Method for simultaneously extracting tanshinone IIA and salvianolic acid B from salvia miltiorrhiza

Abstract

The invention belongs to the field of traditional Chinese medicine compounds, and particularly relates to a method for simultaneously extracting tanshinone IIA and salvianolic acid B from salvia miltiorrhiza. According to the specific technical scheme, the preparation method comprises the following steps: putting salvia miltiorrhiza powder into a container, adding an ethyl lactate solution, mixing, oscillating and shaking uniformly, then carrying out ultrasonic treatment and centrifugation, and taking supernate, so as to obtain a mixed solution of tanshinone IIA and salvianolic acid B; according to the ultrasonic-assisted green solvent ethyl lactate extraction method obtained through optimization, tanshinone IIA and salvianolic acid B with large polarity difference in salvia miltiorrhiza can be extracted at the same time. The extraction method disclosed by the invention is simple in operation steps, and the yield of active ingredients and the utilization rate of raw materials are relatively ideal. The method has the advantages of being simple in process, safe, low in cost, environmentally friendly, convenient to operate, good in reproducibility, short in period, capable of avoiding degradation of effective components and the like.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicine compounds, and particularly relates to a method for simultaneously extracting tanshinone IIA and salvianolic acid B from salvia miltiorrhiza. Background Art

[0002] Salvia miltiorrhiza (Salvia miltiorrhiza), a plant of the Salviaceae family, is widely used to treat cardiovascular and cerebrovascular diseases. Its main bioactive components include lipophilic tanshinones and hydrophilic salvianolic acids. Tanshinone IIA has shown significant therapeutic benefits in a variety of cardiovascular, inflammatory, neurodegenerative, and cancer treatments. Salvianolic acid B is a hydrophilic phenolic compound with anti-inflammatory, anti-apoptotic, and anti-fibrotic properties.

[0003] Due to the significant difference in solubility between tanshinone and salvianolic acid, existing techniques typically extract these two substances separately. Tanshinone IIA is typically extracted using a 95% ethanol solution under reflux, but this method carries risks such as combustion, explosion, and environmental pollution. Salvianolic acid B is often extracted using an aqueous solution under reflux, but its high-temperature instability leads to degradation, thus compromising extraction efficiency.

[0004] As a green solvent, ethyl lactate is widely used in the food and pharmaceutical fields due to its biodegradability, low toxicity and environmentally friendly properties. Ultrasonic-assisted extraction is considered to be a simple, inexpensive and green extraction method. During the extraction process, ultrasound can destroy the plant cell walls, and under the synergistic effect of mechanical and cavitation, the active ingredients are released and diffused. Ultrasonic-assisted extraction can improve extraction efficiency while reducing energy consumption. Ultrasonic devices are divided into water bath type and probe type. Among them, the probe type has high ultrasonic power, good wall breaking effect, short extraction time, low solvent consumption, and can significantly increase the yield of natural products. However, when treated with ultrasound, the sample temperature will rise sharply, making it difficult to extract active compounds that are unstable at high temperatures.

[0005] Therefore, if an efficient, low-cost and green method is developed that can simultaneously extract tanshinone IIA and salvianolic acid B, it will provide a new feasible approach for the resource utilization of Salvia miltiorrhiza. Summary of the Invention

[0006] The purpose of the present invention is to provide a method for simultaneously extracting tanshinone IIA and salvianolic acid B from salvia miltiorrhiza.

[0007] To achieve the above-mentioned purpose of the invention, the technical solution adopted by the present invention is: a method for simultaneously extracting tanshinone IIA and salvianolic acid B from Salvia miltiorrhiza, using ethyl lactate solution as the extracting solution and ultrasonic treatment as an auxiliary.

