CN116148388A - Detection method of adulterated Artemisia mongolica in mugwort medicinal materials and decoction pieces - Google Patents
Detection method of adulterated Artemisia mongolica in mugwort medicinal materials and decoction pieces Download PDFInfo
- Publication number
- CN116148388A CN116148388A CN202310124253.9A CN202310124253A CN116148388A CN 116148388 A CN116148388 A CN 116148388A CN 202310124253 A CN202310124253 A CN 202310124253A CN 116148388 A CN116148388 A CN 116148388A
- Authority
- CN
- China
- Prior art keywords
- sample
- solution
- chromatogram
- reference substance
- sample solution
- 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.)
- Pending
Links
Images
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
-
- 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
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
-
- 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
- G01N30/86—Signal analysis
- G01N30/8624—Detection of slopes or peaks; baseline correction
- G01N30/8631—Peaks
- G01N30/8634—Peak quality criteria
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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)
- Engineering & Computer Science (AREA)
- Quality & Reliability (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
A detection method of adulterated Artemisia mongolica in mugwort medicinal materials and decoction pieces comprises the following steps: s1, preparing a reference substance solution; s2, preparing a sample solution; s3, chromatogram: analyzing the reference substance solution and the sample solution obtained in the steps S1 and S2 by a gas chromatograph to obtain a reference substance solution chromatogram and a sample solution chromatogram; s4, adulteration analysis: comparing the chromatogram of the sample with the chromatogram of the reference sample, and simultaneously meeting the following two conditions, namely, taking the sample as adulteration: s5, verifying. The method is simple and easy to operate, the detection process is quick and effective, the method is stable and reliable, whether the tarragon medicinal materials or decoction pieces are adulterated or not can be effectively identified, the specificity is high, the precision, the repeatability and the stability are good, the detection limit is low, and remarkable social and economic benefits are achieved.
Description
Technical Field
The invention relates to the technical field of drug adulteration detection, in particular to a detection method of adulterated Artemisia mongolica in mugwort medicinal materials and decoction pieces.
Background
The folium Artemisiae Argyi is dry leaf of Artemisia princeps Vant. Picking when flowers are not opened in summer, removing impurities, and sun drying. Has effects of warming channel, stopping bleeding, dispelling cold, and relieving pain, and can be used for treating hematemesis, epistaxis, menorrhagia, infertility due to cold womb, etc.; it is one of the traditional Chinese medicines commonly used in clinic to treat skin itch. However, artemisia genus belonging to Artemisia princeps is one of the most widely distributed species in Compositae plants, and according to the record of China Saint. Among them, many kinds of mugwort leaf are similar, and most of mugwort leaf are used as mugwort leaf in place of production, which causes confusion of mugwort leaf medicinal material varieties to a certain extent.
Mongolian wormwood Artemisia mongolica, also called Mongolian wormwood (Beijing plant), narrow She Hao (Jiangsu), and herba Artemisiae Annuae (Liaoning), is produced in Heilongjiang, hebei, shanxi, henan, jiangsu and other places. The phenomenon that the leaves of the Mongolian artemisia are used as the mugwort leaves exists in the production place, however, experimental researches show that the Mongolian artemisia and the mugwort volatile oil components have larger difference, the content of flavonoid components in the Mongolian artemisia is obviously lower than that of the mugwort leaves, and if the Mongolian artemisia is used as the mugwort leaves, the clinical curative effect is affected, and the use purpose is not achieved. At present, the distinction between the mugwort and the Mongolian artemisia is mainly based on the character and smell difference of the mugwort and the Mongolian artemisia, but the artemisia has great leaf shape change in different growth periods, and the leaf shapes of different parts of the same plant are different, so that the character identification is difficult, and misjudgment is easy to cause. Therefore, the research on a chemical identification method with good precision, repeatability and stability based on chemical components is particularly important.
Disclosure of Invention
Aiming at the situation, the invention aims to overcome the defects of the prior art and provide a detection method of the adulterated tarragon in the mugwort leaf medicinal material and decoction pieces, which can effectively solve the problems of difficult identification of the tarragon and the mugwort leaf and inaccurate identification result.
