Method for simultaneously determining free thyroid hormones T3, rT3, T4 and cortisol in saliva

A technology for thyroid hormone and cortisol, applied in the field of biochemical analysis, can solve the problems of inability to remove thyroid hormone, limit the scope of application, and cannot be used for accurate detection of free thyroid hormone, etc.

Active Publication Date: 2021-09-14
杭州凯莱谱精准医疗检测技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, free thyroid hormones T3, rT3, T4 and cortisol cannot be detected at the same time, and it is not suitable for saliva samples, which limits its application range
[0008] CN107576624B discloses a method for detecting cortisol in saliva, which removes salivary mucin in saliva by filtering with a composite microfiltration membrane, but this method cannot remove cortisol and thyroid hormone in the bound state in saliva, and residual The bound hormones will affect the detection results of free hormones, so it cannot be used to accurately detect the free thyroid hormones T3, rT3, T4 and cortisol in saliva
[0009] Due to the very low content of free thyroid hormones T3, rT3, T4 and cortisol in saliva, there is currently no simple and efficient detection and analysis method that can simultaneously and accurately detect free thyroid hormones T3, rT3, T4 and cortisol in saliva Content of 4 free hormones

Method used

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  • Method for simultaneously determining free thyroid hormones T3, rT3, T4 and cortisol in saliva
  • Method for simultaneously determining free thyroid hormones T3, rT3, T4 and cortisol in saliva
  • Method for simultaneously determining free thyroid hormones T3, rT3, T4 and cortisol in saliva

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Sample preparation, pretreatment and detection

[0049] The following preparations are based on the use of solid standard products. If a commercially purchased standard stock solution is used, the preparation method needs to be adjusted according to the actual concentration of the standard stock solution used.

[0050] 1. Sample preparation

[0051] 1. Preparation of standard series solutions

[0052] 1.1 Preparation of primary stock solution of four hormones

[0053] Preparation of primary stock solutions of thyroid hormones T3, T4, rT3 and cortisol: according to Table 1, accurately weigh an appropriate amount of standard products, add an appropriate volume of methanol, and prepare primary stock solutions of the four hormones.

[0054] Table 1. Preparation of primary stock solution

[0055]

[0056] 1.2 Preparation of secondary stock solution of four hormones

[0057] Appropriate volumes of the primary stock solutions of the four hormones were respecti...

Embodiment 2

[0135] The influence of embodiment 2 different ultrafiltration membranes on detection result

[0136] Since the ultrafiltration membrane has a certain adsorption effect on the free thyroid hormones T3, T4, rT3 and cortisol in the saliva sample, which affects the detection results, after the sample preparation is completed according to the steps in Example 1 in this example, a low-concentration mass is taken. Control (L) samples were pretreated according to the sample pretreatment steps in Example 1, wherein different ultrafiltration membranes were used for ultrafiltration centrifugation, and the effects of different ultrafiltration membranes on the free state of thyroid hormone T3 and T4 in saliva samples were investigated. , rT3 and cortisol adsorption, each group was repeated 3 times, and the average value was taken. The test results are shown in Table 20.

[0137] Table 20. The influence of different ultrafiltration membranes on the detection results

[0138]

[0139] I...

Embodiment 3

[0140] Embodiment 3 The impact of ultrafiltration membrane molecular weight limit value on detection result

[0141] After completing the sample preparation according to the steps in Example 1, take a low-concentration quality control (L) sample and perform sample pretreatment according to the sample pretreatment steps in Example 1, wherein in the ultrafiltration centrifugation process, select ultra The filter membrane is subjected to ultrafiltration and centrifugation. After the pretreatment is completed, the liquid chromatography tandem mass spectrometry conditions in Example 1 are used for detection, and the different molecular weight limits of the ultrafiltration membrane are investigated. The free thyroid hormones T3, T4, and rT3 in the saliva sample and the influence of the detection accuracy of cortisol, each group was repeated 3 times, and the average value was taken, and the detection results are shown in Table 21.

[0142] Table 21. The influence of different molecul...

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Abstract

The invention provides a method for simultaneously determining free thyroid hormones T3, T4, rT3 and cortisol in saliva, which comprises the following steps of performing ultrafiltration and centrifugation, selecting a filter membrane with a proper molecular weight cutoff limit range, removing protein and combined thyroid hormones and cortisol in the saliva, adding anti-adsorption reagents such as methanol according to a proper proportion, and overcoming the problem of serious adsorption in the detection process of free T3, rT3, T4 and cortisol in saliva. The four free thyroid hormones T3, rT3 and T4 and cortisol in saliva can be simultaneously and accurately detected at one time by utilizing a high performance liquid chromatography-tandem mass spectrometry analysis technology. The method has the advantages of high throughput, simplicity and convenience in operation, high treatment efficiency, low consumable cost, high accuracy and high sensitivity, and is convenient for clinical popularization and application.

Description

technical field [0001] The invention belongs to the technical field of biochemical analysis, in particular to a method for simultaneously measuring free thyroid hormones T3, rT3, T4 and cortisol in saliva. Background technique [0002] Thyroid hormone is an important endocrine hormone in the human body. The main function of thyroid hormone is to increase the metabolic rate of the body, increase oxygen consumption, promote growth and development, and promote the differentiation and maturation of the nervous system. Thyroid hormones include thyroxine (T4), triiodothyronine (T3), and trans-triiodothyronine (rT3). Thyroid hormone is an important marker for evaluating thyroid function, and it is the main hormone of the thyroid gland to target organs. The total concentration of thyroid hormone in serum reflects the state of thyroid secretion. [0003] Cortisol is an important glucocorticoid in the body, responsible for regulating mood, health, maintaining immune cells and inflamm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/88
CPCG01N30/02G01N30/88G01N2030/8813
Inventor 刘鹏云刘妍妍孔子青刘华芬
Owner 杭州凯莱谱精准医疗检测技术有限公司
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