Establishment method of hydrangea crispa drug characteristic chromatogram and characteristic chromatogram and application thereof

Abstract

The invention relates to the technical field of medicine analysis, and particularly provides a method for establishing a hydrangea serrata medicine characteristic chromatogram as well as the characteristic chromatogram and application of the hydrangea serrata medicine characteristic chromatogram. The method not only solves the problem of disordered base sources of the sparassis crispa and clematis armandii decoction pieces and standard decoction, but also solves the problems of low extraction rate of effective components and difficulty in detection of formula granules and standard decoction conventional methods. The establishment method of the drug characteristic spectrum of the sparassis crispa can be used for quality control of the sparassis crispa formula granules, common control of active ingredients such as caffeic acid and ferulic acid can be achieved at the same time, basic sources of the sparassis crispa and standard decoction are effectively identified, the content of ferulic acid is limited, and overall quality control of the sparassis crispa formula granules is achieved. The application of the preparation method of the sparassis crispa formula granules and decoction piece screening in practice can effectively ensure that pharmacodynamic substances between the sparassis crispa formula granules and a standard decoction are consistent with the standard decoction, so that the quality of the prepared sparassis crispa formula granules is higher and more stable.

Description

Technical Field

[0001] The present invention relates to the technical field of drug analysis, and in particular to a method for establishing a characteristic spectrum of a hydrangea chinensis drug, and the characteristic spectrum and application thereof. Background Art

[0002] Hydrangea vine is the dried vine stem of Clematis montana Buch.-Ham. under the 2020 edition of the Chinese Pharmacopoeia. Like Akebia affine, Hydrangea vine has the effects of diuresis, relieving stranguria, clearing the heart and eliminating vexation, and promoting menstruation and lactation. It is often used clinically to treat stranguria, edema, irritability and red urine, sores in the mouth and tongue, amenorrhea, oligolactosis, and damp-heat pain. Studies have found that decoctions of different varieties of Akebia affine have different degrees of diuretic effects on water-loaded mice, but Hydrangea vine has the best effect, followed by Akebia affine. Hydrangea vine contains triterpenes, sterols, flavonoids, lignans and phenolic components, and has good antibacterial and anti-inflammatory effects. Under the Chinese Pharmacopoeia Akebia affine material, there are only properties, microscopic identification, thin layer identification and extract determination, which makes it difficult to distinguish the quality differences and quality advantages and disadvantages of Hydrangea vine and Akebia affine. But in actual work, the two are quite different. The surface of Akebia affine is brown or yellowish brown, the diameter is generally 0.5-2.5cm, hard, not easy to break, and has a small pith. The diameter of Hydrangea chinensis is 1-3cm, the surface is yellowish green to dark purple-red, hard, brittle, easy to break, and has a large pith.

[0003] In the Chinese medicinal materials market, the Chinese medicinal materials and pieces of Hydrangea chinensis only need to meet the current edition of the Chinese Pharmacopoeia to be circulated, resulting in the frequent mixing of the two sources of Sichuan Akebia medicinal materials. According to the traditional method of taking, the pieces are boiled in water. On the one hand, the quality varies due to the confusion of the source of the medicinal materials; on the other hand, the Chinese medicinal pieces lose their original properties after being prepared into Chinese medicine formula granules, making it difficult to identify the source. In recent years, the phenomenon of "same source, different effect" in Chinese medicine formula granules has been prominent, and the lack of standards for the determination of the effective ingredients of Hydrangea chinensis and the identification of characteristic spectra is not conducive to the promotion and application of Chinese medicine formula granules. The formula granules with two sources of Sichuan Akebia urgently need to solve the problem of confusion in the source of raw materials from the source. Therefore, the quality standards of Hydrangea chinensis and Chinese medicine formula granules urgently need to be optimized and improved. Summary of the invention

[0004] In view of this, the present invention proposes a method for establishing a characteristic spectrum of a hydrangea chinensis drug, a characteristic spectrum and an application thereof.

