Search

CN-122017082-A - Method for detecting 2-hydroxy naringenin in plant

CN122017082ACN 122017082 ACN122017082 ACN 122017082ACN-122017082-A

Abstract

The invention belongs to the technical field of analysis and detection, and particularly relates to a detection method of 2-hydroxy naringin in plants. The detection method comprises the steps of extracting a plant sample to obtain a liquid to be detected, detecting the liquid to be detected by adopting high performance liquid chromatography-tandem mass spectrometry to obtain the peak area of 2-hydroxy naringenin in the plant sample, and calculating the content of 2-hydroxy naringenin in the plant sample according to the peak area of 2-hydroxy naringenin and a preset standard curve, wherein the detection condition of the ultra-high performance liquid chromatography comprises that a mobile phase A is water, a mobile phase B is acetonitrile, and the ion source temperature of the ultra-high performance liquid chromatography is 500 ℃. The method has good linear relation (the linear relation is good (r is more than 0.999)), and has good precision and repeatability, and can accurately determine the content of the 2-hydroxy naringenin in plants.

Inventors

  • TANG CHAQIN
  • LIU HAORAN
  • DUAN LINGXIAO
  • LI FANG
  • LI YUANLI
  • HU RONG
  • XU YUQIN
  • MIAO HUIMIN
  • YANG XINGFU

Assignees

  • 江苏农林职业技术学院
  • 溧阳市天目湖茶叶研究所

Dates

Publication Date
20260512
Application Date
20260323

Claims (8)

  1. 1. The method for detecting the 2-hydroxy naringin in the plants is characterized by comprising the following steps of: extracting a plant sample to obtain a liquid to be detected; detecting the liquid to be detected by adopting ultra-high performance liquid chromatography-tandem mass spectrometry to obtain the peak area of 2-hydroxynaringenin in a plant sample; Calculating to obtain the content of 2-hydroxynaringenin in the plant sample according to the peak area of the 2-hydroxynaringenin and a preset standard curve; the detection conditions of the ultra-high performance liquid chromatography comprise that the mobile phase A is water, and the mobile phase B is acetonitrile; The ion source temperature of the ultra-high performance liquid chromatography is 500 ℃.
  2. 2. The method according to claim 1, wherein the flow rate of mobile phases A and B of the ultra-high performance liquid chromatography is 1mL/min, the chromatographic column is ATLANTIS PREMIER BEH Z-HILIC column, and the column temperature is 40 ℃.
  3. 3. The method according to claim 1, wherein the mass spectrometry conditions of the ultra performance liquid chromatograph include simultaneous use of positive ion and negative ion modes, ion spray voltage of +5500V/-4500V, gas curtain gas of 35 psi, ion source gas 1 of 50L/min, ion source gas 2 of 60L/min, collision gas of medium, scanning type of multi-reaction monitoring, inlet voltage of + -10V, collision cell outlet voltage of + -10V.
  4. 4. The method according to claim 1, wherein the gradient elution procedure is: 0-1 min, wherein the volume percentage of the mobile phase A is 10%; 1-4 min, wherein the volume percentage of the mobile phase A is increased from 10% to 25% at a constant speed; 4-4.1 min, wherein the volume percentage of the mobile phase A is increased from 25% to 50% at a constant speed; 4.1-5.5 min, wherein the volume percentage content of the mobile phase A is 50%; 5.5-5.6 min, wherein the volume percentage content of the mobile phase A is reduced from 50% to 10% at a constant speed; 5.6-7.5 min, wherein the volume percentage content of the mobile phase A is 10%.
  5. 5. The method according to claim 1, wherein the extractant for extraction is an aqueous methanol solution.
  6. 6. The method according to claim 5, wherein the aqueous methanol solution has a volume concentration of 70%.
  7. 7. The method according to claim 1, wherein the mass spectrum data of the 2-hydroxynaringenin is m/z 287.1,93 of the 2-hydroxynaringenin, the retention time is 1.2min, the declustering voltage is-50V, and the collision energy is-30 eV.
  8. 8. The method of claim 1, wherein the plant comprises aerial parts of arabidopsis and/or poplar tissue.