In order to achieve the above purpose, the technical scheme of the invention is that the detection method of the adulterated Mongolian artemisia in the artemisia argyi medicinal material and decoction pieces comprises the following steps:
s1, preparing a reference substance solution: taking a chamazulene reference substance and an ethyl acetate solvent to prepare a reference substance solution with the concentration of 10-30 mug/mL;
s2, preparing a sample solution: cutting a mugwort leaf medicinal material or decoction piece sample into fragments of 0.4-0.6cm, placing 2-3 g of the fragments in a round bottom flask, adding 200-500 ml of water, uniformly mixing, connecting a volatile oil tester, adding water from the upper end of the tester to fill a scale part, overflowing the volatile oil tester into the flask, adding 1-3 ml of ethyl acetate, connecting a reflux condenser pipe, heating to boil, continuously heating for 5 hours, then cooling to room temperature, separating ethyl acetate liquid, placing into a 10ml measuring flask, washing the tester and the condenser pipe with ethyl acetate for several times, transferring into the same 10ml measuring flask, diluting the ethyl acetate to the scale of 10ml, and shaking uniformly to obtain a sample solution;
s3, chromatogram: taking 1 mu L of the reference substance solution and the sample solution obtained in the steps S1 and S2 respectively, and analyzing by a gas chromatograph to obtain a reference substance solution chromatogram and a sample solution chromatogram; the gas chromatographic analysis conditions are as follows: the chromatographic column stationary phase is 5% phenyl-95% methyl polysiloxane, the carrier gas flow rate is 0.8-1.2 ml/min, and the gradient temperature rise is carried out: the initial temperature is 60 ℃, the temperature is firstly increased to 120 ℃ at the rate of 3 ℃ per minute, and then the temperature is increased to 280 ℃ at the rate of 15 ℃ per minute; the sample inlet temperature is 240 ℃, the sample inlet amount is 1-2 ul, and the FID detector;
s4, adulteration analysis: comparing the chromatogram of the sample with the chromatogram of the reference sample, and simultaneously meeting the following two conditions, namely, taking the sample as adulteration:
(1) Chromatographic peaks appear in the sample solution chromatogram at positions consistent with the retention time of the chromatographic peaks of the chamazulene in the control solution chromatogram;
(2) The chromatographic peak area of the sample solution is larger than that of the reference solution;
s5, verifying: when S4 judges that the Mongolian artemisia is adulterated, a gas chromatography-mass spectrometry method is adopted for verification, namely the reference substance solution and the sample solution obtained in the steps S1 and S2 are respectively filtered by a 0.22 mu m filter membrane, then are injected into a gas chromatography-mass spectrometry analyzer to obtain an ion flow diagram and a mass spectrogram of the reference substance solution and the sample solution, and when the mass spectrograms of the reference substance solution and the sample solution are consistent, the Mongolian artemisia is adulterated to the sample to be detected; the chromatographic conditions of the gas chromatography-mass spectrometry combined analysis are as follows: chromatographic column stationary phase: 5% phenyl-95% methylpolysiloxane; carrier gas flow rate: 1ml/min; gradient heating: the initial temperature is 60 ℃, the temperature is firstly increased to 120 ℃ at the rate of 3 ℃ per minute, and then the temperature is increased to 280 ℃ at the rate of 15 ℃ per minute; sample inlet temperature: 240 ℃.
The method is simple, easy to operate, quick and effective in detection process, stable and reliable, can effectively identify whether the Artemisia mongolica is adulterated in the Artemisia argyi medicinal materials or decoction pieces, has strong specificity, good precision, repeatability and stability, low detection limit and obvious social and economic benefits.
Drawings
FIG. 1 is a gas chromatogram of the chamomile azulene control of the present invention;
FIG. 2 is a gas chromatogram of 3 samples (AC 1-AC 3) of folium Artemisiae Argyi (AC 1, AC2, AC3 in order from top to bottom);
FIG. 3 shows a gas chromatogram of 3 batches of mugwort leaf decoction piece samples (AP 1-AP 3) (AP 1, AP2 and AP3 are sequentially from top to bottom);
FIG. 4 is a gas chromatograph-mass spectrum total ion flow diagram of the samples of the chamazulene control and the folium artemisiae argyi decoction pieces (AP 3) of the invention (the chamazulene control and the AP3 are sequentially arranged from top to bottom);
FIG. 5 is the mass spectrum of the chamomile azulene control and the mugwort leaf decoction pieces AP3 of the present invention (from top to bottom: the mass spectrum of the chamomile azulene control, the mass spectrum of the chromatographic peak with consistent retention time of mugwort leaf decoction pieces AP3 and chamomile azulene);
FIG. 6 is a gas chromatogram of the present invention when different amounts of Mongolian wormwood are adulterated (from top to bottom: adulterated 0% Mongolian wormwood sample, adulterated 5% Mongolian wormwood sample, adulterated 10% Mongolian wormwood sample, adulterated 20% Mongolian wormwood sample);
FIG. 7 shows a sample solution extraction method of the present invention, wherein the sample solution 1 (ethyl acetate ultrasound) gas chromatogram, the sample solution 2 (ethyl acetate reflux) gas chromatogram, and the sample solution 3 (volatile oil extraction) gas chromatogram are shown in the sequence from top to bottom.
Detailed Description
The following describes in detail the embodiments of the present invention with reference to the drawings and examples.
Example 1
A detection method of adulterated Artemisia mongolica in mugwort medicinal materials and decoction pieces comprises the following steps:
s1, preparing a reference substance solution: taking a chamazulene control substance and an ethyl acetate solvent to prepare a control substance solution with the concentration of 10 mug/mL;
s2, preparing a sample solution: cutting mugwort leaf medicinal materials or decoction pieces into pieces of 0.4-0.6cm, placing 2g into a round bottom flask, adding 200ml of water, mixing, connecting a volatile oil tester, adding water from the upper end of the tester to fill the scale part, overflowing into the flask, adding 1ml of ethyl acetate, connecting a reflux condenser pipe, heating to boil, heating for 5 hours, then cooling to room temperature, separating ethyl acetate liquid, placing into a 10ml measuring flask, washing the tester and condenser pipe with ethyl acetate for several times, transferring into the same 10ml measuring flask, diluting with ethyl acetate to the scale of 10ml, shaking uniformly to obtain a sample solution;
s3, chromatogram: and (3) respectively analyzing the reference substance solution and the sample solution obtained in the steps S1 and S2 by a gas chromatograph to obtain a reference substance solution chromatogram and a sample solution chromatogram, wherein a chromatographic column stationary phase of the gas chromatographic analysis is 5% phenyl-95% methylpolysiloxane, the carrier gas flow rate is 0.8-1.2 ml/min, and the gradient temperature is raised: the initial temperature is 60 ℃, the temperature is firstly increased to 120 ℃ at the rate of 3 ℃ per minute, and then the temperature is increased to 280 ℃ at the rate of 15 ℃ per minute; the sample inlet temperature is 240 ℃, the sample inlet amount is 1-2 ul, and the FID detector;
s4, adulteration analysis: comparing the chromatogram of the sample with the chromatogram of the reference sample, and simultaneously meeting the following two conditions, namely, taking the sample as adulteration:
(1) Chromatographic peaks appear in the sample solution chromatogram at positions consistent with the retention time of the chromatographic peaks of the chamazulene in the control solution chromatogram;
(2) The chromatographic peak area of the sample solution is larger than that of the reference solution;
s5, verifying: and (4) when the Mongolian artemisia is judged to be adulterated, verifying by adopting a gas chromatography-mass spectrometry method, namely, injecting the reference substance solution and the sample solution obtained in the steps (S1) and (S2) into a gas chromatography-mass spectrometry analyzer to obtain an ion flow diagram and a mass spectrum diagram of the reference substance solution and the sample solution, and determining that the sample to be detected is adulterated.
