CN113156019A - Method for detecting multi-component content of liquorice heart-fire-purging decoction - Google Patents
Method for detecting multi-component content of liquorice heart-fire-purging decoction Download PDFInfo
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Abstract
A method for detecting multi-component content of Glycyrrhrizae radix decoction for purging stomach-fire comprises preparing test solution, preparing reference solution, and detecting by ultra-high performance liquid chromatography; wherein the test solution is prepared by collecting extractive solution of Glycyrrhrizae radix heart fire clearing decoction, centrifuging, collecting supernatant, volatilizing solvent, dissolving in 50% methanol solution, diluting to desired volume, and filtering with microporous membrane to obtain test solution. The method for detecting the multi-component content of the licorice heart-fire-purging decoction has the advantages that the separation degree of 8 effective components is high, the adjacent two components can be separated from each other by a base line, the method is high in accuracy and good in linearity, the precision and the intermediate precision are good, the solution stability is good, the sample solution is stable within 12 hours, the method is simple to operate, the working efficiency and the detection precision are greatly improved, and the product quality of the licorice heart-fire-purging decoction and related preparations can be controlled more comprehensively and effectively.
Description
Technical Field
The invention relates to the field of traditional Chinese medicine quality detection, and in particular relates to a quantitative detection method for multi-component content of licorice heart-fire-purging decoction.
Background
The licorice heart-fire-purging decoction is a famous formula of a traditional Chinese medicine prescription, is from Shang Han Lun, and comprises 20g of licorice, 15 g of scutellaria, 13 g of pinellia, 12g of Chinese date, 5 g of coptis and 15 g of dried ginger. In the prescription, the liquorice is used for tonifying middle-jiao and spleen-stomach, so that qi of spleen and stomach is restored, qi and blood are generated and the functions of the liquorice are also managed, the coptis chinensis and the scutellaria baicalensis clear heat and dry dampness, so that the spleen and the stomach are not abused by damp heat, the pinellia ternate and the rhizoma zingiberis are used for promoting qi circulation in middle-jiao, so that damp heat is free from internal mechanisms, the Chinese date is used for tonifying middle-jiao and qi, and is used with the liquorice to treat diseases and strengthen body resistance and eliminate evil, healthy qi is recovered and not abused as evil, and symptoms are caused by abuse, so that all medicines are combined to achieve bitter cold and purge but not severe, warm and unblocked but not disperse healthy qi, and the liquorice is used for tonifying and orderly to harmonize middle-jiao and consolidate the root. Has the effects of tonifying qi, regulating stomach function, relieving distension and fullness and arresting vomiting. Can be used for treating typhoid fever and abdominal fullness, stomach qi deficiency, abdominal thunder, diarrhea, food stagnation, nausea, retching, vexation, and Huoxian disease. It is often used clinically for acute and chronic gastroenteritis, Behcet's syndrome, etc.
At present, a method for monitoring the contents of a plurality of effective components of the licorice heart-fire-purging decoction by only one method does not exist, and the quality detection method of the licorice heart-fire-purging decoction only measures the content of a single component according to a high performance liquid chromatography, if a plurality of components need to be controlled, a plurality of methods need to be applied to respectively measure, the detection work is complicated, the energy consumption is large, and the quality of the licorice heart-fire-purging decoction cannot be comprehensively and objectively evaluated.
Disclosure of Invention
The invention aims to provide a method for measuring the multi-component content of liquorice heart-fire-purging decoction, which can objectively, comprehensively and accurately evaluate the quality of the liquorice heart-fire-purging decoction and has important significance for controlling the quality of the liquorice heart-fire-purging decoction and ensuring the clinical curative effect.
The aim of the invention is realized by the following technical measures:
a method for detecting the multi-component content of a licorice heart-fire-purging decoction is characterized by comprising the steps of preparing a test solution, preparing a reference solution and detecting by using an ultra-high performance liquid chromatography; wherein the test solution is prepared by collecting extractive solution of Glycyrrhrizae radix heart fire purging decoction, centrifuging, collecting supernatant, volatilizing solvent, dissolving in methanol solution to desired volume, and filtering with microporous membrane to obtain test solution.
