Method for measuring 9 fat-soluble vitamins in blood serum by UPLC

A technology of fat-soluble vitamins and ultra-high performance liquid phase, applied in the field of analysis and detection, to save time, avoid consumption of organic reagents and equipment loss, and save costs

Active Publication Date: 2014-11-19
杭州凯莱谱精准医疗检测技术有限公司
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  • Description
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  • Method for measuring 9 fat-soluble vitamins in blood serum by UPLC
  • Method for measuring 9 fat-soluble vitamins in blood serum by UPLC
  • Method for measuring 9 fat-soluble vitamins in blood serum by UPLC

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Experimental program
Comparison scheme
Effect test

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);

...

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 e...

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Abstract

The invention discloses a method for measuring 9 fat-soluble vitamins in blood serum by UPLC (Ultra Performance Liquid Chromatography). The 9 fat-soluble vitamins respectively are vitamin A, alpha-tocopherol, gamma-tocopherol, delta-tocopherol, alpha-carotene, beta-carotene, a coenzyme Q10, lycopene and lutein. A UPLC instrument is adopted to detect the 9 fat-soluble vitamins in the preprocessed blood serum and by comparing a response value of a sample with a value of a standard substance, the content of the 9 fat-soluble vitamins is calculated. The method disclosed by the invention has the advantages that the sample is saved and 200 microliters of sample can implement analysis on the 9 fat-soluble vitamins; time is saved, nine target components with different optimal adsorption wavelengths are integrated into one scheme and separation analysis can be completed in 12.5 minutes; the cost is saved and consumption of a great amount of organic reagents and loss of an instrument are avoided.

Description

technical field [0001] The invention relates to a method for measuring nine kinds of fat-soluble vitamins in serum by ultra-high performance liquid chromatography, and belongs to the technical field of analysis and detection. Background technique [0002] Fat-soluble vitamins generally refer to a class of vitamins that are soluble in fat and fat solvents. The present invention mainly analyzes nine kinds of vitamins which are crucial to the normal growth, metabolism and function regulation of the human body, including vitamin A, α-tocopherol, γ-tocopherol, δ-tocopherol, α-carotene, β-carotene lutein, coenzyme Q10, lycopene and lutein. [0003] Among them, vitamin A is essential for good vision, normal development of embryos and babies, immune system function, growth, bone formation, reproduction and wound healing; α-tocopherol, γ-tocopherol, δ -Tocopherols are three common forms of vitamin E, they are highly active free radical scavengers, protect lipid membranes from oxida...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 徐凤仙于嘉屏
Owner 杭州凯莱谱精准医疗检测技术有限公司
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