Example 2
A detection method of adulterated Artemisia mongolica in mugwort medicinal materials and decoction pieces comprises the following steps:
s1, preparing a reference substance solution: taking a chamazulene control and an ethyl acetate solvent to prepare a control solution with the concentration of 20 mug/mL (namely 20 mug per 1mL of chamazulene-containing solution);
s2, preparing a sample solution: cutting mugwort leaf medicinal materials or decoction piece samples into fragments of 0.4-0.6cm, placing 2.5g in a round bottom flask, adding 300ml of water, uniformly mixing, connecting a volatile oil tester, adding water from the upper end of the tester to fill the scale part, overflowing into the flask, adding 2ml of ethyl acetate, connecting a reflux condenser pipe, heating to boil, continuously heating for 5 hours, then cooling to room temperature, separating ethyl acetate liquid, placing into a 10ml measuring flask, washing the tester and the condenser pipe with ethyl acetate in a fractional manner, transferring into the same 10ml measuring flask, diluting the ethyl acetate to the scale of 10ml, and shaking uniformly to obtain a sample solution;
s3, chromatogram: analyzing the reference substance solution and the sample solution obtained in the steps S1 and S2 by a gas chromatograph to obtain a reference substance solution chromatogram and a sample solution chromatogram;
s4, adulteration analysis: comparing the chromatogram of the sample with the chromatogram of the reference sample, and simultaneously meeting the following two conditions, namely, taking the sample as adulteration:
(1) Chromatographic peaks appear in the sample solution chromatogram at positions consistent with the retention time of the chromatographic peaks of the chamazulene in the control solution chromatogram;
(2) The chromatographic peak area of the sample solution is larger than that of the reference solution;
s5, verifying: and (4) when the Mongolian artemisia is judged to be adulterated, verifying by adopting a gas chromatography-mass spectrometry method, namely, injecting the reference substance solution and the sample solution obtained in the steps (S1) and (S2) into a gas chromatography-mass spectrometry analyzer to obtain an ion flow diagram and a mass spectrum diagram of the reference substance solution and the sample solution, and determining that the sample to be detected is adulterated.
Example 3
A detection method of adulterated Artemisia mongolica in mugwort medicinal materials and decoction pieces comprises the following steps:
s1, preparing a reference substance solution: taking a chamazulene control substance and an ethyl acetate solvent to prepare a control substance solution with the concentration of 30 mug/mL;
s2, preparing a sample solution: cutting folium Artemisiae Argyi or decoction pieces into pieces of 0.4-0.6cm, placing 3g into round bottom flask, adding 500ml of water, mixing, connecting volatile oil tester, adding water from the upper end of tester to fill scale part, overflowing into flask, adding ethyl acetate 3ml, connecting reflux condenser, heating to boil, heating for 5 hr, standing at room temperature, separating ethyl acetate liquid, placing into 10ml measuring flask, washing tester and condenser with ethyl acetate, transferring into the same 10ml measuring flask, diluting with ethyl acetate to 10ml scale, shaking to obtain sample solution;
s3, chromatogram: analyzing the reference substance solution and the sample solution obtained in the steps S1 and S2 by a gas chromatograph to obtain a reference substance solution chromatogram and a sample solution chromatogram;
s4, adulteration analysis: comparing the chromatogram of the sample with the chromatogram of the reference sample, and simultaneously meeting the following two conditions, namely, taking the sample as adulteration:
(1) Chromatographic peaks appear in the sample solution chromatogram at positions consistent with the retention time of the chromatographic peaks of the chamazulene in the control solution chromatogram;
(2) The chromatographic peak area of the sample solution is larger than that of the reference solution;
s5, verifying: and (4) when the Mongolian artemisia is judged to be adulterated, verifying by adopting a gas chromatography-mass spectrometry method, namely, injecting the reference substance solution and the sample solution obtained in the steps (S1) and (S2) into a gas chromatography-mass spectrometry analyzer to obtain an ion flow diagram and a mass spectrum diagram of the reference substance solution and the sample solution, and determining that the sample to be detected is adulterated.
In the present invention, the gas chromatography-mass spectrometry analysis is also preferably used as a supplemental verification of the gas chromatography analysis, further excluding the interference.
The detection method of the invention can effectively identify whether the Artemisia mongolica is adulterated in the Artemisia argyi medicinal material or the decoction piece, has strong specificity, good precision, repeatability and stability, low detection limit, and obtains the same or similar result through repeated experiments, taking example 2 as an example, and the related experimental data are as follows:
1. sample processing
The mugwort leaf medicinal material sample and the Mongolian artemisia herb medicinal material sample are self-collected, and mugwort leaf decoction piece samples are purchased in the market. 3 batches of mugwort medicinal material samples (AC 1-AC 3), 3 batches of mugwort decoction piece samples (AP 1-AP 3) and 8 batches of Artemisia mongolica medicinal material samples (MC 1-MC 8) are collected.