[0008] Preferably, the method comprises the following steps: placing Danshen powder in a container, adding ethyl lactate solution, mixing and shaking, then ultrasonicating, centrifuging, and collecting the supernatant to obtain a mixed solution of tanshinone IIA and salvianolic acid B. Preferably, the ethyl lactate solution is an aqueous solution with a pH of 2 and a volume fraction of 55-65%; the solid-liquid ratio of Danshen powder to ethyl lactate solution is 1:15-1:25 mg / L; the ultrasonic irradiation power is 200-350 W; the ultrasonic irradiation time is 20-36 minutes; the extraction temperature is 30-40°C, the cavitation time is 0.5-1.5 seconds, and the buffer time is 0.5-1.5 seconds.

[0009] The present invention has the following beneficial effects: Currently, there are no reports on the simultaneous extraction of tanshinone IIA and salvianolic acid B using ultrasound-assisted ethyl lactate. The optimized ultrasound-assisted green solvent ethyl lactate extraction method of the present invention allows for the simultaneous extraction of tanshinone IIA and salvianolic acid B, which have significantly different polarities, from Danshen. The extraction method of the present invention features simple steps, and achieves ideal yields of active ingredients and raw material utilization.

[0010] The present invention uses the bio-based solvent ethyl lactate as the extraction solvent. This solvent is biodegradable, easily recyclable, and environmentally friendly, improving extraction yield while reducing environmental pollution. The present invention uses a low-temperature coolant circulation pump to precisely control the reaction temperature, effectively preventing high-temperature degradation of the active ingredient and further achieving highly selective extraction of specific compounds.

[0011] Compared with traditional extraction methods (Soxhlet extraction), the method provided by the present invention can significantly shorten extraction time, improve extraction yield, reduce energy consumption, and reduce costs. Furthermore, the method provided by the present invention has high reproducibility and stability, providing a reference for green extraction processes for active compounds with widely varying polarity or active compounds that are unstable at high temperatures.

[0012] In general, the present invention has the advantages of simple process, safety, low cost, green and environmental protection, convenient operation, good reproducibility, short cycle, and avoidance of degradation of active ingredients. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 Schematic diagram of the extraction device of the present invention;

[0014] Figure 2 HPLC charts of tanshinone IIA and salvianolic acid B;

[0015] Figure 3 Schematic diagram of PBD experimental results;

[0016] Figure 4 Schematic diagram of the response surface optimization of the total yield of tanshinone IIA and salvianolic acid B, showing the interaction between the volume fraction of ethyl lactate and ultrasonic irradiation power;

[0017] Figure 5 Schematic diagram of the response surface optimization of the total yield of tanshinone IIA and salvianolic acid B, showing the interaction between the volume fraction of ethyl lactate and ultrasonic irradiation time;

[0018] Figure 6 Schematic diagram of the interaction between ultrasonic power and ultrasonic irradiation time for response surface optimization of the total yield of tanshinone IIA and salvianolic acid B;

[0019] Figure 7 Schematic diagram of the effects of different extraction solvents on the extraction rates of tanshinone IIA and salvianolic acid B. DETAILED DESCRIPTION

[0020] The present invention provides a method for simultaneously extracting tanshinone IIA and salvianolic acid B from salvia miltiorrhiza, which specifically comprises the following steps:

[0021] Weigh salvia miltiorrhiza powder into a container and add ethyl lactate solution, preferably an aqueous solution with a pH of 2 and a volume fraction of 55-65%. Preferably, the solid-liquid ratio of salvia miltiorrhiza powder to ethyl lactate solution is 1:15 to 1:25 mg / L. Mix and shake until evenly mixed, then perform ultrasonic treatment. The ultrasonic treatment conditions are preferably: ultrasonic irradiation power of 200-350 W, ultrasonic irradiation time of 20-36 min, and reaction temperature of 30-40°C. After the treatment is completed, centrifuge and collect the supernatant to obtain a mixed solution of tanshinone IIA and salvianolic acid B.

