CN112305106B - American ginseng extraction analysis method - Google Patents
American ginseng extraction analysis method Download PDFInfo
- Publication number
- CN112305106B CN112305106B CN202011146938.6A CN202011146938A CN112305106B CN 112305106 B CN112305106 B CN 112305106B CN 202011146938 A CN202011146938 A CN 202011146938A CN 112305106 B CN112305106 B CN 112305106B
- Authority
- CN
- China
- Prior art keywords
- american ginseng
- water
- analyzing
- methanol
- ginsenoside
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/14—Preparation by elimination of some components
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The application relates to an American ginseng extraction analysis method, which comprises the steps of mixing American ginseng powder with methanol, carrying out vortex treatment, adding methyl tertiary butyl ether, carrying out vortex treatment, and finally adding water, carrying out vortex treatment, wherein the ratio of the American ginseng powder to the methanol to the methyl tertiary butyl ether to the water is 0.3g to 1mL to 5.8-6mL to 8-8.2mL; standing the mixture, centrifuging, and separating the mixture into upper lipid extract and lower ginsenoside extract; respectively taking lipid extract and ginsenoside extract, respectively freeze-drying, and respectively redissolving; analyzing the re-dissolved lipid extract solution by utilizing liquid chromatography-mass spectrometry in a positive and negative ion mode, and analyzing the ginsenoside extract solution by utilizing liquid chromatography-mass spectrometry in a negative ion mode. The separation of ginsenoside and lipid is realized, 76 ginsenoside components are extracted, 433 lipid components are obtained, and phospholipid, glycerolipid, sphingolipid, sterol lipid and fatty acid are respectively obtained.
Description
Technical Field
The application belongs to the technical field of pharmaceutical analytical chemistry, and particularly relates to an American ginseng extraction and analysis method.
Background
The disclosure of this background section is only intended to increase the understanding of the general background of the application and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art already known to those of ordinary skill in the art.
The American ginseng is dried root of Panax quinquefolium L. Of Araliaceae, has effects of invigorating qi, nourishing yin, clearing heat, and promoting salivation, and is mainly used for treating deficiency of qi and yin, dysphoria due to deficiency heat, cough, asthma, phlegm blood, internal heat, diabetes, and dry throat. Since the Qing dynasty, american ginseng has been used in China for more than 300 years and has been cultivated for more than 40 years. Compared with ginseng, american ginseng is more mild and is not easy to cause excessive internal heat. American ginseng is not only a superior traditional Chinese medicine, but also a good tonic product, and has wide application in the fields of medicine, health care, eating and the like.
American ginseng has rich chemical components. It has been pointed out that ginsenoside is the main effective component. Ginsenoside consists of aglycone and glycosyl, and has strong hydrophilicity due to the existence of sugar ligand. In recent years, lipid compounds (fat-soluble components) have received increasing attention. The lipid not only is a component of cell membranes, but also plays a role in signal molecules and energy storage, and is important for the growth and development of American ginseng. Studies have indicated that plants are rich in lipids, but few of the lipid-based components of American ginseng have been studied.
Disclosure of Invention
Aiming at the problems in the prior art, the application aims to provide an American ginseng extraction and analysis method.
In order to solve the technical problems, the technical scheme of the application is as follows:
a method for extracting and analyzing radix Panacis Quinquefolii comprises mixing radix Panacis Quinquefolii powder with methanol, performing vortex treatment, adding methyl tert-butyl ether, performing vortex treatment, adding water, performing vortex treatment, standing the obtained mixture, centrifuging, and separating the mixture into upper lipid extract and lower ginsenoside extract;
respectively taking lipid extract and ginsenoside extract, respectively freeze-drying, and respectively redissolving;
analyzing the re-dissolved lipid extract solution by utilizing liquid chromatography-mass spectrometry in a positive and negative ion mode, and analyzing the ginsenoside extract solution by utilizing liquid chromatography-mass spectrometry in a negative ion mode.
The separation of ginsenoside and lipid is realized, 76 ginsenoside components can be effectively extracted, 433 lipid components can be obtained, and can be classified into phospholipid, glycerolipid, sphingolipid, sterol lipid and fatty acid.
Through the specific sequential addition sequence of methanol, methyl tertiary butyl ether and water, ginsenoside and lipid can be effectively separated, and the extraction quantity and extraction types of the ginsenoside and the lipid can be effectively increased.
