Method for determining content of syringaldehyde in sandalwood aqueous extract and identification method of sandalwood aqueous extract

Abstract

The invention relates to the technical field of traditional Chinese medicine analysis, in particular to a method for determining the content of syringaldehyde in a sandalwood aqueous extract and a sandalwood identification method. The content of syringaldehyde in the sandalwood aqueous extract can be accurately, rapidly and conveniently determined by providing proper chromatographic conditions and determination methods, the identification method of the sandalwood aqueous extract is established based on the content difference of syringaldehyde in the sandalwood aqueous extract and counterfeit products, the adopted mobile phase A is acetonitrile, the adopted mobile phase B is acetonitrile, the adopted mobile phase C is acetonitrile, and the adopted mobile phase D is acetonitrile. The mobile phase B is an aqueous solution of phosphoric acid with the volume percentage of 0.08%-0.12%, isocratic elution is adopted, and the volume percentage of the mobile phase A is 6%-10%. The determination and identification method has good specificity and durability, the peak purity of syringaldehyde in an obtained chromatogram is high, the detection precision is high, and an important reference is provided for quality control and clinical medicine of the sandalwood aqueous extract.

Description

Technical Field

[0001] The present application relates to the technical field of drug analysis and traditional Chinese medicine detection, and in particular to a method for determining the content of syringaldehyde in sandalwood water extract and a method for identifying the sandalwood water extract. The sandalwood water extract includes a standard sandalwood decoction and sandalwood traditional Chinese medicine formula granules. Background Art

[0002] Water extraction is an important way to extract the effective ingredients of traditional Chinese medicine. By soaking the Chinese medicinal materials in water, the effective ingredients in the medicinal materials are dissolved in water by the solubilizing effect of water, thereby obtaining a water extract of the Chinese medicinal materials. This method has the advantages of being simple and easy to use, having high extraction efficiency, and having no organic solvent residues, and is widely used in clinical practice. Standard decoctions of Chinese herbal medicine slices are single-flavor Chinese herbal medicine slice water decoctions prepared by standardized processes under the guidance of traditional Chinese medicine theory and based on clinical application, with reference to modern extraction methods. As a standard substance and standard system, they can be used to standardize clinical medication, standardize new forms of slices including formula granules that are currently widely used in clinical practice, and ensure the accuracy of medication and consistency of dosage. Therefore, the use of modern analytical technology to establish the quality standard of standard decoctions of Chinese herbal medicine slices can provide an important reference for the establishment of quality standards for modern Chinese medicine preparations using water extraction, such as corresponding formula granules and classic prescriptions.

[0003] Sandalwood, a traditional Chinese medicinal material, is the dried heartwood of the Santalum album L. tree, a member of the Santalaceae family. Its pungent and warm properties enter the spleen, stomach, and lung meridians, promoting heart-warming, appetite-stimulating, and analgesic benefits. Sandalwood has a long history as a traditional Chinese medicine. Modern pharmacological research has demonstrated its gastrointestinal function, anticancer, antitumor, and antioxidant properties. It is primarily used clinically to treat pediatric abdominal distension and stomachache. Currently, limited research exists on sandalwood quality standards, and content determination methods often focus on volatile oils. However, in addition to volatile oils, sandalwood also contains a significant number of water-soluble components, such as sesquiterpenes, monoterpenes, lignin, and phenolic acids. Therefore, it is necessary to develop a method for determining the content of sandalwood water extracts to address the current research gap in the study of these water-soluble components.

[0004] Sandalwood is a valuable traditional Chinese medicine. Unscrupulous vendors use cheaper heartwood from other trees to pass it off as sandalwood. For example, some vendors use cypress heartwood to pass it off as sandalwood. Because the medicinal properties of cypress and sandalwood differ significantly, this counterfeiting has severely disrupted the sandalwood market, making effective identification methods crucial. Summary of the Invention

[0005] Based on this, one or more embodiments of the present application provide a method for determining the syringaldehyde content in sandalwood water extract and a method for identifying sandalwood water extract, which can accurately determine the syringaldehyde content in sandalwood water extract or the test object, and the detection method is simple and time-saving.

[0006] A method for determining the content of syringaldehyde in sandalwood aqueous extract, comprising the following steps:

[0007] Take the test sample, extract it with an extraction solvent to obtain a test sample solution, and the test sample is sandalwood aqueous extract;

[0008] Take the test sample solution for liquid chromatography analysis to obtain the content of syringaldehyde in the sandalwood aqueous extract;

[0009] Among them, the conditions for the liquid chromatography analysis include:

[0010] (1) Mobile phase A is acetonitrile, and mobile phase B is an aqueous solution of phosphoric acid with a volume percentage of 0.08% - 0.12%;

[0011] (2) Isocratic elution is adopted, and the volume percentage of mobile phase A is 6% - 10%; the total volume percentage of mobile phase A and mobile phase B is 100%;

[0012] (3) The time for isocratic elution is 20 min - 30 min.

[0013] Furthermore, when the volume percentage of mobile phase A is 7% - 9%, a better peak shape can be obtained.

[0014] In one embodiment, the conditions for the liquid chromatography analysis further include at least one of the following:

[0015] (1) The chromatographic column is a C18 chromatographic column;

[0016] (2) The column temperature of the chromatographic column is 38°C - 42°C;

[0017] (3) The flow rate for isocratic elution is 0.23 mL / min - 0.27 mL / min;

[0018] (4) The injection volume is 1 μL - 3 μL;

[0019] (5) The detection wavelength is 300 nm - 320 nm.

[0020] In one embodiment, the C18 chromatographic column has a column length of 100 mm - 150 mm, a diameter of 2 mm - 2.2 mm, and a particle size of the packing of 1.6 μm - 1.8 μm.

[0021] In one embodiment, the extraction solvent is an aqueous solution of organic alcohol with a volume percentage of 60% - 80%;

[0022] Furthermore, the organic alcohol is methanol.

[0023] In one embodiment, the extraction method is ultrasonic and / or heat reflux extraction;

[0024] Further, the power of the ultrasonic is 200W - 400W and the frequency is 30kHz - 50kHz.

[0025] The temperature of the heat reflux extraction is 60°C - 80°C.

[0026] Further, the extraction method is ultrasonic; further, the power of the ultrasonic is 200W - 400W and the frequency is 30kHz - 50kHz; still further, the time of the ultrasonic is 20min - 40min, preferably 25min - 35min, which has a higher extraction efficiency.