3 samples of mugwort leaf medicinal materials (AC 1-AC 3) and 3 samples of mugwort leaf decoction pieces (AP 1-AP 3) were detected by the detection method of example 2.
1.1 preparation of control solution: taking chamazulene and adding ethyl acetate to prepare a reference substance solution containing 20 mug of chamazulene (namely, 20 mug/mL concentration) per 1 mL;
1.2 preparation of test solutions: cutting folium Artemisiae Argyi materials or decoction pieces into pieces of 0.4-0.6cm, placing 2.5g into round bottom flask, adding 300ml of water, mixing, connecting volatile oil tester, adding water from the upper end of tester to fill scale part, overflowing into flask, adding ethyl acetate 2ml, connecting reflux condenser, heating to boil, heating for 5 hr, cooling to room temperature, separating ethyl acetate liquid, placing into 10ml measuring flask, washing tester and condenser with ethyl acetate, transferring into the same 10ml measuring flask, diluting with ethyl acetate to 10ml scale, shaking to obtain sample solution;
1.3 determination of control solutions and test solutions:
respectively sucking 1 μl of each of the reference solution and the sample solution, performing gas chromatography, and recording chromatogram;
and respectively passing the reference substance solution and the sample solution through 0.22 μm filter membranes, respectively performing gas chromatography-mass spectrometry analysis, and recording mass spectrograms.
Wherein, the gas chromatography conditions are as follows: a chromatography column of Agilent HP-5 19091J (column length 30m, inner diameter 0.32mm, film thickness 0.25 μm) was used, flow rate 1ml/min, gradient temperature increase: the initial temperature is 60 ℃, the temperature is firstly increased to 120 ℃ at the rate of 3 ℃ per minute, and then the temperature is increased to 280 ℃ at the rate of 15 ℃ per minute; sample inlet temperature 240 ℃, FID detector.
The chromatographic conditions for the gas chromatography-mass spectrometry combined analysis are as follows: a Agilent HP-5MS (column length 30m, inner diameter 0.32mm, film thickness 0.25 μm) column was used, flow rate 1ml/min, gradient heating: the initial temperature is 60 ℃, the temperature is firstly increased to 120 ℃ at the rate of 3 ℃ per minute, and then the temperature is increased to 280 ℃ at the rate of 15 ℃ per minute; the sample inlet temperature was 240 ℃.
Electron bombardment of an ion source (EI) by using a Siemens flight TSQ8000EVO-NOVPI gas chromatograph-mass spectrometer; the temperature of the mass spectrum ion source is 250 ℃; the temperature of the ion transmission tube is 250 ℃; the data is collected by full scanning, and the scanning range is 40-600 amu.
The gas phase chromatograms of the chamazulene control substances are shown in fig. 1, the gas phase chromatograms of 3 batches of folium artemisiae argyi decoction pieces and 3 batches of folium artemisiae argyi medicinal material samples are shown in fig. 2 and 3, the retention time and the peak area are shown in table 1, the fact that the decoction piece sample (AP 3) has chromatographic peaks at the positions with consistent retention time of the chamazulene and the peak area is larger than the chromatographic peak-peak area of the control substances, and the fact that the Mongolian artemisia is adulterated is prompted to have a certain quality risk; other batches of samples showed chromatographic peaks at the positions where the retention times of the chamazulene were consistent, but the peak areas were smaller than the chromatographic peak areas of the control solution. Further verified by gas chromatography-mass spectrometry, as shown in fig. 4, the result shows that the decoction piece sample (AP 3) has chromatographic peaks at the positions where the retention time of the chamazulene is consistent, the peak area is larger than that of the control solution, and the mass spectrum is consistent with that of the chamazulene control (see fig. 5). Therefore, a batch of mugwort pieces (AP 3) can be determined to be adulterated with the mongolian artemisia.
Table 1 3 folium Artemisiae Argyi decoction pieces and 3 folium Artemisiae Argyi medicinal materials gas chromatography detection results
2. Sample solution preparation condition selection
2.1 experiment using folium Artemisiae Argyi decoction pieces (AP 3) as raw materials, preparing test solution according to the following three methods respectively:
1. cutting folium Artemisiae Argyi decoction pieces (AP 3) into pieces of 0.4-0.6cm, mixing, adding ethyl acetate 25ml into 2.5g, ultrasonic extracting for 30min, and filtering to obtain sample solution 1;
2. cutting folium Artemisiae Argyi decoction pieces (AP 3) into pieces of 0.4-0.6cm, mixing, adding ethyl acetate 25ml into 2.5g, heating and refluxing for 30min, and filtering to obtain sample solution 2;
3. cutting folium Artemisiae Argyi decoction pieces (AP 3) into pieces of 0.4-0.6cm, mixing, collecting 2.5g, placing into round bottom flask, adding 300ml water, and connecting with volatile oil tester. Water was added from the upper end of the measuring instrument to fill the scale portion, and the mixture was overflowed into a flask, and 2ml of ethyl acetate was further added thereto, followed by connection to a reflux condenser. Heating to boiling, heating for 5 hr, cooling to room temperature, collecting ethyl acetate solution, placing into 10ml measuring flask, washing the tester and condenser tube with ethyl acetate, transferring into the same measuring flask, diluting with ethyl acetate to scale, and shaking to obtain sample solution 3;
taking 1 μl of each of the sample solution 1, the sample solution 2 and the sample solution 3, injecting into a gas chromatograph, and recording the chromatograms.
