Sinomenine microwave extraction process based on eutectic solvent

Abstract

The invention belongs to the technical field of traditional Chinese medicine active component extraction, and relates to an extraction process of traditional Chinese medicine active components, in particular to a microwave extraction process of sinomenine based on a deep-eutectic solvent. The process comprises the following steps: S1, drying a caulis sinomenii medicinal material, crushing, and sieving with a 40-mesh sieve to obtain caulis sinomenii powder; s2, a deep eutectic solvent is prepared, the deep eutectic solvent is formed by mixing choline chloride, water and one or more of glucose, lactic acid and glycerin, and the mass fraction of water in the deep eutectic solvent is 30%-60%; s3, adding the caulis sinomenii powder into a deep eutectic solvent according to a solid-liquid ratio of 1: 20g / mL, and performing microwave extraction for 12-21 minutes under the power of 400-500w to obtain sinomenine. The process adopts microwave-assisted DESs to extract sinomenine, so that the sinomenine yield is increased, and the process is environment-friendly.

Description

Technical Field

[0001] The invention belongs to the technical field of extraction of active components of traditional Chinese medicines, relates to an extraction process of active components of traditional Chinese medicines, and particularly relates to a microwave extraction process of sinomenine based on a low eutectic solvent. Background Art

[0002] Sinomenine is an alkaloid extracted from the dried stems of Sinomenium sinomenium and Sinomenium pubescens. Sinomenine has analgesic, anti-inflammatory, immunosuppressive and anti-tumor effects. Sinomenine hydrochloride preparations have been used in the clinical treatment of rheumatism, rheumatoid joint fluid, ankylosing spondylitis and anti-arrhythmic diseases with significant effects.

[0003] At present, ethanol extraction and acid-water extraction are mostly used for the extraction of sinomenine. Among them, the acid-water extraction method has the characteristics of simple operation process, low energy consumption, simple solvent recovery and high extraction rate, and is more widely used in industry. For example, the document "Study on the Extraction Process of Sinomenine from Caulis Sinensis" [J] (Kang Hongyu, Ren Liping. Anhui Agricultural Science, 2010, 38 (09): 4578 + 4582.) reported a process for extracting sinomenine from Caulis Sinensis using an acid-base method, and the final average yield of this method was 3.7%. However, in the extraction and purification process, a large amount of acid and alkali is required, the solvent is highly toxic, and a lot of solid waste is generated. Patent CN202111287996.5 reports a method for extracting sinomenine from Caulis Sinensis using a mixed solvent. The process uses water-containing n-butanol and n-heptane or n-octane to prepare a mixed solvent, and extracts multiple times by stirring at room temperature, and repeatedly extracts to prepare sinomenine. This method also does not avoid the use of toxic reagents and cannot reflect the concept of green environmental protection.

[0004] Deep eutectic solvents (DESs) are a new type of green solvents with the advantages of simple preparation, low or non-toxicity, low price, easy availability, biodegradability, and recyclability. They have been widely used in the extraction of active ingredients from various Chinese herbal medicines. Microwave extraction, as a new modern extraction technology, has the advantage of high extraction efficiency. However, based on the concept of green extraction, there is no related report on the use of microwave-assisted DESs solvents in the extraction of sinomenine. Summary of the invention

[0005] The object of the present invention is to provide a microwave extraction process of sinomenine based on a low eutectic solvent, which uses microwave-assisted DESs to extract sinomenine, which improves the yield of sinomenine and is environmentally friendly.

[0006] The technical solution adopted by the present invention to solve its technical problem is:

[0007] A microwave extraction process of sinomenine based on a low eutectic solvent, the process comprising the following methods:

[0008] S1, drying the Caulis Sinomenii Herba and crushing it, passing it through a 40-mesh sieve to obtain Caulis Sinomenii Herba powder;

[0009] S2. preparing a low eutectic solvent, wherein the low eutectic solvent is a mixture of choline chloride, water and one or more of glucose, lactic acid and glycerol, wherein the mass fraction of water in the low eutectic solvent is 30%-60%;

[0010] S3. Add a low eutectic solvent to the Sinomenine Caulis powder at a solid-liquid ratio of 1:20 g / mL, and extract by microwave at a power of 400w-500w for 12-21min to obtain Sinomenine.