Description

Method for detecting 2-hydroxy naringenin in plant Technical Field The invention belongs to the technical field of analysis and detection, and particularly relates to a detection method of 2-hydroxy naringin in plants. Background Flavonoids are a class of natural polyphenol compounds widely existing in plants such as tea leaves, fruits and the like, and have health effects of resisting oxidation, resisting inflammation, protecting cardiovascular and the like, and almost all natural flavonoids exist in the plants in the form of O-glycoside or C-glycoside. 2-hydroxy naringenin (CAS number 58124-18-8) is a 2-hydroxy flavanone compound of flavonoids, and has molecular formula of C 15H12O6 and molecular weight of 288.25, which is produced by naringenin under the action of flavone-2-hydroxylase. 2-hydroxy naringenin is a key precursor for synthesizing vitexin (apigenin-8-C-glucoside, CAS number: 3681-93-4) and isovitexin (apigenin-6-C-glucoside, CAS number: 38953-85-4), and is an important substrate for the action of C-glycosyltransferase. However, 2-hydroxynaringenin has tautomerism (formula I) in the reaction system, and the chemical structure is unstable under acidic conditions, and the 2-hydroxyl group of the 2-hydroxynaringenin is easy to spontaneously remove and convert into apigenin (CAS: 520-36-5), so that the 2-hydroxynaringenin cannot be effectively used as a precursor of a C-glycosylation reaction. Moreover, the content in plants is usually extremely low and difficult to detect, and literature reports on detection and quantitative analysis thereof are rare; Formula I. Disclosure of Invention In view of the above, the invention provides a method for detecting 2-hydroxy naringin in plants, and the method provided by the invention can realize qualitative and accurate quantification of the 2-hydroxy naringin. In order to achieve the above object, the present invention provides the following technical solutions: the invention provides a method for detecting 2-hydroxy naringin in plants, which comprises the following steps: extracting a plant sample to obtain a liquid to be detected; detecting the liquid to be detected by adopting high performance liquid chromatography-tandem mass spectrometry to obtain the peak area of 2-hydroxynaringenin in a plant sample; Calculating to obtain the content of 2-hydroxynaringenin in the plant sample according to the peak area of the 2-hydroxynaringenin and a preset standard curve; the detection conditions of the ultra-high performance liquid chromatography comprise that the mobile phase A is water, and the mobile phase B is acetonitrile; The ion source temperature of the ultra-high performance liquid chromatography is 500 ℃. Preferably, the flow rate of mobile phases A and B of the ultra-high performance liquid chromatography is 1mL/min, the chromatographic column is ATLANTIS PREMIER BEH Z-HILIC column, and the column temperature is 40 ℃. Preferably, the mass spectrum conditions of the ultra-high performance liquid chromatography comprise the simultaneous use of positive ions and negative ions, an ion spray voltage of +5500V/-4500V, a gas curtain gas of 35psi, an ion source gas 1 of 50L/min, an ion source gas 2 of 60L/min, a collision gas of medium, a multi-reaction monitoring of a scanning type, an inlet voltage of +10V and an outlet voltage of a collision cell of +10V. Preferably, the gradient elution is performed by the following steps: 0-1 min, wherein the volume percentage of the mobile phase A is 10%; 1-4 min, wherein the volume percentage of the mobile phase A is increased from 10% to 25% at a constant speed; 4-4.1 min, wherein the volume percentage of the mobile phase A is increased from 25% to 50% at a constant speed; 4.1-5.5 min, wherein the volume percentage content of the mobile phase A is 50%; 5.5-5.6 min, wherein the volume percentage content of the mobile phase A is reduced from 50% to 10% at a constant speed; 5.6-7.5 min, wherein the volume percentage content of the mobile phase A is 10%. Preferably, the extractant for extraction is an aqueous methanol solution. Preferably, the aqueous methanol solution has a volume concentration of 70%. Preferably, the mass spectrum data of the 2-hydroxy naringenin is m/z 287.1,93 of the 2-hydroxy naringenin, the retention time is 1.2min, the declustering voltage is-50V, and the collision energy is-30 eV. Preferably, the plant comprises aerial parts of arabidopsis and/or poplar. The invention provides a method for detecting 2-hydroxy naringenin in a plant, which comprises the following steps of extracting a plant sample to obtain a liquid to be detected, detecting the liquid to be detected by adopting high performance liquid chromatography-tandem mass spectrometry to obtain the peak area of 2-hydroxy naringenin in the plant sample, and calculating the content of 2-hydroxy naringenin in the plant sample according to the peak area of 2-hydroxy naringenin and a preset standard curve, wherein the detection condition of the ultr