In some embodiments of the application, the ratio of American ginseng powder, methanol, methyl tertiary butyl ether, water is 0.3g:1mL:5.8-6mL:8-8.2mL; preferably 0.3 g/1 mL/6 mL/8 mL.
In some embodiments of the application, the time of the mixing vortex treatment of the American ginseng powder and methanol is 1.5-2.5 minutes; preferably 2min.
In some embodiments of the application, the swirling treatment with methyl tertiary butyl ether is for a period of 1.5 to 2.5 minutes; preferably 2min.
In some embodiments of the application, the time of the swirling treatment after the addition of water is 1.5-2.5min; preferably 2min.
In some embodiments of the application, the mixture is allowed to stand for a period of 0.5 to 1.5 minutes and then centrifuged for 12 to 16 minutes at 10000 to 13000rpm; preferably, the mixture is allowed to stand for 1min and then centrifuged for 15min at 12000rpm.
In some embodiments of the present application, the lipid extract is reconstituted with a reconstitution solution a and a reconstitution solution B, wherein the reconstitution solution a is a mixed solution of dichloromethane and methanol, and the reconstitution solution B is a mixed solution of isopropanol, acetonitrile and water; preferably, the volume ratio of dichloromethane to methanol in the redissolved solution is 2:1; preferably, the volume ratio of isopropanol, acetonitrile and water in the redissolved solution is 14:5:1.
In some embodiments of the present application, the redissolution of ginsenoside extract is a mixed solution of acetonitrile and water; preferably, the volume ratio of acetonitrile to water in the redissolved solution is 1:19.
In some embodiments of the application, the liquid chromatography-mass spectrometry conditions in positive and negative ion modes are: column ACQUITY UPLC@BEH C8 column (2.1X100 mm,1.7 μm, waters); mobile phase A, acetonitrile/water (3:2, v/v), containing 10mM ammonium acetate, mobile phase B was isopropanol/acetonitrile (9:1, v/v), containing 10mM ammonium acetate, flow rate 0.26mL/min; elution gradient: 0min,30% b;3min,55% b;8min,60% B;15min,75% B;17min,97% B, 5min;22.1min,30% B, 5min; the total elution time is 27min, the column temperature is 45 ℃, and the temperature of the sample injection chamber is 10 ℃; mass spectrometry conditions: mass number scanning range m/z400-1200 (positive ion mode), 50-1200 (negative ion mode), capillary voltage 3500V (positive ion mode), 3000V (negative ion mode); the atomization air pressure is 2.0Bar, the drying air flow rate is 8.0L/min, and the drying air temperature is 200 ℃; the transmission time was 80 μs, the voltage amplitude of impact Chi Shepin was 750Vpp, and the pre-pulse waiting time was 8 μs.
In some embodiments of the application, the liquid chromatography-mass spectrometry analysis conditions in negative ion mode are: column Zorbax SB-Aq column (2.1X100 mm,1.8 μm, agilent); mobile phase C was an aqueous solution containing 0.1% formic acid, mobile phase D was acetonitrile, flow rate 0.3mL/min, elution gradient: 0min,5% b;5min,20% B;14min,32% B;16min,36% B, hold for 3min;22min,45% B;25min,100% B, hold for 5min;30.1min,5% B, holding for 5min, total elution time 35min, column temperature 40 ℃, and sample injection chamber temperature 10 ℃; mass spectrometry conditions: the mass number scanning range m/z is 50-1500; capillary voltage 3000V; the atomization air pressure is 2.0Bar, the drying air flow rate is 8.0L/min, the drying air temperature is 200 ℃, the transmission time is 80 mu s, the collision Chi Shepin voltage amplitude is 750Vpp, and the waiting time before pulse is 8 mu s.
In some embodiments of the present application, the lipid extract obtained by liquid chromatography-mass spectrometry in positive and negative ion mode comprises 433 lipid components, wherein the phospholipid accounts for 35.8%, the glycerolipid accounts for 31.4%, the sphingolipid accounts for 12.7%, the sterol accounts for 2.8%, and the fatty acid accounts for 17.3%.
In some embodiments of the present application, the ginsenoside extract obtained by liquid chromatography-mass spectrometry in negative ion mode contains 76 ginsenoside components.