[0011] DESs are made of natural ingredients such as glucose, mannitol, and amino acids, mixed in a certain molar ratio. Due to the strong intermolecular interactions (hydrogen bonds, electrostatic forces, etc.) between the components, the melting point of DESs is lower than the melting point of each component itself. Compared with traditional organic solvents, DESs solvents have the following advantages: First, they are more environmentally friendly. The main components of DESs are usually composed of natural products or biodegradable compounds, which have lower toxicity and biodegradability, reducing harm to the environment and human body. Second, DESs have higher selectivity and efficiency in the extraction process. Due to the unique physical properties of DESs, it has a stronger affinity with the target compound, which can achieve the separation and extraction of multiple components and improve the extraction efficiency.

[0012] Microwave-assisted extraction technology uses microwave radiation to heat the water molecules in the sample, thereby achieving rapid and efficient extraction of the target components. Compared with traditional extraction methods, microwave-assisted extraction technology has the following significant advantages: First, rapid heating. Microwave radiation can quickly convert water molecules in the sample into thermal energy, achieve rapid heating, and shorten the extraction time; second, energy saving and environmental protection. The microwave-assisted extraction process has high energy utilization efficiency and does not require a preheating process, which can reduce the use of extraction solvents, reduce energy consumption and solvent emissions; third, high extraction efficiency. Microwave radiation can achieve efficient extraction at a lower temperature, which helps to protect heat-sensitive components and reduce the occurrence of chemical decomposition and oxidation reactions; fourth, easy operation. Microwave-assisted extraction does not require complex equipment and steps, and is simple and convenient to operate.

[0013] Preferably, the deep eutectic solvent comprises choline chloride, water and lactic acid, and the weight ratio of choline chloride to lactic acid is 2:1-1:2.

[0014] Preferably, the deep eutectic solvent consists of choline chloride, lactic acid and water, and the weight ratio of choline chloride to lactic acid is 2:1.

[0015] Preferably, in S3, the microwave extraction time is 20 min-21 min, and the microwave power is 423 W-425 W.

[0016] Preferably, the low eutectic solvent consists of choline chloride, lactic acid and water, the weight ratio of choline chloride to lactic acid is 2:1, and the mass fraction of water is 57%-58%.

[0017] The beneficial effects of the present invention are:

[0018] The present invention uses the rhizome of Caulis Sinomenii as raw material, uses microwave-assisted DESs to extract sinomenine, and investigates the effects of microwave extraction power, time and solvent water content on the extraction effect of sinomenine through single factor experiment and response surface experimental design. By optimizing the above factors, the optimal extraction process parameters are determined.

[0019] Through literature research, the research on the extraction process of sinomenine from Caulis Sinensis mainly focuses on the traditional extraction method, and the traditional ethanol solvent is also used. For example, the document "Determination of sinomenine content in Caulis Sinensis roots by HPLC method and optimization of its extraction process" [J]. (Chen Xi. Anhui Agricultural Science, 2010, 38 (29): 16236-16238.) reported that 70% ethanol was used to optimize the ultrasonic extraction process of sinomenine, and the final sinomenine yield was 1.73%; the document "Star point design-response surface method optimization of microwave extraction process of sinomenine from Caulis Sinensis" [J]. (Li Yinglei, Chen Jianhua, Wang Yin, et al. Shizhen Traditional Chinese Medicine, 2013, 24 (03): 659-661.) reported that 75% ethanol was used to optimize the microwave extraction process of sinomenine, and the final sinomenine yield was 0.803%. The methods in the above documents are all classic organic solvent extraction methods, which do not achieve the purpose of green extraction. The present invention screened a variety of DES solvents, determined a DES solvent type, and optimized the process conditions for microwave extraction of sinomenine with the solvent, with a final yield of 2.0%, which is higher than the traditional extraction method. More importantly, the solvent used in the present invention is green. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is the screening result of sinomenine extracted by 10 groups of different DESs solvents;

[0021] Figure 2 This is the trend graph of microwave time on sinomenine extraction;

[0022] Figure 3 It is the trend graph of microwave power on sinomenine extraction;

[0023] Figure 4 is the trend graph of solvent water content on sinomenine extraction;

[0024] Figure 5 The results of the interaction of various factors based on response surface optimization are: a interaction between microwave power and time, b interaction between microwave time and water content, and c interaction between microwave power and water content;

[0025] Figure 6 This is the chromatogram of sinomenine standard;

[0026] Figure 7 This is the chromatogram of the Qingfengteng sample;

[0027] Figure 8 This is the standard curve for HPLC determination of sinomenine. DETAILED DESCRIPTION

[0028] The technical solution of the present invention is further described in detail below through specific embodiments. It should be understood that the implementation of the present invention is not limited to the following embodiments, and any form of modification and / or change made to the present invention will fall within the protection scope of the present invention.

