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Assay kit for detecting 9 fat-soluble vitamins in blood serum by UPLC

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

Active Publication Date: 2014-11-19
SHANGHAI DIAN CLINICAL TESTING CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no relevant reports on the methods and kits for simultaneous detection of these nine substances.

Method used

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  • Assay kit for detecting 9 fat-soluble vitamins in blood serum by UPLC
  • Assay kit for detecting 9 fat-soluble vitamins in blood serum by UPLC
  • Assay kit for detecting 9 fat-soluble vitamins in blood serum by UPLC

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] The components in the analysis kit are shown in Table 5.

[0072] Table 5 Preparation of 9 kinds of vitamin analysis kit components

[0073]

[0074] The preparation method of above-mentioned kit:

[0075] ,include

[0076] (1) Eluent:

[0077] Eluent A: prepare 0.1% v / v trifluoroacetic acid aqueous solution;

[0078] Eluent B: ethanol;

[0079] Eluent C: prepare 0.1% v / v trifluoroacetic acid methanol solution;

[0080] (2) Standard mother liquor:

[0081] Weigh 8.8 mg of vitamin A, 10 mg of δ-tocopherol, 10 mg of γ-tocopherol, and 9.11 mg of α-tocopherol, and add 1 mL of 2,6-di-tert-butyl-4-methylphenol containing 0.4 g / L Ethanol solution, weigh α-carotene 10mg, β-carotene 17.12mg (purity 99.9%), lycopene 10mg (purity 90%), coenzyme Q 105mg (purity 98%), lutein 20mg (purity 98%) ) in 2mL, 10mL, 1mL, 1mL, and 10mL of n-hexane, respectively, to obtain their respective standard product mother solutions; respectively pipette 40 μL, 80μL, 40μL, and 20μL standard ...

Embodiment 2

[0089] The above-mentioned 9 kinds of fat-soluble vitamins in the pretreated serum are detected by ultra-high performance liquid chromatography, and the content of the above-mentioned 9 kinds of fat-soluble vitamins is calculated by comparing the response value of the sample with the standard product. The specific chromatographic conditions are:

[0090] (1) Column model: BEH C18 1.7μm*2.1μm*50mm;

[0091] (2) mobile phase:

[0092] Mobile phase A: eluent A;

[0093] Mobile phase B: eluent B;

[0094] Mobile phase C: eluent C;

[0095] The mobile phase gradient is shown in Table 2, and the mobile phase changes uniformly according to the gradient retention time in Table 2;

[0096] (3) Flow rate: The initial flow rate is 0.2mL / min, and it rises to 0.3mL / min when the constant speed is increased to 4.51 minutes, then decreases at a constant speed at 8.51 minutes, and then drops to 0.2mL / min at 8.6 minutes;

[0097] (4) Column temperature: 30°C;

[0098] (5) Use an adjustable...

Embodiment 3

[0103] Embodiment 3: Performance verification.

[0104] 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 2 .

[0105] Table 6 Results of intra-batch precision test

[0106]

[0107] Table 7 Results of batch-to-batch precision test

[0108]

[0109] 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 image 3 .

[0110] 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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Abstract

The invention discloses an assay kit for detecting 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. The kit comprises the following reagents: an eluent A, an eluent B, an eluent C, standard substance mother liquor, a diluent 1, a diluent 2, an extract liquor and control materials QC (L), QC (M) and QC (H). Compared with the prior art, the kit disclosed by the invention has the advantages that the kit disclosed by the invention is suitable for an analysis method for detecting the 9 fat-soluble vitamins in the blood serum by the UPLC; the method can save a sample, time and cost.

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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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 徐凤仙于嘉屏
Owner SHANGHAI DIAN CLINICAL TESTING CENT
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