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Detection method of 17-hydroxyprogesterone and androstenedione

A detection method and androstenedione technology, applied in the field of analysis, can solve the problems of less sampling, difficulty, shortening the sample analysis time, etc., and achieve the effect of short time consumption, few steps, and improvement of liquid residue hanging on the wall

Active Publication Date: 2021-07-16
GUANGZHOU KINGMED DIAGNOSTICS CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The manual operation is simplified by the automated pipetting platform, which not only improves the efficiency and detection throughput, but also improves the reproducibility of the results, thereby realizing the accurate quantification of 17-hydroxyprogesterone and androstenedione, and the application of parallel liquid chromatography , which greatly shortens the sample analysis time, and the sample volume is small, which can solve the problem of difficult blood collection for children

Method used

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  • Detection method of 17-hydroxyprogesterone and androstenedione
  • Detection method of 17-hydroxyprogesterone and androstenedione
  • Detection method of 17-hydroxyprogesterone and androstenedione

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment 1

[0065] Instruments and equipment: AB Sciex API 3200MD for liquid mass spectrometry; American Hamilton eight-channel automatic sampling platform, American Agilent Bravo automatic pipetting platform.

[0066] 1. Method parameters

[0067] (1) Reagent configuration

[0068] Stock solution preparation: 17-OHP (17-hydroxyprogesterone) and A4 (androstenedione) stock solutions with a concentration of 1 mg / mL and 17-OHP-d with a concentration of 1 mg / mL were prepared with methanol 8 (2,2,4,6,6,21,21,21), A4-2,3,4- 13 Internal standard stock solution of C3;

[0069] Curve working solution preparation: 35% (v / v) methanol aqueous solution was used as a blank matrix to prepare a series of standard working curves of standards (17-OHP and A4) with a volumetric flask, and the concentration range of the curve was 17-OHP: 0.50-1000nmol / L, A4: 0.44-100 nmol / L.

[0070] Preparation of internal standard working solution: Dilute internal standard stock solution with 35% (v / v) methanol water t...

Embodiment 2

[0129] Embodiment 2 interference experiment

[0130] Mass spectrometry has the advantage of detecting multiple steroid hormone indicators at the same time. Most of the current methods focus on how to detect multiple steroid hormones in a short period of time. However, steroid hormones have similar results and close chemical properties. There are many kinds of steroid hormones in the human body. Isomers, the chromatographic peak times are often relatively close. When quickly detecting multiple steroid indicators, generally only mass spectrometry separation can be achieved, and chromatographic separation cannot be achieved, so that some structural analogs or isomers interfere. The risk of detection results, such as: the chromatographic peak baseline is too high, the chromatographic resolution is not good, etc., which leads to poor precision of the results and cannot achieve accurate quantification.

[0131] 17-OHP has three endogenous interference substances (isomers) in the hum...

Embodiment 3

[0134] The liquid phase condition of present embodiment is different from embodiment 1, and the liquid phase condition of present embodiment is:

[0135]

[0136] The detection result of present embodiment 3 is as Image 6 shown. Different from the chromatographic conditions in Example 1, the isomers of 17-hydroxyprogesterone can be separated by chromatography and mass spectrometry, and the separation degree is good. The liquid phase gradient conditions used in Example 3 cannot well realize the chromatographic separation of 17-hydroxyprogesterone and 21-hydroxyprogesterone, because 21-hydroxyprogesterone is an isomer of 17-hydroxyprogesterone, Mass spectrometry cannot be distinguished, and incomplete liquid phase separation will inevitably affect the precise quantification of 17-hydroxyprogesterone. At present, the main difference between the liquid phase gradient used in the method in Example 3 and that in Example 1 is that the initial Although the organic phase ratio of t...

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Abstract

The invention relates to a method for detecting 17-hydroxyprogesterone and androstenedione. The method comprises the following steps: preparing a to-be-detected sample solution, namely transferring a to-be-detected serum sample into a container by using an automatic sampling platform, adding an internal standard working solution into the container by using an automatic pipetting platform, mixing, adding an extracting solution into the container by using the automatic pipetting platform, mixing, centrifuging, transferring supernate into another container by using the automatic pipetting platform, and concentrating, after concentration, adding the reconstitution fluid by using the automatic pipetting platform, mixing, centrifuging, taking supernate as a sample solution to be detected, injecting the solution into a liquid chromatograph-mass spectrometer, and carrying out parallel liquid chromatography determination and mass spectrometry determination. According to the method, the 17-hydroxyprogesterone and the isomers or the structural analogues thereof in the serum are separated for the first time, the anti-interference capability is high, the result specificity is high, the accuracy is high, sample preparation is carried out through the automatic pipetting platform, the parallel liquid chromatography-mass spectrometer is combined to be applied to sample detection and analysis, the sample preparation time can be shortened by 5 times, the instrument analysis flux is doubled, and the method has the advantages of low sample consumption, wide linear range and the like.

Description

technical field [0001] The invention relates to the field of analysis, in particular to a method for detecting 17-hydroxyprogesterone and androstenedione. Background technique [0002] 17-Hydroxyprogesterone (17-OHP) is an endogenous progesterone produced by the adrenal glands and gonads. It is a specific indicator for screening congenital adrenal hyperplasia. It is currently mainly used for neonatal congenital adrenal hyperplasia. The primary and secondary screening of congenital adrenal hyperplasia (CAH). Serum 17-hydroxyprogesterone is the main diagnostic index of CAH. Combined with serum androstenedione, it can also be used to monitor the drug treatment effect of congenital adrenal hyperplasia. It is used to assist in the investigation of female hirsutism, acne, infertility, polycystic ovary syndrome, etc., and the growth and development monitoring of children with short stature and abnormal gonad development. [0003] Androstenedione (A4) is an androgen, which is mainl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/72
CPCG01N30/02G01N30/06G01N30/72G01N2030/065
Inventor 赵蓓蓓佘旭辉陈秀如董衡陈静宜韦兰清蔡舒婷
Owner GUANGZHOU KINGMED DIAGNOSTICS CENT
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