Chemical derivatization-ultra-high performance liquid chromatography-tandem mass spectrometry method for simultaneously detecting 18 steroid hormones in serum
An ultra-high performance liquid phase and steroid hormone technology, which is applied in the field of chemical derivatization-ultra-high performance liquid chromatography-tandem mass spectrometry, and can solve the problems such as the inability to effectively improve the detection sensitivity of ESI-MS, the low quantitative coverage of hormones, and the low reactivity. , to achieve good ionization efficiency, shorten detection time, and increase detection throughput.
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Embodiment 1
[0067] The chemical derivation-ultra high performance liquid chromatography-tandem mass spectrometry method provided in this example simultaneously detects 18 steroid hormones, including the following steps:
[0068] 1. Materials
[0069] The experimental samples of the methodological study came from the serum samples of Wuhan Kangshengda Medical Laboratory Co., Ltd. in June 2012.
[0070] 2. Instruments and reagent consumables
[0071] Instruments: Shimadzu 8050 triple quadrupole mass spectrometer, Shimadzu 30A ultra-high performance liquid chromatograph, ultrapure water instrument (Millipore ultrapure water machine), electronic analytical balance (FA1004, Shanghai Sunny Hengping Scientific Instrument Co., Ltd.), High-speed refrigerated centrifuge (GL-20M high-speed refrigerated centrifuge), vortex mixer (US SCILOGEX company), GM-12 water bath nitrogen blowing instrument (Beijing Chengmeng Weiye Technology Co., Ltd.), pipette gun (Dalong Xingchuang Experimental Instrument C...
Embodiment 2
[0100] The chemical derivation-ultra-high performance liquid chromatography-tandem mass spectrometry method provided in this example simultaneously detects 18 steroid hormones, the steps of which are the same as those in Example 1, the only difference being:
[0101] The mobile phase gradient elution parameters are:
[0102]
[0103] In Example 2, compared with Example 1, the MRM mass spectrometry response signals and sensitivities of 18 steroid hormones decreased, specifically, the sensitivity (based on the lowest detection limit of quantification) decreased by 8.5-15.0%.
experiment example
[0108] The sensitivity comparative analysis of the detection methods of 18 steroid hormones in Example 1 and Comparative Example 1 was carried out.
[0109] The results of the sensitivity comparison experiment are shown in Table 3.
[0110] Table 3 Sensitivity comparison experiment results
[0111]
[0112] Comparative analysis of the sensitivity comparison experiment results of Example 1 and Comparative Example 1 in Table 1 shows that the sensitivity of chemical derivation-ultra high performance liquid chromatography-tandem mass spectrometry in Example 1 of the application to detect 18 steroid hormones is significantly improved. (Based on the lowest detection limit of quantitation) increased by 2-454 times.
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