Further, the use amount of the licorice heart-fire clearing decoction extract is 100ml, the supernatant obtained after centrifugation is 5ml, the volume fraction of the methanol solution is 50%, the constant volume is a 5ml volumetric flask, and the aperture of the microporous filter membrane is 0.22 micron.
Further, the licorice heart-fire clearing soup is prepared by the following method:
(1) respectively placing Glycyrrhrizae radix, Scutellariae radix, rhizoma Pinelliae, fructus Jujubae, Coptidis rhizoma, and Zingiberis rhizoma in universal pulverizer, pulverizing respectively, sieving with No. 1 sieve and No. 2 sieve, and collecting powder that has passed through No. 1 sieve but not No. 2 sieve;
(2) taking 20 parts by weight of licorice powder, 15 parts by weight of scutellaria powder, 13 parts by weight of pinellia powder, 12 parts by weight of jujube powder, 5 parts by weight of coptis powder and 15 parts by weight of dried ginger powder, uniformly mixing, putting the mixture into a reflux extraction device, extracting for three times, adding 800 parts of water with the pH value adjusted to 4.0 by 0.1mol/L hydrochloric acid for the first time, adding 640 parts of water with the pH value adjusted to 7.0 by 0.1mol/L sodium hydroxide solution for the second time, adding 640 parts of water with the pH value adjusted to 8.0 by 0.1mol/L sodium hydroxide solution for the third time, wherein the extraction time is 1 hour for the first time, 0.5 hour for the second time, 0.5 hour for the third time, the extraction temperature is 90-95 ℃, combining the three extracting solutions, filtering, and concentrating the filtrate under reduced pressure until the relative density is 1.08-1.12 (25 ℃), thus obtaining the extracting solution of licorice heart-eliminating decoction; the temperature obtained by the decompression concentration is 60-80 ℃, and the vacuum degree is-0.05-0.08 MPa.
Further, the reference solution is prepared by dissolving and diluting zingerone, berberine hydrochloride, liquiritigenin, berberine hydrochloride, wogonoside, baicalein, 6-gingerol and ammonium glycyrrhizinate respectively with 50% methanol by volume fraction to obtain 1mg/ml solution, and using the solution as zingerone reference stock solution, berberine hydrochloride reference stock solution, liquiritigenin reference stock solution, berberine hydrochloride reference stock solution, wogonin reference stock solution, baicalein reference stock solution, 6-gingerol reference stock solution and ammonium glycyrrhizinate reference stock solution for later use; and precisely measuring the reference substance stock solutions respectively, placing the reference substance stock solutions in the same volumetric flask, diluting with 50% methanol solution by volume fraction, and fixing the volume to scale to prepare a composite reference substance solution, wherein the concentrations of zingerone, berberine hydrochloride, liquiritigenin, berberine hydrochloride, wogonoside, baicalein, 6-gingerol and ammonium glycyrrhizinate in the composite reference substance solution are all 0.5 mg/ml.
Further, the detection of the ultra-high performance liquid chromatography is that 3 mul of the prepared reference solution and the prepared sample solution are precisely absorbed and injected into the ultra-high performance liquid chromatograph, and the calculation is carried out by an external standard method; the column temperature of the ultra-high performance liquid chromatograph is 35 ℃, the flow rate is 0.35ml/min, and the wavelength range is 190-400 nm.
Further preferably, the flow term of the ultra high performance liquid chromatography includes a flow term a and a flow term B, wherein the flow term a is a trifluoroacetic acid aqueous solution with a volume fraction of 0.1%, and the flow term B is a mixture of methanol: acetonitrile 50: 50, gradient elution is adopted.
