Detection method of Panax notoginseng content in Dan'e Fukang decoction

Abstract

The present invention belongs to the technical field of quality detection, and specifically relates to a method for detecting the content of Panax notoginseng in Dan'e Fukang decoction. The detection method is: using water as a solvent to prepare a test solution, injecting it into an ultra-high performance liquid chromatograph to determine the content; the chromatographic conditions are: using octadecylsilane bonded silica gel as a filler; using acetonitrile and water as mobile phases for gradient elution; the detection wavelength is 203nm, the column temperature is 20-35°C; the flow rate is 0.4-0.6mL / min; the theoretical plate number is calculated based on the ginsenoside Rb1 peak and should not be less than 700000. The method of the present invention has good linearity, repeatability, stability and recovery rate, etc., and can well control the quality of Dan'e Fukang decoction. The method of the present invention optimizes the mobile phase elution procedure, which can not only completely separate the target peak, but also save analysis time.

Description

Technical Field

[0001] The invention belongs to the technical field of quality detection, and particularly relates to a method for detecting the content of Panax notoginseng in Dan'e Fukang decoction. Background Art

[0002] Dan'e Fukang Decoction is a traditional Chinese medicine preparation. It has the effects of promoting blood circulation and removing blood stasis, soothing the liver and regulating qi, regulating menstruation and relieving pain, softening and removing hard masses, etc. It is a commonly used medicine for the clinical treatment of endometriosis. Dan'e Fukang Decoction is dark brown and viscous semi-fluid. The whole formula is composed of 10 herbs: Salvia miltiorrhiza, Curcuma zedoaria, Bupleurum yunnanensis, Panax notoginseng, Paeonia lactiflora, Angelica sinensis, Trillium, Cyperus rotundus, Corydalis yanhusuo and Licorice.

[0003] In the national drug standards of the State Food and Drug Administration, high performance liquid chromatography is used to detect the contents of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 in Dan'e Fukang decoction, wherein the test sample is prepared by methanol extraction, lead acetate solution precipitation, chloroform and water-saturated n-butanol extraction, ammonia test solution washing and other operations to remove interference. This method has complex steps and is time-consuming. The detected ginsenoside Rg1 and impurity peaks are not separated, which can easily cause the loss of the detected components and increase the experimental error. In addition, the detection method has poor repeatability and reproducibility and a long analysis time.

[0004] Therefore, it is necessary to optimize the preparation method of the test solution of Dan'e Fukang decoction and the chromatographic conditions for detection, and to establish an accurate, stable and easy-to-operate method for detecting the content of Panax notoginseng in Dan'e Fukang decoction. Summary of the invention

[0005] The purpose of the present invention is to provide a method for detecting the content of Panax notoginseng in Dan'e Fukang decoction, so as to solve the problems in the prior art that the preparation method of the test solution is too complicated, time-consuming and labor-intensive; the detection target peak cannot be completely separated and the analysis time is long.

[0006] The technical solution adopted by the present invention to solve the above technical problems is to provide a method for detecting the content of Panax notoginseng in Dan'e Fukang decoction. The detection method is: using water as a solvent to prepare a test solution, injecting it into an ultra-high performance liquid chromatograph to determine the content; the chromatographic conditions are: using octadecylsilane bonded silica gel as a filler; using acetonitrile and water as mobile phases for gradient elution; the detection wavelength is 203nm, the column temperature is 20-35℃; the flow rate is 0.4-0.6mL / min; the theoretical plate number calculated based on the ginsenoside Rb1 peak should be no less than 700,000.

[0007] Among them, in the above-mentioned method for detecting the content of Panax notoginseng in Dan'e Fukang decoction, the preparation of the test solution is to take 2-5g of Dan'e Fukang decoction and place it in a 50mL volumetric flask, add water and ultrasonicate for 30-90 minutes, then add water to make up the volume, shake well, and filter.

[0008] Preferably, 2 g of Dan'e Fukang decoction is placed in a 50 mL volumetric flask, and water is added for ultrasonication for 30 minutes, and then water is added to make up the volume, shaken well, and filtered.

[0009] In order to make the preparation method of the test sample simpler and more time-saving, while ensuring the effective extraction of the active ingredients of Panax notoginseng in Dan'e Fukang decoction, the inventors conducted a large number of experimental studies and found that the use of water as a solvent combined with ultrasonic treatment can extract the active ingredients in Panax notoginseng in a short time. Moreover, using water as a solvent can not only reduce the pollution of organic solvents and the harm to human health, but also save costs and simplify steps.

