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Method for simultaneously detecting three cannabinol compounds in hemp with SPE-HPLC (solid phase extraction-high performance liquid chromatography)

A compound and cannabidiol technology, applied in the field of plant component detection, can solve the problems of time-consuming, labor-intensive, and capital-intensive detection, and achieve the effects of suitable analysis conditions, less solvent consumption, and good linear relationship.

Inactive Publication Date: 2018-01-16
DAQING BRANCH OF HEILONGJIANG ACAD OF SCI
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, most of these detection methods are for the detection of a single cannabinoid compound. When it is necessary to detect multiple cannabinoid compounds in hemp, it is necessary to use their own pretreatment technology and analysis method, which is time-consuming and labor-intensive, and consumes a lot of money.

Method used

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  • Method for simultaneously detecting three cannabinol compounds in hemp with SPE-HPLC (solid phase extraction-high performance liquid chromatography)
  • Method for simultaneously detecting three cannabinol compounds in hemp with SPE-HPLC (solid phase extraction-high performance liquid chromatography)
  • Method for simultaneously detecting three cannabinol compounds in hemp with SPE-HPLC (solid phase extraction-high performance liquid chromatography)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: A method of using SPE-HPLC to simultaneously detect three cannabinoid compounds in hemp according to the present invention has the following steps:

[0042] One, determine high performance liquid chromatography condition: described high performance liquid chromatography condition is:

[0043] C18 reverse phase column: 4.6×150mm, 5μm; column temperature: 40°C; detection wavelength: 220nm; elution method: isocratic elution; mobile phase: methanol: 0.1% formic acid = 80:20; flow rate: 1mL / min; Injection volume: 20 μL.

[0044] 2. Prepare mixed standard solution:

[0045] (1) Preparation of mixed standard mother solution: take 0.5mL of THC, CBN, and CBD standard solutions with a mass concentration of 1mg / mL in a 5mL volumetric flask and constant volume with methanol to prepare THC, CBN, and CBD with the same mass concentration. CBD mixed standard mother liquor;

[0046] (2) Preparation of mixed standard solution: take 50μL, 50μL, 40μL, 100μL, 200μL, 400μL, 60...

experiment example 1

[0056] Experimental example 1: Experiment of the adsorption rate of three cannabinol compounds in the sample by the solid phase column:

[0057] Get the solid-phase extraction pretreatment sample that embodiment 1 step 4 obtains as test sample, adopt the high performance liquid chromatography condition of embodiment 1 step 1 to detect after getting 20 μ L of membrane filtration, detection result is as follows Figure 6 As shown, measure the peak area of ​​each component in the sample, qualitatively with retention time, quantify according to the linear regression equation determined in step 3, calculate the concentration of three kinds of cannabinol compounds in the sample, calculate thus without solid phase extraction The contents of the three cannabinoids in the hemp samples were: CBD: 1781 μg / g; CBN: 984 μg / g; THC: 4693 μg / g.

[0058] The adsorption rate ω=(A-a)÷A×100% of the three cannabinol compounds on the solid-phase small column, wherein A is the content of the cannabin...

experiment example 2

[0063] Experimental example 2: the precision experiment of SPE-HPLC method of the present invention

[0064] Extract 1 part of hemp leaf sample, pretreat according to the method of Example 1 of the present invention and continuously inject the obtained test sample three times (N-1, N-2, N-3), and measure three kinds of cannabinoids The content data of the compound are shown in Table 2:

[0065] Table 2:

[0066]

[0067] It can be seen from Table 2 that the RSD values ​​of the precision experiments are all within 1%, indicating that the method of the present invention for simultaneously detecting three cannabinoid compounds in hemp by SPE-HPLC has good precision.

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Abstract

The invention belongs to the field of plant ingredient detection and particularly relates to a method for simultaneously detecting three cannabinol compounds in hemp with SPE-HPLC (solid phase extraction-high performance liquid chromatography). The SPE-HPLC is applied to detection of the cannabinol compounds in the hemp and can simultaneously detect the three cannabinol compounds in the hemp and perform qualitative and quantitative analysis on the compounds. According to the method, firstly, the HPLC conditions are determined, and a standard curve of the three cannabinol compounds is drawn anda linear regression equation is determined by detecting a mixed standard solution; a hemp sample is subjected to SPE for impurity removal, finally, high performance liquid detection is performed on the sample, and the content of the three cannabinol compounds in the hemp is obtained through calculation. To-be-detected cannabinol compounds and impurities are separated through SPE, influence of impurities on detection is reduced, and a good basis is provided for liquid detection. The detection method has a good linear relation in a corresponding application range and good precision, repeatability and accuracy.

Description

technical field [0001] The invention belongs to the field of detection methods of plant components, and in particular relates to a method for simultaneously detecting three cannabinoid compounds in hemp by using SPE-HPLC. Background technique [0002] As one of the oldest crops on earth, hemp has been used by humans as early as 10,000 years ago. Hemp bast fiber is one of the best natural fibers. It is longer than cotton fiber, has stronger toughness and moisture absorption, and has better heat insulation effect. It is a superior raw material for textile and papermaking. With the development of modern science and technology, the active ingredients in hemp have gradually attracted attention, among which cannabinoids have attracted more attention due to their large number and biologically active ingredients. Cannabinoids mainly include tetrahydrocannabinol (THC), cannabidiol (CBD), cannabinol (CBN) and cannabigerol (CBC). [0003] Due to the wide variety of cannabinol compoun...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/74
Inventor 张旭高宝昌田媛石雨崔宝玉
Owner DAQING BRANCH OF HEILONGJIANG ACAD OF SCI
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