[0022] After obtaining the mixed solution, it can be eluted with a macroporous resin and separated by silica gel column chromatography to obtain tanshinone IIA and salvianolic acid B, respectively. The elution and separation conditions can be as follows: An appropriate amount of pretreated macroporous adsorption filler is loaded into the glass column, ensuring uniform distribution throughout the column. The column is treated with deionized water until the effluent is free of visible impurities and bubbles. The mixed solution of tanshinone IIA and salvianolic acid B is gently dripped onto the column along the inner wall. The column is eluted with 2 column volumes (BV) of deionized water at a flow rate of 1 mL / min to remove unadsorbed components. Subsequently, continuous elution is performed at a flow rate of 1 mL / min, first using 5 BV of 20% ethanol, then 5 BV of 90% ethanol, and finally 5 BV of 100% ethanol. The 20% ethanol solution primarily elutes salvianolic acid B, while the 90% ethanol solution primarily elutes the tanshinone IIA fraction.

[0023] The following will be combined with the accompanying drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of them. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art. The data obtained are the average values ​​obtained after at least three repetitions, and the data obtained in each repetition are valid data.

[0024] Example 1: Single factor experiment to test the effect of reaction parameters on extraction effect

[0025] 1. Extraction device settings

[0026] The whole set of extraction equipment is as follows Figure 1 As shown, an ultrasonic cell disruptor with a low-temperature coolant circulation system 6, model BILON-1000Y, includes a control unit 1 for controlling operating parameters such as ultrasonic time and power. It also includes a main unit 2, which is electrically connected to a 6mm φ horn 3 for focusing energy (amplifying the displacement or velocity of mechanical vibration particles and concentrating the ultrasonic energy on a smaller area). The drill bit of the horn 3 is inserted into the reactant material 5 inside a double-jacketed beaker 4. The outer wall of the double-jacketed beaker 4 is provided with a condensate inlet and a condensate outlet for circulating liquid into the outer wall of the double-jacketed beaker 4 for temperature control. The condensate inlet and condensate outlet are respectively connected to the coolant inlet and outlet of a low-temperature coolant circulation pump 6. The low-temperature coolant circulation pump 6 is electrically connected to the control unit 1, and its temperature parameters are controlled by the control unit 1 to ensure that the reaction system is maintained at the desired temperature for constant temperature reaction. Unless otherwise specified, the reaction equipment used is the same as herein.

[0027] 2. Parameter control method and extraction rate calculation

[0028] Before the reaction, turn on the low-temperature coolant circulation pump 6 and preheat for 5 minutes to bring the reaction system temperature to the ultrasonic temperature set by the control unit 1. Adjust the ultrasonic parameters, then extend the drill bit of the horn 3 below the liquid level of the reaction system in the double-jacketed beaker 4, keeping the distance between the needle tip and the bottom of the beaker within 1 cm. Adjust the ultrasonic power, extraction time, extraction temperature, liquid-to-solid ratio, pH, and ethyl lactate volume fraction. Unless otherwise specified, each variable was changed individually during each adjustment, while all other conditions remained unchanged. The remaining conditions, excluding these variables, were: ultrasonic power 300 W, extraction time 30 min, cavitation time 1.5 s, buffer time 1.5 s, extraction temperature 35°C, solid-to-liquid ratio of Salvia miltiorrhiza powder to ethyl lactate solution 1:20 mg / L, ethyl lactate solution volume fraction 65%, and pH 2. The cavitation time refers to the duration of a single drill cycle, and the buffer time refers to the duration of a single drill cycle. These two cycles alternate: the drill operates for 1.5 seconds, stops for 1.5 seconds, then operates for another 1.5 seconds, then stops for another 1.5 seconds, for a full 30 minutes. This intermittent ultrasonic operation produces a cavitation effect, in which tiny bubbles in the liquid vibrate, grow, and eventually collapse and close under the influence of ultrasound, releasing a large amount of energy. Continuous ultrasound can lead to excessive aggregation of cavitation bubbles, forming a bubble cloud, which hinders the effective transmission of ultrasonic energy and reduces cavitation efficiency.

