A method for determining characteristic chromatogram of sandalwood medicinal material, decoction piece, standard decoction and formula granule and application thereof

Abstract

The application provides a characteristic chromatogram determination method of sandalwood medicinal materials, decoction pieces, standard decoction and formula granules and application thereof. The method is that test sample solution and control sample solution are detected and analyzed by liquid chromatography, and the characteristic chromatogram is obtained. The chromatographic conditions are that acetonitrile is used as mobile phase A, water, 0.1-0.2vol% phosphoric acid or 0.1-0.2vol% formic acid is used as mobile phase B, gradient elution is carried out, and the characteristic chromatogram obtained under the detection wavelength of 300-350nm has larger chromatographic peak information and stable chromatogram baseline. The characteristic chromatogram has seven corresponding characteristic peaks, and the verbascoside peak and the isoacteoside peak are successfully identified. The verbascoside and the isoacteoside are identified in the characteristic chromatogram of sandalwood products for the first time. The determination method can effectively identify sandalwood and ensure the uniformity and stability of sandalwood quality.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of quality control of traditional Chinese medicine preparations, and particularly relates to a characteristic spectrum determination method for sandalwood medicinal materials, decoction pieces, standard decoctions and formula granules and application thereof. BACKGROUND

[0002] Sandalwood has a long history of medicinal use, and was first recorded in the Han Dynasty herbal work Shanghan Zabing, which listed sandalwood as Xiaoping and recorded it as “Zi Zhen Tan Mu”. However, sandalwood is now a collective term for plants in the genus Santalum, and is not the sandalwood used today. In Shanghan Zabing, it is recorded that “taste is salty, slightly cold. Mainly treats evil toxin and wind toxin.”, which is different from the modern Pharmacopoeia of the People's Republic of China, which records “promotes qi and warms the middle, relieves stomachache. Used for cold stagnation of qi, chest and diaphragm discomfort, chest pain, stomachache, vomiting and loss of appetite.”. In Ben Cao Gang Mu, it is recorded that “Bai Xuan Tan: pungent, warm, non-toxic.”, “Zi Tan: salty, slightly cold, non-toxic.”, which is ambiguous, and the record of Zi Tan is consistent with that in Shanghan Zabing, and the record of Bai Xuan Tan is consistent with the nature and taste in the 2020 edition of the Pharmacopoeia of the People's Republic of China. In Ben Cao Cong Xin, it is recorded that “Bai Tan Xiang: pungent and warm. Regulates spleen and lung. Benefits chest and diaphragm. Treats vomiting due to dysphagia. Relieves stomachache. Repels ghosts and kills insects. Promotes appetite and food intake.”, which is basically consistent with the nature, taste and function and indications in the 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0003] During the Han Dynasty to the Tang Dynasty, the varieties of sandalwood were confusing, and sandalwood is not a native plant in China. In the entry of Zi Zhen Tan Mu in Tang Xin Xi Ben Cao, it is clearly pointed out that “this plant comes from the Kunlun Panpan country, and only does not grow in China, and is widespread in the world.”. According to the historical data that can be verified, sandalwood appeared as a medicinal plant in Shanghan Zabing in the Han Dynasty, and through the examination of ancient herbal works, it can be confirmed that there is no native sandalwood in China. Through the study of herbal examination, it is found that after sandalwood was introduced to China for several hundred years, there are signs of introduction and cultivation in Minnan and Southwest China, and even there are special cultivation methods.

[0004] It can be seen that sandalwood medicinal materials have undergone many changes in different dynasties, and it is found through the present examination that the mixed use of sandalwood and Zi Tan and Huang Tan does exist. However, at present, there is no method that can evaluate and control the quality of sandalwood as a whole. SUMMARY

[0005] Therefore, the purpose of the present application is to provide a characteristic spectrum determination method for sandalwood medicinal materials, decoction pieces, standard decoctions and formula granules and application thereof. The method uses high performance liquid chromatography to establish a method for determining the characteristic spectrum of sandalwood medicinal materials, decoction pieces, standard decoctions and formula granules, which can effectively separate the chromatographic components of sandalwood, and can be used for overall evaluation and control of the quality of sandalwood.

[0006] In order to achieve this purpose, the present application adopts the following technical solutions:

[0007] In a first aspect, the present application provides a method for determining the characteristic chromatogram of sandalwood medicinal material, decoction piece, standard decoction and formula granule, comprising:

[0008] The test sample solution and the control solution are detected by liquid chromatography;

[0009] The control in the control solution comprises verbascoside and / or isovertoside;

[0010] The conditions of the liquid chromatography are as follows: the mobile phase A is acetonitrile, the mobile phase B is selected from water, 0.1-0.2 vol% phosphoric acid or 0.1-0.2 vol% formic acid, and the detection wavelength is 300-350 nm.

[0011] Preferably, gradient elution is adopted in the process of the liquid chromatography detection, and the program of the gradient elution is as follows:

[0012] 0-8 min: 9-16.5 vol% mobile phase A, 91-83.5 vol% mobile phase B;

[0013] 8-40 min: 16.5 vol% mobile phase A, 83.5 vol% mobile phase B;

[0014] 40-55 min: 16.5-30 vol% mobile phase A, 83.5-70 vol% mobile phase B;

[0015] 55-60 min: 30 vol% mobile phase A, 70 vol% mobile phase B.

[0016] Preferably, the filler of the chromatographic column in the liquid chromatography is octadecylsilane-bonded silica gel; the column temperature is 20-35°C; and the flow rate is 0.1-0.8 mL / min.

[0017] Preferably, the column temperature is 20-35°C, and the flow rate is 0.1-0.8 mL / min.

[0018] Preferably, the detection wavelength is 326 nm.

[0019] Preferably, the amount of the test sample solution and the control solution is 1-5 μL.

[0020] Preferably, the test sample solution is obtained by mixing the test sample with 30-80 vol% methanol, and then ultrasonic extraction or reflux extraction.

[0021] Preferably, the ratio of the test sample to 30-80 vol% methanol is 0.1 g:(10-50) mL.

[0022] Preferably, the time of the ultrasonic extraction or the reflux extraction is not less than 30 min.

[0023] or

[0024] Preferably, the test sample solution is obtained by mixing the test sample with water, decocting, filtering, mixing the residue of the filtrate after evaporation with 30-80 vol% methanol, and then obtaining.

[0025] Preferably, the control sample solution is obtained by mixing the verbascoside and / or isoacteoside with 30-80 vol% methanol.

[0026] Preferably, the concentration of the control sample in the control sample solution is 40-100 μg / mL.

[0027] Preferably, the characteristic spectrum of the sandalwood medicinal material, decoction piece, standard decoction, and formula granule obtained by the determination method presents 7 corresponding characteristic peaks, and the verbascoside peak and / or isoacteoside peak are identified.

[0028] In a second aspect, the present application provides an application of the above determination method in detecting the quality of sandalwood medicinal material, decoction piece, standard decoction, and formula granule.