One or more of the technical schemes of the application has the following beneficial effects:
the American ginseng extraction analysis method utilizes the sequence of methanol, methyl tertiary butyl ether and water to carry out vortex treatment on American ginseng powder, so that ginsenoside and lipid in components contained in American ginseng are fully separated, more types of ginsenoside and lipid are obtained, and compared with the existing method, more types of lipid components including phospholipids, glycerolipids, sterol lipids, sphingolipids and fatty acids are extracted from the extracted lipid components.
The analysis method has the advantages of less interference of extracted ginsenoside matrix and increased detection sensitivity.
Drawings
The accompanying drawings, which are included to provide a further understanding of the application and are incorporated in and constitute a part of this specification, illustrate embodiments of the application and together with the description serve to explain the application.
FIG. 1 is a diagram of an extraction condition optimization model;
FIG. 2 is a representative spectrum of lipid extracts and ginsenoside extracts.
Detailed Description
It should be noted that the following detailed description is illustrative and is intended to provide further explanation of the application. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the present application. As used herein, the singular is also intended to include the plural unless the context clearly indicates otherwise, and furthermore, it is to be understood that the terms "comprises" and/or "comprising" when used in this specification are taken to specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof. The application will be further illustrated by the following examples
Example 1
The method for extracting and analyzing ginsenoside and lipid in American ginseng at one time comprises the following steps:
30mg of American ginseng is weighed into a 2mL centrifuge tube, 100. Mu.L of methanol is firstly added, the mixture is vortexed for 2min, then 600. Mu.L of MTBE is added, the mixture is vortexed for 2min, and finally 800. Mu.L of water is added, and the mixture is vortexed for 2min. Standing for 1min for layering. And then centrifuged for 15min at 12000rpm. And respectively taking supernatant and supernatant of the upper layer and the lower layer, and freeze-drying. And then redissolved. The upper layer was reconstituted using 50. Mu.L of methylene chloride/methanol solution (2:1, v/v) and 150. Mu.L of isopropanol/acetonitrile/water solution (14:5:1, v/v). The lower layer was reconstituted using 200. Mu.L acetonitrile/water (1:19, v/v). And (3) re-dissolving, filling into a liquid phase vial, and performing liquid chromatography-mass spectrometry analysis.
Lipid analysis conditions:
the liquid chromatography-mass spectrometry analysis conditions under positive and negative ion modes are as follows: column ACQUITY UPLC@BEH C8 column (2.1X100 mm,1.7 μm, waters); mobile phase A, acetonitrile/water (3:2, v/v), containing 10mM ammonium acetate, mobile phase B was isopropanol/acetonitrile (9:1, v/v), containing 10mM ammonium acetate, flow rate 0.26mL/min; elution gradient: 0min,30% b;3min,55% b;8min,60% B;15min,75% B;17min,97% B, 5min;22.1min,30% B, 5min; the total elution time is 27min, the column temperature is 45 ℃, and the temperature of the sample injection chamber is 10 ℃; mass spectrometry conditions: mass number scanning range m/z400-1200 (positive ion mode), 50-1200 (negative ion mode), capillary voltage 3500V (positive ion mode), 3000V (negative ion mode); the atomization air pressure is 2.0Bar, the drying air flow rate is 8.0L/min, and the drying air temperature is 200 ℃; the transmission time was 80 μs, the voltage amplitude of impact Chi Shepin was 750Vpp, and the pre-pulse waiting time was 8 μs.
Ginsenoside analysis conditions:
the liquid chromatography-mass spectrometry analysis conditions in the negative ion mode were: column Zorbax SB-Aq column (2.1X100 mm,1.8 μm, agilent); mobile phase C was an aqueous solution containing 0.1% formic acid, mobile phase D was acetonitrile, flow rate 0.3mL/min, elution gradient: 0min,5% b;5min,20% B;14min,32% B;16min,36% B, hold for 3min;22min,45% B;25min,100% B, hold for 5min;30.1min,5% B, holding for 5min, total elution time 35min, column temperature 40 ℃, and sample injection chamber temperature 10 ℃; mass spectrometry conditions: the mass number scanning range m/z is 50-1500; capillary voltage 3000V; the atomization air pressure is 2.0Bar, the drying air flow rate is 8.0L/min, the drying air temperature is 200 ℃, the transmission time is 80 mu s, the collision Chi Shepin voltage amplitude is 750Vpp, and the waiting time before pulse is 8 mu s.