In order to make the detected components reach baseline separation and meet the content detection requirement, the gradient elution procedure of the ultra-high performance liquid chromatography is carried out as follows:
| time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 80 | 20 |
| 20 | 75 | 25 |
| 30 | 65 | 35 |
| 40 | 40 | 60 |
| 55 | 50 | 50 |
| 60 | 80 | 20 |
The specific components of the flowing item A and the flowing item B are matched with a specific gradient elution program, so that the paradol, the berberine hydrochloride, the liquiritigenin, the berberine hydrochloride, the wogonoside, the baicalein, the 6-gingerol and the ammonium glycyrrhizinate can achieve baseline separation within 1 hour, the content detection requirement is met, the content of 8 components can be simultaneously monitored by using one detection method, 0.1% trifluoroacetic acid in the flowing item A ensures that a detected substance is in an acidic environment, the phenomenon that a tailing peak appears in a chromatographic peak of the detected substance of an ultrahigh pressure liquid chromatography is ensured, the accuracy of the method is further improved, and the content of 8 components in the licorice heart-purging soup can be monitored with high sensitivity and high efficiency.
Further, the method for detecting the content of the multiple components of the licorice heart-fire-purging decoction can be applied to content detection of extracts and preparations taking the licorice heart-fire-purging decoction as a component.
The invention has the following beneficial effects:
the invention relates to a method for detecting the multi-component content of licorice heart-fire clearing decoction, which solves the technical problems that the licorice heart-fire clearing decoction has too many components according to the formula, the separation degree of 8 components such as zingerone, berberine hydrochloride, liquiritigenin, berberine hydrochloride, wogonin, baicalein, 6-gingerol, ammonium glycyrrhetate and the like is not high and much interference is generated in the detection process, and 8 effective components cannot be simultaneously monitored by using a content analysis method, the separation degree of 8 effective components is high in the detection method, the adjacent two components can be separated by a base line, the method has high accuracy, the sample recovery rate is 95-108 percent, the RSD of 6 sample recovery samples is less than 4 percent, the linearity is good, the linear regression R is more than 0.999 in the detection concentration range of a reference substance and a sample, the precision and the middle precision are good, the RSD is within 5 percent, the solution stability is good, and the sample solution is stable within 12 hours, the method is simple to operate, greatly improves the working efficiency and the detection precision, and can more comprehensively and effectively control the product quality of the liquorice heart-fire-purging decoction and related preparations.
Drawings
FIG. 1: is the ultra-high performance liquid chromatogram of the composite reference substance in example 1 of the invention.
FIG. 2: is the ultra-high performance liquid chromatogram of the test solution in example 1 of the invention.
FIG. 3: is the linear regression chart of zingerone.
FIG. 4: is the linear regression chart of coptisine hydrochloride.
FIG. 5: is a glycyrrhizic acid linear regression graph.
FIG. 6: is a linear regression chart of berberine hydrochloride.
FIG. 7: is a wogonoside linear regression chart.
FIG. 8: is the baicalein linear regression chart of the invention.
FIG. 9: is a 6-gingerol linear regression diagram.
FIG. 10: is a linear regression chart of ammonium glycyrrhetate.
Detailed Description
The present invention is described in detail below by way of examples, it being necessary to note that the following examples are provided only for illustrating the present invention and are not to be construed as limiting the scope of the present invention, and modifications or substitutions of the method, steps or conditions of the present invention may be made without departing from the spirit and spirit of the present invention.
Example 1
A method for detecting the content of multiple components of licorice heart-fire-purging decoction comprises the following steps:
1. preparation of a test solution:
(1) respectively placing Glycyrrhrizae radix, Scutellariae radix, rhizoma Pinelliae, fructus Jujubae, Coptidis rhizoma, and Zingiberis rhizoma in universal pulverizer, pulverizing respectively, sieving with No. 1 sieve and No. 2 sieve, and collecting powder that has passed through No. 1 sieve but not No. 2 sieve;
(2) taking the pulverized medicinal powder, 200g of licorice powder, 150g of scutellaria powder, 130g of pinellia powder, 120g of jujube powder, 50g of coptis powder and 150g of dried ginger powder, putting the mixture into a suitable container, uniformly mixing, putting the mixture into a reflux extraction device, adding water for three times, adding 8000g of water with the pH value adjusted to 4.0 by using 0.1mol/L hydrochloric acid for the first time, adding 6400g of water with the pH value adjusted to 7.0 by using 0.1mol/L sodium hydroxide solution for the second time, adding 6400g of water with the pH value adjusted to 8.0 by using 0.1mol/L sodium hydroxide solution for the third time, extracting for 1 hour for the first time, extracting for 0.5 hour for the second time and extracting for 0.5 hour for the third time at the extraction temperature of 95 ℃, combining the three extracting solutions, filtering, and concentrating the filtrate under reduced pressure until the relative density is 1.11(25 ℃), thus obtaining the extracting solution of licorice heart-fire-eliminating decoction; the temperature is 70 ℃ after the decompression concentration, and the vacuum degree is-0.07 MPa.
