Method and kit for simultaneously detecting multiple steroid hormones in blood sample

A technology for steroid hormones and blood samples, applied in the field of a method and kit for simultaneous detection of multiple steroid hormones in blood samples, can solve the problems of a large amount of blood required for detection, affecting the detection results, and complicated operations, and achieve the results. Reliable, simple sample handling, high throughput results

Active Publication Date: 2020-05-19
JINAN YING SHENG BIOTECH
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Problems solved by technology

At present, due to the complex components of plasma/serum and the low content of hormones, impurities seriously affect the accuracy of test results during quantitative analysis. Therefore, in order to reduce the matrix effect during detection, HPLC-MS/MS can only be used for one or The inventors found that the qualitative analysis of the two hormon

Method used

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  • Method and kit for simultaneously detecting multiple steroid hormones in blood sample
  • Method and kit for simultaneously detecting multiple steroid hormones in blood sample
  • Method and kit for simultaneously detecting multiple steroid hormones in blood sample

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Experimental program
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Embodiment

[0088] (1) Prepare the sample to be tested

[0089] Add methanol solution containing internal standard sodium bicarbonate to the serum sample, vortex and oscillate, load the sample on a 96-well sample processing plate (provided by Shandong Yingsheng Biotechnology Co., Ltd.), and positive pressure after elution with ethyl acetate or tert-butyl methyl ether, Obtain the treatment solution, dry it with nitrogen, redissolve the sample in 40% v / v methanol aqueous solution, and filter through a 96-hole filter plate (provided by Shandong Yingsheng Biotechnology Co., Ltd.) to obtain the sample to be tested; the serum, containing The volume ratio of internal standard sodium bicarbonate methanol solution, ethyl acetate or tert-butyl methyl ether is 20:5:70, and the volume ratio of serum to 40% methanol aqueous solution is 4:1.

[0090] (2) Ultra-high performance liquid chromatography-mass spectrometry detection

[0091] The sample to be tested enters the chromatographic column for separ...

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Abstract

The invention provides a method and a kit for simultaneously detecting multiple steroid hormones in a blood sample. According to the method, a biological sample is pretreated through methanol deproteinization combination of sodium bicarbonate and a solid-phase support extraction method, chromatographic separation and mass spectrometric detection are performed, a pair of qualitative ions and a pairof quantitative ions are selected for each hormone, and the relative retention time and the qualitative ion pair are used as qualitative basis to determine the content of the sodium bicarbonate in the biological sample. And making a standard curve for quantification by using a standard substance. Meanwhile, three levels of quality control products are applied to investigate the accuracy and effectiveness of the method, and detection result distortion is avoided. The purpose of simultaneously detecting eighteen hormones in a serum sample by applying an HPLC-MS/MS technology is achieved for thefirst time, the influence of interferents is reduced, the method is easy, convenient and rapid to operate, high in flux and low in cost, the steroid hormone level in a human body is effectively monitored in real time, a sufficient basis is provided for clinical disease diagnosis, and the method is worthy of popularization and application.

Description

technical field [0001] The invention belongs to the technical field of steroid hormone detection substance analysis, in particular to a method and a kit for simultaneously detecting multiple steroid hormones in blood samples. Background technique [0002] The information disclosed in the Background of the Invention is only intended to increase the understanding of the general background of the invention, and is not necessarily to be taken as an acknowledgment or any form of suggestion that the information constitutes the prior art that is already known to those skilled in the art. [0003] Steroid hormones, also known as steroid hormones, mainly include adrenocortical hormones and sex hormones. They are trace chemical messenger molecules produced by endocrine glands and some tissues with endocrine functions that regulate tissue, organ and cell functions. They also belong to cyclopentane in structure. Polyhydrophenanthrene derivatives can be converted from cholesterol. [00...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/88
CPCG01N30/06G01N30/88
Inventor 冯振景叶松弭兆元
Owner JINAN YING SHENG BIOTECH
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