As shown in fig. 7, as can be seen from the figure, no chamazulene chromatographic peak appears in the chromatograms of the sample solution 1 and the sample solution 2, but only in the sample solution 3, which means that the ethyl acetate ultrasonic and reflux method cannot extract chamazulene, so that the method for extracting volatile oil is selected for optimizing the extraction conditions of the sample in the next step.
2.2 preparing folium Artemisiae Argyi sample solution with different water addition amount, different extraction solvent amount and different extraction time when extracting volatile oil, and performing gas chromatography analysis to obtain the Matricaria azulene peak area results shown in tables 2-4.
TABLE 2 sample chamazulene chromatographic peak areas with different water addition
TABLE 3 Matricaria azulene chromatographic peak areas with different amounts of extractive solvents
TABLE 4 different extraction times Matricaria azulene chromatographic peak areas
As can be seen from Table 2 above, the extraction efficiency was slightly lower when the water addition amount was 200ml, and the extraction efficiency was equivalent when the water addition amount was 300ml to 500ml, so that 300ml was used as the water addition amount in example 2; table 3 shows that the extraction efficiency is relatively low with ethyl acetate in an amount of 1ml, the peak area of the chamazulene is minimal, the ethyl acetate in an amount of 2ml or 3ml, there is no difference in the peak area of the chamazulene, so that example 2 selects ethyl acetate in an amount of 2ml; table 4 shows that the peak area of chamazulene is maximum with an extraction time of 5 hours, so that the extraction time of the present invention is 5 hours.
3. Methodology investigation
3.1 investigation of specificity
Respectively taking mugwort medicinal materials AC 1-AC 3 and Mongolian artemisia medicinal materials MC 1-MC 3, shearing into fragments of 0.4-0.6cm, respectively taking 2.5g of fragments, putting into round-bottom flasks, adding 300ml of water, and connecting with a volatile oil tester. Water was added from the upper end of the measuring instrument to fill the scale portion, and the mixture was overflowed into a flask, and 2ml of ethyl acetate was further added thereto, followed by connection to a reflux condenser. Heating to boiling, heating for 5 hr, cooling to room temperature, collecting ethyl acetate solution, placing into 10ml measuring flask, washing the tester and condenser tube with ethyl acetate, transferring into the same measuring flask, diluting to scale with ethyl acetate, shaking, filtering with 0.45 μm microporous membrane, and performing gas chromatography according to the method described in the above 1.3, and the results are shown in Table 5. The results show that both the mugwort leaf and the Mongolian artemisia contain the chamazulene, but the chamazulene content in the Mongolian artemisia is extremely high and is about 200-500 times of that of the mugwort leaf, which indicates that the mugwort leaf and the Mongolian artemisia can be distinguished by detecting the peak area of the chamazulene.
Table 5 area of Matricaria azulene peak in Artemisia princeps Pampanini
3.2 linear relationship investigation
Taking a chamazulene control and ethyl acetate to prepare a solution containing 1mg/ml of chamazulene per 1ml of the chamazulene control as a chamazulene stock solution. The working control solutions of Matricaria azulene (1. Mu.g/ml, 10. Mu.g/ml, 20. Mu.g/ml, 50. Mu.g/ml, 1000. Mu.g/m) were prepared by stepwise dilution of the stock solutions of Matricaria azulene. Analysis was performed according to the gas chromatography conditions described in 1.3 above, with the concentration of chamazulene as the abscissa and the peak area as the ordinate, and a standard curve was drawn, and the linear regression equation was y= 1.7751x-0.5801, r=1, which indicated that the concentration of chamazulene was good in the concentration range of 1 to 1000 μg/ml.
3.3 concentration of control solution
According to the general rule of detection of medicinal materials and decoction pieces in four portions 0212 of Chinese pharmacopoeia, the impurity content is usually less than 3%. Considering that the mugwort leaf shape is greatly changed, the character identification is difficult, the mixed growth risk of mugwort and mongolian artemisia is generated, the limit of the mixed mongolian artemisia in mugwort is 10%, 8 batches of mongolian artemisia samples (MC 1-MC 8) are mixed, 0%,5%,10% and 20% of the mixed mongolian artemisia samples in mugwort are respectively prepared, the analysis by a gas chromatograph is carried out according to the method 1.3, the area of the azulene peak of the chamomile is measured, the result is shown in figure 6, and the data are shown in Table 6. The peak area of chamazulene at 10% adulteration was finally referenced. According to the linear relation between the chromatographic peak area and the concentration of the chamazulene, the concentration of the chamazulene when the Mongolian artemisia is doped by 10 percent is calculated to be 18.4 mug/ml. Therefore, the concentration of the chamazulene control solution is preferably 20. Mu.g/ml.
TABLE 6 Peak areas for samples of different adulteration amounts
3.4 precision investigation
The chamazulene control solution (20 mug/ml) was taken and continuously injected for 6 times under the gas chromatography condition described in 1.3, the peak area of the chamazulene is shown in Table 7, and the result shows that the detection method of the invention has good precision.
TABLE 7 precision investigation results
3.5 repeatability investigation
6 parts of the same sample (AP 3) are taken, treated according to the method described in 1.2, and measured under the gas chromatography condition described in 1.3, and the test results are shown in Table 8, and the results show that the detection method provided by the invention has good repeatability.