[0010] Furthermore, the power of the ultrasound is 480-520W; the frequency of the ultrasound is 35-45kHz.

[0011] Preferably, the power of the ultrasound is 500W; the frequency of the ultrasound is 40Hz.

[0012] Further, the gradient elution conditions with acetonitrile and water as the mobile phase are:

[0013] The time was 0 to 11 min, the mobile phase was 16% acetonitrile, and the mobile phase was 84% ​​water;

[0014] The time is 11 to 15 minutes, the mobile phase is 16 to 21% acetonitrile, and the mobile phase is 84 to 79% water;

[0015] The time is 15 to 24 minutes, the mobile phase is 21 to 31% acetonitrile, and the mobile phase is 79 to 69% water;

[0016] The time is 24 to 26 minutes, the mobile phase is 31 to 40% acetonitrile, and the mobile phase is 69 to 60% water;

[0017] The time was 26 to 29 min, the mobile phase was 40% acetonitrile, and the mobile phase was 60% water;

[0018] The time is 29 to 30 minutes, the mobile phase is 40 to 16% acetonitrile, and the mobile phase is 60 to 84% water;

[0019] The time is 30 to 33 minutes, the mobile phase is 16% acetonitrile, and the mobile phase is 84% ​​water.

[0020] The inventors investigated various mobile phase gradient elution conditions and found that under the gradient elution conditions described in the present invention, Panax notoginseng saponin R1, ginsenoside Rg1 and ginsenoside Rb1 can all be completely separated, and the peak shapes are all sharp and upright; and the analysis time using this gradient elution method is short, and the choice of mobile phase does not contain buffer salts, which can extend the life of the chromatographic column.

[0021] Furthermore, the column temperature was 25°C; and the flow rate was 0.4 mL / min.

[0022] The beneficial effects of the present invention are:

[0023] The method of the present invention adopts an ultra-high performance liquid chromatograph, and determines the contents of notoginseng saponin R1, ginsenoside Rg1 and ginsenoside Rb1 in Dan'e Fukang decoction by optimizing the preparation method of the test sample and the detection chromatographic conditions, and has good linearity, repeatability, stability and recovery rate, and can well control the quality of Dan'e Fukang decoction. The method of the present invention simplifies the preparation process of the test sample, uses water as a solvent combined with ultrasonic treatment, can extract the effective components in Panax notoginseng in a short time, saves time and labor, is more environmentally friendly, and saves costs; the method of the present invention optimizes the mobile phase elution procedure, can completely separate the target peak, and saves analysis time. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is the chromatogram of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 in Dan'e Fukang decoction measured in Example 1 (① is notoginsenoside R1, ② is ginsenoside Rg1, ③ is ginsenoside Rb1).

[0025] Figure 2 This is the chromatogram of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 in Dan'e Fukang decoction measured in Comparative Example 1 (① is notoginsenoside R1, ② is ginsenoside Rg1, ③ is ginsenoside Rb1).

[0026] Figure 3 This is an enlarged view of the impurity peak before the ginsenoside Rg1 peak in Dan'e Fukang decoction measured in Comparative Example 1.

[0027] Figure 4 This is the chromatogram of the mixed reference substance of notoginsenosides measured in comparative example 2 (① is notoginsenoside R1, ② is ginsenoside Rg1, ③ is ginsenoside Rb1).

[0028] Figure 5 The chromatograms of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 in the Dan'e Fukang decoction sample were measured for Comparative Example 2 (① is notoginsenoside R1, ② is ginsenoside Rg1, ③ is ginsenoside Rb1).

[0029] Figure 6 Chromatograms of the separation of notoginsenoside R1, ginsenoside Rg1, and ginsenoside Rb1 using different mobile phases (from top to bottom, acetonitrile-water, acetonitrile-0.1% phosphoric acid; ① is notoginsenoside R1, ② is ginsenoside Rg1, and ③ is ginsenoside Rb1).

[0030] Figure 7Chromatograms of the separation of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 at different column temperatures (① is notoginsenoside R1, ② is ginsenoside Rg1, ③ is ginsenoside Rb1, and the column temperatures from top to bottom are 20℃, 25℃, 30℃ and 35℃ respectively).

[0031] Figure 8 The standard curve of notoginseng saponin R1 was investigated for linear relationship.

[0032] Fig. 9 The standard curve of notoginseng saponin Rg1 was investigated for linear relationship.

[0033] Fig.10 The standard curve of notoginseng saponin Rb1 was examined for linear relationship.