[0029] In the present invention, unless otherwise specified, all parts and percentages are by weight, and the equipment and raw materials used can be purchased from the market or are commonly used in the art. The methods in the following embodiments are all conventional methods in the art unless otherwise specified.

[0030] The reagents used in the following examples, unless otherwise specified, can be purchased from conventional biochemical reagent stores.

[0031] The present invention mainly protects a microwave extraction process of sinomenine based on a low eutectic solvent, which comprises the following methods:

[0032] S1, drying the Caulis Sinomenii Herba and crushing it, passing it through a 40-mesh sieve to obtain Caulis Sinomenii Herba powder;

[0033] S2. preparing a low eutectic solvent, wherein the low eutectic solvent is a mixture of choline chloride, water and one or more of glucose, lactic acid and glycerol, wherein the mass fraction of water in the low eutectic solvent is 30%-60%;

[0034] S3. Add a low eutectic solvent to the Caulis Sinomenii powder at a solid-liquid ratio of 1:20 g / mL, and perform microwave extraction at a power of 400w-500w for 12-21 min to obtain a sinomenine extract.

[0035] Example A microwave extraction process of sinomenine based on a deep eutectic solvent

[0036] 1. Preparation and screening of DESs

[0037] In this experiment, two hydrogen bond acceptors (HBAs) and five hydrogen bond donors (HBDs) were selected to form 10 DESs, as shown in Table 1. The specific preparation operation is as follows: according to the formula and ratio of DESs in Table 1, appropriate amounts of HBA and HBD were weighed and placed in a 500 mL beaker, heated to 70°C on a magnetic stirrer to form a stable, transparent solution, and then sealed for later use.

[0038] Table 1 Formulas and ratios of different solvent types

[0039]

[0040] Ten kinds of DESs were prepared according to the above method. Single factor experiments were carried out under four fixed conditions: extraction time (15min), microwave power (300W), solid-liquid ratio (1:20g / mL), and solvent water content (40%). HPLC analysis was performed with sinomenine content as an indicator. The chromatographic peak of sinomenine in the sample was determined by comparing the elution time with that of sinomenine standard. The peak area of ​​the chromatograms of the ten solvents was used as an evaluation index to screen the DESs with the best extraction effect. The results are shown in Figure 2. Figure 1 shown.

[0041] Determination of extraction solvent: By comparing the peak areas of sinomenine extracted with 10 different DESs solvents, it can be seen that the extraction rate of sinomenine is the highest when choline chloride:lactic acid is used in a ratio of 2:1. Therefore, choline chloride-lactic acid is determined to be the best DESs type and used as the extraction solvent for subsequent experiments.

[0042] 2. Microwave Extraction Technology of Sinomenine

[0043] After determining the DESs with the best extraction effect, microwave power, microwave time, and solvent water content were selected as the investigation factors of the single factor experiment to investigate the effects of different microwave powers, different microwave times, and different solvent water contents on the extraction rate of sinomenine. The experiment was carried out under the experimental conditions of fixed factors of solid-liquid ratio of 1:20g / mL, 1g of sinomenine powder, and temperature of 50℃.

[0044] 2.1 Single factor experiment on microwave extraction of Sinomenium sinomenium

[0045] 2.1.1 Effect of different microwave times on the extraction of sinomenine

[0046] Weigh 1.0 g of Sinomenium sinensis powder into a 50 mL four-necked flask, add DESs with a water content of 40% at a solid-liquid ratio of 1:20 g / mL, and extract at different microwave extraction times (12, 15, 18, 21, 24 min) and a fixed microwave power of 400 W to obtain an extract.