[0005] The technical solution of the present invention is implemented as follows: The present invention provides a method for establishing a characteristic spectrum of a hydrangea vine drug, comprising: taking a sample to prepare a hydrangea vine test solution, using ultra-high performance liquid chromatography to obtain a characteristic spectrum of the test solution and a reference peak of a reference solution, and the chromatographic conditions of the high performance liquid chromatography are:

[0006] Chromatographic column: chromatographic column with octadecylsilane bonded silica gel as filler;

[0007] Detection wavelength: 321-331nm;

[0008] Mobile phase A: acetonitrile, mobile phase B: 0.05-0.2% phosphoric acid solution;

[0009] Column temperature: 25-35°C;

[0010] Flow rate: 0.2-0.3ml / min;

[0011] The number of theoretical plates calculated based on the ferulic acid peak should be no less than 10,000;

[0012] The reference solution had a ferulic acid concentration of 10 μg / mL and a caffeic acid concentration of 20 μg / mL;

[0013] Gradient elution.

[0014] In some embodiments, the method of gradient elution is:

[0015] 0-6min, the volume ratio of mobile phase A to mobile phase B was 3-9%:97-91%;

[0016] 6-9min, the volume ratio of mobile phase A to mobile phase B is 9-10%:91-90%;

[0017] 9-11min, the volume ratio of mobile phase A to mobile phase B is 10-15%:90-85%;

[0018] 11-15min, the volume ratio of mobile phase A to mobile phase B is 15-25%:85-75%;

[0019] 15-20min, the volume ratio of mobile phase A to mobile phase B is 25-30%:75-70%;

[0020] 20-25min, the volume ratio of mobile phase A to mobile phase B is 30-35%:70-65%.

[0021] In some embodiments, the test sample is at least one of hydrangea chinensis medicinal materials, hydrangea chinensis decoction pieces, hydrangea chinensis water extract and hydrangea chinensis formula granules.

[0022] In some embodiments, when the test sample is hydrangea vine medicinal material or hydrangea vine decoction pieces, the preparation method of the test sample includes: taking the test sample powder, adding water to heat and reflux, cooling, shaking, filtering and taking the filtrate to obtain; when the test sample is hydrangea vine water extract or hydrangea vine formula granules, the preparation method of the test sample includes: taking the test sample powder, adding solvent, weighing quantitatively, ultrasonic treatment, cooling and weighing again, making up the lost weight with solvent, shaking, filtering and taking the filtrate to obtain.

[0023] In some embodiments, the method further includes preparing a reference solution, injecting reference solutions of different concentrations using a high performance liquid chromatogram, and plotting a standard curve of the reference solution with the injection concentration as the horizontal axis and the peak area as the vertical axis.

[0024] The second aspect of the present invention is to provide a characteristic spectrum of a hydrangea vine drug, which is established by the above-mentioned establishment method. The characteristic spectrum includes 8 characteristic peaks, with peak No. 3 where caffeic acid is located as the reference peak S1, and the relative retention times of peak No. 1, peak No. 2, peak No. 4 and the reference peak S1 are 0.77±0.08, 0.96±0.10, and 1.14±0.11, respectively; with peak No. 6 where ferulic acid is located as the reference peak S2, the relative retention times of peak No. 5, peak No. 7, peak No. 8 and the reference peak S2 are 0.91±0.09, 1.06±0.11, and 1.34±0.13, respectively.

[0025] The third aspect of the present invention is to provide an application of a characteristic spectrum of a hydrangea vine drug in a quality control method for hydrangea vine formula granules, comprising: obtaining the characteristic spectrum of the hydrangea vine formula granules, and screening out the formula granules whose substance content of peak No. 6 in the characteristic spectrum is in the range of 0.08 mg / g-0.51 mg / g, namely, the target formula granules.

[0026] In some embodiments, the quality control method of hydrangea vine formula granules also includes screening out formula granules whose ratio of peak area No. 3 to the total area of ​​8 characteristic peaks is not less than 0.064, namely, the target formula granules.

[0027] The above-mentioned quality control method can simultaneously realize the common control of active ingredients such as caffeic acid and ferulic acid, and limit the content of ferulic acid, so as to achieve the overall quality control of the hydrangea chinensis formula granules. The method of the present invention is applicable to a wide range of detection samples, for example, hydrangea chinensis medicinal materials, hydrangea chinensis decoction pieces, hydrangea chinensis standard decoctions and hydrangea chinensis formula granules can all achieve effective detection. The quality control method of the present invention can accurately, objectively and effectively control the quality of hydrangea chinensis and its formula granules, so that it meets the quality control requirements of clinical medication and ensures the effectiveness of clinical medication.