[0029] After the extraction, the filtrate was obtained by vacuum filtration. The filtrate was diluted four times with methanol before testing. The diluted solution was extracted with a syringe, filtered through a 0.22μm microporous membrane, and then analyzed by HPLC. Specifically: a Shimadzu LC-20AD high-performance liquid chromatography system was used with a Kromasil-C18 column (250mm×4.6mm, 5μm) for chromatographic quantification of tanshinone IIA and salvianolic acid B. The standard solutions of tanshinone IIA and salvianolic acid B were first diluted with methanol to the required concentration, filtered through a 0.22μm filter membrane, and then detected by HPLC. When the detection time was 0-8min, the detection wavelength was 286nm, and the mobile phase was acetonitrile and water in a volume ratio of 40:60 (v / v). When the detection time was 8-20min, the wavelength was adjusted to 270nm, and the mobile phase was acetonitrile and water in a mobile phase ratio of 95:5 (v / v). The flow rate was set to 0.8mL / min throughout the process, and the column temperature was 30℃. Under the specified analytical conditions, the retention times of tanshinone IIA and salvianolic acid B were 16.98 min and 4.90 min, respectively. Figure 2 As shown. Calibration curves of tanshinone IIA and salvianolic acid B were established respectively, and the linear regression equations of the calibration curves were: Y 丹参酮IIA =1235.4X+552.91(R 2 =0.9995,n=7),Y 丹酚酸B =5012.4X+394.16(R 2=0.9998,n=7).

[0030] The calculation formula for the extraction rate (mg / g) of tanshinone IIA and salvianolic acid B is:

[0031] Y=4C·V / M

[0032] Where Y is the extraction yield of tanshinone IIA or salvianolic acid B (mg / g); C is the concentration of tanshinone IIA or salvianolic acid B in the sample determined by HPLC (mg / mL); V is the volume of the extraction solvent (mL); and M is the mass of the sample (g).

[0033] 3. Effect of ethyl lactate volume fraction on extraction rate

[0034] Weigh 0.5 g of Danshen powder and add ethyl lactate solution (55%, 60%, 65%, 70%, 75%, and 80% by volume) at pH 2. Mix and shake thoroughly. Extract tanshinone IIA and salvianolic acid B under ultrasonic irradiation at 300 W for 30 minutes at 35°C. After extraction, centrifuge the supernatant, and determine the extraction yields of tanshinone IIA and salvianolic acid B by HPLC. The results are shown in Table 1.

[0035] Table 1 Comparison of the effects of ethyl lactate volume fraction on the extraction rates of tanshinone IIA and salvianolic acid B

[0036] Ethyl lactate volume fraction Tanshinone IIA (mg / g) Salvianolic acid B (mg / g) 55％ 0.96 124.70 60％ 1.02 140.10 65％ 1.11 151.02 70％ 1.11 139.73 75％ 1.12 129.63 80％ 1.12 100.94

[0037] 4. Effect of pH of ethyl lactate on extraction rate

[0038] The other conditions remained unchanged, and hydrochloric acid was used to adjust the pH of ethyl lactate to 1, 2, 3, 4, and 5. The results are shown in Table 2.

[0039] Table 2 Comparison of the effects of ethyl lactate solution pH on the extraction rates of tanshinone IIA and salvianolic acid B

[0040] pH of ethyl lactate solution Tanshinone IIA (mg / g) Salvianolic acid B (mg / g) 1 0.98 112.14 2 1.11 151.09 3 1.13 140.87 4 1.16 129.34 5 1.18 121.45

[0041] 5. Effect of extraction temperature on extraction rate

[0042] The other conditions remained unchanged, and the extraction temperatures were controlled at 25, 30, 35, 40, 50, and 60°C. The results are shown in Table 3.