Table 8 repeatability results
3.6 stability investigation
Folium artemisiae argyi decoction pieces (AP 3) were taken, treated as described in 1.2 above, and analyzed under the gas chromatography conditions described in 1.3 for 0,2,4,6,8, 10 and 12 hours, respectively, and the test results are shown in Table 9, which shows that the test sample solution was stable within 12 hours from the preparation.
TABLE 9 stability results
3.7 limit of detection experiment
The detection limit of the gas chromatography detection method is determined according to the signal to noise ratio of more than 3:1, and the detection limit of the chamazulene in the gas chromatography detection method is 1ng, which indicates that the detection limit of the detection method established by the invention is low.
3.8 recovery experiments
Taking a proper amount of a sample (MC 3, chamazulene content 1.0 mg/g) of a medicinal material of the Artemisia mongolica with known content, shearing the sample into fragments of 0.4-0.6cm, uniformly mixing the fragments, taking 1.25g of the fragments, placing the fragments into a volatile oil extraction device, adding 1006.8mg of a chamazulene reference substance solution, preparing a sample-adding test substance solution according to the method 1.2, carrying out parallel treatment on 6 parts, carrying out gas chromatography detection according to the chromatographic condition 1.3, calculating a sample-adding recovery rate, and obtaining a result shown in Table 10, wherein the recovery rate is more than 90%, and the established method is good in recovery rate.
TABLE 10 recovery measurement results
Note that: recovery = (measured crude chamazulene content-chamazulene content in sample)/amount of added chamazulene control
Meanwhile, the invention performs the experiment on the embodiment 2, and simultaneously performs the same screening and verification experiments on the embodiments 1 and 3, and the results which are the same as and similar to those of the embodiment 2 are obtained, so that the invention is not listed here, and the effects of the invention are good, and the curative effect is stable and reliable.
The method is simple and easy to operate, and the experiment shows that the detection method is based on the chemical components of the Artemisia mongolica, namely the chamomile azulene with higher content and lower content in the Artemisia argyi, is a rapid and accurate detection method of the Artemisia mongolica doped in the Artemisia argyi, has the advantages of rapid and effective detection process, stable and reliable method, strong specificity, good precision, repeatability and stability, low detection limit, 1ng detection limit, and can effectively identify whether the Artemisia argyi is doped in the Artemisia argyi medicinal materials or decoction pieces, effectively fill the technical blank of detecting the Artemisia argyi doped in the Artemisia argyi based on the chemical components, provide powerful technical guarantee for medicine quality supervision, and is a great innovation in the detection method of the Artemisia argyi doped in the Artemisia argyi, and has remarkable social and economic benefits.
It is noted that the above-mentioned embodiments are merely preferred embodiments of the present invention, and the present invention is not limited thereto, and that any person skilled in the art can make modifications or variations within the scope of the present invention without departing from the scope of the present invention.
Claims (6)
1. The detection method of the adulterated Artemisia mongolica in the mugwort medicinal materials and decoction pieces is characterized by comprising the following steps of:
s1, preparing a reference substance solution: taking a chamazulene reference substance and an ethyl acetate solvent to prepare a reference substance solution with the concentration of 10-30 mug/mL;
s2, preparing a sample solution: cutting a mugwort leaf medicinal material or decoction piece sample into fragments of 0.4-0.6cm, placing 2-3 g of the fragments in a round bottom flask, adding 200-500 ml of water, uniformly mixing, connecting a volatile oil tester, adding water from the upper end of the tester to fill a scale part, overflowing the volatile oil tester into the flask, adding 1-3 ml of ethyl acetate, connecting a reflux condenser pipe, heating to boil, continuously heating for 5 hours, then cooling to room temperature, separating ethyl acetate liquid, placing into a 10ml measuring flask, washing the tester and the condenser pipe with ethyl acetate for several times, transferring into the same 10ml measuring flask, diluting the ethyl acetate to the scale of 10ml, and shaking uniformly to obtain a sample solution;
s3, chromatogram: taking 1 mu L of the reference substance solution and the sample solution obtained in the steps S1 and S2 respectively, and analyzing by a gas chromatograph to obtain a reference substance solution chromatogram and a sample solution chromatogram;
s4, adulteration analysis: comparing the chromatogram of the sample with the chromatogram of the reference sample, and simultaneously meeting the following two conditions, namely, taking the sample as adulteration:
(1) Chromatographic peaks appear in the sample solution chromatogram at positions consistent with the retention time of the chromatographic peaks of the chamazulene in the control solution chromatogram;
(2) The chromatographic peak area of the sample solution is larger than that of the reference solution;
s5, verification: and when S4, judging that the Mongolian artemisia is adulterated, verifying by adopting a gas chromatography-mass spectrometry method, namely, respectively passing the reference substance solution and the sample solution obtained in the steps S1 and S2 through a 0.22 mu m filter membrane, then injecting the reference substance solution and the sample solution into a gas chromatography-mass spectrometry analyzer to obtain an ion flow diagram and a mass spectrogram of the reference substance solution and the sample solution, and when the mass spectrograms of the reference substance solution and the sample solution are consistent, determining that the sample to be detected is adulterated.