Representative spectrograms of lipid extracts in American ginseng are shown in part A of figure 2 and part B of figure 2. Representative spectrogram of ginsenoside extract is shown in section C of figure 2.
433 lipids were found in the lipid analysis of American ginseng, wherein the phospholipid accounts for 35.8%, the glycerolipid accounts for 31.4%, the sphingolipid accounts for 12.7%, the sterol accounts for 2.8%, and the fatty acids account for 17.3%.
76 ginsenoside is identified in the ginsenoside extract.
Comparative example 1
The volume of methanol was 100. Mu.L, the volume of methyl t-butyl ether was 1000. Mu.L, the volume of water was 400. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 2
The volume of methanol was 100. Mu.L, the volume of methyl t-butyl ether was 800. Mu.L, the volume of water was 600. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 3
The volume of methanol was 100. Mu.L, the volume of methyl t-butyl ether was 400. Mu.L, the volume of water was 1000. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 4
The volume of methanol was 150. Mu.L, the volume of methyl t-butyl ether was 800. Mu.L, the volume of water was 550. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 5
The volume of methanol was 150. Mu.L, the volume of methyl t-butyl ether was 1000. Mu.L, the volume of water was 400. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 6
The volume of methanol was 133. Mu.L, the volume of methyl t-butyl ether was 567. Mu.L, the volume of water was 800. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 7
The volume of methanol was 175. Mu.L, the volume of methyl t-butyl ether was 1000. Mu.L, the volume of water was 325. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 8
The volume of methanol was 200. Mu.L, the volume of methyl t-butyl ether was 500. Mu.L, the volume of water was 800. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 9
The volume of methanol was 250. Mu.L, the volume of methyl t-butyl ether was 1000. Mu.L, the volume of water was 250. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 10
The volume of methanol was 250. Mu.L, the volume of methyl t-butyl ether was 800. Mu.L, the volume of water was 450. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 11
The volume of methanol was 250. Mu.L, the volume of methyl t-butyl ether was 300. Mu.L, the volume of water was 950. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 12
The volume of methanol was 275. Mu.L, the volume of methyl tert-butyl ether was 625. Mu.L, the volume of water was 600. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 13
The volume of methanol was 300. Mu.L, the volume of methyl t-butyl ether was 400. Mu.L, the volume of water was 800. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 14
The volume of methanol was 325. Mu.L, the volume of methyl t-butyl ether was 750. Mu.L, the volume of water was 425. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 15
The volume of methanol was 350. Mu.L, the volume of methyl t-butyl ether was 450. Mu.L, the volume of water was 700. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 16
The volume of methanol was 400. Mu.L, the volume of methyl t-butyl ether was 700. Mu.L, the volume of water was 400. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 17
The volume of methanol was 400. Mu.L, the volume of methyl t-butyl ether was 400. Mu.L, the volume of water was 700. Mu.L, and the other operating conditions were the same as in example 1.
Comparative example 18
The volume of methanol was 200. Mu.L, the volume of methyl t-butyl ether was 650. Mu.L, the volume of water was 650. Mu.L, and the other operating conditions were the same as in example 1.
By Design-expert software, 18 experimental conditions were designed as shown in table 1, corresponding numbers 1 to 18 in table 1 represent comparative examples 1 to 18, respectively, and the 18 th experimental condition was repeated 6 times (18 to 23) to examine the reproducibility of the experimental optimization process.
Table 1 optimization of experimental conditions
Experimental results: the peak area and the number of peaks were used to evaluate the extraction efficiency. RSD of peak number and peak area of 6 duplicate samples (numbers 18-23) were calculated. The RSD of the number of peaks and the peak area for the lipid extract was 2.9% and 2.8%, and the RSD of the number of peaks and the peak area for the ginsenoside extract was 3.7% and 3.9%, reflecting the reliability of the analytical experiment procedure. Based on the number of peaks and the peak area of the ginsenoside extract and the lipid extract under 18 experimental conditions, a model was established as shown in fig. 1. The optimal volumes of methanol, MTBE and water were optimized to be 100. Mu.L, 589.487. Mu.L and 810.513. Mu.L, respectively, so that the volume ratio of methanol, MTBE and water was about 1:6:8 (v/v/v) in example 1.
