Kit for detecting multiple steroid hormones in biological fluid and use method of kit
A technology for steroid hormones and biological fluids, applied in the fields of analytical chemistry and clinical testing, can solve the problems of low detection sensitivity, long detection time, and low detection efficiency, and achieve the effects of eliminating low sensitivity, high accuracy and easy operation.
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Embodiment 1
[0113] The present embodiment adopts kit of the present invention and using method thereof to treat 9 kinds of steroid hormones (cortisol, cortisone, 11-deoxycortisol, corticosterone, dehydroepiandrosterone, testosterone, 17-hydroxyprogesterone, androsne diketone and progesterone) were verified for accuracy.
[0114] 1. Kit and its composition
[0115] (1) Standard product working solution: prepare cortisol, cortisone, 11-deoxycortisol, corticosterone, dehydroepiandrosterone, testosterone, 17-hydroxyprogesterone, androstenedione and progesterone respectively with methanol Stock solution 1mg / mL, then dilute the above stock solution with 50% methanol solution into a standard working solution, a total of 5 concentration points, the concentration of steroid hormones at each point is as follows:
[0116] Cortisol 10ng / mL, 20ng / mL, 100ng / mL, 500ng / mL, 2500ng / mL;
[0117] Cortisone 2ng / mL, 4ng / mL, 20ng / mL, 100ng / mL, 500ng / mL;
[0118] 11-deoxycortisol 0.1ng / mL, 0.2ng / mL, 1ng / mL, 5...
Embodiment 2
[0178] In this embodiment, the kit and its usage method are the same as those in Embodiment 1, the difference is that this embodiment includes a pretreatment method for biological fluid samples.
[0179] In this embodiment, the contents of 9 steroid hormones in 13 plasma samples were detected.
[0180] Biological fluid samples were collected at the Seventh People's Hospital of Dalian, and all volunteers collected blood on an empty stomach between 7:00 and 8:00 in the morning. Whole blood was collected in vacuum blood collection tubes containing anticoagulants, centrifuged at 3000rpm for ten minutes, and the upper layer of plasma was collected and stored at -80°C for later use.
[0181] Pretreatment method for biological fluid samples: add 20 μL internal standard working solution to 200 μL plasma sample, add 1 mL liquid-liquid extractant, vortex for 3 minutes, centrifuge at 15000 rpm for 10 minutes, take the supernatant and dry it with nitrogen at 40 °C, add 90 50 μL of % acet...
Embodiment 3
[0186] In this embodiment, the kit and its usage method are the same as in Embodiment 2, except that the biological fluid sample is different, and the biological fluid sample in this embodiment is serum.
[0187] In this embodiment, the contents of 9 steroid hormones in 13 serum samples were detected.
[0188] Samples were collected at Dalian Central Hospital, and all volunteers collected blood on an empty stomach between 7:00 am and 8:00 am. Whole blood was collected in vacuum blood collection tubes without anticoagulant, centrifuged at 3000rpm for ten minutes after standing for coagulation, and the upper serum layer was collected and stored at -80°C for later use.
[0189] See Table 13 for the contents of 9 steroid hormones in the serum samples of 13 cases.
[0190] Table 1413 Serum Samples of 9 Steroid Hormone Contents
[0191]
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