2. The method for detecting the adulterated tarragon in the mugwort medicinal materials and the decoction pieces according to claim 1, which is characterized by comprising the following steps:
s1, preparing a reference substance solution: taking a chamazulene control substance and an ethyl acetate solvent to prepare a control substance solution with the concentration of 10 mug/mL;
s2, preparing a sample solution: cutting mugwort leaf medicinal materials or decoction pieces into pieces of 0.4-0.6cm, placing 2g into a round bottom flask, adding 200ml of water, mixing, connecting a volatile oil tester, adding water from the upper end of the tester to fill the scale part, overflowing into the flask, adding 1ml of ethyl acetate, connecting a reflux condenser pipe, heating to boil, heating for 5 hours, then cooling to room temperature, separating ethyl acetate liquid, placing into a 10ml measuring flask, washing the tester and condenser pipe with ethyl acetate for several times, transferring into the same 10ml measuring flask, diluting with ethyl acetate to the scale of 10ml, shaking uniformly to obtain a sample solution;
s3, chromatogram: taking 1 mu L of the reference substance solution and the sample solution obtained in the steps S1 and S2 respectively, and analyzing by a gas chromatograph to obtain a reference substance solution chromatogram and a sample solution chromatogram;
s4, adulteration analysis: comparing the chromatogram of the sample with the chromatogram of the reference sample, and simultaneously meeting the following two conditions, namely, taking the sample as adulteration:
(1) Chromatographic peaks appear in the sample solution chromatogram at positions consistent with the retention time of the chromatographic peaks of the chamazulene in the control solution chromatogram;
(2) The chromatographic peak area of the sample solution is larger than that of the reference solution;
s5, verification: and when S4, judging that the Mongolian artemisia is adulterated, verifying by adopting a gas chromatography-mass spectrometry method, namely, respectively passing the reference substance solution and the sample solution obtained in the steps S1 and S2 through a 0.22 mu m filter membrane, then injecting the reference substance solution and the sample solution into a gas chromatography-mass spectrometry analyzer to obtain an ion flow diagram and a mass spectrogram of the reference substance solution and the sample solution, and when the mass spectrograms of the reference substance solution and the sample solution are consistent, determining that the sample to be detected is adulterated.
3. The method for detecting the adulterated tarragon in the mugwort medicinal materials and the decoction pieces according to claim 1, which is characterized by comprising the following steps:
s1, preparing a reference substance solution: taking a chamazulene control substance and an ethyl acetate solvent to prepare a control substance solution with the concentration of 20 mug/mL;
s2, preparing a sample solution: cutting mugwort leaf medicinal materials or decoction piece samples into fragments of 0.4-0.6cm, placing 2.5g in a round bottom flask, adding 300ml of water, uniformly mixing, connecting a volatile oil tester, adding water from the upper end of the tester to fill the scale part, overflowing into the flask, adding 2ml of ethyl acetate, connecting a reflux condenser pipe, heating to boil, continuously heating for 5 hours, then cooling to room temperature, separating ethyl acetate liquid, placing into a 10ml measuring flask, washing the tester and the condenser pipe with ethyl acetate in a fractional manner, transferring into the same 10ml measuring flask, diluting the ethyl acetate to the scale of 10ml, and shaking uniformly to obtain a sample solution;
s3, chromatogram: taking 1 mu L of the reference substance solution and the sample solution obtained in the steps S1 and S2 respectively, and analyzing by a gas chromatograph to obtain a reference substance solution chromatogram and a sample solution chromatogram;
s4, adulteration analysis: comparing the chromatogram of the sample with the chromatogram of the reference sample, and simultaneously meeting the following two conditions, namely, taking the sample as adulteration:
(1) Chromatographic peaks appear in the sample solution chromatogram at positions consistent with the retention time of the chromatographic peaks of the chamazulene in the control solution chromatogram;
(2) The chromatographic peak area of the sample solution is larger than that of the reference solution;
s5, verification: and when S4, judging that the Mongolian artemisia is adulterated, verifying by adopting a gas chromatography-mass spectrometry method, namely, respectively passing the reference substance solution and the sample solution obtained in the steps S1 and S2 through a 0.22 mu m filter membrane, then injecting the reference substance solution and the sample solution into a gas chromatography-mass spectrometry analyzer to obtain an ion flow diagram and a mass spectrogram of the reference substance solution and the sample solution, and when the mass spectrograms of the reference substance solution and the sample solution are consistent, determining that the sample to be detected is adulterated.
4. The method for detecting the adulterated tarragon in the mugwort medicinal materials and the decoction pieces according to claim 1, which is characterized by comprising the following steps:
s1, preparing a reference substance solution: taking a chamazulene control substance and an ethyl acetate solvent to prepare a control substance solution with the concentration of 30 mug/mL;
s2, preparing a sample solution: cutting folium Artemisiae Argyi or decoction pieces into pieces of 0.4-0.6cm, placing 3g into round bottom flask, adding 500ml of water, mixing, connecting volatile oil tester, adding water from the upper end of tester to fill scale part, overflowing into flask, adding ethyl acetate 3ml, connecting reflux condenser, heating to boil, heating for 5 hr, standing at room temperature, separating ethyl acetate liquid, placing into 10ml measuring flask, washing tester and condenser with ethyl acetate, transferring into the same 10ml measuring flask, diluting with ethyl acetate to 10ml scale, shaking to obtain sample solution;
s3, chromatogram: taking 1 mu L of the reference substance solution and the sample solution obtained in the steps S1 and S2 respectively, and analyzing by a gas chromatograph to obtain a reference substance solution chromatogram and a sample solution chromatogram;
s4, adulteration analysis: comparing the chromatogram of the sample with the chromatogram of the reference sample, and simultaneously meeting the following two conditions, namely, taking the sample as adulteration:
(1) Chromatographic peaks appear in the sample solution chromatogram at positions consistent with the retention time of the chromatographic peaks of the chamazulene in the control solution chromatogram;
(2) The chromatographic peak area of the sample solution is larger than that of the reference solution;
s5, verification: and when S4, judging that the Mongolian artemisia is adulterated, verifying by adopting a gas chromatography-mass spectrometry method, namely, respectively passing the reference substance solution and the sample solution obtained in the steps S1 and S2 through a 0.22 mu m filter membrane, then injecting the reference substance solution and the sample solution into a gas chromatography-mass spectrometry analyzer to obtain an ion flow diagram and a mass spectrogram of the reference substance solution and the sample solution, and when the mass spectrograms of the reference substance solution and the sample solution are consistent, determining that the sample to be detected is adulterated.