Methodology investigation
Method reproducibility
6 samples were processed in parallel for continuous analysis. The RSD of each peak was calculated and the number of peaks and the cumulative peak area in the different RSD ranges (0-10%, 10-20%,20-30%, > 30%) were counted. For lipid extracts, in positive ion mode, 92.6% peak RSD is less than 20% and peak area ratio is 98.9%; in the negative ion mode, the RSD of 92.1% peak is less than 20%, the peak area ratio is 96.6%, and in the negative ion mode, the RSD of 94.5% peak is less than 20%, and the peak area ratio is 98.2%.
Precision within the day
6 samples were treated in parallel and analyzed at 24 h. The RSD of each peak was calculated and the number of peaks and the cumulative peak area in the different RSD ranges (0-10%, 10-20%,20-30%, > 30%) were counted. For lipid extracts, 93.2% of peaks have an RSD of less than 20% and a peak area ratio of 98.9% in positive ion mode; in the negative ion mode, 93.4% of the peaks have an RSD of less than 20% and a peak area ratio of 97.5%. For ginsenoside extract, in negative ion mode, 96.9% of peak has RSD less than 20%, and peak area ratio is 99.3%.
Precision of daytime
18 samples were treated in parallel and analyzed on 3 days each, 6 per day. The RSD of each peak was calculated and the number of peaks and the cumulative peak area in the different RSD ranges (0-10%, 10-20%,20-30%, > 30%) were counted. For lipid extracts, in positive ion mode, 92.0% peak RSD is less than 20% and peak area ratio is 98.6%; in the negative ion mode, the RSD of 92.0% peak is less than 20%, and the peak area is 99.0%. For ginsenoside extract, in negative ion mode, 97.4% of peak has RSD less than 20%, and peak area ratio is 99.4%.
The results show that the method has good stability and meets the requirements of ginsenoside and lipid analysis.
Comparative example 19
The method for extracting and analyzing ginsenoside and lipid in American ginseng at one time comprises the following steps:
30mg of American ginseng is weighed into a 2mL centrifuge tube, 600. Mu.L of MTBE is added first, vortexed for 2min, then 100. Mu.L of methanol is added, vortexed for 2min, and finally 800. Mu.L of water is vortexed for 2min. Standing for 1min for layering. And then centrifuged for 15min at 12000rpm. And respectively taking supernatant and supernatant of the upper layer and the lower layer, and freeze-drying. And then redissolved. The upper layer was reconstituted using 50. Mu.L of methylene chloride/methanol solution (2:1, v/v) and 150. Mu.L of isopropanol/acetonitrile/water solution (14:5:1, v/v). The lower layer was reconstituted using 200. Mu.L acetonitrile/water (1:19, v/v). And (3) re-dissolving, filling into a liquid phase vial, and performing liquid chromatography-mass spectrometry analysis.
The rest of the analysis was the same as in example 1.
Comparative example 20
The method for extracting and analyzing ginsenoside and lipid in American ginseng at one time comprises the following steps:
30mg of American ginseng is weighed into a 2mL centrifuge tube, 800. Mu.L of water is firstly added, then 100. Mu.L of methanol is added, the mixture is vortexed for 2min, and finally 600. Mu.L of MTBE is added, the mixture is vortexed for 2min and the mixture is vortexed for 2min. Standing for 1min for layering. And then centrifuged for 15min at 12000rpm. And respectively taking supernatant and supernatant of the upper layer and the lower layer, and freeze-drying. And then redissolved. The upper layer was reconstituted using 50. Mu.L of methylene chloride/methanol solution (2:1, v/v) and 150. Mu.L of isopropanol/acetonitrile/water solution (14:5:1, v/v). The lower layer was reconstituted using 200. Mu.L acetonitrile/water (1:19, v/v). And (3) re-dissolving, filling into a liquid phase vial, and performing liquid chromatography-mass spectrometry analysis.
The rest of the analysis was the same as in example 1.
Comparative example 19 and comparative example 20 were different in the order of addition of methanol, methyl t-butyl ether, and water, and example 1 was distinguished from comparative example 19 and comparative example 20 in that the layered extraction of example 1 was the best. The lipid layer and the ginsenoside layer are layered clearly, and no turbidity exists. Comparative example 19 there was a layer of milky turbidity in the middle of the lipid layer and the ginsenoside layer. Comparative example 20 a thick layer of powdered solid of American ginseng was provided between the lipid layer and the ginsenoside layer.
