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A method for simultaneously detecting vitamin D and vitamin K

A technology for vitamins and substances to be tested, applied in the field of detection, can solve the problems of unfavorable health of inspectors, long time, complicated operation process of vitamin D3 and vitamin K2, etc., and achieves high accuracy, high detection accuracy, and improved detection efficiency. Effect

Active Publication Date: 2022-02-18
BY HEALTH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In view of this, the present invention provides a method for simultaneous detection of vitamin D and vitamin K, which is used to solve the problems of cumbersome operation process, long time and unfavorable detection personnel's health in the prior art detection of vitamin D3 and vitamin K2

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  • A method for simultaneously detecting vitamin D and vitamin K
  • A method for simultaneously detecting vitamin D and vitamin K
  • A method for simultaneously detecting vitamin D and vitamin K

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] In this example, vitamin D3 stock solution and vitamin K2 stock solution were prepared.

[0046] Preparation of vitamin D3 stock solution: Accurately weigh 3.06 mg of vitamin D3 standard substance and place it in a 500ml brown volumetric flask, dissolve it with absolute ethanol, constant volume, and shake well to obtain a vitamin D3 stock solution, and measure the absorbance with a UV spectrophotometer , the measured concentration was 6.4091 μg / ml. Preparation of vitamin K2 stock solution: Accurately weigh 10.30 mg of vitamin K2 standard product and place it in a 100ml brown volumetric flask, dissolve it with ethyl acetate, constant volume, and shake well to obtain vitamin K2 stock solution, and measure the absorbance with a UV spectrophotometer , the measured concentration was 100 μg / ml. Both the vitamin D3 stock solution and the vitamin K2 stock solution were stored in a -18°C refrigerator for future use.

Embodiment 2

[0048] In this example, the standard working solution was prepared.

[0049]Precisely draw 10.00ml of vitamin D3 stock solution and 2.50ml of vitamin K2 stock solution into a 25ml brown volumetric flask, add absolute ethanol to the volume, and shake well to obtain the standard working solution. 5μl, 8μl, 10μl, under the following liquid chromatography conditions: the chromatographic column is Agilent C18 250×4.6mm, 5μm; the column temperature is 40°C; the detection wavelength is 264nm; the mobile phase includes mobile phase A and mobile phase B, mobile phase Phase A is 95% methanol-water solution, mobile phase B is isopropanol; mobile phase flow rate is 1mL / min; gradient elution is adopted, 0.01-20.00min, and the volume ratio of mobile phase A to mobile phase B is 98:2; 20.01~38.00min, the volume ratio of mobile phase A and mobile phase B is 50:50; 38.01~42.00min, the volume ratio of mobile phase A and mobile phase B is 0:100; 42.01~45.00min Mobile phase A and mobile phase Th...

Embodiment 3

[0051] Accurately weigh 1.1000g (1g~3g) of the substance to be tested, put it in a 10ml (or 25ml) brown volumetric flask, add n-hexane: absolute ethanol (20:80) mixed solution, and place it in a 60°C ultrasonic instrument Carry out ultrasonic extraction for 30 minutes, take it out, place it at room temperature, adjust the volume to the scale with a mixed solution of n-hexane: absolute ethanol (20:80), shake well and pass through a 0.45 μm organic filter membrane to obtain the test solution, and perform liquid chromatography detection The injection volume of the sample was 20 μl (10 μl~30 μl), and the conditions of the liquid chromatography were the same as those of the liquid chromatography of Example 2.

[0052] The content of vitamin D3 or vitamin K2 in the analyte is calculated according to the formula X=V×C / M×K.

[0053] In the formula: X is the content of vitamin D3 or vitamin K2 in the test object, the unit is μg / grain;

[0054] C is the concentration of vitamin D3 or v...

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Abstract

The invention belongs to the technical field of detection, in particular to a method for simultaneously detecting vitamin D and vitamin K. The invention provides a method for simultaneously detecting vitamin D and vitamin K, comprising the following steps: a) extracting vitamin D3 and vitamin K2 from the test object with a mixed solution of n-hexane and absolute ethanol, and b) using liquid chromatography The method is used to detect the vitamin D3 and vitamin K2 of the analyte, obtain the liquid chromatography data, and obtain the content of the vitamin D3 and vitamin K2 of the analyte according to the liquid chromatography standard working curve of the vitamin D3 and vitamin K2. Experimental results show that the method of the present invention can detect vitamin D3 and vitamin K2 at the same time, and the detection accuracy is high. The detection of vitamin D3 and vitamin K2 by the method of the present invention only takes 8.5 hours for each batch of samples, which greatly improves the detection efficiency.

Description

technical field [0001] The invention belongs to the technical field of detection, in particular to a method for simultaneously detecting vitamin D and vitamin K. Background technique [0002] At present, vitamin D and vitamin K are detected separately. For the detection of vitamin D3, the sample processing takes 4 hours, and the operation is complicated and cumbersome, including sample saponification, extraction, concentration, washing, constant volume and filtration, and the sample processing time is 6 hours. , it takes 10 hours to detect the vitamin D3 of a batch of samples; for the detection of vitamin K2, it takes 1.5 hours for sample processing, 7 hours for the sample on the machine, and 8.5 hours for the detection of vitamin K2 of a batch of samples; the vitamin D3 of a batch of samples is completed It takes a total of 18.5 hours for the detection of vitamin K2, which is not conducive to the efficient and rapid detection of products. Moreover, in the detection of vita...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/82
CPCG01N33/82
Inventor 刘光兰郑良陈宏壁
Owner BY HEALTH CO LTD
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