(3) Taking 100ml of the extract of the licorice heart-fire-purging decoction, centrifuging, taking 5ml of supernatant after centrifugation, volatilizing the solvent, adding 50% methanol by volume fraction to dissolve, metering to a volumetric flask of 5ml, and filtering by using a 0.22 micron microporous membrane to obtain a test solution.
2. Preparation of control solutions:
dissolving zingerone, berberine hydrochloride, liquiritigenin, berberine hydrochloride, wogonin, baicalein, 6-gingerol and ammonium glycyrrhizinate reference substances with 50% methanol by volume fraction, and diluting to obtain 1mg/ml solution as zingerone reference substance stock solution, berberine hydrochloride reference substance stock solution, liquiritigenin reference substance stock solution, berberine hydrochloride reference substance stock solution, wogonin reference substance stock solution, baicalein reference substance stock solution, 6-gingerol reference substance stock solution and ammonium glycyrrhizinate reference substance stock solution for use; and precisely measuring the reference substance stock solutions respectively, placing the reference substance stock solutions in the same volumetric flask, diluting with 50% methanol solution by volume fraction, and fixing the volume to scale to prepare a composite reference substance solution, wherein the concentrations of zingerone, berberine hydrochloride, liquiritigenin, berberine hydrochloride, wogonoside, baicalein, 6-gingerol and ammonium glycyrrhizinate in the composite reference substance solution are all 0.5 mg/ml.
3. Detecting by ultra-high performance liquid chromatography:
precisely sucking 3 μ l of the prepared reference solution and test solution, injecting into an ultra-high performance liquid chromatograph, calculating the contents of zingerone, berberine hydrochloride, liquiritigenin, berberine hydrochloride, wogonin, baicalein, 6-gingerol and ammonium glycyrrhizinate in the sample of example 1 by an external standard method, wherein the ultra-high performance liquid chromatogram of the composite reference product is shown in figure 1, and as can be seen from the figure, the retention time is 7.473min for zingerone, 17.345min for berberine hydrochloride, 19.959min for liquiritigenin, 25.821min for small plica of hydrochloric acid, 32.206min for wogonin, 34.224min for baicalein, 38.257min for 6-gingerol and 41.212min for ammonium glycyrrhizinate; wherein the Glycyrrhrizae radix comprises 2 medicinal materials, which are glycyrrhizin and ammonium glycyrrhizinate respectively; 2 medicinal materials of scutellaria baicalensis are respectively wogonoside and baicalein; 2 coptis medicinal materials are respectively berberine hydrochloride and berberine hydrochloride; 2 ginger medicinal materials are gingerol and 6-gingerol respectively. The ultra-high performance liquid chromatogram of the sample is shown in figure 2, and it can be seen that retention time is 7.233min for zingerone, 17.977min for coptisine hydrochloride, 20.263min for glycyrrhizin, 25.582min for paulline hydrochloride, 32.251min for wogonin, 33.634min for baicalein, 38.281min for 6-gingerol, and 41.245 ammonium glycyrrhizinate; the detection retention time of 8 components of the test solution corresponds to the retention time of 8 components of the composite reference substance one by one.