[0027] Further, take 0.2g of the test sample and add 20mL - 30mL of the extraction solvent for extraction, which has a higher extraction efficiency.

[0028] In one embodiment, it further includes the steps of preparing a reference substance solution, performing liquid chromatography analysis on the reference substance solution, and formulating a reference substance standard curve equation;

[0029] The conditions for the liquid chromatography analysis are the same as those described above for the liquid chromatography analysis;

[0030] The reference substance solution includes syringaldehyde.

[0031] Further, calculate the content of syringaldehyde in the test sample according to the reference substance standard curve equation.

[0032] In one embodiment, the reference substance standard curve equation includes: y = 0.2838x + 0.0034, r = 0.9999.

[0033] In one embodiment, the aqueous extract of sandalwood is prepared by a method comprising the following steps:

[0034] Take sandalwood decoction pieces, add 8 - 12 times the volume of water for the first decoction, filter and collect the first filtrate, add 6 - 10 times the volume of water to the remaining filter residue for the second decoction, filter and collect the second filtrate, and combine the first filtrate and the second filtrate;

[0035] The first decoction includes boiling at 400W - 600W and then decocting at 100W - 300W for 20 - 40min;

[0036] The second decoction includes boiling at 400W - 600W and then decocting at 100W - 300W for 20 - 30min.

[0037] One or more embodiments of the present application further provide a method for identifying sandalwood aqueous extract, comprising the following steps:

[0038] Determine the content of syringaldehyde in the sandalwood aqueous extract by the method described in any of the above technical solutions;

[0039] Take the test sample, add an extraction solvent for extraction to obtain a test sample solution;

[0040] Take the test sample solution for liquid chromatography analysis, and calculate the content of syringaldehyde in the test sample;

[0041] The conditions for the liquid chromatography analysis are defined as defined in any of the above technical solutions;

[0042] Compare the content of syringaldehyde in the sandalwood aqueous extract and the content of syringaldehyde in the test sample to achieve identification.

[0043] In one embodiment, the sandalwood aqueous extract contains 0.182 mg / g to 0.3927 mg / g of syringaldehyde.

[0044] The present application performs liquid chromatography analysis on the sandalwood aqueous extract by establishing appropriate chromatographic conditions, realizing the accurate determination of syringaldehyde in the sandalwood aqueous extract. The results obtained by this determination method have good reproducibility, high precision, and high accuracy. The chromatographic peak of syringaldehyde in the liquid chromatography has a good peak shape, without the influence of impurity peaks, and is easy to identify.

[0045] The detection method of the present application is simple in operation, consumes less reagents, and is fast, providing a rapid and accurate detection means for the detection of water-soluble components in the sandalwood aqueous extract.

[0046] The detection method of the present application can achieve the authenticity identification of the sandalwood aqueous extract by measuring the content of syringaldehyde in the test substance, providing an important basis for the quality control of the sandalwood aqueous extract. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] In order to more clearly illustrate the specific embodiments of the present application or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the specific embodiments or the prior art. Obviously, the following drawings are some embodiments of the present application. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0048] Figure 1 It is the specificity inspection result of the method for determining the content of syringaldehyde in the sandalwood standard decoction in Example 1 of the present application;

[0049] Figure 2 It is the peak purity inspection result of the method for determining the content of syringaldehyde in the sandalwood standard decoction in Example 1 of the present application;

[0050] Figure 3 It is the inspection result of the linear relationship of the method for determining the content of syringaldehyde in the standard decoction of Santali Albi Lignum in Example 1 of this application;

[0051] Figure 4 It is the inspection result of the durability of the method for determining the content of syringaldehyde in the standard decoction of Santali Albi Lignum in Example 1 of this application, and it is a comparison chart of chromatographic detection results under different chromatographic column conditions;

[0052] Figure 5 It is the inspection result of the durability of the method for determining the content of syringaldehyde in the standard decoction of Santali Albi Lignum in Example 1 of this application, and it is a comparison chart of chromatographic detection results under different column temperature conditions;

[0053] Figure 6 It is the inspection result of the durability of the method for determining the content of syringaldehyde in the standard decoction of Santali Albi Lignum in Example 1 of this application, and it is a comparison chart of chromatographic detection results under different flow rate conditions;

[0054] Figure 7 It is the inspection result of the durability of the method for determining the content of syringaldehyde in the standard decoction of Santali Albi Lignum in Example 1 of this application, and it is a comparison chart of chromatographic detection results under different mobile phase conditions;

[0055] Figure 8 It is a comparison chart of chromatographic detection results for identifying the standard decoction of Santali Albi Lignum and the reference substance of syringaldehyde in Example 2 of this application, and BM-GT-1 is the standard decoction of Cupressus funebris Endl.;

[0056] Figure 9 It is a comparison chart of chromatographic detection results of the method for determining the content of syringaldehyde in the standard decoction of Santali Albi Lignum in Comparative Example 1 of this application under different chromatographic conditions. Specific Embodiments

[0057] The present application will be further described below in conjunction with the embodiments, examples and drawings. It should be understood that these examples are only used to illustrate the present application and not to limit the scope of the present application. In addition, it should be understood that after reading the content taught in the present application, those skilled in the art can make various changes or modifications to the present application, and these equivalent forms also fall within the protection scope of the appended claims of the present application.

[0058] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the technical field to which this application belongs. The terms used in the specification of this application are only for the purpose of describing specific embodiments and are not intended to limit this application.

[0059] Terms

[0060] Unless otherwise stated or there is a contradiction, the terms or phrases used herein have the following meanings:

[0061] As used herein, the alternative scopes of the terms "and / or", "or / and", and "and / or" include any one of two or more related listed items, as well as any and all combinations of the related listed items. The any and all combinations include combinations of any two related listed items, any more related listed items, or all related listed items.

[0062] In this application, terms such as "further" and "especially" are used for descriptive purposes, indicating differences in content, but should not be construed as limiting the scope of protection of this application.

[0063] In this application, among the technical features described in an open-ended manner, it includes a closed technical solution composed of the listed features, as well as an open technical solution containing the listed features.

[0064] In this application, regarding numerical intervals (i.e., numerical ranges), unless otherwise specified, the selectable numerical values are considered continuous within the above numerical intervals, and include the two numerical endpoints of the numerical range (i.e., the minimum value and the maximum value), as well as each numerical value between these two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to integers within the numerical interval, it includes the two endpoint integers of the numerical range, as well as each integer between the two endpoints. In addition, when multiple ranges are provided to describe features or characteristics, these ranges can be combined. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all sub-ranges subsumed therein.

