Determination of nine fat-soluble vitamins in serum by ultra-high performance liquid chromatography
A technology of fat-soluble vitamins and ultra-high performance liquid phase, applied in the field of analysis and detection, to save time, cost, and samples
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Embodiment 1
[0042] 1. Materials
[0043] (1) Instruments: ultra-high performance liquid chromatography (UPLC, American Waters Company, H-class type); medical high-speed refrigerated centrifuge (Beijing Baiyang Medical Equipment Co., Ltd.); ultrapure water instrument (Chengdu Youpu Ultrapure Technology Co., Ltd.); multi-tube vortex mixer (Hangzhou Aosheng Instrument Co., Ltd.); fast mixer (Jiangyan Kangtai Medical Equipment Factory); nitrogen blowing instrument (MD200, Hangzhou Aosheng Instrument Co., Ltd.); adjustable Liquid container (Thermo Scientific 5-50 μL, 10-100 μL, 20-200 μL, 100-1000 μL); glassware, beaker, graduated cylinder, etc.
[0044] (2) Reagent consumables: methanol (HPLC grade, Tedia High Purity Solvent Company.Inc); ethanol (HPLC grade, Anaqua Chemicals Supply); n-hexane (HPLC grade, Scharlab S.L); trifluoroacetic acid (Sigma-Aldrich); Pure water; 2,6-di-tert-butyl-4-methylphenol (Sigma-Aldrich); BSA (Sigma-Aldrich); BEH-C18 chromatographic column (Waters Company);
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Embodiment 2
[0065] Embodiment 2: Performance verification.
[0066] 1. Separation spectrum of the standard product: 9 kinds of fat-soluble vitamins are completely separated, the peak shape is relatively symmetrical, and there is basically no interference from other peaks, indicating that good separation can be obtained under this condition. See the chromatogram figure 1 .
[0067] Table 4 Results of intra-assay precision test
[0068]
[0069] Table 5 Results of batch-to-batch precision test
[0070]
[0071] 2. Separation spectrum of serum samples: 9 kinds of fat-soluble vitamins in serum samples are completely separated, and there are some miscellaneous peaks, but they have no effect on the target peak. For the chromatograms corresponding to 9 kinds of fat-soluble vitamins in serum, see figure 2 .
[0072] 3. Precision test: 6 batches of normal human serum samples were taken and processed once, and the concentrations of 9 fat-soluble vitamins were quantitatively determined by ...
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