CN-119044383-B - Method for detecting content of multi-index chemical components in Phellinus linteus

Abstract

The invention provides a method for detecting the content of multi-index chemical components in Phellinus linteus, and relates to the technical field of analysis and detection. The method for simultaneously measuring the contents of multi-index chemical components (up to 14) in the phellinus linteus medicinal material is established based on HPLC-MS/MS, 70-100% lower alcohol is adopted as an extraction solvent, the HPLC-MS/MS detection shows that the response of chromatographic peaks is stronger, and after 0.08-0.12vol% formic acid is added into a mobile phase, the peak tailing phenomenon can be well improved, and the simultaneous measurement of the contents of the multi-index chemical components of the phellinus linteus Huang Yaocai is realized. The detection method provided by the invention is simple and convenient, high in efficiency, high in sensitivity and strong in specificity, has good precision, repeatability and stability, can reflect the content difference of the chemical components detected in the Phellinus linteus medicinal materials in different batches, and can provide a reference for quality evaluation of the Phellinus linteus medicinal materials.

Inventors

• HE JUN
• ZHAO PENG
• OUYANG HUIZI
• Zhu Yamang
• SHANG YE
• LV ZHENGUO

Assignees

• 天津中医药大学

Dates

Publication Date

20260508

Application Date

20240905

Claims (8)

1. 1. A method for detecting the content of multi-index chemical components in Phellinus linteus comprises the following steps: Carrying out ultrasonic extraction on Phellinus linteus powder to be detected by using an extracting agent to obtain a sample solution to be detected, wherein the extracting agent comprises lower alcohol or lower alcohol aqueous solution, the volume fraction of the lower alcohol in the extracting agent is 70-100%, and the lower alcohol is methanol; Performing high performance liquid chromatography tandem mass spectrometry detection on the sample solution to be detected to obtain a detection result of the content of multi-index chemical components in the Phellinus linteus; the multi-index chemical components comprise nobiletin, tangerines, phenylalanine, betulinic acid, protocatechuic aldehyde, hesperidin, milk trealine, caffeic acid, sang Huangfen A, osmyl ketone, amygdalin, salvianolic acid B and rutin; the high performance liquid chromatography tandem mass spectrometry detection comprises high performance liquid chromatography separation and mass spectrometry detection; The high performance liquid chromatography separation conditions comprise a CORTECS C 18 chromatographic column, 2.1X 50mm and 2.7 mu m, a 0.08-0.12 vol% formic acid aqueous solution of a mobile phase A, acetonitrile of a mobile phase B, and gradient elution, wherein the gradient elution is performed in the following steps of 0-2 min, the volume fraction of the mobile phase B is increased from 15% to 40%, 2-9 min, the volume fraction of the mobile phase B is increased from 40% to 75%, and 9-10 min, and the volume fraction of the mobile phase B is increased from 75% to 100%.
2. 2. The detection method according to claim 1, wherein the solid-to-liquid ratio of the Phellinus linteus powder to be detected to the extractant is 0.19-0.21 g:10mL.
3. 3. The detection method according to claim 1, wherein the power of ultrasonic extraction is 280-320 w, the frequency is 30-50 khz, and the time is 0.5-1.5 h.
4. 4. The detection method according to claim 1, wherein the column temperature of the high performance liquid chromatography separation is 28-32 ℃.
5. 5. The detection method according to claim 1, wherein the sample injection amount of the high performance liquid chromatography is 1.8-2.2 μl.
6. 6. The detection method according to claim 1, 4 or 5, wherein the mobile phase flow rate of the high performance liquid chromatography separation is 0.28-0.32 ml/min.
7. 7. The method according to claim 1, wherein the mass spectrum detection conditions comprise an ion source being an electrospray ion source, a detection mode being multi-reaction ion monitoring, a scanning mode being a positive and negative ion scanning mode, a gas temperature of 320 ℃, a gas flow rate of 10L/min, an atomizer pressure of 40psig, a fragmentation voltage of 66-219V, and a collision energy of 8-40V.
8. 8. The method according to claim 1 or 7, wherein the quantitative ion pairs of nobiletin, hesperetin, phenylalanine, betulinic acid, protocatechuic aldehyde, hesperidin, cow's milk base, caffeic acid, sang Huangfen a, dicranopterone, amygdalin, salvianolic acid B and naringin are 403.1/373.3、395.0/365.0、166.1/120.1、474.1/60.1、153.0/109.0、137.0/108.0、645.1/301.1、245.0/159.1、179.0/135.0、219.0/135.1、177.0/134.1、456.1/323.1、717.1/321.0 and 579.2/271.0 in order.