[0029] In a third aspect, the present application provides a quality control method of sandalwood medicinal material, decoction piece, standard decoction, and formula granule, comprising:

[0030] obtaining the characteristic spectrum of the test sample according to the above determination method;

[0031] comparing with the standard characteristic spectrum obtained by the above determination method; if the characteristic spectrum of the test sample presents 7 corresponding characteristic peaks, and the RSD value of the relative retention time of each peak in the characteristic spectrum of the sample and the relative retention time of each peak in the standard characteristic spectrum is ≤10.0%, the sample is determined as a qualified product.

[0032] Compared with the prior art, the present application has the following beneficial effects:

[0033] The present application provides a characteristic spectrum determination method of sandalwood medicinal material, decoction piece, standard decoction, and formula granule, which detects and analyzes the test sample solution and the control sample solution by liquid chromatography. The chromatographic conditions are as follows: acetonitrile is used as the mobile phase A, water, 0.1-0.2 vol% phosphoric acid, or 0.1-0.2 vol% formic acid is used as the mobile phase B, gradient elution is performed, and the characteristic spectrum obtained at a detection wavelength of 300-350 nm has a large amount of chromatographic peak information and a stable chromatogram baseline. The characteristic spectrum presents 7 corresponding characteristic peaks, and the verbascoside peak and isoacteoside peak are successfully identified. The verbascoside and isoacteoside are identified for the first time in the characteristic spectrum of sandalwood products.

[0034] The method has the characteristics of comprehensive detection and simple operation, can effectively identify medicinal materials, decoction pieces, standard decoctions and formula granules, and can ensure the uniformity and stability of the quality of medicinal materials, decoction pieces, standard decoctions and formula granules. BRIEF DESCRIPTION OF DRAWINGS

[0035] Figure 1 The liquid chromatogram obtained by taking sandalwood formula granules as the test sample under different mobile phases;

[0036] Figure 2 The liquid chromatogram obtained by taking sandalwood formula granules as the test sample under different detection wavelengths;

[0037] Figure 3 The liquid chromatogram obtained by taking sandalwood formula granules as the test sample under different flow rates;

[0038] Figure 4 The liquid chromatogram obtained by taking sandalwood formula granules as the test sample under different column temperatures;

[0039] Figure 5 The liquid chromatogram obtained by taking sandalwood formula granules as the test sample under different extraction solvents;

[0040] Figure 6 The chromatographic peak identification diagram of the characteristic chromatogram of sandalwood formula granules;

[0041] Figure 7 The liquid chromatogram obtained by taking sandalwood formula granules as the test sample after different instrument investigations;

[0042] Figure 8 The liquid chromatogram obtained by taking sandalwood formula granules as the test sample after different chromatographic column investigations;

[0043] Figure 9 The characteristic chromatogram of sandalwood formula granules of different batches;

[0044] Figure 10 The control chromatogram of sandalwood formula granules;

[0045] Figure 11 The characteristic chromatogram of sandalwood medicinal materials of different batches;

[0046] Figure 12 The control chromatogram of sandalwood medicinal materials;

[0047] Figure 13 The characteristic chromatogram of sandalwood decoction pieces of different batches;

[0048] Figure 14 The control chromatogram of sandalwood decoction pieces;

[0049] Figure 15 The characteristic chromatogram of sandalwood standard decoctions of different batches;

[0050] Figure 16 The standard decoction of sandalwood is used as a control map. DETAILED DESCRIPTION

[0051] The technical solutions of the present application will be described clearly and completely below in combination with the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, but not all the embodiments. Based on the embodiments in the present application, all the other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0052] In view of the fact that there is no method for overall evaluation and control of the quality of sandalwood in the prior art, the present application provides a method for determining the characteristic map of sandalwood medicinal materials, decoction pieces, standard decoction and formula granules, which comprises:

[0053] The test sample solution and the control solution are detected and analyzed by liquid chromatography.

[0054] The control in the control solution comprises verbascoside and / or isoferulic acid.

[0055] In the present application, the test sample solution is the corresponding solution of sandalwood medicinal materials, decoction pieces, standard decoction and formula granules. In some embodiments of the present application, when the test sample is sandalwood formula granules, the test sample solution is prepared by mixing the test sample with 30-80 vol% methanol, preferably 50-70 vol% methanol, and most preferably 70 vol% methanol. In the present application, based on 0.2 g of the test sample, the amount of 30-80 vol% methanol added is 10-50 mL, preferably 20-30 mL, and more preferably 25 mL. According to the present application, after the test sample is mixed with 30-80 vol% methanol, the resulting mixed solution is extracted by ultrasonic extraction or reflux extraction to obtain the test sample solution. To ensure complete extraction, the time for ultrasonic extraction or reflux extraction is preferably not less than 30 min, and more preferably 30 min. Since there is little difference between reflux extraction and ultrasonic extraction for the test sample, the preferred extraction method in the present application is ultrasonic extraction, and the power of the ultrasonic extraction is 500-600 W and the frequency is 30-40 kHz.

[0056] In some specific embodiments of the present application, the preparation method of the test sample solution of sandalwood formula granules is as follows:

[0057] An appropriate amount of the product (i.e., sandalwood formula granules) is finely ground, about 0.2 g, placed in a conical flask with a stopper, 25 mL of 70 vol% methanol is added, ultrasonic treatment (power 600 W, frequency 40 kHz) is performed for 30 min, cooled, shaken uniformly, filtered, and the filtrate is obtained.

[0058] When the test sample is any one of the sandalwood medicinal material, decoction piece, standard decoction, the test sample solution is obtained by mixing the test sample with water, decocting, filtering, and mixing the residue after the filtrate is evaporated to dryness with 30-80 vol% methanol, preferably 70 vol% methanol.

[0059] In some embodiments of the present application, the preparation method of the test sample solution of the sandalwood medicinal material, decoction piece, standard decoction is as follows:

[0060] Take 1.0 g of the powder (pass through a No. 3 sieve), add 50 mL of water, decoct for 30 min, cool, filter, evaporate the filtrate to dryness, dissolve the residue with 25 mL of 70 vol% methanol, filter, and take the filtrate to obtain the solution.

[0061] In the present application, the control in the control solution includes verbascoside and / or isoacteoside, preferably includes verbascoside and isoacteoside. In the present application, the control solution is preferably obtained by mixing verbascoside and / or isoacteoside with 30-80 vol% methanol, preferably 40-70 vol% methanol, most preferably 70 vol% methanol. The concentration of the control in the control solution is 40-100 μg / mL, preferably 50-80 μg / mL.

[0062] In some embodiments of the present application, the control solution is preferably prepared as follows:

[0063] Take the appropriate amount of verbascoside and isoacteoside control, accurately weigh and determine, add 70 vol% methanol to prepare a mixed solution containing 50 μg of verbascoside and isoacteoside per 1 mL, as the control solution.

[0064] In the prior art, most of the compounds isolated from sandalwood medicinal materials are mainly sesquiterpenes, in addition to monoterpenes and lignans and other chemical components, but sesquiterpenes, monoterpenes and other components are lipophilic components, which limits most research to the study of lipophilic components of sandalwood medicinal materials, such as methanol components, which are also relatively lipophilic components. In the present application, 30-80 vol% methanol, most preferably 70 vol% methanol, is used to study both hydrophilic and lipophilic components. Through the development of chromatographic conditions, two new components, verbascoside and isoacteoside, are identified. Both of them are phenylethanoid glycosides, which are hydrophilic compounds and have a high content in sandalwood. Therefore, they are used as controls.

[0065] In the present application, the determination method further comprises the step of detecting and analyzing the negative control solution by liquid chromatography. The negative control solution is prepared according to the preparation method of the test sample solution, with the difference that it lacks the product, i.e. the negative control solution is 30-80 vol% methanol.

[0066] In the present application, the filler of the chromatographic column in the determination method is octadecylsilane-bonded silica gel, the column temperature is 20-35°C, preferably 25-30°C; the flow rate is 0.1-0.8 mL / min, preferably 0.15-0.25 mL / min, more preferably 0.2 mL / min; the mobile phase A is acetonitrile, the mobile phase B is selected from water, 0.1-0.2 vol% phosphoric acid or 0.1-0.2 vol% formic acid, the detection wavelength is 300-350 nm, preferably 326 nm. The theoretical plate number calculated according to the peak of verbascoside should be not less than 3000.

[0067] The gradient elution is used in the process of the liquid chromatography detection, and the program of the gradient elution is as follows:

[0068] 0-8 min: 9-16.5 vol% mobile phase A, 91-83.5 vol% mobile phase B;

[0069] 8-40 min: 16.5 vol% mobile phase A, 83.5 vol% mobile phase B;

[0070] 40-55 min: 16.5-30 vol% mobile phase A, 83.5-70 vol% mobile phase B;

[0071] 55-60 min: 30 vol% mobile phase A, 70 vol% mobile phase B.

[0072] In some specific embodiments of the present application, the chromatographic column in the liquid chromatography has octadecylsilane-bonded silica gel as the filler (the column length is 150 mm, the inner diameter is 2.1 mm, and the particle size is 1.8 μm); acetonitrile is used as the mobile phase A, 0.1 vol% phosphoric acid is used as the mobile phase B, and the gradient elution is carried out according to the above program; the flow rate is 0.2 mL / min; the column temperature is 25°C; and the detection wavelength is 326 nm. The theoretical plate number calculated according to the peak of verbascoside should be not less than 3000.

[0073] In the present application, preferably, the amount of the test solution and the control solution is 1-5 μL, which can be 1 μL, 2 μL, 3 μL, 4 μL or 5 μL, preferably 1 μL, which is added into the liquid chromatograph, and the detection and analysis are carried out according to the above chromatographic conditions. It is found that the determination method obtains 7 corresponding characteristic peaks in the standard characteristic chromatogram of sandalwood medicinal materials, decoction pieces, standard decoction and formula granules, and successfully identifies the peaks of verbascoside and isoferulic acid for the first time.

[0074] The application carries out methodological investigation on the above-mentioned determination method, including precision test, repeatability investigation, intermediate precision investigation (different instrument investigation, different personnel and time investigation), durability investigation (chromatographic column durability investigation, stability investigation), and the results show that the determination method has good precision, repeatability and durability.

[0075] The application also adopts the prepared determination method to determine the characteristic chromatogram of different batches of sandalwood medicinal materials, decoction pieces, standard decoction and formula granules, calculates the relative retention time and relative peak area, and the results show that the characteristic peaks can be detected and the peaks are relatively high, therefore, 7 peaks with good repeatability are selected as the characteristic peaks.

[0076] In the application, when the test sample is sandalwood formula granules, it is provided that 7 characteristic peaks should be present in the test sample chromatogram, wherein peak 4 (S) and peak 5 should correspond to the retention time of the reference peaks of the corresponding control samples, the peak corresponding to the retention time of the verbascoside reference peak is peak 4 (S), the relative retention time of the remaining characteristic peaks to peak 4 (S) is calculated, and the relative retention time should be within ±10% of the specified value, and the specified value is: 0.46 (peak 1), 0.51 (peak 2), 0.76 (peak 3), 1.39 (peak 6), and 2.27 (peak 7).

[0077] When the test sample is sandalwood medicinal materials, it is provided that 7 characteristic peaks should be present in the test sample chromatogram, wherein peak 4 (S) and peak 5 should correspond to the retention time of the reference peaks of the verbascoside and isoacteoside control samples, the peak corresponding to the retention time of the verbascoside reference peak is peak 4 (S), the relative retention time of peak 1, peak 2, peak 3, peak 6 and peak 7 to peak 4 (S) is calculated, and the relative retention time should be within ±10% of the specified value, and the specified value is: 0.46 (peak 1), 0.51 (peak 2), 0.76 (peak 3), 1.39 (peak 6), and 2.27 (peak 7).

[0078] When the test sample is sandalwood decoction pieces, it is provided that 7 characteristic peaks should be present in the test sample chromatogram, wherein peak 4 (S) and peak 5 should correspond to the retention time of the reference peaks of the verbascoside and isoacteoside control samples, the peak corresponding to the retention time of the verbascoside reference peak is peak 4 (S), the relative retention time of the remaining peaks to peak 4 (S) is calculated, and the relative retention time should be within ±10% of the specified value, and the specified value is: 0.46 (peak 1), 0.51 (peak 2), 0.76 (peak 3), 1.39 (peak 6), and 2.27 (peak 7).

[0079] When the test sample is sandalwood standard decoction, it is required that 7 characteristic peaks should be presented in the test sample chromatogram, wherein peak 4 (S) and peak 5 should correspond to the retention time of the reference peak of verbascoside and isovertoside, the peak corresponding to the reference peak of verbascoside is peak 4 (S), and the relative retention time of the remaining peaks to peak 4 (S) should be within the range of ±10% of the specified value, wherein the specified value is: 0.46 (peak 1), 0.51 (peak 2), 0.76 (peak 3), 1.39 (peak 6), and 2.27 (peak 7).

[0080] The present application also adopts the traditional Chinese medicine chromatographic fingerprint similarity evaluation system (2012 version) to synthesize 3 batches of sandalwood formula granules, 5 batches of sandalwood medicinal materials, 5 batches of sandalwood decoction pieces and 5 batches of sandalwood standard decoction, and establishes the control chromatogram of the sandalwood formula granules, sandalwood medicinal materials, sandalwood decoction pieces and sandalwood standard decoction.

[0081] Based on the above description, the present application also provides an application of the above-mentioned determination method in detecting the quality of sandalwood medicinal materials, decoction pieces, standard decoction and formula granules.

[0082] The present application also provides a quality control method of sandalwood medicinal materials, decoction pieces, standard decoction and formula granules, which comprises:

[0083] The test sample of sandalwood medicinal materials, decoction pieces, standard decoction and formula granules is subjected to the above-mentioned determination method to obtain the characteristic chromatogram of the test sample;

[0084] The characteristic chromatogram of the test sample is compared with the standard characteristic chromatogram obtained by the above-mentioned determination method; if 7 corresponding characteristic peaks are presented in the characteristic chromatogram of the test sample, and the RSD value of the relative retention time of each peak in the characteristic chromatogram of the sample to the relative retention time of each peak in the standard characteristic chromatogram is ≤10.0%, the sample is determined to be qualified.

[0085] In order to further illustrate the present application, the following examples are used for detailed description. The experimental raw materials used in the following examples of the present application are all general commercially available products. Unless otherwise specified, the "%" involved in the following description all represents volume percentage.

[0086] Example 1

[0087] 1 sandalwood formula granules

[0088] The high-performance liquid chromatographic characteristic chromatogram of the sandalwood formula granules is established, which can be used as a quality control method of sandalwood medicinal materials.

[0089] 1.1 materials, reagents and instruments

[0090] Instrument 1 (Shimadzu LC-40), instrument 2 (Agilent 1290 type), instrument 3 (Waters);

[0091] Electronic balance: ME204E / 02, MS205DM, XP26 (Mettler-Toledo Instruments Co., Ltd.);

[0092] Ultrapure water machine: CelluSafe 1810A (Shanghai Moore Scientific Instruments Co., Ltd.);

[0093] Ultrasonic cleaner: KQ5200DB type (600W, 40KHz; Kunshan Ultrasonic Instruments Co., Ltd.);

[0094] Chromatographic column: Chromatographic column 1 (Agilent SB-C18), chromatographic column 2 (Agilent XDB-C18), chromatographic column 3 (Agilent TC-C18);

[0095] Acetonitrile and phosphoric acid are chromatographically pure; water is ultrapure water, and other reagents are analytically pure;

[0096] Verbascoside (Aescin) reference substance (China Food and Drug Inspection Research Institute, batch number: 111530-201914, content is 95.2%);

[0097] Isofraxidin (Isoverbascoside) reference substance (Sichuan Weiqicheng Biological Technology Co., Ltd., batch number: wkq20050603, content is 98.0%);

[0098] Sandalwood formula granules: TX-01, TX-02, TX-03 (provided by Sichuan Xinglvc Pharmaceutical Technology Development Co., Ltd.).

[0099] 1.2 Chromatographic condition test

[0100] Octadecylsilane-bonded silica gel as filler (column length 150 mm, inner diameter 2.1 mm, particle size 1.8 μm); acetonitrile as mobile phase A, 0.1 vol% phosphoric acid as mobile phase B, gradient elution according to the provisions in Table 1 below; flow rate is 0.2 mL per minute; column temperature is 25℃; detection wavelength is 326 nm. The theoretical plate number should not be less than 3000 calculated according to the verbascoside peak.

[0101] Table 1 Proposed mobile phase gradient

[0102] Time (min) Mobile phase A (vol%) Mobile phase B (vol%) 0～8 9→16.5 91→83.5 8～40 16.5 83.5 40～55 16.5→30 83.5→70 55～60 30 70

[0103] 1.3 Preparation of reference solution

[0104] Take the verbascoside and isofraxidin reference substances, accurately weigh, add 70 vol% methanol to prepare a mixed solution containing 50 μg of verbascoside and isofraxidin per 1 mL, as the reference reference solution.

[0105] 1.4 Preparation of test solution

[0106] Take the product, grind finely, about 0.2g, put into a conical flask with a plug, add 70vol% methanol 25mL, ultrasonic treatment (power 600W, frequency 40kHz) for 30min, cool, shake well, filter, take the filtrate, and obtain.

[0107] 1.5 Determination method

[0108] Take 1μL of the control solution and test solution respectively, inject into the liquid chromatograph, and determine, and obtain.

[0109] 1.6 Chromatographic conditions and system suitability test

[0110] 1.6.1 Selection of mobile phase

[0111] On the basis of the above proposed experimental conditions, the separation effects of three different mobile phases were investigated, which were acetonitrile-water, acetonitrile-0.1vol% phosphoric acid, acetonitrile-0.1vol% formic acid, and the results are shown in Figure 1 .

[0112] The results show that the chromatographic peak baseline is relatively stable under the gradient elution conditions of each mobile phase solution, and the chromatographic peak is more, and the chromatographic peak shape and separation degree are better under the condition of acetonitrile-0.1vol% phosphoric acid, so the gradient elution of acetonitrile-0.1vol% phosphoric acid solution is selected as the mobile phase of the determination method of the characteristic chromatogram of sandalwood granules.

[0113] It should be noted that the present application also investigates acetonitrile-0.2vol% phosphoric acid and acetonitrile-0.2vol% formic acid, which are not embodied in Figure 1 for the sake of avoiding complexity, but the results are the same, and can all achieve the separation purpose, but lower concentration of acid is generally selected in the experiment to protect the instrument and chromatographic column.

[0114] 1.6.2 Selection of wavelength

[0115] On the basis of the above proposed experimental conditions, the test solution was scanned by using a diode array detector, and the chromatograms of the test solution at 210nm, 230nm, 254nm, 280nm, 300nm, 326nm, 350nm were extracted, as shown in Figure 2 .

[0116] The results show that the separation purpose can be achieved at the detection wavelength of 300-350nm, among which the chromatographic peak information is larger and the chromatogram baseline is more stable at the detection wavelength of 326nm, so the detection wavelength is preferably determined as 326nm.

[0117] 1.6.3 Flow rate investigation

[0118] On the basis of the above experimental conditions, the flow rate of 0.15 mL / min, 0.2 mL / min, 0.25 mL / min were investigated respectively. The results are shown in Figure 3 .

[0119] The results showed that the flow rate of 0.15 mL / min, 0.20 mL / min, 0.25 mL / min, the overall peak shape of chromatogram was good, but the flow rate of 0.20 mL / min, the separation effect of chromatographic peak was the best. Considering the integrity, the flow rate of 0.20 mL / min was selected for subsequent study.

[0120] 1.6.4 Column temperature investigation

[0121] On the basis of the above experimental conditions, the column temperature of 20℃, 30℃, 35℃ were investigated respectively. The results are shown in Figure 4 .

[0122] The results showed that the column temperature of 25℃, the overall peak shape and resolution of chromatogram were good, so the column temperature of 25℃ was selected for subsequent study.

[0123] 1.6.5 System suitability test results

[0124] Octadecylsilane-bonded silica gel as filler (column length of 150 mm, inner diameter of 2.1 mm, particle size of 1.8 μm); acetonitrile as mobile phase A, 0.1 vol% phosphoric acid as mobile phase B, gradient elution according to the provisions in Table 2; flow rate of 0.2 mL / min; column temperature of 25℃; detection wavelength of 326 nm. The theoretical plate number should not be less than 3000 according to the peak of verbascoside.

[0125] Table 2 Mobile phase gradient

[0126] Time (min) Mobile phase A (vol%) Mobile phase B (vol%) 0～8 9→16.5 91→83.5 8～40 16.5 83.5 40～55 16.5→30 83.5→70 55～60 30 70

[0127] 1.7 Investigation of test solution

[0128] 1.7.1 Investigation of extraction solvent

[0129] On the basis of the above experimental conditions, the extraction solvent of methanol, 30% methanol, 50% methanol, 70% methanol, ethanol, water were investigated respectively. The results are shown in Figure 5 .

[0130] It can be seen that when 70% methanol was used as the extraction solvent, the size of each peak was appropriate, and the baseline was relatively stable.

[0131] 1.8 Characteristic methodological validation

[0132] 1.8.1 Chromatographic peak identification

[0133] Preparation of the test solution: Take an appropriate amount of this product, grind it into a fine powder (about 0.2 g), place it in a stoppered conical flask, add 25 mL of 70 vol% methanol, sonicate (power 600 W, frequency 40 kHz) for 30 min, cool, shake well, filter, and take the filtrate to obtain the test solution.

[0134] Preparation of reference solution: Take appropriate amounts of verbascoside and isocitric acid glycoside reference standards, accurately weigh them, and add 70% methanol to prepare a solution containing 50 μg of each per 1 mL, as the reference solution.

[0135] Preparation of negative control solution: Prepare a negative control solution lacking sandalwood granules by referring to the preparation method of the test solution described above.

[0136] The peaks of the characteristic spectral analysis of sandalwood formulation particles were located. For example... Figure 6 As shown.

[0137] The results showed that peak 4 (S) was verbascoside and peak 5 was heterologous cimicifugoside. In the following methodological investigation, seven characteristic peaks in the sample were examined.

[0138] 1.8.2 Precision Test

[0139] Take the sandalwood formula granules (batch number: TX-01) test solution and inject it 6 times consecutively according to the proposed experimental method, 1 μL each time. Calculate the retention time and peak area of ​​each characteristic peak. As shown in Tables 3 and 4.

[0140] Table 3 Precision Examination - Retention Time

[0141] Serial number Peak 1 Peak 2 Peak 3 Peak 4 (S) Peak 5 Peak 6 Peak 7 1 9.907 10.843 16.215 21.537 28.105 29.962 48.610 2 9.918 10.852 16.225 21.563 28.155 30.003 48.633 3 9.915 10.853 16.220 21.573 28.165 30.010 48.630 4 9.912 10.852 16.223 21.563 28.167 30.018 48.630 5 9.913 10.843 16.202 21.518 28.095 29.927 48.600 6 9.913 10.845 16.218 21.562 28.135 29.973 48.610 RSD (%) 0.04 0.04 0.05 0.10 0.11 0.12 0.03

[0142] Table 4 Precision Examination - Peak Area

[0143] Serial number Peak 1 Peak 2 Peak 3 Peak 4 (S) Peak 5 Peak 6 Peak 7 1 50.232 49.500 51.300 172.677 370.008 253.815 593.137 2 50.287 49.708 51.264 171.077 369.573 258.923 594.873 3 50.344 49.931 51.612 171.873 368.106 253.996 596.052 4 50.541 50.032 51.155 170.938 364.069 258.097 596.678 5 50.574 50.251 50.077 168.922 363.136 261.562 597.388 6 50.542 50.375 50.364 164.195 352.210 255.418 594.882 RSD (%) 0.30 0.66 1.18 1.81 1.83 1.20 0.26

[0144] The results show that the instrument has good precision.

[0145] 1.8.3 Repeatability Test

[0146] Six portions of sandalwood formula granules (batch number: TX-01) were weighed and prepared and measured according to the proposed experimental method. The relative retention times of each characteristic peak were calculated, as shown in Table 5.

[0147] Table 5 Repeatability Tests - Relative Retention Time

[0148] Serial number Peak 1 Peak 2 Peak 3 Peak 4 (S) Peak 5 Peak 6 Peak 7 Peak 8 Repeatability 1 0.46 0.50 0.75 1.00 1.31 1.39 2.26 0.46 Repeatability 2 0.46 0.50 0.75 1.00 1.31 1.39 2.26 0.46 Repeatability 3 0.46 0.50 0.75 1.00 1.31 1.39 2.26 0.46 Repeatability 4 0.46 0.50 0.75 1.00 1.31 1.39 2.26 0.46 Repeatability 5 0.46 0.50 0.75 1.00 1.31 1.39 2.26 0.46 Repeatability 6 0.46 0.50 0.75 1.00 1.31 1.39 2.26 0.46 RSD (%) 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00

[0149] The results show that the method has good repeatability.

[0150] 1.8.4 Intermediate Precision Examination

[0151] 1.8.4.1 Different instrument investigation

[0152] Based on the above experimental conditions, sandalwood formula granules (batch number: TX-01) were weighed to prepare the test solution, which was determined on instrument 1, instrument 2 and instrument 3, respectively. The relative retention time of each characteristic peak was calculated, as shown in Table 6, Figure 7

[0153] Table 6 Relative retention time of instrument durability investigation

[0154] Instrument Peak 1 Peak 2 Peak 3 Peak 4 (S) Peak 5 Peak 6 Peak 7 Instrument 1 0.46 0.50 0.75 1.00 1.31 1.39 2.26 Instrument 2 0.52 0.53 0.72 1.00 1.25 1.31 2.39 Instrument 3 0.62 0.69 0.82 1.00 1.15 1.19 1.80 RSD (%) 15.16 17.82 6.72 0.00 6.54 7.76 14.42

[0155] The results show that when instrument 2 is used for detection, the peak relative retention time RSD is too large; when instrument 1 and instrument 3 are used to detect the test sample, the RSD of the relative retention time of each characteristic peak is 1.58% to 6.73%, indicating that the method has good durability on instrument 1 and instrument 3.

[0156] 1.8.4.2 Different personnel and time investigation

[0157] Based on the above experimental conditions, two samples of sandalwood formula granules (batch number: TX-01) were weighed by different personnel (A, B) at different times (T1, T2) to prepare the test sample and determine. As shown in Table 7.

[0158] Table 7 Relative retention time ratio of personnel and time investigation

[0159] Serial number Peak 1 Peak 2 Peak 3 Peak 4 (S) Peak 5 Peak 6 Peak 7 A 0.46 0.50 0.75 1.00 1.31 1.39 2.26 B 0.46 0.50 0.75 1.00 1.30 1.39 2.26 T1 0.46 0.50 0.75 1.00 1.31 1.39 2.26 T2 0.46 0.50 0.75 1.00 1.30 1.39 2.25 RSD (%) 0.00 0.00 0.00 0.00 0.44 0.00 0.22

[0160] The results show that when the same sample is determined by different personnel at different times, the RSD of the relative retention time of each characteristic peak is 0.00% to 0.44%, indicating that the method has good stability.

[0161] 1.8.5 Durability investigation

[0162] 1.8.5.1 Column durability investigation

[0163] Based on the above experimental conditions, column 1, column 2 and column 3 were investigated, respectively. As shown in Table 8, Figure 8

[0164] Table 8 Column durability investigation

[0165] Serial number Peak 1 Peak 2 Peak 3 Peak 4 (S) Peak 5 Peak 6 Peak 7 Column 1 0.46 0.50 0.75 1.00 1.31 1.39 2.26 Column 2 0.43 0.47 0.74 1.00 1.34 1.44 1.99 Column 3 0.50 0.56 0.80 1.00 1.22 1.33 2.23 RSD (%) 7.58 8.99 4.21 0.00 4.84 3.97 6.85

[0166] The results show that the method is stable when the above three columns are used to detect the sample.

[0167] 1.8.5.2 Stability investigation​​

[0168] On the basis of the above experimental conditions, the same test solution was taken and measured at 0 h, 2 h, 4 h, 8 h, 16 h and 24 h. The results are shown in Table 9.

[0169] Table 9 Stability Investigation

[0170] Serial number Peak 1 Peak 2 Peak 3 Peak 4 (S) Peak 5 Peak 6 Peak 7 Stability 0 h 9.907 10.843 16.215 21.537 28.105 29.962 48.610 Stability 2 h 9.918 10.852 16.225 21.563 28.155 30.003 48.633 Stability 4 h 9.915 10.853 16.220 21.573 28.165 30.010 48.630 Stability 8 h 9.912 10.852 16.223 21.563 28.167 30.018 48.630 Stability 16 h 9.913 10.843 16.202 21.518 28.095 29.927 48.600 Stability 24 h 9.913 10.845 16.218 21.562 28.135 29.973 48.610 RSD (%) 0.04 0.04 0.05 0.10 0.11 0.12 0.03

[0171] The results show that the sample solution is relatively stable within 24 hours.

[0172] In summary, the RSD of the relative retention time of each characteristic peak meets the requirements in the above investigations, indicating that the method is good. The above 7 characteristic peaks are included in the subsequent investigation.

[0173] 1.9 Determination of Characteristic Peaks and Establishment of Reference Chromatogram

[0174] 1.9.1 Verification Results of 3 Batches of Tanxiang Formula Granules

[0175] The proposed method was used to determine the characteristic chromatogram of 3 batches of samples. The results are shown in Table 10, Figure 9

[0176] Table 10 Verification of 3 Batches of Tanxiang Formula Granules - Relative Retention Time

[0177] Serial number Batch number Peak 1 Peak 2 Peak 3 Peak 4 (S) Peak 5 Peak 6 Peak 7 1 TX-01 0.46 0.50 0.76 1.00 1.30 1.39 2.26 2 TX-02 0.46 0.50 0.75 1.00 1.30 1.39 2.26 3 TX-03 0.46 0.50 0.75 1.00 1.30 1.39 2.26 RSD (%) - 0.00 0.00 0.00 0.00 0.00 0.00 0.00

[0178] According to the principles of stability of relative retention time and detection of samples from each batch with relatively high peaks, a total of 7 peaks with good reproducibility were selected as characteristic peaks. The results show that the RSD of the relative retention time of the 7 characteristic peaks of 3 batches of Tanxiang Formula Granules is 0.00%, and the method can effectively detect Tanxiang Formula Granules.

[0179] The characteristic chromatograms of 3 batches of Tanxiang Formula Granules were synthesized using Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 version) to establish a reference chromatogram of Tanxiang Formula Granules characteristic chromatogram, as Figure 10

[0180] 1.9.2 Establishment of Relative Retention Time Limit

[0181] The results of the methodological investigations and verification are shown in Table 11:

[0182] Table 11 Summary of RSD% of Results of Each Item of Methodology - Relative Retention Time

[0183] Methodology Peak 1 Peak 2 Peak 3 Peak 4 (S) Peak 5 Peak 6 Peak 7 Repeatability 0.00 0.00 0.00 0.00 0.40 0.00 0.18 Precision 0.04 0.04 0.05 0.10 0.11 0.12 0.03 Stability 0.04 0.04 0.05 0.10 0.11 0.12 0.03 Intermediate precision 0.00 0.00 0.00 0.00 0.44 0.00 0.22 Chromatographic column robustness 7.58 8.99 4.21 0.00 4.84 3.97 6.85 Different instruments 15.16 17.82 6.72 0.00 6.54 7.76 14.42 3 batch validation 0.00 0.00 0.77 0.00 0.00 0.00 0.00

[0184] ​​​​​​​​​​​​​​​​​​​​​​​From the above table, different instruments and different chromatographic columns have great influence on the relative retention time of each characteristic peak. In order to increase the reproducibility and applicability of the method, the relative retention time of each peak is temporarily set as 10%.

[0185] Finally, it is determined that 7 characteristic peaks should be present in the chromatogram of the test sample, among which peak 4 (S) and peak 5 should correspond to the retention time of the corresponding reference peak, and the peak corresponding to the retention time of the reference peak of verbascoside is the S peak. The relative retention time of the remaining characteristic peaks to the S peak should be within ± 10% of the specified value. The specified value is: 0.46 (peak 1), 0.51 (peak 2), 0.76 (peak 3), 1.39 (peak 6), and 2.27 (peak 7).

[0186] Example 2

[0187] 2.2.1 Materials, reagents and instruments

[0188] 2.2.2 Chromatographic conditions and system suitability test

[0189] 2.2.1 Materials, reagents and instruments

[0190] 2.2.1 Materials, reagents and instruments

[0191] 2.2.1 Materials, reagents and instruments

[0192] 2.2.2 Chromatographic conditions and system suitability test

[0193] 2.2.2 Chromatographic conditions and system suitability test

[0194] 2.3 Preparation of reference solution

[0195] 2.3 Preparation of reference solution

[0196] 2.4 Preparation of test sample solution

[0197] Take the powder (pass through No. 3 sieve) 1.0 g, add water 50 mL, decoct for 30 min, cool, filter, evaporate the filtrate to dryness, dissolve the residue with 70% methanol 25 mL, filter, and take the filtrate.

[0198] 2.5 Determination method

[0199] 2.5 Determination method

[0200] 2.6 Determination of characteristic peaks and establishment of reference chromatogram

[0201] The prepared method was used to determine the characteristic chromatogram of 3 batches of samples, and the relative retention time was calculated. As shown in Table 12. Figure 11

[0202] Table 12 Relative retention time of 5 batches of sandalwood medicinal materials

[0203] Serial number Batch number Peak 1 Peak 2 Peak 3 Peak 4 (S) Peak 5 Peak 6 Peak 7 1 TX-YC-01 0.46 0.50 0.75 1.00 1.30 1.39 2.26 2 TX-YC-02 0.46 0.51 0.75 1.00 1.31 1.39 2.26 3 TX-YC-03 0.46 0.50 0.75 1.00 1.31 1.39 2.26 4 TX-YC-04 0.46 0.51 0.75 1.00 1.31 1.39 2.26 5 TX-YC-05 0.46 0.51 0.75 1.00 1.31 1.39 2.26

[0204] The results showed that the relative retention time of the 7 characteristic peaks presented in the chromatogram of 5 batches of sandalwood medicinal materials was within ±10% of the specified value.

[0205] The final specification is that 7 characteristic peaks should be present in the test sample chromatogram, among which peak 4 (S) and peak 5 should correspond to the retention time of the reference peaks of verbascoside and isoheterosidaloin, and the peak corresponding to the reference peak of verbascoside is the S peak. The relative retention time of peaks 1, 2, 3, 6, and 7 to the S peak should be within ±10% of the specified value, and the specified value is: 0.46 (peak 1), 0.51 (peak 2), 0.76 (peak 3), 1.39 (peak 6), and 2.27 (peak 7).

[0206] The characteristic chromatogram of 5 batches of sandalwood medicinal materials was synthesized using Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 version), and a control chromatogram of sandalwood medicinal materials was established, as shown in Table 12. Figure 12 .

[0207] Example 3

[0208] 3. Sandalwood decoction pieces

[0209] A high-performance liquid chromatographic characteristic chromatogram of sandalwood decoction pieces was established, which can be used as a method for overall quality control of sandalwood decoction pieces.

[0210] 3.1 Materials, reagents and instruments

[0211] The same as 1.1. Materials, reagents and instruments.

[0212] Sandalwood decoction pieces: TX-YP-01, TX-YP-02, TX-YP-03, TX-YP-04, TX-YP-05 (provided by Sichuan Xinglv Pharmaceutical Technology Development Co., Ltd.).

[0213] 3.2 Chromatographic conditions and system suitability test

[0214] The same as 1.2 Chromatographic conditions and system suitability test.

[0215] 3.3 Preparation of reference solution

[0216] The same as 1.3 Preparation of reference solution.

[0217] 3.4 Preparation of test sample solution​

[0218] Take the powder (pass through No. 3 sieve) 1.0 g, add water 50 mL, decoct for 30 min, cool, filter, evaporate the filtrate to dryness, dissolve the residue with 70% methanol 25 mL, filter, take the filtrate, and you get it.

[0219] 3.5 Determination method

[0220] The same as 1.5 determination method.

[0221] 3.6 Determination of characteristic peaks and establishment of control chromatogram

[0222] Using the proposed method, 5 batches of samples of the product were determined by characteristic chromatogram, and the relative retention time was calculated. As shown in Figure 13 , Table 13.

[0223] Table 13 5 batches of sandalwood tablets verification

[0224] Serial number Batch number Peak 1 Peak 2 Peak 3 Peak 4 (S) Peak 5 Peak 6 Peak 7 1 TX-YP-01 0.46 0.50 0.75 1.00 1.31 1.31 2.26 2 TX-YP-02 0.46 0.50 0.75 1.00 1.30 1.30 2.26 3 TX-YP-03 0.46 0.51 0.76 1.00 1.31 1.31 2.26 4 TX-YP-04 0.46 0.51 0.76 1.00 1.30 1.30 2.27 5 TX-YP-05 0.46 0.51 0.76 1.00 1.30 1.30 2.27

[0225] The results showed that the relative retention time of the 7 characteristic peaks presented in the chromatogram of 5 batches of sandalwood tablets was within ± 10% of the specified value.

[0226] The final provision is that 7 characteristic peaks should be presented in the test sample chromatogram, among which peak 4 (S) and peak 5 should correspond to the retention time of the reference peak of verbascoside and isoviolanthoside control, and the peak corresponding to the reference peak of verbascoside is S peak. Calculate the relative retention time of the remaining peaks to S peak, and the relative retention time should be within the specified ± 10% range. The specified value is: 0.46 (peak 1), 0.51 (peak 2), 0.76 (peak 3), 1.39 (peak 6), 2.27 (peak 7).

[0227] Using Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 version) to synthesize the characteristic chromatogram of 5 batches of sandalwood tablets, the control chromatogram of sandalwood tablet characteristic chromatogram was established, as shown in Figure 14 .

[0228] Example 4

[0229] 4 Standard decoction of sandalwood

[0230] The high performance liquid characteristic chromatogram of sandalwood standard decoction was established, which can be used as the overall quality control method of sandalwood standard decoction.

[0231] 4.1 Materials, reagents and instruments

[0232] The same as 1.1. Materials, reagents and instruments.

[0233] Santalum album L. standard decoction: TX-BT-01, TX-BT-02, TX-BT-03, TX-BT-04, TX-BT-05 (provided by Sichuan Xinglv Pharmaceutical Science and Technology Development Co., Ltd.).

[0234] 4.2 Chromatographic conditions and system suitability test

[0235] The same as 1.2 Chromatographic conditions and system suitability test.

[0236] 4.3 Preparation of reference solution

[0237] The same as 1.3 Preparation of reference solution.

[0238] 4.4 Preparation of test solution

[0239] Take the powder (pass through No. 3 sieve) 1.0 g, add water 50 mL, decoct for 30 min, cool, filter, evaporate the filtrate to dryness, dissolve the residue with 70% methanol 25 mL, filter, and take the filtrate.

[0240] 4.5 Determination method

[0241] The same as 1.5 Determination method.

[0242] 4.6 Determination of characteristic peaks and establishment of reference chromatogram

[0243] The proposed method was used to determine the characteristic chromatogram of 5 batches of samples, and the relative retention time was calculated. As shown in Table 14. Figure 15

[0244] Table 14 Verification of 5 batches of Santalum album L. standard decoction

[0245] Serial number Batch number Peak 1 Peak 2 Peak 3 Peak 4 (S) Peak 5 Peak 6 Peak 7 1 TX-BT--01 0.46 0.51 0.76 1.00 1.30 1.39 2.27 2 TX-BT--02 0.46 0.51 0.76 1.00 1.30 1.39 2.27 3 TX-BT--03 0.46 0.51 0.76 1.00 1.30 1.39 2.27 4 TX-BT--04 0.46 0.51 0.76 1.00 1.30 1.39 2.27 5 TX-BT--05 0.46 0.51 0.76 1.00 1.30 1.39 2.27

[0246] The results showed that the relative retention times of the 7 characteristic peaks presented in the chromatograms of the 5 batches of standard decoction were within ±10% of the specified values.

[0247] The final specification: 7 characteristic peaks should be presented in the test solution chromatogram, among which peak 4 (S) and peak 5 should correspond to the relative retention time of the reference peak of verbascoside and isobatrachoside, and the peak corresponding to the reference peak of verbascoside is S peak. The relative retention times of the remaining peaks to S peak should be within ±10% of the specified value, and the specified value is: 0.46 (peak 1), 0.51 (peak 2), 0.76 (peak 3), 1.39 (peak 6), 2.27 (peak 7).

[0248] The Chinese medicine chromatographic fingerprint similarity evaluation system (2012 version) was used to synthesize the characteristic chromatogram of 5 batches of Santalum album L. decoction, and the reference chromatogram of Santalum album L. decoction characteristic chromatogram was established, as shown in Figure 16 . ​

[0249] It should be noted that the present application also carries out chromatographic conditions and system suitability test, test sample preparation method, methodological investigation for the test sample being sandalwood medicinal materials, sandalwood standard decoction and sandalwood decoction pieces, and the conclusions obtained are consistent with the test sample being sandalwood formula granules. In order to avoid complexity, it will not be repeated here.

[0250] The above description of disclosed embodiments enables one of ordinary skill in the art to make or use the application. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein can be applied to other embodiments without departing from the spirit or scope of the application. Thus, the present application is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A characteristic chromatographic determination method for sandalwood medicinal materials, decoction pieces, standard decoction, and formula granules, characterized in that, Comprising: The test sample solution and the control sample solution are detected by liquid chromatography, and analyzed; The control sample in the control sample solution comprises verbascoside and / or isoacteoside; The test sample solution is obtained by mixing the test sample with 30-80 vol% methanol, and then ultrasonic extraction or reflux extraction; Or The test sample solution is obtained by mixing the test sample with water, decocting, filtering, and mixing the residue obtained by evaporating the filtrate with 30-80 vol% methanol; The conditions of the liquid chromatography are as follows: the mobile phase A is acetonitrile, the mobile phase B is selected from water, 0.1-0.2 vol% phosphoric acid or 0.1-0.2 vol% formic acid, and the detection wavelength is 326 nm; Gradient elution is adopted in the process of the liquid chromatography detection, and the program of the gradient elution is as follows: 0-8 min: 9-16.5 vol% mobile phase A, 91-83.5 vol% mobile phase B; 8-40 min: 16.5 vol% mobile phase A, 83.5 vol% mobile phase B; 40-55 min: 16.5-30 vol% mobile phase A, 83.5-70 vol% mobile phase B; 55-60 min: 30 vol% mobile phase A, 70 vol% mobile phase B.

2. The assay method according to claim 1, characterized by The filler of the chromatographic column in the liquid chromatography is octadecylsilane-bonded silica gel; the column temperature is 20-35℃; and the flow rate is 0.1-0.8 mL / min.

3. The assay method according to claim 2, characterized in that, The column temperature is 25℃, and the flow rate is 0.2 mL / min.

4. The assay method according to claim 1, characterized by, The dosages of the test sample solution and the control sample solution are both 1-5 μL.

5. The assay method according to claim 1, characterized by, The control sample solution is obtained by mixing verbascoside and / or isoacteoside with 30-80 vol% methanol; The concentration of the control sample in the control sample solution is 40-100 μg / mL.

6. The assay method according to claim 1, characterized by, The characteristic chromatogram of sandalwood medicinal material, decoction piece, standard decoction, and formula granule obtained by the determination method presents 7 corresponding characteristic peaks, and the peaks of verbascoside and / or isoacteoside are identified.

7. The determination method according to any one of claims 1-6 is applied to the detection of the quality of sandalwood medicinal material, decoction piece, standard decoction, and formula granule.

8. A quality control method of Santali Albae Radix, decoction pieces, standard decoction, and formula granules, characterized in that, Comprising: The test sample solution and the control sample solution are detected by liquid chromatography, and analyzed; The control sample in the control sample solution comprises verbascoside and / or isoacteoside; The test sample solution is obtained by mixing the test sample with 30-80 vol% methanol, and then ultrasonic extraction or reflux extraction; Or The test sample solution is obtained by mixing the test sample with water, decocting, filtering, and mixing the residue obtained by evaporating the filtrate with 30-80 vol% methanol; The conditions of the liquid chromatography are as follows: the mobile phase A is acetonitrile, the mobile phase B is selected from water, 0.1-0.2 vol% phosphoric acid or 0.1-0.2 vol% formic acid, and the detection wavelength is 326 nm; Gradient elution is adopted in the process of the liquid chromatography detection, and the program of the gradient elution is as follows: 0-8 min: 9-16.5 vol% mobile phase A, 91-83.5 vol% mobile phase B; 8-40 min: 16.5 vol% mobile phase A, 83.5 vol% mobile phase B; 40-55 min: 16.5-30 vol% mobile phase A, 83.5-70 vol% mobile phase B; 55-60 min: 30 vol% mobile phase A, 70 vol% mobile phase B. The filler of the chromatographic column in the liquid chromatography is octadecylsilane-bonded silica gel; the column temperature is 20-35℃; and the flow rate is 0.1-0.8 mL / min. The column temperature is 25℃, and the flow rate is 0.2 mL / min. The dosages of the test sample solution and the control sample solution are both 1-5 μL. The control sample solution is obtained by mixing verbascoside and / or isoacteoside with 30-80 vol% methanol; The concentration of the control sample in the control sample solution is 40-100 μg / mL. The characteristic chromatogram of sandalwood medicinal material, decoction piece, standard decoction, and formula granule obtained by the determination method presents 7 corresponding characteristic peaks, and the peaks of verbascoside and / or isoacteoside are identified.

7. The determination method according to any one of claims 1-6 is applied to the detection of the quality of sandalwood medicinal material, decoction piece, standard decoction, and formula granule. Comprising: The test sample solution and the control sample solution are detected by liquid chromatography, and analyzed; The control sample in the control sample solution comprises verbascoside and / or isoacteoside; The test sample solution is obtained by mixing the test sample with 30-80 vol% methanol, and then ultrasonic extraction or reflux extraction; Or The test sample solution is obtained by mixing the test sample with water, decocting, filtering, and mixing the residue obtained by evaporating the filtrate with 30-80 vol% methanol; The conditions of the liquid chromatography are as follows: the mobile phase A is acetonitrile, the mobile phase B is selected from water, 0.1-0.2 vol% phosphoric acid or 0.1-0.2 vol% formic acid, and the detection wavelength is 326 nm; Gradient elution is adopted in the process of the liquid chromatography detection, and the program of the gradient elution is as follows: 0-8 min: 9-16.5 vol% mobile phase A, 91-83.5 vol% mobile phase B; 8-40 min: 16.5 vol% mobile phase A, 83.5 vol% mobile phase B; 40-55 min: 16.5-30 vol% mobile phase A, 83.5-70 vol% mobile phase B; 55-60 min: 30 vol% mobile phase A, 70 vol% mobile phase B. The filler of the chromatographic column in the liquid chromatography is octadecylsilane-bonded silica gel; the column temperature is 20-35℃; and the flow rate is 0.1-0.8 mL / min. The column temperature is 25℃, and the flow rate is 0.2 mL / min. The dosages of the test sample solution and the control sample solution are both 1-5 μL. The control sample solution is obtained by mixing verbascoside and / or isoacteoside with 30-80 vol% methanol; The concentration of the control sample in the control sample solution is 40-100 μg / mL. The characteristic chromatogram of sandalwood medicinal material, decoction piece, standard decoction, and formula granule obtained by the determination method presents 7 corresponding characteristic peaks, and the peaks of verbascoside and / or isoacteoside are identified.

7. The determination method according to any one of claims 1-6 is applied to the detection of the quality of sandalwood medicinal material, decoction piece, standard decoction, and formula granule.

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