[0028] The fourth aspect of the present invention is to provide an application of a characteristic spectrum of a hydrangea vine drug in the identification of the origins of hydrangea vine and akebia schoenensis, comprising respectively obtaining the characteristic spectrum of the corresponding products, calculating the area ratio of peak 3 to peak 6, when the area ratio of peak 3 / peak 6 is 0.59-2.1, the origin of the corresponding product is hydrangea vine, and when the area ratio of peak 3 / peak 6 is ≥7.3, the origin of the corresponding product is akebia schoenensis.

[0029] The above identification method can effectively identify the origin of hydrangea vine and Akebia dasyphylla medicinal materials and products, and ensure the stability of the quality of hydrangea vine medicinal materials, decoction pieces, extracts, standard decoctions and Chinese medicine formula granules from the source. At the same time, the separation degree and peak shape of the characteristic peaks obtained are good, the method has strong repeatability and high sensitivity.

[0030] The fifth aspect of the present invention is to provide an application of a characteristic map of hydrangea chinensis medicine in the screening of hydrangea chinensis decoction pieces, and to screen out decoction pieces with a ratio of the area of ​​peak No. 3 to the total area of ​​8 characteristic peaks ≤0.2, which are the target decoction pieces.

[0031] Compared with the prior art, the present invention has the following beneficial effects:

[0032] The present invention provides a method for establishing a characteristic spectrum of a hydrangea chinensis drug, which can effectively identify the origin of hydrangea chinensis and Akebia dasyphylla medicinal materials and their products, and ensure the stability of the quality of hydrangea chinensis medicinal materials, decoction pieces, extracts, standard decoctions and important formula granules from the source. At the same time, the characteristic peaks obtained by the characteristic spectrum have good separation and peak shape, and the method has strong repeatability and high sensitivity. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0034] Figure 1 The reference characteristic spectra of 15 batches of hydrangea chinensis granules in Example 2 of the present invention;

[0035] Figure 2 The characteristic spectra of samples of each link of the hydrangea vine formula granules in Example 2 of the present invention;

[0036] Figure 3 The characteristic spectrum of the hydrangea chinensis formula granules and the reference substance in Example 2 of the present invention;

[0037] Figure 4 is a standard curve diagram of ferulic acid in the detection method of Example 1 of the present invention;

[0038] Figure 5 This is the comparative characteristic map of Hydrangea quinata and Akebia chinensis;

[0039] Figure 6 It is the characteristic spectrum at a wavelength of 294.2 nm in the detection method of Example 6 of the present invention;

[0040] Figure 7It is the characteristic spectrum when the wavelength is 320nm in the detection method of Example 6 of the present invention;

[0041] Figure 8 It is a control characteristic spectrum of the detection method of Example 7 of the present invention with a flow rate of 0.20-0.30 ml / min;

[0042] Fig. 9 It is a control characteristic spectrum when the column temperature is 25-35° C. in the detection method of Example 8 of the present invention;

[0043] Fig.10 is the reference characteristic spectrum of the chromatographic column ① in the detection method of Example 9 of the present invention;

[0044] Fig.11 is the reference characteristic spectrum of the chromatographic column ② in the detection method of Example 9 of the present invention;

[0045] Fig.12 It is the reference characteristic spectrum of chromatographic column ③ in the detection method of Example 9 of the present invention. DETAILED DESCRIPTION

[0046] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0047] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the embodiments of the present invention belong. If the definitions set forth in this section are contrary to or otherwise inconsistent with the definitions set forth in the patents, patent applications, published patent applications, and other publications incorporated herein by reference, the definitions listed in this section take precedence over the definitions incorporated herein by reference.

[0048] Unless otherwise specified, the methods used in the following examples are all conventional methods. Unless otherwise specified, the materials, reagents and instruments used are all conventional materials, reagents and instruments in the art, and can be obtained by those skilled in the art through commercial channels.