5. The method for detecting the adulterated tarragon in the mugwort medicinal material and the decoction piece according to any one of claims 1 to 4, wherein the gas chromatographic analysis condition in the step S3 is as follows: the chromatographic column stationary phase is 5% phenyl-95% methyl polysiloxane, the carrier gas flow rate is 0.8-1.2 ml/min, and the gradient temperature is raised: the initial temperature is 60 ℃, the temperature is firstly increased to 120 ℃ at the rate of 3 ℃ per minute, and then the temperature is increased to 280 ℃ at the rate of 15 ℃ per minute; the temperature of the sample inlet is 240 ℃, the sample inlet amount is 1-2 ul, and the FID detector.
6. The method for detecting the adulterated tarragon in the mugwort medicinal material and the decoction piece according to any one of claims 1 to 4, wherein the chromatographic conditions of the gas chromatography-mass spectrometry combined analysis in the step S5 are as follows: chromatographic column stationary phase: 5% phenyl-95% methylpolysiloxane; carrier gas flow rate: 1ml/min; gradient heating: the initial temperature is 60 ℃, the temperature is firstly increased to 120 ℃ at the rate of 3 ℃ per minute, and then the temperature is increased to 280 ℃ at the rate of 15 ℃ per minute; sample inlet temperature: 240 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310124253.9A CN116148388A (en) | 2023-02-16 | 2023-02-16 | Detection method of adulterated Artemisia mongolica in mugwort medicinal materials and decoction pieces |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310124253.9A CN116148388A (en) | 2023-02-16 | 2023-02-16 | Detection method of adulterated Artemisia mongolica in mugwort medicinal materials and decoction pieces |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116148388A true CN116148388A (en) | 2023-05-23 |
Family
ID=86352272
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310124253.9A Pending CN116148388A (en) | 2023-02-16 | 2023-02-16 | Detection method of adulterated Artemisia mongolica in mugwort medicinal materials and decoction pieces |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116148388A (en) |
-
2023
- 2023-02-16 CN CN202310124253.9A patent/CN116148388A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113866293A (en) | Method for rapidly identifying and analyzing chemical components in chamomile | |
| CN113777183B (en) | Glossy privet fruit medicinal material and its processed product characteristic spectrum construction method and multi-index component content detection method | |
| CN109655558B (en) | Method for detecting effective part group of periploca forrestii schltr | |
| CN110286169B (en) | Method for simultaneously extracting and respectively purifying 5 chemical components from processed ramulus mori and application thereof | |
| CN110031564B (en) | Quality detection method of natural plant anticoccidial feed additive based on HPLC fingerprint | |
| CN109374786B (en) | Construction method and quality detection method of UPLC (ultra performance liquid chromatography) characteristic spectrum of chrysanthemum morifolium ramat medicinal material | |
| CN111077242A (en) | High performance liquid chromatography analysis method of traditional Chinese medicine formula Dachengqi decoction | |
| CN109307717B (en) | Method for detecting content of polyphenol compounds in cannabis sativa oil | |
| CN111679015A (en) | Bletilla striata small molecule characteristic fingerprint spectrum analysis method | |
| CN115575551B (en) | Bletilla striata detection method | |
| CN109187784B (en) | Headspace solid phase microextraction-gas chromatography-mass spectrometry detection and analysis method for sea buckthorn volatile oil component | |
| CN115078608B (en) | Method for identifying UPLC characteristic patterns of herba abri and herba abri | |
| CN116148388A (en) | Detection method of adulterated Artemisia mongolica in mugwort medicinal materials and decoction pieces | |
| CN114354828B (en) | Characteristic spectrum of herba Lagotis Brevibacterium, construction method thereof and quality detection method of herba Lagotis Brevibacterium | |
| CN114646690B (en) | Detection method for chemical components in immature bitter orange, longstamen onion bulb and cassia twig decoction and fingerprint establishment method | |
| CN113759011B (en) | Method for establishing characteristic spectrum of starwort root and preparation thereof | |
| CN109374787B (en) | Construction method and detection method of radix stemonae tuberose medicinal material UPLC characteristic spectrum | |
| CN109358120B (en) | Method for detecting characteristic spectrum of sophora fruit preparation | |
| CN114720602B (en) | Method for constructing characteristic spectrum of lithospermum through medicinal material or standard decoction thereof and detection method | |
| CN116818918B (en) | Fingerprint construction method of ligustrum robustum and construction method of high-resolution mass spectrum component basic identification and application thereof | |
| CN113325095B (en) | UPLC fingerprint construction method and detection method of garden burnet medicinal material | |
| CN114689750B (en) | Construction method of HPLC fingerprint of n-butanol part of Mongolian almond and kindred species thereof | |
| CN114894922B (en) | Detection and quality control method of gynostemma pentaphylla water extract | |
| CN115236224B (en) | Method for measuring scopoletin content in Chinese honeylocust spine traditional Chinese medicine decoction piece standard decoction | |
| CN115452964B (en) | UPLC characteristic spectrum construction method and quality control method of calyx seu fructus physalis |
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 |