[0043] Table 3 Effect of extraction temperature on the extraction rate of tanshinone IIA and salvianolic acid B

[0044] Extraction temperature ℃ Tanshinone IIA (mg / g) Salvianolic acid B (mg / g) 25 0.97 140.07 30 1.06 149.08 35 1.11 151.81 40 1.10 151.49 50 1.09 150.12 60 1.08 149.23

[0045] 6. Effect of material-liquid ratio on extraction rate

[0046] The other conditions remained unchanged, and the solid-liquid ratios of Danshen powder and ethyl lactate solution were controlled to be 1:10, 1:15, 1:20, 1:25, and 1:30 mg / L, respectively. The results are shown in Table 4.

[0047] Table 4 Comparison of the effects of material-liquid ratio on the extraction rate of tanshinone IIA and salvianolic acid B

[0048] Material-liquid ratio mg / L Tanshinone IIA (mg / g) Salvianolic acid B (mg / g) 1:10 0.98 138.44 1:15 1.04 142.54 1:20 1.11 151.06 1:25 1.11 151.67 1:30 1.10 151.06

[0049] 7. Effect of ultrasonic irradiation power on extraction rate

[0050] The other conditions remained unchanged, and the ultrasonic irradiation powers were controlled to 200, 250, 300, 350, 400 and 450 W, respectively. The results are shown in Table 5.

[0051] Table 5 Comparison of the effects of ultrasonic power on the extraction rates of tanshinone IIA and salvianolic acid B

[0052]

[0053]

[0054] 8. Effect of ultrasonic irradiation time on extraction rate

[0055] The other conditions remained unchanged, and the ultrasonic irradiation time was controlled to be 10, 20, 30, 40, 50 and 60 min respectively. The results are shown in Table 6.

[0056] Table 6 Comparison of the effects of ultrasonic time on the extraction rate of tanshinone IIA and salvianolic acid B

[0057] Ultrasonic time min Tanshinone IIA (mg / g) Salvianolic acid B (mg / g) 10 0.87 118.15 20 1.08 142.43 30 1.11 151.26 40 1.10 150.79 50 1.10 150.44 60 1.10 150.20

[0058] 9. Effect of cavitation time on extraction rate

[0059] The other conditions remained unchanged, and the cavitation time was controlled to be 0.5, 1.0, 1.5, 2 and 2.5 s respectively. The results are shown in Table 7.

[0060] Table 7 Comparison of the effects of cavitation time on the extraction rates of tanshinone IIA and salvianolic acid B

[0061] Cavitation time min Tanshinone IIA (mg / g) Salvianolic acid B (mg / g) 0.5 0.89 128.96 1.0 1.04 140.60 1.5 1.11 151.02 2.0 1.08 149.80 2.5 1.07 145.16

[0062] 9. Impact of buffer time on extraction rate

[0063] The other conditions remain unchanged, and the buffer time is controlled to be 0.5, 1.0, 1.5, 2 and 2.5s respectively. The results are shown in Table 8.

[0064] Table 8 Comparison of the effects of buffer time on the extraction rates of tanshinone IIA and salvianolic acid B

[0065] Buffer time min Tanshinone IIA (mg / g) Salvianolic acid B (mg / g) 0.5 0.94 127.56 1.0 1.08 145.32 1.5 1.11 151.21 2.0 1.10 146.79 2.5 1.08 140.39

[0066] Example 2: Response surface optimization experiment

[0067] 1. Plackett-Burman experiment (PBD experiment)

[0068] According to the results of Example 1, the volume fraction of ethyl lactate (55% to 65%, X1), ultrasonic power (250 to 350 W, X2), ultrasonic irradiation time (10 to 30 min, X3), liquid-to-material ratio (15 to 25 mL / g, X4), reaction temperature (25 to 35 ° C, X5), solvent pH (2 to 4, X6), cavitation time (0.5 to 1.5 s, X7) and buffer time (0.5 to 1.5 s, X8) were selected, and the experimental design and statistical analysis were performed using DesignExpert software (Stat-Ease, Ver 8.0, Minneapolis, USA). The significance analysis of each parameter was performed using the t-test. Under the assumption that there was no interaction between the variables, a first-order polynomial model was used to fit the PBD.