[0034] Fig.11 The chromatograms obtained for the specificity investigation (from top to bottom are the negative sample of Dan'e Fukang decoction (lacking Panax notoginseng), the sample of Dan'e Fukang decoction, and the mixed reference of Panax notoginseng saponins). DETAILED DESCRIPTION

[0035] The present invention will be further described in detail below through specific examples.

[0036] Example 1

[0037] Preparation of reference solution: Accurately weigh appropriate amounts of notoginsenoside R1, ginsenoside Rg1, and ginsenoside Rb1 reference substances, add methanol to make a mixed reference solution containing 40 μg of notoginsenoside R1 reference substance, 150 μg of ginsenoside Rg1 reference substance, and 150 μg of ginsenoside Rb1 reference substance per 1 mL.

[0038] Preparation of test solution: Take 2g of Dan'e Fukang decoction, weigh accurately, put it in a 50mL volumetric flask, add appropriate amount of water, seal it, and treat it with ultrasound for 30 minutes. The ultrasonic power is 500W and the ultrasonic frequency is 40kHz. Take it out, cool to room temperature, add water to the volume, shake well, filter, and take the filtrate.

[0039] Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler (2.1×100m, 1.9μm); acetonitrile and water were used as mobile phases, and gradient elution was performed according to Table 1; the detection wavelength was 203nm, and the column temperature was 25°C. The theoretical plate number calculated based on the ginsenoside Rb1 peak should be no less than 700,000.

[0040] Determination method: Accurately pipette 2 μL of reference solution and test solution respectively, inject into ultra-high performance liquid chromatography, and determine. Figure 1 .

[0041] Each 1g of this product contains notoginseng saponin R1 (C 47 H80 O 18 )、ginsenoside Rg1(C 42 H 72 O 14 ) and ginsenoside Rb1(C 54 H 92 O 23 ) The total amount of the three shall not be less than 2 mg.

[0042] Table 1 Mobile phase gradient conditions used in Example 1

[0043]

[0044]

[0045] Comparative Example 1 was tested using the National Drug Standard WS-10227 (ZD-0227) 2002-2012Z-2016 method

[0046] Preparation of reference solution: Accurately weigh appropriate amounts of ginsenoside Rg1 reference, ginsenoside Rb1 reference and notoginsenoside R1 reference, add methanol to make a mixed solution containing 10.4 mg of ginsenoside Rg, 10.4 mg of ginsenoside Rb and 10.1 mg of notoginsenoside R per 1 mL.

[0047] Preparation of test solution: Take 7.5g of this product, accurately weigh it, put it in a 100mL volumetric flask, add about 90mL of methanol, ultrasonically treat (power 500W, frequency 59kHz) for 30 minutes, let it stand overnight, ultrasonically treat (power 500W, frequency 59kHz) for 10 minutes, cool, dilute to the scale with methanol, shake well, filter, accurately measure 50mL of the filtrate, put it in a 100mL volumetric flask, add 40mL of 5% lead acetate solution, ultrasonically treat (power 500W, frequency 59kHz) for 20 minutes, cool, dilute to the scale with water, shake well, filter, accurately measure 40 mL of the filtrate was evaporated to dryness, 25 mL of water was added to dissolve it, and it was transferred to a separatory funnel. It was washed with chloroform three times, 25 mL each time, and the chloroform solution was discarded. The aqueous solution was extracted with water-saturated n-butanol by shaking for 5 times (30 mL, 25 mL, 15 mL, 15 mL, 15 mL), and the n-butanol solutions were combined and concentrated to about 25 mL, and it was transferred to a separatory funnel. It was washed with ammonia test solution twice while hot, 25 mL each time, and the ammonia solution was discarded. The n-butanol solution was evaporated to dryness, and methanol was added to dissolve the residue, and it was transferred to a 10 mL volumetric flask, and it was diluted to the scale with methanol, shaken well, filtered, and the filtrate was taken.

[0048] Chromatographic conditions and system suitability test: octadecylsilane bonded silica gel is used as filler; acetonitrile is used as mobile phase A, 0.45% sodium dihydrogen phosphate solution is used as mobile phase B, elution is performed according to the gradient specified in Table 2, and the detection wavelength is 203 nm. The theoretical plate number calculated based on the ginsenoside Rg1 peak should not be less than 10,000.

[0049] Accurately pipette 5 μL of the reference solution and the test solution respectively and inject them into the liquid chromatograph for determination.

