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Kit and detection method for accurately determining concentrations of vitamin A and vitamin E in human serum

A technology for accurate determination of human serum, applied in the field of medical testing, can solve the problems of large amount of serum required, expensive solid-phase extraction cartridges, and many organic solvents, and achieves simple processing method, short detection time, and reagent consumption. small amount of effect

Inactive Publication Date: 2020-11-13
NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently reported reverse-phase HPLC assays mostly use solid-phase extraction, liquid-liquid extraction, cloud point extraction, dispersed liquid-phase microextraction, etc., using solid-phase extraction technology to process blood samples can remove more impurities, but solid phase extraction Phase extraction cartridges are relatively expensive, and the cost for routine clinical inspection projects is high, while other pretreatment techniques are cumbersome and time-consuming, require more organic solvents, and require a large amount of serum. During the entire sample analysis process, pretreatment The time required for the process accounts for 2 / 3 of the entire analysis time, and may produce more than 1 / 3 of the error
Moreover, liquid-phase clinical testing methods are all methods established by laboratories themselves, and most of the reagents used are non-kit products. Therefore, there is an urgent need for commercial liquid-phase detection kits

Method used

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  • Kit and detection method for accurately determining concentrations of vitamin A and vitamin E in human serum
  • Kit and detection method for accurately determining concentrations of vitamin A and vitamin E in human serum
  • Kit and detection method for accurately determining concentrations of vitamin A and vitamin E in human serum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] 1. Calibrator mother solution: use absolute ethanol containing 2,6-di-tert-butyl-4-methylphenol (BHT) as the diluent, the concentration of BHT in the diluent is 0.5mg / mL, and then prepare 6 The concentration point of the calibrator mother solution, the six concentration points of the calibrator mother solution are as follows:

[0066] Retinol: 0.25μg / mL, 0.50μg / mL, 1.00μg / mL, 2.50μg / mL, 7.50μg / mL, 15.00μg / mL;

[0067] ±α-tocopherol: 5.00μg / mL, 10.00μg / mL, 20.00μg / mL, 50.00μg / mL, 150.00μg / mL, 300.00μg / mL;

[0068] The mother liquor of the above-mentioned calibrator is a mixed solution of retinol and ±α-tocopherol (that is, the mother liquor contains both retinol and ±α-tocopherol, and contains 2,6-di-tert-butyl-4-methylphenol (BHT) The absolute ethanol is the diluent); the concentration of the six points is the respective concentration of each component of retinol and ±α-tocopherol in the total mixed solution; the concentration of the calibrator working solution = the c...

Embodiment 2

[0094]1. Calibrator mother solution: use absolute ethanol containing 2,6-di-tert-butyl-4-methylphenol (BHT) as the diluent, the concentration of BHT is 0.5mg / mL, and the preparation includes calibration of 6 concentration points The 6 concentration points of the master solution of the product and the mother solution of the calibration product are as follows:

[0095] Retinol: 0.25μg / mL, 0.50μg / mL, 1.00μg / mL, 2.50μg / mL, 7.50μg / mL, 15.00μg / mL;

[0096] ±α-tocopherol: 5.00μg / mL, 10.00μg / mL, 20.00μg / mL, 50.00μg / mL, 150.00μg / mL, 300.00μg / mL;

[0097] The mother solution of the above-mentioned calibrator is a mixed solution of retinol and ±α-tocopherol; the concentration of the six points is the concentration of each component in the total mixed solution; the concentration of the working solution of the calibrator = the concentration of the mother liquor of the calibrator × 20 / 100 ;

[0098] 2. Internal standard: a mixed solution of retinyl acetate and vitamin E acetate, using abs...

Embodiment 3

[0106] 1. Calibrator mother solution: use absolute ethanol containing 2,6-di-tert-butyl-4-methylphenol (BHT) as the diluent, the concentration of BHT is 0.5mg / mL, and the preparation includes calibration of 6 concentration points The 6 concentration points of the master solution of the product and the mother solution of the calibration product are as follows:

[0107] Retinol: 0.25μg / mL, 0.50μg / mL, 1.00μg / mL, 2.50μg / mL, 7.50μg / mL, 15.00μg / mL;

[0108] ±α-tocopherol: 5.00μg / mL, 10.00μg / mL, 20.00μg / mL, 50.00μg / mL, 150.00μg / mL, 300.00μg / mL;

[0109] The mother solution of the above-mentioned calibrator is a mixed solution of retinol and ±α-tocopherol; the concentration of the six points is the concentration of each component in the total mixed solution; the concentration of the working solution of the calibrator = the concentration of the mother liquor of the calibrator × 20 / 100 ;

[0110] 2. Internal standard: a mixed solution of retinyl acetate and vitamin E acetate, using ab...

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Abstract

The invention relates to a kit and a detection method for accurately determining concentrations of vitamin A and vitamin E in human serum. The kit comprises a calibration material mother liquor, an internal standard, quality control materials, a diluent, an extraction liquid, a reconstitution fluid, and mobile phases. The calibration material mother liquor contains 2, 6-di-tert-butyl-4-methylphenol absolute ethyl alcohol as a diluent, the concentration of 2, 6-di-tert-butyl-4-methylphenol is 0.1-1.0 mg / mL, and the calibration material mother liquor has six concentration points; an internal standard solution is a mixed solution of retinol acetate and vitamin E acetate; the quality control materials comprise a quality control material 1 and a quality control material 2; the diluent is a blank matrix solution of bovine protein serum, and the mass percentage content of the bovine protein serum is 1-10%; the extraction liquid is an n-hexane solution containing 2, 6-di-tert-butyl-4-methylphenol; the reconstitution fluid is a methanol aqueous solution with the volume percent of 93%-100%; and a mobile phase A is an aqueous solution, a mobile phase B is a methanol solution, and the mobile phase A and the mobile phase B contain formic acid with the volume percentage concentration of 0-0.2%. The kit has the advantages of high accuracy, short detection time, low reagent consumption, convenient operation and low cost.

Description

technical field [0001] The invention relates to the technical field of medical testing, in particular to a kit and a detection method for accurately measuring the concentrations of vitamin A and E in human serum. Background technique [0002] Vitamin A and vitamin E are fat-soluble vitamins necessary for the human body to maintain normal metabolism and functions. They cannot be synthesized by the human body and must be ingested from food. Vitamin A is mainly found in animal liver, blood and retina of the eyeball, also known as retinol, which has the functions of maintaining normal vision, regulating human growth and development, and increasing body immunity. Cause respiratory, digestive tract and other infectious diseases and eye diseases and other diseases. Vitamin E plays an important role in anti-oxidation, anti-free radicals, improving immune function, anti-aging, and anti-cancer; it can promote the secretion of sex hormones, increase the vitality and quantity of sperm ...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/74
CPCG01N30/06G01N30/74
Inventor 邹继华沈敏潘清清杨晓东屠敏敏邹炳德
Owner NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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