Description

Method for detecting content of multi-index chemical components in Phellinus linteus Technical Field The invention relates to the technical field of analysis and detection, in particular to a method for detecting the content of multi-index chemical components in Phellinus linteus. Background Phellinus linteus is dry fruiting body of Phellinus linteus Inonotus hispidus (Bull.) P.Karst of Phanerochaeteceae, has effects of promoting blood circulation, stopping bleeding, invigorating spleen, relieving diarrhea, invigorating liver yang, improving sleep, and tranquilizing mind, and can be used for treating stranguria, amenorrhea, spleen deficiency diarrhea, liver and kidney yin deficiency, insomnia and dreaminess. Sang Huanghua has complex chemical components, mainly comprises polysaccharides, phenols, flavonoids, terpenes and the like, and has remarkable functions of resisting oxidation, aging, tumor and inflammation and improving sleep. Because of the influence of factors such as geographic conditions, climate, environment and the like, the Phellinus linteus chemical components produced in different producing areas are various, the content of the same components is obviously different, and the problem of different quality is avoided. The related art discloses a method for processing Phellinus linteus by analyzing the content changes of three compounds of Phellinus linteus Huang Zhongyuan catechin, protocatechuic aldehyde and ergosterol by High Performance Liquid Chromatography (HPLC). Although the above related art relates to HPLC detection of the contents of Mulberry Huang Zhongyuan catechin, protocatechuic aldehyde and ergosterol, the detected components are small, and the complete quality control of Phellinus linteus is not possible. Therefore, the establishment of the efficient and accurate multi-component content determination method has important significance for quality evaluation of the Phellinus linteus. Disclosure of Invention In view of the above, the invention aims to provide a method for detecting the content of multi-index chemical components in Phellinus linteus. The detection method provided by the invention can simultaneously realize the accuracy and high-sensitivity detection of the content of the multi-index chemical components in the phellinus linteus, thereby realizing the quality monitoring of the phellinus linteus. In order to achieve the above object, the present invention provides the following technical solutions: the invention provides a method for detecting the content of multi-index chemical components in Phellinus linteus, which comprises the following steps: extracting Phellinus linteus powder to be detected by using an extracting agent to obtain a sample solution to be detected, wherein the extracting agent comprises lower alcohol or lower alcohol aqueous solution, and the volume fraction of the lower alcohol in the extracting agent is 70-100%; Performing high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) detection on the sample solution to be detected to obtain a detection result of the content of multi-index chemical components in the Phellinus linteus; The multi-index chemical components comprise at least 4 of nobiletin, tangerines, phenylalanine, betulinic acid, protocatechuic aldehyde, hesperidin, milk trealine, caffeic acid, sang Huangfen A, osmunda, amygdalin, salvianolic acid B and rutin; the high performance liquid chromatography tandem mass spectrometry detection comprises high performance liquid chromatography separation and mass spectrometry detection; The high performance liquid chromatography separation conditions comprise 0.08-0.12vol% formic acid aqueous solution of mobile phase A, acetonitrile of mobile phase B and gradient elution, wherein the gradient elution is performed in the following steps of 0-2 min, the volume fraction of mobile phase B is increased from 15% to 40%, 2-9 min, the volume fraction of mobile phase B is increased from 40% to 75%, and 9-10 min, and the volume fraction of mobile phase B is increased from 75% to 100%. Preferably, the lower alcohol comprises methanol and/or ethanol. Preferably, the solid-to-liquid ratio of the Phellinus linteus powder to be detected to the extractant is 0.19-0.21 g/10 mL. Preferably, the extraction comprises ultrasonic extraction. Preferably, the power of ultrasonic extraction is 280-320W, the frequency is 30-50 kHz, and the time is 0.5-1.5 h. Preferably, the chromatographic column used for the high performance liquid chromatography separation is a C 18 chromatographic column, and the column temperature is 28-32 ℃. Preferably, the sample injection amount of the high performance liquid chromatography separation is 1.8-2.2 mu L. Preferably, the flow rate of the mobile phase separated by the high performance liquid chromatography is 0.28-0.32 mL/min. Preferably, the mass spectrum detection conditions comprise an ion source being an electrospray ion source, a detection mode being multi-react