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.
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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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