[0050] Table 2 Mobile phase gradient conditions used in Comparative Example 1

[0051] Time (minutes) Acetonitrile (%) 0.45% sodium dihydrogen phosphate solution (%) 0～10 15 84 10～40 15→45 85→55 40～50 45→70 55→30 50～70 70→15 30→85

[0052] Comparative Example 2

[0053] The test solution and reference solution prepared in Example 1 of the present invention were tested under the following conditions:

[0054] Chromatographic column (Ultimate XB-C18 column, 2.1mm×50.0mm, 1.8μm); detection wavelength 203nm; column temperature 25℃; flow rate 0.6mL / min; injection volume 2μL; water and acetonitrile as mobile phases for gradient elution according to the provisions in Table 3. The chromatograms of the mixed reference substance of Panax notoginseng saponins and the test substance of Dan'e Fukang decoction are shown in Figure 3 and Figure 4 .

[0055] Table 3 Mobile phase gradient conditions used in Comparative Example 2

[0056] Time (minutes) Acetonitrile (%) water(%) 0 19 81 4.00 19 81 8.00 35 65 10.00 95 5 11.00 95 5 11.01 19 81 12.00 19 81

[0057] Experimental Example 1 Selection of Chromatographic Conditions

[0058] 1.1 Choice of mobile phase

[0059] Acetonitrile-0.1% phosphoric acid solution and acetonitrile-water were used as mobile phases, and the mobile phase gradient conditions of the present invention were investigated. The test results showed that the use of acetonitrile-water as the mobile phase can separate the target peaks completely without interference, so acetonitrile-water was selected as the mobile phase. Figure 5 .

[0060] 1.2 Selection of column temperature

[0061] Different column temperatures of 20℃, 25℃, 30℃, and 35℃ were investigated. The results showed that the separation was best at 25℃, and each target peak and adjacent peak could be completely separated. Therefore, 25℃ was selected as the column temperature. Figure 6 .

[0062] 1.3 Selection of extraction method

[0063] Take 6 portions of Dan'e Fukang decoction, weigh 2g each, put 3 portions in 50mL volumetric flask, add appropriate amount of water, plug tightly, extract for 60 minutes with ultrasonic treatment (power 500W, frequency 40KHz), take out, cool to room temperature, add water to the scale, shake well; take another 3 portions in a stoppered conical flask, accurately add 50mL water, weigh the weight, heat reflux extraction for 60 minutes, take out, cool, weigh, make up the lost weight with water, shake well. Filter the two extracted samples, take the filtrate for determination, and the results are shown in Table 4.

[0064] Table 4 Extraction method investigation (n=3)

[0065] Extraction method Ultrasound Reflow Total amount of notoginsenosides mg / g 7.7 7.6

[0066] The results showed that the ultrasonic extraction method had a higher extraction efficiency and the ultrasonic treatment method was simpler, so ultrasound (power 500W, frequency 40KHz) was selected as the extraction method for this product.

[0067] Test Example 2 Methodology Verification

[0068] Preparation of reference solution: Weigh appropriate amounts of notoginsenoside R1, ginsenoside Rg1, and ginsenoside Rb1 reference substances, add methanol to prepare a mixed reference solution containing 40 μg of notoginsenoside R1 reference substance, 150 μg of ginsenoside Rg1 reference substance, and 150 μg of ginsenoside Rb1 reference substance per 1 mL.

[0069] 2.1 System suitability test

[0070] The mixed reference solution was injected into the ultra-high performance liquid chromatograph, and 5 injections were made continuously. The peak area was recorded and the RSD value was calculated. The results are shown in Table 5. The injection precision was good, the theoretical plate number of ginsenoside Rb1 was greater than 700,000, the separation of each peak was greater than 1.5, and the tailing factor was greater than 1.5, indicating that the system had good applicability.

[0071] Table 5 Injection precision investigation results (n=5)

[0072]

[0073] 2.2 Linearity

[0074] Preparation of reference stock solution: Take an appropriate amount of notoginsenoside R1 reference substance, accurately weigh it, and add methanol to make a solution containing 1 mg per 1 mL; take an appropriate amount of ginsenoside Rg1 reference substance, accurately weigh it, and add methanol to make a solution containing 2 mg per 1 mL; take an appropriate amount of ginsenoside Rb1 reference substance, accurately weigh it, and add methanol to make a solution containing 2 mg per 1 mL.

[0075] Accurately measure 2 mL of each of the three reference solutions above, place them in a 10 mL volumetric flask, add methanol to dilute to the scale, and shake well to obtain the notoginseng saponin mixed reference stock solution.