[0065] In this application, the weight can be mass units well-known in the chemical industry such as μg, mg, g, kg, etc.

[0066] In this application, "%" represents a percentage, referring to the proportion of volume or mass; the percentage of alcohol refers to the proportion by volume at 20°C. In addition, the following symbols can be used as needed: %(g / g) indicates the number of grams of solute contained in 100 g of the solution; %(mL / mL) indicates the number of milliliters of solute contained in 100 mL of the solution; %(mL / g) indicates the number of milliliters of solute contained in 100 g of the solution; %(g / mL) indicates the number of grams of solute contained in 100 mL of the solution.

[0067] In the liquid chromatogram of this application, the abscissa is time (unit: min), and the ordinate is the signal intensity (unit: mAU or AU).

[0068] The following are some specific examples.

[0069] In the experimental methods of the following examples, unless otherwise specified, they are all conventional methods. The medicinal raw materials, excipients, reagents, etc. used in the following examples, unless otherwise specified, are all commercially available products.

[0070] Example 1

[0071] In this example, the content of syringaldehyde in the standard decoction of Santali Albi Radix was determined, including the following steps:

[0072] 1. Preparation of Santali Albi Radix slices

[0073] The Santali Albi Radix slices were prepared according to the processing regulations of Santali Albi Radix in the Chinese Pharmacopoeia 2020 Edition. The processing method was as follows: Take Santali Albi Radix medicinal materials, remove impurities, pound into slices or saw into small sections, and split into small pieces of 5 mm - 8 mm.

[0074] 2. Preparation of the standard decoction of Santali Albi Radix

[0075] Take 100 g of Santali Albi Radix slices and place them in an electric ceramic pot. Decoct with water twice. For the first decoction, add 10 times the amount of water, soak for 30 minutes, then bring to a boil over high heat (500 W), and then keep it simmering gently over low heat (200 W) for 30 minutes. Filter while it is hot through a 350 - mesh sieve, and quickly cool the filtrate with cold water. For the second decoction, add 8 times the amount of water, bring to a boil over high heat (500 W), and then keep it simmering gently over low heat (200 W) for 25 minutes. Filter while it is hot through a 350 - mesh sieve, and quickly cool the filtrate with cold water. Combine the two filtrates. Transfer the decoction to a 5000 mL round - bottom flask, and concentrate it under reduced pressure and low temperature (temperature: 65 °C) to 100 mL of a fluid extract using a rotary evaporator; dispense it into petri dishes, and after dispensing, transfer it to a vacuum freeze - dryer for freeze - drying. Take it out and seal it to obtain the freeze - dried powder of the standard decoction. The batch numbers of the freeze - dried powders of the standard decoctions prepared from 15 batches of Santali Albi Radix from different origins are shown in Table 1.

[0076] Table 1 Origin information of 15 batches of Santali Albi Radix slices and their freeze - dried powders of the standard decoctions

[0077]

[0078] 3. Establishment of the method for determining the content of syringaldehyde

[0079] 3.1 Chromatographic conditions

[0080] Use a Waters CORTECS T3 chromatographic column (2.1 mm×150 mm, 1.6 μm); use acetonitrile: 0.1% phosphoric acid solution (8:92) as the mobile phase; the flow rate is 0.25 mL / min; the column temperature is 40 °C; the injection volume is 1 μL; the detection wavelength is 310 nm.

[0081] 3.2 Preparation of the reference substance solution

[0082] Accurately weigh 2.394 mg of syringaldehyde reference substance, place it in a 100 mL volumetric flask, add methanol to make a reference substance stock solution containing 23.4612 μg per 1 mL; accurately pipette 1.0 mL of the above reference substance solution into a 10 mL volumetric flask, add methanol to make a reference substance solution containing 2.346 μg per 1 mL.

[0083] 3.3 Investigation on the pretreatment method of the test solution

[0084] Investigate the extraction solvent, extraction method, extraction time and extraction solvent volume of the standard decoction of Santalum album Chinese herbal pieces to determine the best sample pretreatment method.

[0085] (1)Investigation on extraction solvent

[0086] Take an appropriate amount of the standard decoction of Santalum album Chinese herbal pieces (TX-GT-1), grind it finely, take about 0.2 g, accurately weigh it, in parallel for 7 groups, 2 portions in each group, place it in a stoppered conical flask, accurately add 25 mL of water, 30% methanol, 70% methanol, methanol, 30% ethanol, ethanol, 70% ethanol and ethanol respectively, weigh it, ultrasonically treat (power 300 W, frequency 40 kHz) for 30 minutes, let it cool, weigh it again, make up the lost weight with the corresponding solvent, shake well, filter, take the continuous filtrate, inject and analyze according to the chromatographic conditions under item "3.1", record the chromatographic peak area, and calculate the content of syringaldehyde. The experimental results are shown in Table 2.

[0087] Table 2 Investigation results of extraction solvents for the determination of syringaldehyde content in the standard decoction of Santalum album Chinese herbal pieces

[0088]

[0089] According to Table 2, the differences in the syringaldehyde content measured with different extraction solvents are not significant, but when 70% methanol is used as the extraction solvent, the measured syringaldehyde content is the highest. Therefore, 70% methanol is finally selected as the extraction solvent for the determination of syringaldehyde content in the standard decoction of Santalum album Chinese herbal pieces.

[0090] (2)Investigation on extraction method

[0091] Take an appropriate amount of the standard decoction of Santalum album Chinese herbal pieces (TX-GT-1), grind it finely, take about 0.2 g, accurately weigh it, in parallel for 2 groups, 2 portions in each group, place it in a stoppered conical flask, accurately add 25 mL of 70% methanol, weigh it, ultrasonically treat (power 300 W, frequency 40 kHz) for 30 minutes and heat under reflux for 30 minutes respectively, let it cool, weigh it again, make up the lost weight with 70% methanol, shake well, filter, take the continuous filtrate, inject and analyze according to the chromatographic conditions under item "3.1", record the peak area, and calculate the content of syringaldehyde. The experimental results are shown in Table 3.

[0092] Table 3 Investigation results of extraction methods for the determination of syringaldehyde content in the standard decoction of Santalum album

[0093]

[0094] According to Table 3, the content of syringaldehyde during ultrasonic treatment is slightly higher than that during heating under reflux. Considering environmental protection and ease of operation, ultrasonic extraction is selected as the optimal extraction method for the determination of syringaldehyde content in the standard decoction of Santalum album L. Chinese herbal pieces.