[0069]

[0070] Y is the predicted response, β0 is the intercept, β i is the linear regression coefficient, X i is the coded independent variable.

[0071] The results are as follows Figure 3 As shown. Generally, variables with a confidence level greater than 95% (P < 0.05) are considered to be statistically significant factors. The results showed that X1, X2 and X3 had a greater effect on the yield of tanshinone IIA and salvianolic acid B, while the effects of other parameters were not significant. The Pareto chart illustrates the different effects of multiple factors on the extraction rate of tanshinone IIA and salvianolic acid B by plotting the t value of each variable against the corresponding effect. If the t value exceeds the threshold, it indicates that the confidence level exceeds 95%, indicating significance. In addition, the significance coefficient of the influencing factor can also be determined by comparison. In contrast, the t values ​​of ethyl lactate volume fraction, ultrasonic irradiation power and ultrasonic irradiation time all exceeded the threshold, and X1>X3>X2. Therefore, ethyl lactate volume fraction, ultrasonic irradiation power and ultrasonic irradiation time were selected as important parameters for subsequent optimization.

[0072] 2. Box-Behnken design experiment (BBD experiment)

[0073] BBD was used to optimize the important variables selected from the PBD experiment. BBD includes three parameters and three levels (the three levels are coded as -1, 0, and +1, corresponding to low, medium, and high values, respectively). Using 17 experimental data points, the independent variables and their interactions that affect the extraction yield of tanshinone IIA and salvianolic acid B were evaluated. Significant variables included the volume fraction of ethyl lactate (X1, 60-70%), ultrasonic irradiation power (X2, 300-400W), and ultrasonic irradiation time (X3, 20-40min). Design-Expert software (version 8.0) was used to perform regression analysis on the data to facilitate the establishment of a quadratic polynomial model. The generalized quadratic equation was used to analyze the correlation between the observed response values ​​and the three independent variables:

[0074]

[0075] Where Y is the predicted response, X i and X j are coded independent variables, β0, β ii and β ij Regression parameters for intercept, linear, quadratic, and interaction effects, respectively.

[0076] The "F" value and "P" value are used to test the interaction between variables. Generally speaking, the higher the F value and P < 0.05, the more statistically significant the model term is. In this embodiment, the model F value is 284.56, and the P value < 0.05 indicates that it is significant.

[0077] In addition, the coefficient of determination R of the quadratic regression model 2 The higher the value, the higher the feasibility of the model. The expected response to the total extraction rate of tanshinone IIA and salvianolic acid B is expressed by a second-order polynomial equation:

[0078] Extraction rate (%) =

[0079] 152-2.25X1-2.39X2+5.74X3-3.13X1X2+1.42X1X3+3.63X2X3-8.29X1 2 -7.47X2 2 -2.56X3 2

[0080] The total extraction rate of tanshinone IIA and salvianolic acid B was calculated by the extraction rate formula. 2 The adjusted R value (0.9980) was obtained by Design-Expert software. 2 The experimental data and the predicted values ​​are very close, indicating that there is a strong correlation between the experimental data and the predicted values. The coefficient of variance of the experiment is low at 0.45, indicating that the experiment has high reproducibility and reliability.

[0081] The total extraction rates of tanshinone IIA and salvianolic acid B were associated with two experimental variables, respectively, and the third experimental variable was set to zero, so as to study the effects of the three parameters and their interactions on the extraction rate.

[0082] When the ultrasonic irradiation time is kept constant at zero level, the interaction effect between the volume fraction of ethyl lactate X1 and the ultrasonic irradiation power X2 is as follows: Figure 4 The results showed that the total extraction rate of tanshinone IIA and salvianolic acid B increased significantly with the increase of ethyl lactate volume fraction and extraction temperature, but the extraction rate decreased with the further increase of volume fraction or ultrasonic irradiation power. Figure 5 The interaction between the volume fraction of ethyl lactate and the ultrasonic irradiation time is shown. The higher the volume fraction and the longer the ultrasonic irradiation time, the higher the total extraction rate of tanshinone IIA and salvianolic acid B. When the volume fraction of ethyl lactate remains constant, the response surface of the effect of ultrasonic irradiation power and ultrasonic irradiation time on the total yield is shown in Figure 2. Figure 6 shown.