[0076] 1 mL, 1 mL, 1 mL, 1 mL, 2 mL of the mixed reference solution of Panax notoginseng saponins were accurately measured and placed in 100 mL, 50 mL, 10 mL, 5 mL, and 5 mL volumetric bottles, respectively, and diluted to the mark with methanol, shaken, filtered, and 2 μL of methanol (blank) and the above reference solution were injected into the ultra-high performance liquid chromatograph, and the chromatogram was recorded and the peak area was determined. The standard curve of each reference was drawn with the reference concentration as the abscissa and the peak area as the ordinate, and the linear regression equation, correlation coefficient and linear range were calculated. The results are shown in Tables 6, 7, 8 and Figure 7 , Figure 8 , Fig. 9 .

[0077] Table 6 Investigation of linear relationship of notoginseng saponin R1

[0078] Concentration mg / mL 0 0.002152 0.004304 0.02152 0.04304 0.08608 0.2152 Peak area 0 3.10 6.69 35.66 71.58 143.58 348.24

[0079] The linear regression equation is: y=1621.7x+0.7261, and the correlation coefficient r=0.99991.

[0080] Table 7 Investigation of linear relationship of ginsenoside Rg1

[0081] Concentration mg / mL 0 0.004053 0.008106 0.04053 0.08106 0.16212 0.4053 Peak area 0 7.82 15.65 80.51 161.18 322.51 780.55

[0082] The linear regression equation is: y=1928.9x+2.2495, and the correlation coefficient r=0.99990.

[0083] Table 8 Investigation of linear relationship of ginsenoside Rb1

[0084] Concentration mg / mL 0 0.004253 0.008506 0.04253 0.08506 0.17012 0.4253 Peak area 0 5.32 11.05 56.88 113.43 226.83 549.48

[0085] The linear regression equation is: y=1293.9x+1.5639, and the correlation coefficient r=0.99991.

[0086] The experimental results show that in the concentration range of 0-0.2152 mg / mL and the injection volume of 2 μL, the peak area of ​​Panax notoginseng saponin R1 has a good linear relationship with the concentration; in the concentration range of 0-0.4053 mg / mL and the injection volume of 2 μL, the peak area of ​​ginsenoside Rg1 has a good linear relationship with the concentration; in the concentration range of 0-0.4253 mg / mL and the injection volume of 2 μL, the peak area of ​​ginsenoside Rb1 has a good linear relationship with the concentration.

[0087] 2.3 Exclusivity Investigation

[0088] Take the negative sample of this product (lacking Panax notoginseng) and prepare the negative sample solution according to the preparation method of the test solution and measure it. The result shows that there is no interference peak at the retention time position corresponding to the chromatographic peak of the reference substance, indicating that the method of the present invention has good specificity. Fig.10 .

[0089] 2.4 Stability of test solution

[0090] Take this product according to the test solution preparation method to prepare samples, and measure them at 0h, 4h, 8h, 12h, 16h, 20h, 24h, and 36h, and calculate the total amount of notoginseng saponins and RSD value. The results are shown in Table 9. The results show that the test solution is relatively stable within 36h.

[0091] Table 9 Stability study of test solution

[0092]

[0093] 2.5 Repeatability

[0094] Six test sample solutions were prepared from this product, and the total amount of notoginseng saponins and RSD values ​​were calculated according to the method of the present invention. The results are shown in Table 10. The results show that the method of the present invention has good repeatability.

[0095] Table 10 Stability study of test solution (n=6)

[0096] serial number 1 2 3 4 5 6 average value RSD(%) Notoginsenoside content (mg / g) 8.0 7.8 8.0 8.0 8.0 7.9 7.9 1.0

[0097] 2.6 Recovery rate

[0098] Take 1g of this product (6 portions in total), accurately weigh, place in 50mL volumetric bottles, add notoginseng saponin R1 reference solution (1.0758mg), ginsenoside Rg1 reference solution (2.9319mg), ginsenoside Rb1 reference solution (3.3533mg), add appropriate amount of water, plug, ultrasonic treatment (power 500, frequency 40kHz) for 30 minutes, take out, cool to room temperature, add water to scale, shake well, filter, take 2μL of the filtrate and inject it into ultra-high performance liquid chromatography, determine the content, and calculate the recovery rate according to the following formula. The results show that the average recovery rate of notoginseng saponin R1 is 100%, and the RSD value is 1.2%, the average recovery rate of ginsenoside Rg1 is 102%, and the RSD value is 1.4%, and the average recovery rate of ginsenoside Rb1 is 104%, and the RSD value is 1.1%. The results show that the accuracy of the method of the present invention is good. See Table 11, Table 12, and Table 13.