[0095] (3)Investigation of extraction time

[0096] Take an appropriate amount of the standard decoction of Santalum album L. Chinese herbal pieces (TX-GT-1), grind it finely, take about 0.2 g, weigh it precisely, in parallel with 3 groups, 2 portions in each group, place them in a stoppered conical flask, precisely add 25 mL of 70% methanol, weigh it, ultrasonically treat (power 300 W, frequency 40 kHz) for 15 minutes, 30 minutes and 45 minutes respectively, let it cool, weigh it again, make up the lost weight with 70% methanol, shake well, filter, take the subsequent filtrate, inject and analyze according to the chromatographic conditions under item "3.1", record the chromatographic peak area, and calculate the content of syringaldehyde. The experimental results are shown in Table 4.

[0097] Table 4 Results of the investigation of extraction time for the determination of syringaldehyde content in the standard decoction of Santalum album L.

[0098]

[0099] According to Table 4, by comparing the effects of different extraction times on the syringaldehyde content, it is found that the extraction efficiency is the highest when extracting for 30 minutes. Therefore, ultrasonic treatment for 30 minutes is selected as the optimal extraction time for the determination of syringaldehyde content in the standard decoction of Santalum album L. Chinese herbal pieces.

[0100] (4)Investigation of the volume of extraction solvent

[0101] Take an appropriate amount of the standard decoction of Santalum album L. Chinese herbal pieces (TX-GT-1), grind it finely, take about 0.2 g, weigh it precisely, in parallel with 3 groups, 2 portions in each group, place them in a stoppered conical flask, precisely add 15 mL, 25 mL and 50 mL of 70% methanol respectively, weigh it, ultrasonically treat (power 300 W, frequency 40 kHz) for 30 minutes, let it cool, weigh it again, make up the lost weight with 70% methanol, shake well, filter, take the subsequent filtrate, inject and analyze according to the chromatographic conditions under item "3.1", record the peak area, and calculate the content of syringaldehyde. The experimental results are shown in Table 5.

[0102] Table 5 Results of the investigation of the volume of extraction solvent for the determination of syringaldehyde content in the standard decoction of Santalum album L.

[0103]

[0104] According to Table 5, the effect of different amounts of extraction solvent on the content of syringaldehyde was compared. When 25 mL of extraction solvent was used to extract 0.2 g of the standard decoction, the extraction efficiency was the highest. Therefore, 25 mL was selected as the volume of the extraction solvent for the determination of syringaldehyde content in the sandalwood standard decoction.

[0105] (5)Determination of the preparation method of the test solution

[0106] According to the above experimental results, the preparation method of the test solution was finally determined as follows: Take an appropriate amount of this product, grind it finely, accurately weigh 0.2 g, place it in a conical flask, accurately add 25 mL of 70% methanol, weigh it, ultrasonically treat it (power 300 W, frequency 40 kHz) for 30 minutes, let it cool, weigh it again, make up the lost weight with 70% methanol, shake well, filter, and take the subsequent filtrate, that is, obtain.

[0107] 3.4 Methodology verification

[0108] (1)Specificity investigation

[0109] Take an appropriate amount of the sandalwood standard decoction (TX-GT-1), grind it finely, take about 0.2 g, prepare the test solution according to the preparation method of the test solution determined under item "3.3", and take 1 μL each of the syringaldehyde reference solution and the blank solvent, inject them into the liquid chromatograph, and determine according to the chromatographic conditions under item "3.1". The results are shown in Figure 1 .

[0110] According to Figure 1 , the test solution chromatogram has the same chromatographic peak at the retention time corresponding to the reference solution chromatogram, and the blank solvent has no interference, indicating that the method has good specificity.

[0111] (2)Peak purity investigation

[0112] Take an appropriate amount of the sandalwood decoction standard (TX-GT-1), grind it finely, take about 0.2 g, prepare the test solution according to the preparation method of the test sample determined under item "3.3", accurately absorb the test solution and the syringaldehyde reference solution, inject them into the liquid chromatograph, and perform a scan detection at 210 nm to 400 nm with a DAD detector according to the chromatographic conditions under item "3.1", and calculate the peak purity. The results are shown in Figure 2 .

[0113] According to Figure 2 , no impurity peak was detected in the syringaldehyde peak in the sample, and the purity angle was less than the purity threshold, indicating that under this chromatographic condition, the peak purity of syringaldehyde meets the requirements.

[0114] (3)Linearity investigation

[0115] Accurately weigh 2.394 mg of syringaldehyde reference substance, place it in a 100 mL volumetric flask, add methanol to make a linear reference stock solution containing 23.4612 μg per 1 mL. Accurately pipette 1.5 mL, 1 mL, 2.5 mL, 1 mL, 1 mL, and 1 mL of the above stock solution into 2 mL, 2 mL, 10 mL, 10 mL, 150 mL, and 1000 mL volumetric flasks respectively, add methanol to the mark, and prepare reference linear solutions containing 17.5959 μg, 11.7306 μg, 5.8653 μg, 2.3461 μg, 0.1564 μg, and 0.0235 μg per 1 mL respectively. Accurately pipette the above reference linear solutions, inject samples for determination according to the chromatographic conditions under item "3.1", and record the chromatographic peak areas. Take the peak area as the ordinate (y) and the reference substance concentration (μg / mL) as the abscissa (x), and draw a standard curve, as shown in Table 6, Figure 3 。

[0116] Table 6 Results of linear investigation of syringaldehyde

[0117]

[0118] According to Table 6, the regression equation of syringaldehyde is: y = 0.2838x + 0.0034, r = 0.9999, indicating that there is a good linear relationship between the injection concentration and the peak area of syringaldehyde in the range of injection concentration from 0.0235 μg / mL to 23.4612 μg / mL.

[0119] (4) Precision investigation

[0120] Accurately pipette the reference solution with a concentration of 11.7306 μg / mL, repeat the injection 6 times according to the chromatographic conditions under item "3.1", calculate the RSD value based on the peak area of syringaldehyde, and the determination results are shown in Table 7.

[0121] Table 7 Results of precision investigation for the determination of syringaldehyde content in the standard decoction of Santalum album

[0122]

[0123] According to Table 7, for the same test solution, inject samples continuously for 6 needles, and the RSD value of the peak area of syringaldehyde is 0.82%, which is less than 3.0%, indicating good instrument precision.