The above description is only of the preferred embodiments of the present application and is not intended to limit the present application, but various modifications and variations can be made to the present application by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the protection scope of the present application.
Claims (10)
1. A American ginseng extraction analysis method is characterized in that: the method comprises the following specific steps:
mixing radix Panacis Quinquefolii powder with methanol, performing vortex treatment, adding methyl tert-butyl ether, performing vortex treatment, adding water, performing vortex treatment, standing the obtained mixture, centrifuging, and separating the mixture into upper lipid extract and lower ginsenoside extract;
the ratio of the American ginseng powder to the methanol to the methyl tertiary butyl ether to the water is 0.3g to 1mL to 5.8-6mL to 8-8.2mL;
the mixing vortex treatment time of the American ginseng powder and the methanol is 1.5-2.5min; adding methyl tertiary butyl ether for vortex treatment for 1.5-2.5min; adding water, and performing vortex treatment for 1.5-2.5min;
standing the mixture for 0.5-1.5min, centrifuging for 12-16min, and rotating at 10000-13000rpm;
respectively taking lipid extract and ginsenoside extract, respectively freeze-drying, and respectively redissolving; analyzing the re-dissolved lipid extract solution by utilizing liquid chromatography-mass spectrometry in a positive and negative ion mode, and analyzing the ginsenoside extract solution by utilizing liquid chromatography-mass spectrometry in a negative ion mode;
the lipid extract obtained by liquid chromatography-mass spectrometry under positive and negative ion mode contains 433 lipid components, wherein the phospholipid accounts for 35.8%, the glyceride accounts for 31.4%, the sphingolipid accounts for 12.7%, the sterol accounts for 2.8%, and the fatty acid accounts for 17.3%;
the ginsenoside extract obtained by liquid chromatography-mass spectrometry under anion mode contains 76 ginsenoside components;
the liquid chromatography-mass spectrometry analysis conditions under positive and negative ion modes are as follows: the chromatographic column is Waters ACQUITY UPLC @BEH2C8, and the specification is 2.1X100 mm,1.7 μm; the mobile phase A is acetonitrile/water, the volume ratio of acetonitrile/water is 3:2, the mobile phase B is isopropanol/acetonitrile, the volume ratio of isopropanol/acetonitrile is 9:1, and both the mobile phase A and the mobile phase B contain 10mM ammonium acetate; the flow rate is 0.26mL/min; elution gradient: 0min,30% B;3min,55% B;8min,60% B;15min,75% B;17min,97% B, for 5min;22.1min,30% B, for 5min; column temperature is 45 ℃, and sample injection chamber temperature is 10 ℃; mass spectrometry conditions: the mass number scanning range is 400-1200 m/z in positive ion mode, 50-1200 m/z in negative ion mode, 3500V in capillary voltage positive ion mode and 3000V in negative ion mode; the atomization air pressure is 2.0Bar, the drying air flow rate is 8.0L/min, and the drying air temperature is 200 ℃; the transmission time is 80 mu s, the voltage amplitude of collision Chi Shepin is 750Vpp, and the waiting time before pulse is 8 mu s;
the liquid chromatography-mass spectrometry analysis conditions in the negative ion mode were: the chromatographic column is Agilent Zorbax SB-Aq, the specification is 2.1X100 mm,1.8 μm; mobile phase C was an aqueous solution containing 0.1% formic acid, mobile phase D was acetonitrile, flow rate 0.3mL/min, elution gradient: 0min,5% d;5min,20% D;14min,32% D;16min,36% D, hold for 3min;22min,45% D;25min,100% D, hold for 5min;30.1min,5% D, 5min total elution time 35min, column temperature 40 ℃, sample injection chamber temperature 10 ℃; mass spectrometry conditions: the mass number scanning range m/z is 50-1500; capillary voltage 3000V; the atomization air pressure is 2.0Bar, the drying air flow rate is 8.0L/min, the drying air temperature is 200 ℃, the transmission time is 80 mu s, the collision Chi Shepin voltage amplitude is 750Vpp, and the waiting time before pulse is 8 mu s.