[0049] When an amount, concentration or other value or parameter is expressed as a range, a preferred range or a range defined by a series of upper preferred values ​​and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pairing of any range upper limit or preferred value with any range lower limit or preferred value, regardless of whether the range is disclosed separately. For example, when a range of "1 to 5" is disclosed, the described range should be interpreted as including the range "1 to 4", "1 to 3", "1 to 2", "1 to 2 and 4 to 5", "1 to 3 and 5", etc. When a numerical range is described herein, unless otherwise stated, the range is intended to include its end values ​​and all integers and fractions within the range. In the present specification and claims, range definitions can be combined and / or interchanged, and if not otherwise stated, these ranges include all subranges contained therein.

[0050] The instruments and reagents used in the following examples are as follows:

[0051] instrument:

[0052] Agilent 1290 series ultra-high performance liquid chromatograph (1290 quaternary gradient pump, 1290 autosampler, 1290 column oven), American 1290 VWD detector; Waters series ultra-high performance liquid chromatograph (ACQUITY UPLC H-Class Plus), American PDA detector; 1 / 100,000 electronic balance (Sartorius); XMTD-7000 constant temperature water bath (Beijing Guangming Medical Instrument Co., Ltd.); SK8200XS ultrasonic cleaner (Shanghai Kedao Ultrasonic Instrument Co., Ltd.).

[0053] Reagents:

[0054] Ferulic acid reference substance (batch number: 110773-201915; purity 99.4%).

[0055] Caffeic acid reference (batch number: 110885-201703; purity 99.7%).

[0056] The reference medicinal materials of Akebia trifoliata (121409-202003) were purchased from China Food and Drug Inspection Institutes.

[0057] The control medicinal material of Hydrangea chinensis (230121-202111) was purchased from Shanghai Hongyong Biotechnology Co., Ltd.

[0058] Methanol, phosphoric acid (Sinopharm Chemical Reagent Co., Ltd., batch numbers 20240612 and 20240816 respectively), acetonitrile (Thermo Fisher Scientific Inc., batch number: F24057205), experimental water was ultrapure water, acetonitrile was chromatographic grade, and the others were chemically pure.

[0059] The preparation method of the standard decoction of Chuanmutong is as follows: weigh 100 g of Chuanmutong slices, put them in a decoction pot, add 1200 mL of pure water, soak for 30 minutes, boil over high heat, and then simmer for 40 minutes over low heat. After the extract is filtered through a 200-mesh sieve, add 1000 mL of pure water to the residue, boil over high heat, and then simmer for 30 minutes over low heat. Combine the two filtrates, concentrate them on a rotary evaporator to a solid-liquid ratio of about 1:1, set the concentration temperature to 65°C, place the concentrate on a tray, dry it in a low-temperature vacuum, and pulverize it.

[0060] Example 1

[0061] This embodiment provides a method for establishing a characteristic spectrum of hydrangea chinensis medicine

[0062] The chromatographic conditions of the high performance liquid chromatography used in this embodiment are: using octadecyl bonded silica gel as the filler (column length is 100 mm, column inner diameter is 2.1 mm, particle size is 1.6 μm), using acetonitrile as mobile phase A, using 0.1% phosphoric acid solution as mobile phase B, and performing gradient elution according to the provisions in Table 1 below; the flow rate is 200 μm per minute.

[0063] 0.25ml; column temperature 30℃; detection wavelength 331nm. The theoretical plate number calculated based on the ferulic acid peak should not be less than 10000.

[0064] Table 1 Gradient elution program

[0065] Time (minutes) Mobile phase A (%) Mobile phase B (%) 0-6 3→9 97→91 6-9 9→10 91→90 9-11 10→15 90→85 11-15 15→25 85→75 15-20 25→30 75→70 20-25 30→35 70→65

[0066] Preparation of reference solution: Take 0.5g of Hydrangea chinensis reference medicinal material, place it in a stoppered conical bottle, add 20ml of water, heat and reflux for 30 minutes, cool, shake well, filter, and take the filtrate as the reference medicinal material reference solution. Take an appropriate amount of caffeic acid and ferulic acid reference substances, accurately weigh them, add 70% methanol to make a mixed solution containing 10μg of caffeic acid and 5μg of ferulic acid per 1ml, and use it as the reference substance solution.