[0124] (5) Stability investigation

[0125] Take an appropriate amount of the standard decoction of Santalum album L. (TX-GT-1), grind it finely, take about 0.2 g, weigh it accurately, prepare the test solution according to the method for preparing the test sample determined under item "3.3", and inject samples at 0 hour, 2 hours, 4 hours, 8 hours, 12 hours, and 18 hours respectively according to the chromatographic conditions under item "3.1". Determine the peak area of syringaldehyde in the test solution, calculate the RSD value of the peak area, and the determination results are shown in Table 8.

[0126] Table 8 Results of the stability study on the determination of syringaldehyde content in the standard decoction of Santalum album L.

[0127]

[0128] According to Table 8, for the same test solution, samples were injected and determined at 0 hour, 2 hours, 4 hours, 8 hours, 12 hours, and 18 hours respectively. The RSD value of the peak area of syringaldehyde was 2.89%, indicating that the test solution was stable within 18 hours under this method.

[0129] (6) Repeatability study

[0130] Take an appropriate amount of the standard decoction of Santalum album L. (TX-GT-1), grind it finely, take about 0.2 g, weigh it accurately, prepare 6 parallel portions, and prepare 6 test solutions according to the method for preparing the test solution determined under item "3.3". Inject samples for analysis according to the chromatographic conditions under item "3.1", calculate the syringaldehyde content and the RSD value, and the determination results are shown in Table 9.

[0131] Table 9 Results of the repeatability study on the determination of syringaldehyde content in the standard decoction of Santalum album L.

[0132]

[0133] According to Table 9, the same batch of samples was repeatedly determined 6 times, and the RSD value of the syringaldehyde content was 1.32%. According to the "Guidelines for the Validation of Analytical Methods for Drug Quality Standards" in the 2020 Edition of the Chinese Pharmacopoeia, when the content of the analyte in the sample is between 0.01% and 0.1%, the RSD limit for repeatability is <3%, indicating that the repeatability of this analytical method is good.

[0134] (7) Intermediate precision study

[0135] Select different experimenters to determine at different times and with different instruments. Take an appropriate amount of the standard decoction of Santalum album L. (TX-GT-1), grind it finely, take about 0.2 g, prepare 6 parallel portions, weigh them accurately, prepare the test solution according to the method for preparing the test solution determined under item "3.3", and conduct the determination according to the chromatographic conditions under item "3.1". Inject samples respectively, determine the syringaldehyde content in the test solution, calculate the syringaldehyde content and the RSD value, and compare with the results of the repeatability study. The comparison results are shown in Table 10.

[0136] Table 10 Results of intermediate precision for determination of syringaldehyde content in the standard decoction of Santali Albi Radix Pieces

[0137]

[0138] According to Table 10, the same batch of samples was operated by different personnel at different times on different instruments, and repeated determination was carried out 6 times. The RSD value of the syringaldehyde content was 1.63%. Compared with the RSD value of 2.43% for the 6 data in the repeatability test, according to the "Guidelines for Validation of Analytical Methods for Drug Quality Standards" in the 2020 Edition of the Chinese Pharmacopoeia, when the content of the component to be measured in the sample is between 0.01% and 0.1%, the RSD limit for intermediate precision is <4%. Therefore, the intermediate precision of this method is good when operated by different analysts on different dates and different chromatographs.

[0139] (8) Accuracy test

[0140] Accurately weigh 2.178 mg of syringaldehyde reference substance, place it in a 50 mL volumetric flask, and add 70% methanol to make a reference substance solution containing 42.6888 μg per 1 mL. Accurately pipette 5 mL of the above reference substance solution into a 50 mL volumetric flask, and add 70% methanol to make a reference substance solution containing 3.2689 μg per 1 mL.

[0141] Using the standard addition method, according to the ratio of the amount of reference substance added to the component to be measured in the test sample taken of 1:1, take an appropriate amount of the standard decoction of Santali Albi (TX-GT-1), grind it finely, take about 0.1 g, accurately weigh it, in parallel for 6 portions, place it in a stoppered conical flask, accurately add 5 mL of the reference substance solution to each portion, and prepare the test sample solution according to the test sample preparation method determined under item "3.3". Carry out the determination according to the chromatographic conditions under item "3.1", determine the syringaldehyde content in the test sample solution, calculate the recovery rate of standard addition, and the results are shown in Table 11.

[0142] Table 11 Results of recovery rate of standard addition for determination of syringaldehyde content in the standard decoction of Santali Albi

[0143]

[0144] According to Table 11, the recovery rate of syringaldehyde was 92.97%. According to the "Guidelines for Validation of Analytical Methods for Drug Quality Standards" in the 2020 Edition of the Chinese Pharmacopoeia, when the content of the component to be measured in the sample is between 0.01% and 0.1%, the recovery rate limit is 90% - 108%. The range of the recovery rate of standard addition of syringaldehyde was 91.46% - 95.75%, and the average recovery rate of standard addition was 92.97%, with an RSD value of 1.96%, indicating that the accuracy of this method is good.

[0145] (9) Robustness investigation

[0146] ① Investigation with different chromatographic columns

[0147] Comparison of chromatographic columns of different brands: Waters CORTECS T3 (2.1 mm × 150 mm, 1.6 μm); SHIMADZU Shim-pack XR-ODSII (2.1 mm × 100 mm, 1.6 μm); ACE EXCEL Super C18 chromatographic column (2.1 mm × 100 mm, 1.7 μm) on the determination of syringaldehyde content in the standard decoction of Santalum album

[0148] Take an appropriate amount of the standard decoction of Santalum album slices (TX-GT-1), grind it finely, take about 0.2 g, weigh it accurately, prepare the test solution according to the test solution preparation method determined under item "3.3", except for the different chromatographic columns, other conditions are the same as those specified under item "3.1", inject for analysis, and the obtained chromatogram is as Figure 4 , calculate the content of syringaldehyde and the RSD value. The chromatographic columns and the determination results are shown in Table 12.

[0149] Table 12 Results of the durability investigation of different chromatographic columns for the determination of syringaldehyde content in the standard decoction of Santalum album

[0150]

[0151] According to Table 12, the separation effects of chromatographic columns of different brands are good, and the RSD value of the syringaldehyde content is 2.56%, indicating that this method has good durability for chromatographic columns of different brands.

