, and target compounds. Low concentration and other problems

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] A method for simultaneously detecting 11 steroid hormones in serum, comprising the following steps:

[0032] (1) Take 2.0ml of venous blood and put it in an inert separation gel to accelerate coagulation vacuum blood collection tube, add inert separation gel and coagulant, mix and centrifuge to obtain the serum to be tested;

[0033] (2) Take the internal standard solution of steroid hormones and add release agent A to dilute after reconstitution to obtain internal standard working solution. The release agent A includes acetonitrile and methanol, and the mass concentration of release agent A is 80-85%. , The volume ratio of release agent A is 1:200;

[0034] (3) Take 6 parts of the mother solution of steroid hormones of known concentration and add methanol to dilute to obtain standard curve solutions of different concentrations. The mass ratio of ketone, androstenedione, testosterone, 17-hydroxypregnenolone, 17-hydroxyprogesterone, progesterone, and dehydroepiandrosteron...

Embodiment 2

[0047] A method for simultaneously detecting 11 steroid hormones in serum, comprising the following steps:

[0048] (1) Take 2.0ml of venous blood and put it in an inert separation gel to accelerate coagulation vacuum blood collection tube, add inert separation gel and coagulant, mix and centrifuge to obtain the serum to be tested;

[0049] (2) Take the internal standard solution of steroid hormones and add release agent A to dilute after reconstitution to obtain internal standard working solution. The release agent A includes acetonitrile and methanol, and the mass concentration of release agent A is 80-85%. , The volume ratio of release agent A is 1:200;

[0050] (3) Take 12 parts of the mother solution of steroid hormones of known concentration and add methanol to dilute to obtain standard curve solutions of different concentrations. The mass ratio of ketone, androstenedione, testosterone, 17-hydroxypregnenolone, 17-hydroxyprogesterone, progesterone, and dehydroepiandroste...

Embodiment 3

[0061] The operation in this embodiment refers to Example 2, the difference is that the mother liquor is 10 parts, and 10 parts of standard curve samples can be prepared.

[0062] The invention can simultaneously detect steroid hormones in serum. First, protein precipitation was used for sample pretreatment, and the supernatant was collected by centrifugation for solid-phase extraction, and high-performance liquid chromatography combined with triple quadrupole tandem mass spectrometer was used for detection and analysis. The method can effectively remove the interference of matrix substances, can simultaneously identify and quantitatively analyze steroid hormones, has short detection time, high throughput, high detection sensitivity and good specificity.

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Abstract

The invention provides a method for simultaneously detecting 11 steroid hormones in serum, which relates to the technical field of blood analysis. The method comprises the steps of (1) centrifuging venous blood to obtain serum to be detected; (2) adding a releasing agent A into the internal standard solution to obtain an internal standard working solution; (3) adding methanol into the mother liquor for dilution to obtain a standard curve solution; (4) preparing a test solution: preparing a to-be-detected sample, a standard curve sample and a quality control sample; (5) mixing and centrifugingthe sample to be detected, the standard curve sample and the quality control sample; (6) balancing a solid-phase extraction plate; (7) putting the solution on a balanced solid-phase extraction plate;(8) leaching: putting methanol on the solid-phase extraction plate; (9) elution: collecting an eluent, adding methanol, and carrying out sample injection analysis; and (10) constructing a linear regression equation. The method can simultaneously detect steroid hormones in serum, can simultaneously identify and quantitatively analyze the steroid hormones, and is short in detection time, high in flux, high in detection sensitivity and good in specificity.

Description

technical field [0001] The invention relates to the technical field of blood analysis, in particular to a method for simultaneously detecting 11 steroid hormones in serum. Background technique [0002] Ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS / MS) integrates the super separation ability of ultra-high performance liquid chromatography and the characteristics of high specificity and high sensitivity of mass spectrometry. Through the separation of ultra-high performance liquid chromatography, the target compound It is separated from other interfering substances, and the tandem mass spectrometer is further used to selectively detect the ion signal of the target compound, and eliminate the ion signal of the interfering component, so that the detection result is more sensitive and accurate. [0003] Using ultra-high performance liquid chromatography tandem mass spectrometry to detect the concentration of steroid hormones in serum can not only a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/34G01N30/36G01N30/72G01N30/88G01N33/74
CPCG01N30/02G01N30/06G01N30/34G01N30/36G01N30/7233G01N30/88G01N33/743G01N2030/062G01N2030/8822G01N2560/00G01N2030/042
Inventor 薛雪陈虎诚陈效张桉瑜郑国艳
Owner 浙江湖州三体生物科技有限公司
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