2. The method for extracting and analyzing American ginseng according to claim 1, wherein: american ginseng powder, methanol, methyl tertiary butyl ether and water are mixed in a ratio of 0.3g to 1mL to 6mL to 8mL.
3. The method for extracting and analyzing American ginseng according to claim 1, wherein: the mixing vortex treatment time of the American ginseng powder and the methanol is 2min.
4. The method for extracting and analyzing American ginseng according to claim 1, wherein: adding methyl tertiary butyl ether for vortex treatment for 2min;
or, the time of the vortex treatment after adding water is 2min.
5. The method for extracting and analyzing American ginseng according to claim 1, wherein: the mixture was allowed to stand for 1min and then centrifuged for 15min at 12000rpm.
6. The method for extracting and analyzing American ginseng according to claim 1, wherein: the lipid extract is prepared from a redissolution A and a redissolution B, wherein the redissolution A is used for redissolution, and the redissolution B is used for redissolution, and the redissolution A is a mixed solution of dichloromethane and methanol, and the redissolution B is a mixed solution of isopropanol, acetonitrile and water.
7. The method for extracting and analyzing American ginseng according to claim 6, wherein: the volume ratio of dichloromethane to methanol in the redissolution solution A is 2:1.
8. The method for extracting and analyzing American ginseng according to claim 6, wherein: the volume ratio of isopropanol, acetonitrile and water in the redissolution solution B is 14:5:1.
9. The method for extracting and analyzing American ginseng according to claim 1, wherein: the redissolution of ginsenoside extract is mixed solution of acetonitrile and water.
10. The method for extracting and analyzing American ginseng according to claim 9, wherein: the volume ratio of acetonitrile to water in the redissolved solution is 1:19.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011146938.6A CN112305106B (en) | 2020-10-23 | 2020-10-23 | American ginseng extraction analysis method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011146938.6A CN112305106B (en) | 2020-10-23 | 2020-10-23 | American ginseng extraction analysis method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112305106A CN112305106A (en) | 2021-02-02 |
| CN112305106B true CN112305106B (en) | 2023-09-29 |
Family
ID=74327412
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011146938.6A Active CN112305106B (en) | 2020-10-23 | 2020-10-23 | American ginseng extraction analysis method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112305106B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113109476B (en) * | 2021-04-15 | 2023-10-13 | 岛津企业管理（中国）有限公司 | Dual gradient system in liquid chromatograph and matched elution program setting method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103776911A (en) * | 2012-10-22 | 2014-05-07 | 中国科学院大连化学物理研究所 | Method for simultaneous extraction and analysis of metabolite group and lipid group in microtissue |
| CN107589206A (en) * | 2017-10-30 | 2018-01-16 | 云南省烟草农业科学研究院 | A kind of quantitative analysis method of tobacco leaf maleic acid hydrazide and its glucosides based on Liquid Chromatography-Tandem Mass Spectrometry |
| CN107807188A (en) * | 2017-10-30 | 2018-03-16 | 云南省烟草农业科学研究院 | It is a kind of based on the tobacco leaf maleic acid hydrazide of liquid chromatogram high resolution mass spectrum and its quantitative analysis method of glucosides |
| CN110174468A (en) * | 2019-03-26 | 2019-08-27 | 陕西科技大学 | The analysis method of lipid composition in a kind of Fu-brick tea |
-
2020
- 2020-10-23 CN CN202011146938.6A patent/CN112305106B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103776911A (en) * | 2012-10-22 | 2014-05-07 | 中国科学院大连化学物理研究所 | Method for simultaneous extraction and analysis of metabolite group and lipid group in microtissue |
| CN107589206A (en) * | 2017-10-30 | 2018-01-16 | 云南省烟草农业科学研究院 | A kind of quantitative analysis method of tobacco leaf maleic acid hydrazide and its glucosides based on Liquid Chromatography-Tandem Mass Spectrometry |
| CN107807188A (en) * | 2017-10-30 | 2018-03-16 | 云南省烟草农业科学研究院 | It is a kind of based on the tobacco leaf maleic acid hydrazide of liquid chromatogram high resolution mass spectrum and its quantitative analysis method of glucosides |
| CN110174468A (en) * | 2019-03-26 | 2019-08-27 | 陕西科技大学 | The analysis method of lipid composition in a kind of Fu-brick tea |