[0152] ② Investigation at different column temperatures

[0153] Compare different column temperatures, which are 38 °C, 40 °C and 42 °C respectively, on the determination of syringaldehyde content in the standard decoction of Santalum album.

[0154] Take an appropriate amount of the standard decoction of Santalum album slices (TX-GT-1), grind it finely, take about 0.2 g, weigh it accurately, prepare the test solution according to the test solution preparation method determined under item "3.3", except that the column temperatures are 38 °C, 40 °C and 42 °C respectively, other conditions are the same as those specified under item "3.1", inject for analysis, and the obtained chromatogram is as Figure 5 , calculate the content of syringaldehyde and the RSD value. The column temperatures and the determination results are shown in Table 13.

[0155] Table 13 Results of the durability investigation of different column temperatures for the determination of syringaldehyde content in the standard decoction of Santalum album

[0156]

[0157] According to Table 13, the RSD value of the syringaldehyde content in the standard decoction of Santalum album slices measured at different column temperatures (±2 °C) is 2.87%, indicating that this analytical method has good durability within the range of column temperature ±2 °C.

[0158] ③ Investigation at different flow rates

[0159] Compare the effects of different flow rates, namely 0.23 mL / min, 0.25 mL / min, and 0.27 mL / min, on the determination of the syringaldehyde content in the standard decoction of Santali Albi Lignum.

[0160] Take an appropriate amount of the standard decoction of Santali Albi Lignum (TX-GT-1), grind it finely, take about 0.2 g, weigh it accurately, and prepare the test solution according to the method for preparing the test solution determined under item "3.3". Except for the flow rates of 0.23 mL / min, 0.25 mL / min, and 0.27 mL / min respectively, other conditions are the same as those specified under item "3.1". Inject for analysis to obtain the chromatogram as Figure 6 , calculate the syringaldehyde content and the RSD value. The flow rate and the determination results are shown in Table 14.

[0161] Table 14 Results of the durability investigation of different flow rates for the determination of the syringaldehyde content in the standard decoction of Santali Albi Lignum

[0162]

[0163] According to Table 14, the RSD value of the syringaldehyde content in the standard decoction of Santali Albi Lignum measured at different flow rates (0.25 ± 0.02 mL / min) is 0.92%, indicating that the analytical method has good durability within the range of 0.25 ± 0.02 mL / min for the flow rate.

[0164] ④ Investigation of different mobile phase ratios

[0165] Compare the effects of different mobile phase ratios, namely acetonitrile: 0.1% phosphoric acid solution (7:93), acetonitrile: 0.1% phosphoric acid solution (8:92), and acetonitrile: 0.1% phosphoric acid solution (9:91), on the determination of the syringaldehyde content in Santali Albi Lignum.

[0166] Take an appropriate amount of the standard decoction of Santali Albi Lignum (TX-GT-1), grind it finely, take about 0.2 g, and prepare the test solution according to the method for preparing the test solution determined under item "3.3". Except for the different mobile phase ratios, other chromatographic conditions are the same as those specified under item "3.1". Inject for analysis to obtain the chromatogram as Figure 7 , calculate the syringaldehyde content and the RSD value. The mobile phase and the determination results are shown in Table 15.

[0167] Table 15 Results of the durability investigation of different mobile phases for the determination of the syringaldehyde content in the standard decoction of Santali Albi Lignum

[0168]

[0169] According to Table 15, compare the syringaldehyde content values of Santali Albi Lignum with three different mobile phase ratios. The RSD value is 2.26%, indicating that the analytical method has good durability within the range of ±1% for the mobile phase ratio.

[0170] 3.5 Sample Determination

[0171] Take 15 batches of standard decoctions of Santali Albi Lignum, prepare 15 portions of test solution according to the preparation method of test solution under item "3.3", inject and determine according to the chromatographic conditions under item "3.1", record the peak areas, and calculate by the external standard method. The calculation results are shown in Table 16.

[0172] Table 16 Determination Results of Syringaldehyde Content in 15 Batches of Standard Decoctions of Santali Albi Lignum

[0173]

[0174] According to Table 16, the syringaldehyde content measured in 15 batches of standard decoctions of Santali Albi Lignum is 0.1825 mg / g - 0.3405 mg / g, the mean value is 0.2600 mg / g, and the SD is 0.04 mg / g; within the range of 70% - 130% of the mean value, the syringaldehyde content is 0.1820 mg / g - 0.3380 mg / g, and within the range of mean value ± 3 times SD, the syringaldehyde content is 0.1273 mg / g - 0.3927 mg / g.

[0175] Considering the representativeness of 15 batches of samples, take the content of 70% of the mean value of syringaldehyde as the lower limit and the content of mean value + 3 times SD of syringaldehyde as the upper limit, and determine that the content range of syringaldehyde in the standard decoction of Santali Albi Lignum should be 0.1820 mg / g - 0.3927 mg / g.

[0176] Example 2

[0177] This example is a method for identifying standard decoctions of Santali Albi Lignum traditional Chinese medicine by using the determination index of syringaldehyde content. This method includes the following steps:

[0178] 1. Chromatographic Conditions

[0179] Use Waters CORTECS T3 chromatographic column (2.1 mm × 150 mm, 1.6 μm); use acetonitrile: 0.1% phosphoric acid solution (8:92) as the mobile phase; the flow rate is 0.25 mL / min; the column temperature is 40 °C; the injection volume is 1 μL; the detection wavelength is 310 nm.

[0180] 2. Preparation of Reference Solution

[0181] Precisely weigh 2.394 mg of syringaldehyde reference substance, place it in a 100 mL volumetric flask, add methanol to make a reference stock solution containing 23.4612 μg per 1 mL; precisely transfer 1.0 mL of the above reference solution into a 10 mL volumetric flask, add methanol to make a reference solution containing 2.346 μg per 1 mL.

[0182] 3. Preparation of Test Solution

[0183] Take an appropriate amount of the standard decoction of Santalum album Chinese medicinal pieces for identification (batch number: BM-GT-1), grind it finely, take about 0.2 g, weigh it accurately, place it in a conical flask, accurately add 25 mL of 70% methanol, weigh it, ultrasonically treat it (power 300 W, frequency 40 kHz) for 30 minutes, let it cool, weigh it again, make up the lost weight with 70% methanol, shake well, filter, and take the subsequent filtrate, that is obtained.

[0184] 4. Determination method

[0185] Precisely pipette 1 μL each of the reference substance solution and the above-mentioned test solution, inject them into the liquid chromatograph, and determine, then it is obtained.

