Method for simultaneously detecting five kinds of steroid hormones in serum
A technology of steroid hormones and serum, applied in the field of analysis and detection, can solve the problems of high reagent cost, complicated processing, low detection limit, etc., and achieve the effects of strong anti-matrix interference, high detection sensitivity, and reduced cost of consumables
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Embodiment 1
[0080] The detection method of five steroid hormones of the present embodiment comprises the following steps:
[0081] 1. Sample pretreatment
[0082] Take 200 μL sample (standard curve point and quality control synchronous processing) in a 2.0 centrifuge tube, add 400 μL internal standard working solution ( 13 C3-testosterone 4mg / L, d4-cortisol 4mg / L, D5-11-deoxycortisol 16mg / L) and 13 C3-androstenedione stock solution (8 mg / L acetonitrile solution), high-speed shaking and mixing for 3 minutes; add 800uL tert-butyl methyl ether, high-speed shaking and mixing for 3 minutes; centrifuge at 4°C, 14000r for 10 minutes; take 1 mL of supernatant in 1.5 mL In the centrifuge tube, blow dry with nitrogen at 40°C; reconstitute 100uL of the reconstitution solution and shake at high speed for 3min; centrifuge at 10,000r for 5min at 4°C; transfer 80μL of the centrifuged sample to a sample bottle for detection on the machine.
[0083] 2. Enrichment, separation and detection
[0084] The ...
Embodiment 2
[0109] Embodiment 2 methodological verification experiment
[0110] In this example, methodological verification experiments are carried out for the detection methods of the five steroid hormones in Example 1.
[0111] 1. Precision experiment
[0112] 1.1 Intra-assay: The actual serum samples were spiked to the low, medium and high levels respectively, and the obtained samples were subjected to the intra-assay precision experiment; 20 samples of each concentration level were processed in parallel, and each Sample 1 time; calculate the mean value and relative standard deviation (RSD) of the test results.
[0113] 1.2 Inter-assay precision (Inter-assay): Take the pre-prepared low, medium and high levels of quality control products, and conduct inter-assay precision experiments; samples of each level are made 4 parallels, that is, 4 groups are detected The data were continuously measured for 5 days; the mean value and relative standard deviation (RSD) of the test results were c...
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