Chromatographic conditions are as follows: shimadzu Shim-pack XR-ODS II (75 mm. times.2.0 mm i.d., 2.2 μm) was used as a column, the column temperature was 35 ℃, the flow rate was 0.35ml/min, and the wavelength was 201 nm;
flow term A: 0.1% by volume of aqueous trifluoroacetic acid;
flow term B: the volume ratio is methanol: acetonitrile 50: 50.
elution was performed according to the following gradient program:
| time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 80 | 20 |
| 20 | 75 | 25 |
| 30 | 65 | 35 |
| 40 | 40 | 60 |
| 55 | 50 | 50 |
| 60 | 80 | 20 |
According to the peak area of the reference substance and the peak area of the sample in the ultra-high performance liquid chromatogram, the content detection results of zingerone, berberine hydrochloride, liquiritigenin, berberine hydrochloride, wogonoside, baicalein, 6-gingerol and ammonium glycyrrhizinate in example 1 are calculated by an external standard method and are shown in the following table:
the results show that: the detection method can simultaneously detect the content of 8 effective components under one chromatographic condition, wherein the content comprises zingerone, berberine hydrochloride, liquiritigenin, berberine hydrochloride, wogonoside, baicalein, 6-gingerol and ammonium glycyrrhizinate, and the adjacent two component peaks can achieve baseline separation and good separation degree.
Example 2
A linear relation investigation of a detection method for the content of multiple components of the licorice heart-fire-purging decoction comprises the following steps:
taking mixed reference substance solution (containing zingerone, berberine hydrochloride, liquiritigenin, berberine hydrochloride, wogonoside, baicalein, 6-gingerol and ammonium glycyrrhizinate at concentrations of 0.2700, 0.0552, 0.0145, 1.2138, 0.5664, 0.0115, 0.0048 and 0.2190mg/ml respectively), automatically injecting sample at 0.4, 0.8, 1.0, 3.0, 5.0, 8.0 and 10.0 μ L respectively, and measuring chromatographic peak area. Taking the sample amount of the reference solution of zingerone, berberine hydrochloride, liquiritigenin, berberine hydrochloride, wogonin, baicalein, 6-gingerol and ammonium glycyrrhizinate as abscissa and the area of chromatographic peak as ordinate to obtain the standard curve. The results show that: the regression equation for zingerone is: Y2E +0.6x +53766, R20.9993; the regression equation of the coptisine hydrochloride is as follows: 828202x-87170 and R20.9996; the regression equation for glycyrrhizin is: Y-1E +0.6x-444698, R20.9991; the regression equation of berberine hydrochloride is: Y-1E +0.6x-228287，R20.9997; the regression equation of wogonoside is as follows: y ═ 1E +0.6x-87788, R20.9995; the regression equation for baicalein is: Y-2E +0.6x-435630, R20.9996; the regression equation for 6-gingerol is: Y2E +0.6x +307533, R20.9990; the regression equation for ammonium glycyrrhizinate is: 117480x-42069, R20.9993; the method shows that the sampling amounts of the zingerone, the coptisine hydrochloride, the liquiritigenin, the berberine hydrochloride, the wogonin, the baicalein, the 6-gingerol and the ammonium glycyrrhizinate are respectively 0.1080-2.700 mg/ml, 0.0221-0.5520 mg/ml, 0.0058-0.1450 mg/ml, 0.4855-12.1380 mg/ml, 0.2266-5.6640 mg/ml, 0.0046-0.1150 mg/ml, 0.0019-0.0480 mg/ml and 0.0876-2.1900 mg/ml, the linear relation is good, and the detection requirements are met.
Example 3
A detection method for the content of multiple components of a licorice heart-fire-purging decoction has the following precision tests:
and taking the mixed reference substance solution, continuously injecting samples for 6 times respectively according to the established chromatographic conditions, recording the chromatographic peak area and calculating RSD. The results show that the RSD values of the zingerone, the coptisine hydrochloride, the liquiritigenin, the berberine hydrochloride, the wogonin, the baicalein, the 6-gingerol and the ammonium glycyrrhetate are respectively 3.56%, 3.44%, 3.56%, 3.21%, 1.50%, 3.75%, 2.08% and 2.03%, and the detection conditions are met, which indicates that the detection method disclosed by the invention is good in precision and meets the detection requirements.