[0186] 5. Determination results

[0187] There is no chromatographic peak response at the position corresponding to the retention time of the syringaldehyde reference substance chromatographic peak in the standard decoction of the Chinese medicinal pieces for identification with batch number BM-GT-1 (actually the standard decoction of Cupressus funebris Endl.). The chromatographic detection results are shown in Figure 8 . Therefore, the sample with batch number BM-GT-1 is not the standard decoction of Santalum album. This application can be used to identify the standard decoction of Santalum album and its common near-counterfeit, the standard decoction of Cupressus funebris Endl.

[0188] Comparative example 1

[0189] This comparative example examines the influence of different chromatographic conditions (mobile phase ratio, column temperature, etc.) on the determination of the syringaldehyde content in the standard decoction of Santalum album. Take the test solution TX-GT-15 in Example 1 and determine the syringaldehyde in the standard decoction of Santalum album using different chromatographic conditions.

[0190] (1) High-performance liquid chromatograph (Thermo vanquish, Thermo Fisher Scientific), Waters CORTECS T3 chromatographic column (2.1 mm × 150 mm, 1.6 μm); use acetonitrile as mobile phase A and 0.1% phosphoric acid as mobile phase B, and perform gradient elution according to the regulations in Table 17; the flow rate is 0.25 mL / min; the column temperature is 30 °C; the injection volume is 1 μL; the detection wavelength is 310 nm.

[0191] Table 17 Gradient elution table

[0192]

[0193] (2) High-performance liquid chromatograph (Thermo vanquish, Thermo Fisher Scientific), Waters CORTECS T3 chromatographic column (2.1 mm × 150 mm, 1.6 μm); use acetonitrile: 0.1% phosphoric acid solution (5:95) as the mobile phase; the flow rate is 0.25 mL / min; the column temperature is 40 °C; the injection volume is 1 μL; the detection wavelength is 310 nm.

[0194] (3) High performance liquid chromatograph (Thermo vanquish, Thermo Fisher Scientific), Waters CORTECS T3 chromatographic column (2.1 mm × 150 mm, 1.6 μm); using acetonitrile: 0.1% phosphoric acid solution (6:94) as the mobile phase; flow rate of 0.25 mL / min; column temperature of 40 °C; injection volume of 1 μL; detection wavelength of 310 nm.

[0195] (4) High performance liquid chromatograph (Thermo vanquish, Thermo Fisher Scientific), Waters CORTECS T3 chromatographic column (2.1 mm × 150 mm, 1.6 μm); using acetonitrile: 0.1% phosphoric acid solution (10:90) as the mobile phase; flow rate of 0.25 mL / min; column temperature of 40 °C; injection volume of 1 μL; detection wavelength of 310 nm.

[0196] (5) High performance liquid chromatograph (Thermo vanquish, Thermo Fisher Scientific), Waters CORTECS T3 chromatographic column (2.1 mm × 150 mm, 1.6 μm); using acetonitrile: 0.1% phosphoric acid solution (12:88) as the mobile phase; flow rate of 0.25 mL / min; column temperature of 30 °C; injection volume of 1 μL; detection wavelength of 310 nm.

[0197] (6) High performance liquid chromatograph (Thermo vanquish, Thermo Fisher Scientific), Waters CORTECS T3 chromatographic column (2.1 mm × 150 mm, 1.6 μm); using acetonitrile: 0.1% phosphoric acid solution (8:92) as the mobile phase; flow rate of 0.25 mL / min; column temperature of 40 °C; injection volume of 1 μL; detection wavelength of 310 nm.

[0198] The chromatographic detection results of conditions (1) to (6) are as Figure 9 , according to Figure 9 , using other different mobile phase elution programs and ratios (conditions (1) to (5)), the resolution and peak shape of the syringaldehyde chromatographic peak in the obtained chromatogram cannot reach the effect of Example 1 (condition (6)).

[0199] When gradient elution was adopted, there were impurity peaks interfering beside the syringaldehyde chromatographic peak, and the resolution could not meet the requirements (Condition (1)); when eluting with acetonitrile: 0.1% phosphoric acid solution 6:94 and 10:90 respectively, the peak shapes of the syringaldehyde chromatographic peak and the adjacent chromatographic peaks were poor (Conditions (3) and (4)); when eluting with acetonitrile: 0.1% phosphoric acid solution 5:95 and 12:88 respectively, the resolution between the syringaldehyde chromatographic peak and the adjacent chromatographic peaks was poor and the peak shape was also poor (Conditions (2) and (5)). It can be seen that changing the mobile phase elution program and ratio has a great influence on the syringaldehyde chromatographic peak, and a good resolution and peak shape cannot be obtained. In order to more accurately determine the content of syringaldehyde in the sandalwood standard decoction, the chromatographic conditions in Example 1 (Condition (6)) were selected.

[0200] Comparative Example 3

[0201] This comparative example examines the applicability of the determination method of this application to different test samples and examines the determination results of this method for the cut crude drugs.

[0202] 1. Chromatographic conditions

[0203] Waters CORTECS T3 chromatographic column (2.1 mm × 150 mm, 1.6 μm) was used; acetonitrile: 0.1% phosphoric acid solution (8:92) was used as the mobile phase; the flow rate was 0.25 mL per minute; the column temperature was 40 °C; the injection volume was 1 μL; the detection wavelength was 310 nm.

[0204] 2. Preparation of reference substance solution

[0205] Precisely weigh 2.394 mg of syringaldehyde reference substance, place it in a 100 mL volumetric flask, add methanol to make a reference substance stock solution containing 23.4612 μg per 1 mL; precisely transfer 1.0 mL of the above reference substance solution to a 10 mL volumetric flask, add methanol to make a reference substance solution containing 2.346 μg per 1 mL.

[0206] 3. Preparation of test sample solution

[0207] Take an appropriate amount of sandalwood cut crude drugs (batch number: TX-YP-15, passing through No. 3 sieve), take about 2.0 g, precisely weigh it, place it in a conical flask, precisely add 25 mL of 70% methanol, weigh it, ultrasonically treat it (power 300 W, frequency 40 kHz) for 30 minutes, let it cool, weigh it again, make up the lost weight with 70% methanol, shake well, filter, and take the continuous filtrate, that is, obtain it.

[0208] 4. Determination method

[0209] Precisely absorb 1 μL of the reference substance solution and the test sample solution respectively, inject them into the liquid chromatograph, and determine, that is, obtain it.