[0099] Table 11 Notoginsenoside R1 recovery test results (n=6)

[0100]

[0101]

[0102] Table 12 Ginsenoside Rg1 recovery test results (n = 6)

[0103]

[0104] Table 13 Ginsenoside Rb1 recovery test results (n = 6)

[0105]

[0106] 2.7 Intermediate precision study

[0107] Different analysts were investigated at different times and using different instruments to measure the same test product. The results are shown in Table 14. There was no significant difference in the content results, indicating that the intermediate precision of the method of the present invention was good.

[0108] Table 14 Intermediate precision results

[0109]

[0110] 2.8 Content Limit

[0111] According to the method of the present invention, 19 batches of finished products of Dan'e Fukang decoction were tested, and the total amount of notoginsenoside R1, ginsenoside Rg1, and ginsenoside Rb1 was 6.6-12.7 mg / g, with an average value of 9.7 mg / g. The results are shown in Table 15.

[0112] Table 15 Results of determination of notoginsenoside content in 19 batches of Dan'e Fukang decoction (n=3)

[0113]

[0114]

[0115] The test results show that the present invention uses ultra-high performance liquid chromatography to determine the content of Panax notoginseng in Dan'e Fukang decoction by optimizing the preparation method of the test sample and the detection chromatographic conditions, and can well separate Panax notoginseng saponin R1, ginsenoside Rg1 and ginsenoside Rb1, and the method of the present invention has good linearity, repeatability, stability and recovery rate, etc., and the methodology is good. Compared with the existing national drug standard method, Panax notoginseng saponin R1, ginsenoside Rg1 and ginsenoside Rb1 have good separation, and the method of the present invention is simple in steps, saves time and labor, can completely separate the target peaks, and saves analysis time.

Claims

1. A method for detecting the content of Panax notoginseng in Dan'e Fukang decoction, characterized in that: The detection method is as follows: water is used as a solvent to prepare the test solution, and the solution is injected into an ultra-high performance liquid chromatograph to determine the content of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1; the chromatographic conditions are as follows: octadecylsilane bonded silica gel is used as a filler; acetonitrile and water are used as mobile phases for gradient elution; the detection wavelength is 203 nm, the column temperature is 20-35°C; the flow rate is 0.4-0.6 mL / min; the theoretical plate number calculated based on the ginsenoside Rb1 peak should be no less than 700,000; The gradient elution conditions with acetonitrile and water as the mobile phase were: The time was 0 to 11 min, the mobile phase was 16% acetonitrile, and the mobile phase was 84% ​​water; The time is 11 to 15 minutes, the mobile phase is 16 to 21% acetonitrile, and the mobile phase is 84 to 79% water; The time is 15 to 24 minutes, the mobile phase is 21 to 31% acetonitrile, and the mobile phase is 79 to 69% water; The time is 24 to 26 minutes, the mobile phase is 31 to 40% acetonitrile, and the mobile phase is 69 to 60% water; The time was 26 to 29 min, the mobile phase was 40% acetonitrile, and the mobile phase was 60% water; The time is 29 to 30 minutes, the mobile phase is 40 to 16% acetonitrile, and the mobile phase is 60 to 84% water; The time was 30 to 33 min, the mobile phase was 16% acetonitrile, and the mobile phase was 84% ​​water; The preparation of the test solution is to take 2-5g of Dan'e Fukang decoction and put it in a 50mL volumetric flask, add water and ultrasonicate for 30-90 minutes, then add water to make up the volume, shake well, and filter.

2. The method for detecting the content of Panax notoginseng in Dan'e Fukang decoction according to claim 1, characterized in that: Take 2 g of Dan'e Fukang decoction and put it in a 50 mL volumetric flask. Add water and ultrasonicate for 30 minutes, then add water to make up the volume, shake well, and filter.

3. The method for detecting the content of Panax notoginseng in Dan'e Fukang decoction according to claim 1 or 2, characterized in that: The power of the ultrasound is 480-520W; the frequency of the ultrasound is 35-45kHz.

4. The method for detecting the content of Panax notoginseng in Dan'e Fukang decoction according to claim 1, characterized in that: The column temperature was 25°C; the flow rate was 0.4 mL / min.

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