Example 4
A method for detecting the content of multiple components of licorice heart-fire-purging decoction comprises the following steps:
the sample solution is precisely absorbed, and sample injection is repeated for 7 times at 0, 2, 4, 6, 8, 10 and 12 hours according to the chromatographic conditions established above, and the results show that the RSD values of the zingerone, the coptisine hydrochloride, the liquiritigenin, the berberine hydrochloride, the wogonin, the baicalein, the 6-gingerol and the ammonium glycyrrhetate are respectively 3.52%, 3.12%, 4.70%, 0.97%, 3.02%, 4.10%, 2.78% and 3.01%, which shows that the stability of the sample solution is good in 12 hours, and the sample injection does not cause detection errors when the sample solution is prepared for 12 hours.
Example 5
A method for detecting the content of multiple components of licorice heart-fire-purging decoction comprises the following steps:
and preparing 6 test solution samples in parallel, injecting samples according to the established chromatographic conditions, recording chromatographic peak areas and calculating RSD. The results show that the RSD values of the zingerone, the coptisine hydrochloride, the liquiritigenin, the berberine hydrochloride, the wogonin, the baicalein, the 6-gingerol and the ammonium glycyrrhetate are respectively 4.32%, 4.13%, 3.80%, 0.83%, 1.64%, 2.82%, 4.19% and 4.13%, and the detection conditions are met, which indicates that the detection method has high intermediate precision and meets the detection requirements in terms of reproducibility.
Example 6
A method for detecting the content of multiple components of licorice heart-fire-purging decoction comprises the following steps:
precisely sucking 1.0mL of 6 test sample solutions, respectively adding 1.0mL of each reference solution, diluting to 10mL with 50% methanol, introducing sample according to the above-established chromatographic conditions, and calculating sample recovery rate. The average sample adding recovery rates of the zingerone, the coptisine hydrochloride, the liquiritigenin, the berberine hydrochloride, the wogonoside, the baicalein, the 6-gingerol and the ammonium glycyrrhizinate are 104.66%, 106.81%, 102.65%, 107.34%, 97.36%, 105.14%, 102.56% and 97.50% respectively, and the RSDs are 2.24%, 2.33%, 3.10%, 1.98%, 2.14%, 3.23%, 2.57% and 3.25% respectively, which shows that the detection method has high detection accuracy.
Claims (8)
1. A method for detecting the multi-component content of a licorice heart-fire-purging decoction is characterized by comprising the steps of preparing a test solution, preparing a reference solution and detecting by using an ultra-high performance liquid chromatography; wherein the test solution is prepared by collecting extractive solution of Glycyrrhrizae radix heart fire purging decoction, centrifuging, collecting supernatant, volatilizing solvent, dissolving in methanol solution to desired volume, and filtering with microporous membrane to obtain test solution.
2. The method for detecting the content of the multiple components of the licorice heart-fire clearing decoction according to claim 1, wherein the dosage of the licorice heart-fire clearing decoction extracting solution is 100ml, the supernatant obtained after centrifugation is 5ml, the volume fraction of the methanol solution is 50%, the volumetric flask with the constant volume of 5ml is provided, and the pore diameter of the microfiltration membrane is 0.22 micron.
3. The method for detecting the content of the multiple components of the licorice heart-fire clearing decoction according to claim 2, wherein the extract of the licorice heart-fire clearing decoction is prepared by the following method:
(1) respectively placing Glycyrrhrizae radix, Scutellariae radix, rhizoma Pinelliae, fructus Jujubae, Coptidis rhizoma, and Zingiberis rhizoma in universal pulverizer, pulverizing respectively, sieving with No. 1 sieve and No. 2 sieve, and collecting powder that has passed through No. 1 sieve but not No. 2 sieve;
(2) taking 20 parts by weight of licorice powder, 15 parts by weight of scutellaria powder, 13 parts by weight of pinellia powder, 12 parts by weight of jujube powder, 5 parts by weight of coptis powder and 15 parts by weight of dried ginger powder, uniformly mixing, putting the mixture into a reflux extraction device, extracting for three times, adding 800 parts of water with the pH value adjusted to 4.0 by 0.1mol/L hydrochloric acid for the first time, adding 640 parts of water with the pH value adjusted to 7.0 by 0.1mol/L sodium hydroxide solution for the second time, adding 640 parts of water with the pH value adjusted to 8.0 by 0.1mol/L sodium hydroxide solution for the third time, wherein the extraction time is 1 hour for the first time, 0.5 hour for the second time, 0.5 hour for the third time, the extraction temperature is 90-95 ℃, combining the three extracting solutions, filtering, and concentrating the filtrate under reduced pressure until the relative density is 1.08-1.12 (25 ℃), thus obtaining the extracting solution of licorice heart-eliminating decoction; the temperature obtained by the decompression concentration is 60-80 ℃, and the vacuum degree is-0.05-0.08 MPa.
4. The method according to claim 3, wherein the reference solution is prepared by dissolving and diluting zingerone, coptisine hydrochloride, liquiritigenin, berberine hydrochloride, wogonin, baicalein, 6-gingerol and ammonium glycyrrhetate with 50% methanol to obtain 1mg/ml solution as reference stock solution of zingerone, coptisine hydrochloride, liquiritigenin, berberine hydrochloride, wogonin, scutellarin, 6-gingerol and ammonium glycyrrhetate, respectively, and storing the reference stock solution of zingerone, berberine hydrochloride, wogonin, baicalein, 6-gingerol and ammonium glycyrrhetate; and precisely measuring the reference substance stock solutions respectively, placing the reference substance stock solutions in the same volumetric flask, diluting with 50% methanol solution by volume fraction, and fixing the volume to scale to prepare a composite reference substance solution, wherein the concentrations of zingerone, berberine hydrochloride, liquiritigenin, berberine hydrochloride, wogonoside, baicalein, 6-gingerol and ammonium glycyrrhizinate in the composite reference substance solution are all 0.5 mg/ml.
5. The method for detecting the contents of the multiple components of the licorice heart-fire clearing decoction according to claim 4, wherein the detection by the ultra-high performance liquid chromatography is carried out by precisely sucking 3 μ l of the prepared reference solution and the prepared test solution into the ultra-high performance liquid chromatography, and calculating by an external standard method; the column temperature of the ultra-high performance liquid chromatograph is 35 ℃, the flow rate is 0.35ml/min, and the wavelength range is 190-400 nm.
6. The method for detecting the contents of the multiple components of the licorice heart-fire clearing decoction according to claim 5, wherein the flow term of the ultra-high performance liquid chromatography comprises a flow term A and a flow term B, the flow term A is a trifluoroacetic acid aqueous solution with a volume fraction of 0.1%, and the flow term B is a mixture of methanol: acetonitrile 50: 50, gradient elution is adopted.
7. The method for detecting the contents of the multiple components of the licorice heart-fire clearing decoction according to claim 6, wherein the gradient elution procedure of the ultra-high performance liquid chromatography is performed as follows:
8. The method for detecting the multi-component content of the licorice heart-fire-purging decoction according to claim 7, wherein the method for detecting the multi-component content of the licorice heart-fire-purging decoction can be applied to content detection of extracts and preparations taking the licorice heart-fire-purging decoction as a component.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN202110465440.4A CN113156019B (en) | 2021-04-28 | 2021-04-28 | Method for detecting multi-component content of liquorice heart-fire-purging decoction |
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| WO2024171115A1 (en) * | 2023-02-15 | 2024-08-22 | Evithe Limited | Methods of preparation of zingerone, compositions comprising zingerone, and uses therefor |
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| WO2024171115A1 (en) * | 2023-02-15 | 2024-08-22 | Evithe Limited | Methods of preparation of zingerone, compositions comprising zingerone, and uses therefor |
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