[0047] Figure 2 The figure is a trend chart showing the effect of microwave extraction time on the amount of sinomenine extracted. Figure 2It can be seen that the microwave extraction time has a significant effect on the extraction of sinomenine. In the period of 12-18 minutes, as the extraction time increases, the content of sinomenine shows an upward trend, reaching the highest at 18 minutes; while in the period of 18-24 minutes, as the extraction time increases, the content of sinomenine shows a downward trend. Therefore, 18 minutes is determined to be the optimal extraction time for microwave-assisted extraction.

[0048] 2.1.2 Effect of different microwave powers on the extraction of sinomenine

[0049] 1.0 g of Sinomenium sinensis powder was weighed into a 50 mL four-necked flask, and DESs with a water content of 40% was added at a solid-liquid ratio of 1:20 g / mL. The mixture was extracted for 15 min at different microwave powers (250, 300, 400, 500, and 600 W) to obtain an extract.

[0050] Figure 3 The trend diagram of the effect of microwave power on the extraction amount of sinomenine. Figure 3 It can be seen that microwave power has a significant effect on sinomenine extraction. When the microwave power is in the range of 250-400W, with the increase of microwave power, the sinomenine content shows an upward trend, reaching the highest at 400W; while in the power range of 400-600W, with the increase of microwave power, the sinomenine content shows a downward trend. Therefore, 400W microwave extraction power is determined to be the optimal extraction power.

[0051] 2.1.3 Effect of different solvent water content on the extraction of sinomenine

[0052] 1.0 g of Sinomenium sinensis powder was weighed into a 50 mL four-necked flask, and the mixture was extracted for 15 min at a fixed microwave power of 400 W at a solid-liquid ratio of 1:20 g / mL at different water contents (20, 30, 40, 50, 60%) to obtain an extract.

[0053] Figure 4 The following is a trend chart showing the effect of water content in DESs on the amount of sinomenine extracted. Figure 4 It can be seen that the water content has a significant effect on the extraction of sinomenine. When the water content is in the range of 30-50%, the content of sinomenine shows an upward trend with the increase of water content, and the content reaches the highest value at 50%; while in the range of 50-60%, the content of sinomenine shows a downward trend with the increase of water content. Therefore, 50% is determined to be the optimal solvent water content.

[0054] 2.2 Microwave response surface optimization experiment of Sinomenium sinomenium based on DESs

[0055] On the basis of the single factor experimental results, taking sinomenine content as the response value, selecting microwave time, microwave power, and solvent water content as three factors, and based on the Box-Behnken design principle, using Design-Expert.V8.0.6.1 experimental design software to design the experiment, further optimize the microwave extraction process of sinomenine in Caulis Sinomenii. See Table 2 for details.

[0056] Table 2 Factor levels of DESs-assisted microwave extraction of sinomenine based on Box-Behnken design

[0057]

[0058] 2.2.1 Response surface experiment results and variance analysis

[0059] The response surface optimization experiment was carried out according to the response surface experiment plan. The experimental results are shown in Table 3.

[0060] Table 3 Response surface experimental design and results

[0061]

[0062] The data in Table 3 were processed using Design-Expert 8.0.6 software to calculate the regression coefficients, and the quadratic multinomial regression model for microwave-assisted extraction of sinomenine from Caulis Sinomenii was obtained:

[0063] Sinomenine content = 19.68-0.25*A+0.60*B+1.45*C-0.10*A*B+0.60*A*C+0.70*B*C+0.26*A 2 -1.84*B 2 -2.14*C 2 ,

[0064] In the equation, A represents the microwave extraction time, B represents the microwave power, and C represents the water content. The correlation coefficient of the quadratic multinomial regression equation is R 2 The above response surface experimental data were subjected to variance analysis, and the results are shown in Table 4.

[0065] Table 4 Analysis of variance in response surface experiments

[0066]

[0067] Note: **The difference is extremely significant, P<0.01; *The difference is significant, P<0.05.

[0068] The P value and F value are parameters used to determine the test results and the model fit. The smaller the P value, the more significant the result (a P value less than 0.01 indicates that the difference is extremely significant, and a P value less than 0.05 indicates that the difference is significant). The larger the F value, the better the model fit. As shown in Table 4, the F of the regression model is 38.01, P < 0.0001, indicating that the model has a high significance; the lack of fit term F = 2.26, P > 0.2234, indicating that the model is not significant and has good feasibility. In addition, it can be seen that B, C, B in the model 2 and C 2 The differences were extremely significant, indicating that microwave power and water content had a significant effect on the extraction of sinomenine; the A, AC, BC, and A 2 There is a significant difference, indicating that microwave extraction time has a great influence on the extraction of sinomenine. 2 =0.9799, model fit index (R 2 Adj)=0.9542, indicating that the model has good stability, and the model prediction value has a high degree of agreement with the experimental measurement value, which can be used to analyze and predict the extraction rate of sinomenine. Further data fitting and variance analysis of the model showed that the influencing factors from strong to weak are water content, microwave power, and microwave extraction time.

[0069] 2.2.2 Analysis of interaction effects of various factors in response surface diagram

[0070] The above regression equation was plotted using Design-Expert 8.0.6 software. The results are shown in Figure 5 ac.

[0071] Depend on Figure 5 It can be seen that there is an interactive effect between the factors of microwave power, water content and microwave extraction time. Overall, the response surface trends are all convex, indicating that the extraction rate has a strong correlation with the above three factors. With the increase of microwave power, water content and microwave extraction time, the content of sinomenine shows a trend of first increasing and then decreasing. The change of contour color and the slope of the response surface show that microwave power and water content are the main influencing factors, followed by microwave extraction time. In addition, when water content interacts with microwave power, it can be clearly observed that the color of the contour line changes significantly and the slope of the response surface becomes steeper, indicating that the difference in the influence of these two interactions on the extraction rate of sinomenine is statistically significant.

[0072] In summary, the influence of various factors on the extraction of sinomenine from Caulis Sinomenii is as follows: water content > microwave power > microwave extraction time. According to the Box-Behnken response surface methodology, the optimal extraction process is: DESs (choline chloride: lactic acid = 2:1), microwave extraction time 20.70 min, microwave power 423.33 W, water content 57.67%. Under this process condition, the content of sinomenine extracted from Caulis Sinomenii can reach 20.08 mg / g.

[0073] 2.2.3 Confirmatory test

[0074] In order to verify the accuracy and reliability of the process optimization experiment, a confirmation experiment was carried out on the response surface optimization conditions. In order to facilitate the experimental operation, reduce errors, and improve the feasibility of actual operation, the optimal process parameters optimized by the response surface were adjusted to microwave extraction time of 21min, microwave power of 425W, and water content of 58%. The experiment was repeated 3 times, and the results showed that the actual value was not much different from the theoretical value, which verified the feasibility of the process.

[0075] 2.3 Determination of sinomenine content

[0076] 2.3.1 Chromatographic conditions

[0077] Mobile phase: acetonitrile (A)-0.2% phosphoric acid water (B), flow rate 1.0 mL / min, UV detection wavelength 224 nm, column temperature 25°C, injection volume 10 μl. Chromatographic column: Medium Spectrum Red RD-C18 (4.6×250 mm, 5 μm).

[0078] Linear gradient elution, elution program see Table 5.

[0079] Table 5 Elution schedule

[0080]

[0081] 2.3.2 Standard curve drawing

[0082] Take 10 mg of sinomenine standard in a 10 mL volumetric flask, dilute with methanol to obtain a 1 mg / mL standard stock solution, and dilute to obtain a series of standard solutions with mass concentrations of 0.01, 0.05, 0.2, 0.3, and 0.4 mg / mL. Analyze by HPLC, record the peak area of ​​the sample, establish a regression equation with the peak area value (Y) and the standard concentration (X), and draw a standard curve.

[0083] 2.3.3 Preparation of test solution

[0084] Prepare the extract according to the above experimental method, filter and concentrate the extract. Take 1mL of the extract and dilute it to a 10mL volumetric flask, then pass the dilution through a 0.45μm microporous filter membrane to obtain the test solution. Store it in a refrigerator at 4℃ for content determination.

[0085] 2.3.4 Analysis of content determination results

[0086] 1 Liquid chromatography analysis

[0087] According to the above chromatographic conditions, the sinomenine standard and the Caulis Sinomenii samples were injected separately. By comparing the retention time and ultraviolet absorption characteristics of the sinomenine standard and the Caulis Sinomenii samples, the chromatographic peak of sinomenine in the sample was determined. The results are as follows Figure 6 and Figure 7 .

[0088] 2 Analysis of standard curve drawing results

[0089] According to the standard curve drawing method under "2.3.2", prepare a series of standard samples with gradient mass concentrations, draw a standard curve with the standard concentration as the X-axis and the chromatographic peak area as the Y-axis. The result is as follows: Figure 8 The final regression equation is Y = 8*10 6 X-7146.7, correlation coefficient R 2 It is 0.9999, indicating that the linearity is good in the concentration range of 0.01-0.4 mg / g.

[0090] 3 Calculation formula of sinomenine content

[0091] The content of sinomenine in each sample was calculated by the following formula.

[0092]

[0093] In the formula: Y is the sample chromatographic peak area;

[0094] D is the dilution factor of the liquid sample;

[0095] V is the volume of extraction solvent, mL;

[0096] M is the dosage of Caulis Sinomenii, g.

[0097] 3. Experimental conclusion

[0098] The invention uses the rhizome of Caulis Sinomenii as raw material and adopts DESs solvent combined with microwave-assisted method to extract sinomenine.

[0099] Based on single factor and response surface experiments, the extraction process parameters were optimized and the conclusions were:

[0100] 1. Among the 10 different DESs solvents, the solvent with the best extraction effect and its ratio are choline chloride: lactic acid = 2:1.

[0101] 2. The relationship between the influence of different factors on the extraction of sinomenine from Caulis Sinomenii is: water content > microwave power > microwave extraction time.

[0102] 3. The best process for extracting sinomenine from Caulis Sinomenii by microwave-assisted DESs is: DESs (choline chloride: lactic acid = 2:1), microwave extraction time 20.70 min, microwave power 423.33 W, water content 57.67%. Under this process condition, the content of sinomenine extracted from Caulis Sinomenii can reach 20.08 mg / g.

[0103] The method of the invention realizes the green and efficient extraction of sinomenine from Caulis Sinomenii, and is environmentally friendly while increasing the yield of sinomenine.

[0104] In this specification, each embodiment is described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the embodiments can be referred to each other. For the device disclosed in the embodiment, since it corresponds to the method disclosed in the embodiment, the description is relatively simple, and the relevant parts can be referred to the method part.

[0105] The above is a detailed introduction to the microwave extraction process of sinomenine based on a low eutectic solvent provided by the present invention. Specific examples are used herein to illustrate the principles and implementation methods of the present invention. The description of the above embodiments is only used to help understand the method of the present invention and its core idea. It should be pointed out that for ordinary technicians in this technical field, without departing from the principles of the present invention, the present invention can also be improved and modified, and these improvements and modifications also fall within the scope of protection of the claims of the present invention.

Claims

1. A microwave extraction process of sinomenine based on a deep eutectic solvent, characterized in that The process includes the following methods: S1, drying the Caulis Sinomenii Herba and crushing it, passing it through a 40-mesh sieve to obtain Caulis Sinomenii Herba powder; S2. preparing a low eutectic solvent, wherein the low eutectic solvent is a mixture of choline chloride, water and one or more of glucose, lactic acid and glycerol, wherein the mass fraction of water in the low eutectic solvent is 30%-60%; S3. Add a low eutectic solvent to the Sinomenine Caulis powder at a solid-liquid ratio of 1:20 g / mL, and extract by microwave at a power of 400w-500w for 12-21min to obtain Sinomenine.

2. The microwave extraction process of sinomenine based on a deep eutectic solvent according to claim 1, characterized in that: The deep eutectic solvent comprises choline chloride, water and lactic acid, and the weight ratio of choline chloride to lactic acid is 2:1-1:

2.

3. The microwave extraction process of sinomenine based on a deep eutectic solvent according to claim 1, characterized in that: The deep eutectic solvent consists of choline chloride, lactic acid and water, and the weight ratio of choline chloride to lactic acid is 2:

1.

4. The microwave extraction process of sinomenine based on a deep eutectic solvent according to claim 1, characterized in that: In S3, the microwave extraction time was 20 min-21 min, and the microwave power was 423 W-425 W.

5. The microwave extraction process of sinomenine based on a deep eutectic solvent according to claim 1, characterized in that: The low eutectic solvent consists of choline chloride, lactic acid and water, the weight ratio of choline chloride to lactic acid is 2:1, and the mass fraction of water is 57%-58%.

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