[0210] 5. Determination results

[0211] The determination results of the content of syringaldehyde in the Santali Albi Lignum decoction pieces with batch number TX-YP-15 are shown in Table 18. The content of syringaldehyde in the Santali Albi Lignum decoction pieces is not within the range specified in the standard decoction.

[0212] Table 18 Determination results of the content of syringaldehyde in Santali Albi Lignum decoction pieces (batch number: TX-YP-15)

[0213]

[0214] According to Table 18, when directly detecting the decoction pieces, the true content of syringaldehyde cannot be well characterized. This may be because syringaldehyde is a water-soluble component. It can be seen that the determination method established for the water extract in this application has good specificity.

[0215] All the documents mentioned in this application are cited as references in this application, just as if each document is cited separately as a reference. Unless it conflicts with the application purpose and / or technical solution of this application, the cited documents involved in this application are cited for all contents and all purposes. When this application involves cited documents, the definitions of relevant technical features, terms, nouns, phrases, etc. in the cited documents are also cited together. When this application involves cited documents, the examples and preferred methods of the relevant technical features cited can also be used as references and incorporated into this application, but only to the extent that this application can be implemented. It should be understood that when the cited content conflicts with the description in this application, this application shall prevail or be amended adaptively according to the description in this application.

[0216] The technical features of the above-described embodiments and examples can be combined in any suitable manner. For the sake of brevity of description, not all possible combinations of the technical features in the above-described embodiments and examples are described. However, as long as there is no contradiction in the combination of these technical features, it should be considered to be within the scope described in this specification.

[0217] The above-described embodiments only express several implementation manners of this application, but should not be construed as limiting the scope of the patent application. It should be noted that for those of ordinary skill in the art, without departing from the concept of this application, several modifications and improvements can still be made, and these all belong to the protection scope of this application. In addition, it should be understood that after reading the above teachings of this application, those skilled in the art can make various changes or modifications to this application, and the equivalent forms obtained also fall within the protection scope of this application. It should also be understood that the technical solutions obtained by those skilled in the art through logical analysis, reasoning or limited experiments based on the technical solutions provided in this application are all within the protection scope of the appended claims of this application. Therefore, the protection scope of this patent application shall be subject to the appended claims, and the specification and drawings can be used to explain the content of the claims.

Claims

1. A method for determining the content of syringaldehyde in the aqueous extract of sandalwood, characterized in that, It includes the following steps: Take the test sample, add an extraction solvent for extraction to obtain a test sample solution; the test sample is the aqueous extract of sandalwood; Take the test sample solution for liquid chromatography analysis to obtain the content of syringaldehyde in the test sample; Among them, the conditions for the liquid chromatography analysis include: (1) Mobile phase A is acetonitrile, and mobile phase B is an aqueous solution of phosphoric acid with a volume percentage of 0.08% - 0.12%; (2) Isocratic elution is adopted, and the volume percentage of mobile phase A is 6% - 10%; the sum of the volume percentages of mobile phase A and mobile phase B is 100%; (3) The time for isocratic elution is 20 min - 30 min.

2. The method for determining the content of syringaldehyde in the aqueous extract of sandalwood as described in claim 1, wherein, The conditions for the liquid chromatography analysis further include at least one of the following: (1) The chromatographic column is a C18 chromatographic column; (2) The column temperature of the chromatographic column is 38°C - 42°C; (3) The flow rate for isocratic elution is 0.23 mL / min - 0.27 mL / min; (4) The injection volume is 1 μL - 3 μL; (5) The detection wavelength is 300 nm - 320 nm.

3. The method for determining the content of syringaldehyde in the aqueous extract of sandalwood according to claim 2, wherein The C18 chromatographic column has a column length of 100 mm - 150 mm, a diameter of 2 mm - 2.2 mm, and a particle size of the packing of 1.6 μm - 1.8 μm.

4. The method for determining the content of syringaldehyde in the aqueous extract of sandalwood according to any one of claims 1 to 3, characterized in that, The extraction solvent is an aqueous solution of organic alcohol with a volume percentage of 60% - 80%; Furthermore, the organic alcohol is methanol.

5. The method for determining the content of syringaldehyde in the aqueous extract of sandalwood according to any one of claims 1 to 3, characterized in that, The extraction method is ultrasonic and / or heat reflux extraction; Furthermore, the power of the ultrasonic is 200 W - 400 W, and the frequency is 30 kHz - 50 kHz, The temperature for heat reflux extraction is 60°C - 80°C.

6. The method for determining the content of syringaldehyde in the aqueous extract of sandalwood as described in claim 1, characterized in that, It also includes the steps of preparing a reference solution, taking the reference solution for liquid chromatography analysis, and formulating a reference standard curve equation; The conditions for the liquid chromatography analysis are the same as those in any one of claims 1 - 5; The reference solution contains syringaldehyde.

7. The method for determining the content of syringaldehyde in the aqueous extract of sandalwood as described in claim 6, characterized in that, The reference standard curve equation includes: y = 0.2838x + 0.0034, r = 0.9999.

8. The method for determining the content of syringaldehyde in the aqueous extract of sandalwood according to any one of claims 1 to 3, 6 to 7, characterized in that, The aqueous extract of sandalwood is prepared by a method including the following steps: Take sandalwood decoction pieces, add 8 - 12 times the volume of water for the first decoction, filter and collect the first filtrate, add 6 - 10 times the volume of water to the remaining filter residue for the second decoction, filter and collect the second filtrate, and combine the first filtrate and the second filtrate; The first decoction includes boiling at 400 W - 600 W and then decocting at 100 W - 300 W for 20 - 40 min; The second decoction includes boiling at 400 W - 600 W and then decocting at 100 W - 300 W for 20 - 30 min.

9. Identification method of sandalwood aqueous extract, characterized in that, It includes the following steps: Determine the content of syringaldehyde in the aqueous extract of sandalwood by the method described in any one of claims 1 - 8; Take the test sample, add an extraction solvent for extraction to obtain a test sample solution; Take the test sample solution for liquid chromatography analysis and calculate the content of syringaldehyde in the test sample; The conditions for the liquid chromatography analysis are defined as in any one of claims 1 - 8; Compare the content of syringaldehyde in the aqueous extract of sandalwood and the content of syringaldehyde in the test sample to achieve identification.

10. The identification method according to claim 9, characterized in that, The sandalwood aqueous extract contains 0.182 mg / g to 0.3927 mg / g of syringaldehyde.

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