Non-Patent Citations (6)
| Title |
|---|
| High-mass-resolution MALDI mass spectrometry imaging reveals detailed spatial distribution of metabolites and lipids in roots of barley seedlings in response to salinity stress;Lenin D. Sarabia等;《Metabolomics》;20180419;第14卷;1-16 * |
| In-depth profiling, characterization, and comparison of the ginsenosides among three different parts (the root, stem leaf, and flower bud) of Panax quinquefolius L. by ultra-high performance liquid chromatography/quadrupole-Orbitrap mass spectrometry;Hongda Wang等;《Analytical and Bioanalytical Chemistry》;20191115;第411卷;7817–7829 * |
| NanoESI-MS-based lipidomics to discriminate between cultivars, cultivation ages, and parts of Panax ginseng;So-Hyun Kim等;《Anal Bioanal Chem》;20160122;1-13 * |
| Nontargeted metabolomic analysis and "Commercial-homophyletic" comparison-induced biomarkers verification for the systematic chemical differentiation of five different parts of Panax ginseng;Shi Qiu等;《Journal of Chromatography A》;20160513;第1453卷;78-87 * |
| Systematic profiling and comparison of the lipidomes from Panaxginseng, P. quinquefolius, and P. notoginseng by ultrahigh performancesupercritical fluid chromatography/high-resolution massspectrometry and ion mobility-derived collision cross section measur;Xiaojian Shi等;《Journal of Chromatography A》;20180314;第1548卷;64-75 * |
| 超高效液相色谱-飞行时间-串联质谱法用于大白菜叶片中的脂质成分分析;郑姝宁等;《色谱》;20170208(第02期);169-177 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112305106A (en) | 2021-02-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Cai et al. | A capsule review of recent studies on the application of mass spectrometry in the analysis of Chinese medicinal herbs | |
| Du et al. | Isolation of dammarane saponins from Panax notoginseng by high-speed counter-current chromatography | |
| CN106526002B (en) | Ginseng-astragalus blood-sugar lowering preparation content determining method and its application in global quality control | |
| CN107271577A (en) | A kind of analysis of effective component method of stilbene Siberian cocklebur warm kidney medicine for eliminating bursa | |
| CN112305106B (en) | American ginseng extraction analysis method | |
| CN105911131A (en) | Method for detecting lipid molecules in salmon | |
| CN104713953B (en) | A kind of assay method of medicine finger-print | |
| Li et al. | Extraction and in vitro screening of potential acetylcholinesterase inhibitors from the leaves of Panax japonicus | |
| CN105663195A (en) | Extracting method of ginsenoside | |
| CN106706786A (en) | Method for determining content of six ginsenoside ingredients of folium ginseng | |
| CN113820419B (en) | Method for determining effective components in beautiful millettia root wine | |
| CN106918655A (en) | A kind of polygala UPLC assay methods | |
| CN111487350B (en) | Quality detection method of salvia miltiorrhiza-panax notoginseng medicine pair | |
| CN111289648B (en) | Method for establishing traditional Chinese medicine compound preparation fingerprint and fingerprint thereof | |
| Kakigi et al. | Analysis of terpene lactones in a Ginkgo leaf extract by high-performance liquid chromatography using charged aerosol detection | |
| CN106483260A (en) | A kind of pill of Eight Treasures capsule quality standard control method | |
| CN113759011B (en) | Method for establishing characteristic spectrum of starwort root and preparation thereof | |
| CN110398551B (en) | Establishment method of ginseng fruit medicinal material fingerprint spectrum and standard fingerprint spectrum thereof | |
| CN110907581B (en) | Mass spectrum detection method for concentration of seven-component blood plasma or tissue of compound salvia miltiorrhiza preparation | |
| CN110927292A (en) | Characteristic spectrum based on gleditsia sinensis extract and construction method and application thereof | |
| CN109481531B (en) | Qi-benefiting and blood-nourishing composition as well as preparation method and application thereof | |
| Rodin et al. | Modern methods of identifying and determining ginsenosides | |
| KR0161748B1 (en) | Process for preparing ginseng extract containing improved saponin contents | |
| CN107064402B (en) | A kind of nourishing qi and blood, tonifying kidney and strengthening yang pharmaceutical preparation detection method | |
| CN115337332B (en) | Preparation method of fried crisp pseudo-ginseng for gynecological reconstruction pill and gynecological reconstruction pill |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |