CN110702822A

CN110702822A - Method for detecting content of pseudo-ginseng in pseudo-ginseng traumatic rheumatism ointment

Info

Publication number: CN110702822A
Application number: CN201910137818.0A
Authority: CN
Inventors: 陈溪; 吴志红; 陈贤安
Original assignee: Baiyunshan Hejigong Pharmaceutical Factory Guangzhou Baiyunshan Pharmaceutical Holdings Co Ltd
Current assignee: Baiyunshan Hejigong Pharmaceutical Factory Guangzhou Baiyunshan Pharmaceutical Holdings Co Ltd
Priority date: 2019-02-25
Filing date: 2019-02-25
Publication date: 2020-01-17
Anticipated expiration: 2039-02-25
Also published as: CN110702822B

Abstract

The invention relates to a method for detecting the content of pseudo-ginseng in pseudo-ginseng traumatic rheumatism ointment, which comprises the following steps: preparing a reference substance solution; preparing a test solution: extracting Notoginseng radix ointment for treating traumatic injury and rheumatism with water and dichloromethane; standing the extract obtained in the previous step, separating a water layer, and adding petroleum ether for extraction; standing the extract obtained in the previous step, separating a water layer, and adding n-butanol for extraction; washing the extract obtained in the previous step with ammonia test solution to obtain n-butanol solution; evaporating the n-butanol solution to dryness to obtain residue, dissolving with methanol, and making into sample solution; and (5) detecting by chromatography. The application adopts a proper test sample preparation process, can extract the notoginsenoside R1 from the panax notoginseng traumatic rheumatism ointment for liquid chromatography analysis, has less formula matrix residue, less notoginsenoside R1 loss and less interference components in a test sample solution, and can obtain an accurate content value under a proper washing gradient. In addition, the notoginsenoside R1 is used for representing the content of the pseudo-ginseng, has strong specificity, is not interfered by chemical components of other medicinal materials, and has good repeatability and stability.

Description

Method for detecting content of pseudo-ginseng in pseudo-ginseng traumatic rheumatism ointment

Technical Field

The invention relates to the technical field of component detection of traditional Chinese medicine preparations, in particular to a method for detecting the content of pseudo-ginseng in pseudo-ginseng traumatic rheumatism ointment.

Background

The pseudo-ginseng ointment for treating traumatic injury and rheumatism is a Chinese medicinal compound preparation, is composed of 25 Chinese medicaments such as pseudo-ginseng, Chinese angelica tail, drynaria rhizome, himalayan teasel root, peach seed, doubleteeth pubescent angelica root, clematis root and the like, has the effects of promoting blood circulation and removing blood stasis, relaxing tendons and activating collaterals, relieving swelling and pain, dispelling wind and removing dampness, and is mainly used for treating diseases such as soft tissue injury, rheumatism and lumbago. The product has reasonable formulation and good therapeutic effect.

As an important traditional Chinese medicine compound preparation, the quality detection of the panax notoginseng traumatic injury and rheumatism ointment is very important. However, the quality detection standard of the panax notoginseng traumatic injury rheumatism ointment is only the standard of the sixth volume (WS3-B-1114-92) of the traditional Chinese medicine prescription preparation in the ministerial standards, the standard can only detect the liquorice and the rhubarb in the panax notoginseng traumatic injury rheumatism ointment, and the standard can only carry out qualitative tests and does not comprise quantitative tests. In addition, the formula of the panax notoginseng ointment for treating traumatic injury and rheumatism is complex, the extraction difficulty of active ingredients is high, and no scheme for controlling the quality of the panax notoginseng ointment by using the content of a certain active ingredient exists at present.

Therefore, it is highly desirable to provide a method for quantitatively detecting the traumatic injury and rheumatism ointment of panax notoginseng.

Disclosure of Invention

Based on the above, the main purpose of the invention is to provide a method for detecting the content of pseudo-ginseng in pseudo-ginseng traumatic injury rheumatism ointment, wherein the content of pseudo-ginseng can well reflect the quality of the pseudo-ginseng traumatic injury rheumatism ointment, and the method is a good quality control method of the pseudo-ginseng traumatic injury rheumatism ointment.

The purpose of the invention is realized by the following technical scheme:

a detection method for the content of pseudo-ginseng in pseudo-ginseng traumatic injury rheumatism ointment comprises the following steps:

preparation of a reference solution: dissolving notoginsenoside R1 in methanol to obtain control solution;

preparing a test solution:

(1) extracting Notoginseng radix ointment for treating traumatic injury and rheumatism with water and dichloromethane;

(2) standing the extracting solution obtained in the step (1), separating a water layer, adding petroleum ether into the water bath, and heating and refluxing the mixture;

(3) standing the reflux liquid obtained in the step (2), separating a water layer, and adding n-butanol for extraction;

(4) washing the extract obtained in the step (3) with ammonia test solution to obtain n-butanol solution;

(5) evaporating the n-butanol solution obtained in the step (4) to dryness to obtain residue, and dissolving with methanol to obtain a test solution;

and (3) chromatographic detection: and sucking the reference substance solution and the test solution, and injecting the reference substance solution and the test solution into a liquid chromatograph for measurement.

In some embodiments, in step (1), the amount ratio of the panax notoginseng traumatic injury and rheumatism ointment to the water to the dichloromethane is (10-15) g: (7-15) ml: (20-30) ml, and heating and refluxing the extraction by using a water bath;

in the step (3), the extraction frequency of the n-butanol is not less than 3 times;

in the step (4), the washing times of the ammonia test solution are not higher than 3.

In some embodiments, in step (1), the amount ratio of the panax notoginseng traumatic injury and rheumatism ointment to the water to the dichloromethane is (12-13) g: (9-11) ml: (23-27) ml;

in the step (3), the n-butanol is extracted for 4 to 5 times;

in the step (4), the washing times of the ammonia test solution are 1-2 times.

In one embodiment, the chromatographic detection conditions include:

stationary phase: octadecyl bonded silica gel is used as a filling agent;

mobile phase: acetonitrile is taken as a mobile phase A, and water is taken as a mobile phase B;

gradient elution: 0min to 60min, 15.5 to 16.5 percent → 17.5 to 18.5 percent of mobile phase A, and 83.5 to 84.5 percent → 81.5 to 82.5 percent of mobile phase B; 60min-70min, 17.5% -18.5% of mobile phase A and 81.5% -82.5% of mobile phase B.

In one embodiment, the chromatographic detection conditions include:

stationary phase: welch XB-C₁₈(4.6mm×250mm)、Welch PG-C₁₈(4.6 mm. times.250 mm) or Shimadzu inert sustainin C₁₈(4.6mm×250mm)；

gradient elution: 0min-60min, 16% → 18% mobile phase a, 84% → 82% mobile phase B; 60min-70min, 18% mobile phase A, 82% mobile phase B.

In some of these embodiments, the chromatographic detection employs a wavelength of 200nm to 205 nm.

In some of these embodiments, the chromatographic detection is at a wavelength of 203 nm.

In some of these embodiments, the chromatographic assay uses no less than 5000 theoretical plates.

In some of these examples, the step of preparing the control solution comprises dissolving 0.08mg to 0.12mg of notoginsenoside R1 control per 1ml of methanol.

In some of these examples, the step of preparing the control solution comprises dissolving 0.1mg of notoginsenoside R1 control per 1ml of methanol.

In some embodiments, the sample loading amount of the control solution and the test solution is 8-13. mu.l.

Compared with the prior art, the invention has the following beneficial effects:

by adopting a proper test sample preparation process, the notoginsenoside R1 can be extracted from the panax notoginseng traumatic rheumatism ointment with a complex formula for liquid chromatography analysis, so that the obtained test sample solution has less residue of the formula matrix, less loss of the notoginsenoside R1 and less interference components, and an accurate content measurement value can be obtained by matching with a proper washing gradient. In addition, the content of pseudo-ginseng is represented by pseudo-ginseng saponin R1, the specificity is strong, the interference of chemical components of other medicinal materials is avoided, and the repeatability and stability are good.

Drawings

FIG. 1 is a control profile of example 1;

FIG. 2 is a test sample profile of example 1;

FIG. 3 is a negative control map of example 1 lacking Panax notoginseng;

FIG. 4 is a standard curve of notoginsenoside R1 of example 1;

FIG. 5 is a control profile of example 2;

FIG. 6 is a test sample profile of example 2;

FIGS. 7 and 8 are detection maps obtained in comparative example 1;

FIGS. 9 and 10 are detection maps obtained in comparative example 2;

FIGS. 11 and 12 are detection maps obtained in comparative example 3.

Detailed Description

In order that the invention may be more fully understood, reference will now be made to the following description. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

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 invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

The formula of the pseudo-ginseng traumatic injury and rheumatism ointment provided by the invention is as follows:

the pseudo-ginseng traumatic injury and rheumatism ointment has the functions and main functions that: promoting blood circulation, removing blood stasis, relaxing muscles and tendons, dredging collaterals, relieving swelling and pain, dispelling pathogenic wind, and removing dampness. Can be used for treating soft tissue contusion, rheumatic lumbago, tendon strain, and soft tissue injury caused by fracture and dislocation.

The panax notoginseng-deficient negative control refers to ointment prepared under the condition that panax notoginseng raw materials are deficient relative to the prescription of panax notoginseng traumatic rheumatism ointment.

Example 1

The embodiment provides a method for detecting the content of pseudo-ginseng in pseudo-ginseng traumatic injury rheumatism ointment, which comprises the following steps:

1. preparation of control solutions

Precisely weighing appropriate amount of notoginsenoside R1 reference substance, and adding methanol to obtain reference substance solution containing 0.1mg per 1 ml.

2. Preparation of test solution

(1) Taking 12.5g of pseudo-ginseng traumatic injury and rheumatism ointment, precisely weighing, adding 10ml of water and 25ml of dichloromethane, heating in a water bath, and refluxing until the pseudo-ginseng traumatic injury and rheumatism ointment is dissolved;

(2) placing in a separating funnel while hot, standing, collecting water layer, adding petroleum ether (60-90 deg.C) 25ml (removing impurities), heating in water bath, and refluxing for 10 min;

(3) placing in separating funnel, standing to separate water layer, extracting with water saturated n-butanol for 4 times (20ml, 10ml), and mixing the obtained n-butanol solutions;

(4) washing with ammonia solution 30ml each time for 2 times, and reserving the ammonia solution;

(5) evaporating the n-butanol solution washed by ammonia solution, dissolving the residue with methanol, transferring to 10ml measuring flask, adding methanol to scale, shaking, filtering, and collecting the filtrate.

3. Preparation of negative control solution for lack of pseudo-ginseng

According to the preparation method of the test solution, the panax notoginseng lacking negative control sample is taken to prepare the panax notoginseng lacking negative control solution.

4. Measuring by high performance liquid chromatography (China pharmacopoeia 2015 edition general rule 0512)

Precisely sucking 10 μ l of each of the reference solution, the test solution and the negative reference solution, injecting into liquid chromatograph, and measuring.

The detection conditions of the liquid chromatogram comprise:

(1) stationary phase: octadecyl bonded silica gel as filler (Welch PG-C)₁₈4.6 × 250mm stainless steel columns);

(2) mobile phase: acetonitrile is taken as a mobile phase A, and water is taken as a mobile phase B;

(3) elution gradient: see table 1;

TABLE 1

Time of day	Mobile phase A (%)	Mobile phase B (%)
			0-60min	16→18	84→82
60-70min	18	82

(4) Detection wavelength: the detection wavelength is 203 nm;

(5) the number of theoretical plates: not less than 5000;

(6) column temperature: 35 ℃;

(7) flow rate: 1.0 ml/min.

The detection results are shown in fig. 1, fig. 2 and fig. 3. FIG. 1 is a map of a control, FIG. 2 is a map of a test sample, and FIG. 3 is a negative control of Panax notoginseng. The signal peak of retention time 64.990 in fig. 1 is the control peak of notoginsenoside R1, the corresponding peak area is the standard peak area (as 310670 in the figure), the signal peak of retention time 65.266 in fig. 2 is the signal peak of notoginsenoside R1 in the panax notoginseng traumatic injury ointment, the corresponding peak area is the test sample peak area (as 290949 in the figure), and the calculation formula is as follows:

the average content of notoginsenoside R1 in Table 1 can be obtained by calculation.

Using the method of example 1, different batches of the panax notoginseng traumatic rheumatism ointment were tested, and the results are shown in the following Table 1:

TABLE 1

Serial number	Batch number	Average content of notoginsenoside R1 (mg/g ointment)
			1	17050901	0.100
2	17050902	0.079
			3	170621	0.089
4	170624	0.087
			5	170718	0.089
6	180202	0.068
			7	180305	0.071
8	180312	0.075

The tracking test of different batches of the panax notoginseng traumatic rheumatism ointment using the method of example 1 is shown in the following table 2:

TABLE 2 Notoginsenoside R1 content (mg/g cream)

Serial number	Batch number		0 month	Accelerated for 1 month	Accelerated for 2 months	Accelerated for 3 months	Accelerated for 6 months
								1	180202	0.068	0.067	0.066	0.066	0.56
2	180305	0.071	0.070	0.068	0.066	0.51
							3	180312	0.075	0.074	0.074	0.073	0.58

5. Investigation of linear relationships

Precisely weighing notoginsenoside R₁10.47mg (95.0%) of a reference substance is placed in a 10ml measuring flask, a proper amount of methanol is added, ultrasonic treatment is carried out to dissolve the reference substance, the methanol is added to the scale and is shaken up to be used as a reference substance stock solution, 0.1ml, 0.25ml, 0.5ml, 1.0ml, 2.5ml and 4ml of stock solutions are precisely measured respectively and are placed in a 10ml measuring flask, the methanol is added to the scale and is shaken up to be used as a reference substance solution; and precisely sucking 10 mu l of each of the reference solution and the stock solution, injecting the solution into a liquid chromatograph, and recording a chromatogram and a peak area under the detection conditions, wherein the results are shown in the table.

TABLE 3 results of linear range investigation

From the data obtained in the above table, notoginsenoside R₁Concentration (C) is the abscissa, for the corresponding mean peak surfaceTaking product (A) as ordinate, and performing linear regression analysis to obtain notoginsenoside R₁The regression equation is: a is 3298501.7C +1580.6 and the correlation coefficient r is 1.000. The result shows that the notoginsenoside R₁The concentration is within the range of 0.01-0.39 mg/ml, and the good linear relation is presented. The standard curve is shown in FIG. 4.

6. Stability test

Respectively and precisely sucking the same reference substance solution (notoginsenoside R)₁0.1026mg/ml) and 10 mul of each of the test solutions were injected at regular intervals for 1 time, and the chromatogram and the peak area were recorded by the method of this example, as shown in the following table.

TABLE 4 stability test results for test solutions and control solutions

The test result shows that the reference solution and the test solution are stable within 24 hours.

7. Precision test

Precisely sucking the same reference substance solution (notoginsenoside R)₁0.1026mg/ml) 10. mu.l, and 6 times of continuous sample injection, and the chromatogram and peak area were recorded by the method of this example, as shown in the following Table.

TABLE 5 results of precision test

Test results show that the method has good precision.

8. Repeatability test

The test method of the test solution comprises preparing 6 test solutions from 20180305 batch of radix Notoginseng ointment for treating traumatic injury and rheumatism, and determining the content according to the test method, the results are shown in the following table.

TABLE 6 results of the repeatability tests

Test results show that the method has good repeatability.

9. Recovery test

Collecting 20180305 batch of Notoginseng radix ointment for treating traumatic injury and rheumatism 6.25g (notoginsenoside R)₁Content of 0.073mg/g), precisely weighing 9 parts, dividing into 3 groups, precisely adding reference substance solution (notoginsenoside R) into each group at ratio of 0.5:1, 1:1, and 1.5:1₁0.10032mg/ml)0.22ml, 0.45ml and 0.67ml, test solutions were prepared and the contents were measured according to the method of this example, and the results are shown in the following table.

TABLE 7 recovery test results

Test results show that the method has better recovery rate.

10. System durability test

Observing the change of the theoretical plate number and the separation degree of 2 indexes of the recent impurity peak under the condition of small determined conditions of flow rate, column temperature and flow, and the like₁The reliability of the method when the content is determined, the results are shown in the following table:

TABLE 8 notoginsenoside R₁Durability of assay method

TABLE 9 notoginsenoside R₁Parameters relevant for the assay durability study

And (4) conclusion: the above results show that under the condition of small variation of the determination conditions of flow rate, column temperature and flow, the notoginsenoside R in the sample is subjected to₁The content is measured, the number of theoretical plates is not less than 5000, and notoginsenoside R₁The separation degree of the peak and the nearest impurity peak is more than 1.5, and the method has good durability.

Example 2

1. preparation of control solutions

2. Preparation of test solution

(1) Taking 10g of pseudo-ginseng traumatic injury and rheumatism ointment (recorded as a test sample 3-2), precisely weighing, adding 7ml of water and 20ml of dichloromethane, heating in a water bath and refluxing until the pseudo-ginseng traumatic injury and rheumatism ointment is dissolved;

(2) placing in a separating funnel while hot, standing to separate water layer, adding 20ml petroleum ether (60-90 deg.C), heating in water bath and refluxing for 10 min;

(3) placing in separating funnel, standing to separate water layer, extracting with water saturated n-butanol for 5 times (20ml, 10ml), and mixing n-butanol solutions;

(4) washing with ammonia solution 30ml each time for 1 time;

3. Measuring by high performance liquid chromatography (China pharmacopoeia 2015 edition general rule 0512)

Precisely sucking 10 μ l of each of the reference solution and the sample solution, injecting into a liquid chromatograph, and measuring to obtain the final product, wherein the detection conditions of the liquid chromatograph include:

(1) stationary phase: octadecyl bonded silica gel is used as a filling agent;

(3) elution gradient: same as example 1, table 1;

(4) detection wavelength: the detection wavelength is 203 nm;

(5) the number of theoretical plates: not less than 5000.

The detection results are shown in fig. 5 and 6. FIG. 5 is a reference map and FIG. 6 is a test sample map. The signal peak of retention time 64.875 in fig. 5 is the control peak of notoginsenoside R1, the corresponding peak area is the standard peak area (319137 in the figure), the signal peak of retention time 64.962 in fig. 6 is the signal peak of notoginsenoside R1 in the notoginseng traumatic injury ointment, the corresponding peak area is the test sample peak area (302149 in the figure), and the calculation formula refers to example 1.

Example 3

This example is a variation of example 1, including in the preparation of the test sample solution, relative to example 1:

in the step (1), 15g of pseudo-ginseng traumatic injury and rheumatism ointment is precisely weighed, 15ml of water and 30ml of dichloromethane are added;

in step (3), extraction was performed 3 times (20ml ) with water-saturated n-butanol.

The effect of this example is the same as example 2.

Comparative example 1

This comparative example is that of example 1, and the main differences with respect to example 1 include the preparation of the test solution, which is as follows:

adding 50ml of ethanol into the panax notoginseng traumatic injury and rheumatism ointment, uniformly stirring, placing the ointment on a water bath for heating for about 20 minutes, cooling, removing coagula on the surface layer, filtering, recovering the ethanol, adding 20ml of hydrochloric acid (1mol/L) into residues for dissolving, filtering, placing filtrate in a separating funnel, adding 80ml of chloroform for extracting, placing chloroform in an evaporation dish, volatilizing, adding 2ml of absolute ethanol into residues for dissolving to obtain a sample solution. The rest of the procedure was the same as in example 1.

The measurement was repeated twice, and the results are shown in fig. 7 and 8, respectively, and it can be seen from fig. 7 and 8 that: in the fig. 7 and 8, no characteristic peak of notoginsenoside R1 appears in 72min to 75min, which proves that the method has low extraction efficiency. Compared with the example 1, the impurity removal difficulty is high, the time is long, the loss in the extraction and separation process is large, the effective extraction and separation of the notoginsenoside R1 cannot be carried out, and the content measurement is difficult to carry out.

Comparative example 2

(1) precisely weighing 8g of radix Notoginseng traumatic injury rheumatism ointment, adding 20ml of water and 15ml of dichloromethane, heating and stirring until the radix Notoginseng traumatic injury rheumatism ointment is dissolved;

(2) placing in a separating funnel while hot, standing to separate water layer, adding petroleum ether (60-90 deg.C) 25ml, heating in water bath and refluxing for 10 min;

(3) placing in separating funnel, standing to separate water layer, extracting with water saturated n-butanol for 2 times (20ml ), and mixing n-butanol solutions;

(4) washing with ammonia solution 30ml for 4 times;

The measurement was repeated twice, and the results are shown in FIGS. 9 and 10. As can be seen from the results of fig. 9 and 10: in the fig. 9 and 10, no characteristic peak of notoginsenoside R1 appears in 72min to 75min, which proves that the method has low extraction efficiency. Compared with the example 1, the method has the advantages of less sampling amount, less n-butanol extraction times, more ammonia test solution washing times, incapability of effectively extracting and separating the notoginsenoside R1 and difficulty in content measurement.

Comparative example 3

This comparative example is that of example 1, the main differences with respect to example 1 include that the high performance liquid chromatography employs an elution gradient of: 0-12min, 19% of mobile phase A, 81% of mobile phase B and 12-60min, wherein 19% to 36% of mobile phase A and 81% to 64% of mobile phase B are adopted.

The results of the two repeated measurements are shown in fig. 11 and 12, and it can be seen from the results of fig. 11 and 12 that: a plurality of miscellaneous peaks exist near the characteristic peaks of notoginsenoside R1 from 30min to 40min and 46min to 53min, the separation effect is not ideal, and the gradient is proved to be not suitable for the content determination of notoginsenoside R1 in the panax notoginseng traumatic injury and rheumatism ointment.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

1. A detection method for the content of pseudo-ginseng in pseudo-ginseng traumatic rheumatism ointment is characterized by comprising the following steps:

preparing a test solution:

2. The method for detecting the content of pseudo-ginseng in pseudo-ginseng traumatic-injury and rheumatism ointment according to claim 1, wherein in the step (1), the using amount ratio of the pseudo-ginseng traumatic-injury and rheumatism ointment to water to dichloromethane is (10-15) g: (7-15) ml: (20-30) ml, and heating and refluxing the extraction by using a water bath;

3. The method for detecting the content of pseudo-ginseng in pseudo-ginseng traumatic-injury and rheumatism ointment according to claim 2, wherein in the step (1), the using amount ratio of the pseudo-ginseng traumatic-injury and rheumatism ointment to the water to the dichloromethane is (12-13) g: (9-11) ml: (23-27) ml;

in the step (3), the n-butanol is extracted for 4 to 5 times;

in the step (4), the washing times of the ammonia test solution are 1-2 times.

4. The method for detecting the content of pseudo-ginseng in pseudo-ginseng traumatic rheumatism ointment according to any one of claims 1 to 3, wherein the chromatographic detection conditions comprise:

stationary phase: octadecyl bonded silica gel is used as a filling agent;

5. The method for detecting the content of pseudo-ginseng in pseudo-ginseng traumatic rheumatism ointment according to claim 4, wherein the chromatographic detection conditions comprise:

6. The method for detecting the content of pseudo-ginseng in pseudo-ginseng traumatic rheumatism ointment according to any one of claims 1 to 3, wherein the chromatographic detection conditions comprise: the wavelength is 200nm-205nm, the column temperature is 33-37 ℃, and the flow rate is 0.98ml/min-1.02 ml/min.

7. The method for detecting the content of pseudo-ginseng in pseudo-ginseng traumatic rheumatism ointment according to claim 6, wherein the chromatographic detection conditions comprise: the wavelength was 203nm, the column temperature was 35 ℃ and the flow rate was 1 ml/min.

8. The method for detecting the content of panax notoginseng in panax notoginseng traumatic rheumatism ointment according to any one of claims 1 to 3, wherein in the step of preparing the control solution, 0.08mg to 0.12mg of notoginsenoside R1 control is dissolved in every 1ml of methanol.

9. The method for detecting the content of pseudo-ginseng in pseudo-ginseng traumatic rheumatism ointment according to claim 8, wherein in the step of preparing the reference substance solution, 0.1mg of notoginsenoside R1 reference substance is dissolved in every 1ml of methanol.

10. The method for detecting the content of pseudo-ginseng in pseudo-ginseng traumatic rheumatism ointment according to any one of claims 1 to 3, wherein the loading amount of the control solution and the test solution is 8 to 13 μ l.