[0083] Using BBD software, the optimal conditions predicted were: 65% ethyl lactate volume fraction, 350W ultrasonic irradiation power, and 36 minutes of ultrasonic irradiation. Under these optimized conditions, the extraction yields of tanshinone IIA and salvianolic acid B were 1.14 mg / g and 154.59 mg / g, respectively.

[0084] 3. Optimization parameter verification

[0085] Three validation tests were carried out using the optimized parameters (ethyl lactate volume fraction 65%, ultrasonic irradiation power 350 W, ultrasonic irradiation time 36 min, liquid-solid ratio 1:25 mL / g, pH = 2, reaction temperature 35 °C, cavitation time 1.5 s, and buffer time 1.5 s) to evaluate the reliability of the model.

[0086] The results showed that the yield of tanshinone IIA was 1.14±0.03 mg / g, and the yield of salvianolic acid B was 154.42±0.61 mg / g. The measured values ​​were very close to the predicted values, proving the reliability of the model.

[0087] 4. Repeatability verification

[0088] Using the optimized parameters, ultrasonic extraction was performed on five samples of Salvia miltiorrhiza from the same source. The results showed that the extraction rates of tanshinone IIA and salvianolic acid B were highly reproducible, with relative standard deviations of 3.42% and 4.18%, respectively.

[0089] Example 3: Comparison of extraction results with ethanol solution

[0090] According to the optimized parameters of Example 2, while keeping other conditions unchanged, the volume fraction of ethyl lactate was set to 50% to 80%, or the ethyl lactate solution was replaced with ethanol solution, and the volume fraction of ethanol solution was also set to 50% to 80%, and the extraction rates of tanshinone IIA and salvianolic acid B in the Danshen samples were measured. The results are shown in Figure 2. Figure 7 shown.

[0091] The results showed that with the increase of ethyl lactate volume fraction, the extraction rate of tanshinone IIA increased from 0.92 to 1.14 mg / g, and the extraction rate of salvianolic acid B increased from 108.94 to 154.75 mg / g. When the volume fraction of ethyl lactate was 60% to 75%, the extraction rate was significantly higher than that of ethanol solution.

[0092] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various deformations, modifications, and substitutions made to the technical solutions of the present invention by ordinary technicians in this field should fall within the scope of protection determined by the claims of the present invention.

Claims

1. A method for simultaneously extracting tanshinone IIA and salvianolic acid B from Salvia miltiorrhiza, characterized in that: Ethyl lactate solution was used as the extraction solution, and ultrasonic treatment was used as an auxiliary.

2. The method according to claim 1, wherein: The method comprises the following steps: placing salvia miltiorrhiza powder in a container, adding ethyl lactate solution, mixing and shaking, then performing ultrasonic treatment, centrifuging, and collecting the supernatant to obtain a mixed solution of tanshinone IIA and salvianolic acid B.

3. The method according to claim 2, wherein: The ethyl lactate solution is an aqueous solution with a pH of 2 and a volume fraction of 55 to 65%.

4. The method according to claim 2, wherein: The solid-liquid ratio of salvia miltiorrhiza powder and ethyl lactate solution is 1:10-1:30 mg / L.

5. The method according to claim 2, wherein: The ultrasonic irradiation power is 200-350W.

6. The method according to claim 2, wherein: The ultrasonic irradiation time is 20 to 36 minutes.

7. The method according to claim 2, characterized in that: The extraction temperature is 30-40℃.

8. The method according to claim 2, wherein: The cavitation time is 0.5 to 1.5 seconds.

9. The method according to claim 2, wherein: The buffer time is 0.5 to 1.5 seconds.

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