[0067] Preparation of test solution:

[0068] When the test sample is a medicinal material or a decoction piece, take about 0.5 g of the medicinal material or decoction piece powder (passed through a No. 3 sieve), accurately weigh it, place it in a stoppered conical flask, add 20 ml of water, and heat it.

[0069] When the test sample is a formula granule, take an appropriate amount of the granule, grind it finely, take an appropriate amount of this product, grind it finely, take about 0.3g, weigh it accurately, put it in a stoppered conical bottle, accurately add 15ml of 70% methanol, weigh it, ultrasonically treat it (power 300W, frequency 40kHz) for 30 minutes, let it cool, weigh it again, make up the lost weight with 70% methanol, shake it well, filter it, and take the filtrate. Among them, if the test sample is a hydrangea vine water extract, and the hydrangea vine water extract is a hydrangea vine extract or a hydrangea vine water extract concentrate, first dry it under reduced pressure (drying temperature is 65℃) into a standard decoction or formula granule, and then prepare the test sample solution.

[0070] Determination method: Accurately pipette 2 μl of reference solution and test solution respectively, inject into ultra-high performance liquid chromatography, and determine.

[0071] Example 2

[0072] The present embodiment provides a method for preparing a formula granule of Hydrangea chinensis, comprising: taking 8300 g of Hydrangea chinensis slices, extracting twice, adding 12 times the amount of water for one decoction, soaking for 30 minutes, boiling over high heat, and then decocting over low heat for 40 minutes, filtering the extract through a 200-mesh filter bag, adding 10 times the amount of water to the residue, boiling over high heat, and then decocting over low heat for 30 minutes, combining the two filtrates, and concentrating under reduced pressure at 70°C to an extract with a density of 1.00-1.05 (50°C-60°C); transferring the concentrated extract into a spherical concentrate Continue to concentrate in a concentrator or scraper concentrator. When the extract is concentrated to a relative density of 1.05-1.15 (30℃-40℃), weigh an appropriate amount of maltodextrin, stir in a dry raw material tank, heat to 60℃-80℃ for insulation, stir evenly, carry out belt vacuum drying, set the drying temperature to below 100℃, crush, sieve, and collect the material; add an appropriate amount of maltodextrin to the above extract powder, mix evenly, dry granulate, and make 1000g of granules, and package with polyester / aluminum / polyethylene medicinal composite film: 200g / bag, 1g / bag.

[0073] In this embodiment, 15 batches of hydrangea chinensis slices and hydrangea chinensis granules prepared therefrom were used as test samples and the detection method of Example 1 was used for determination. The "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System" (2012.1 Edition) was used according to the obtained liquid phase spectrum, and the S1 liquid phase spectrum was used as the reference spectrum. The reference spectrum was obtained by median calculation, and the common peaks were identified. A total of 8 common peaks were identified, such as Figure 1 shown.

[0074] At the same time, samples of various production links in the process research of hydrangea chinensis Chinese medicine formula granules were collected, and the HPLC spectra of each sample were obtained according to the method in Example 1, such as Figure 2As shown, S1 is the reference medicinal material of Hydrangea chinensis; S2 is the standard decoction of Hydrangea chinensis; S3 is the extract of Hydrangea chinensis production; S4 is the concentrated solution of Hydrangea chinensis production; S5 is the dry powder of Hydrangea chinensis production; S6 is the formula granules of Hydrangea chinensis. The changes of the eight common peaks of the samples in each link are analyzed. According to the analysis of the liquid phase spectrum of the samples in each link, it can be seen that the characteristic spectrum of each link in the preparation of the formula granules of Hydrangea chinensis has not changed significantly, and each characteristic peak is very stable.

[0075] Take the hydrangea vine formula granules and the mixed solution of the chemical components caffeic acid and ferulic acid reference substances, and integrate the 8 characteristic peaks in the characteristic spectrum of the test solution according to the above chromatographic conditions. The results are as follows Figure 3 As shown, peak 3 is caffeic acid and peak 6 is ferulic acid.

[0076] Example 3

[0077] The present embodiment provides a method for establishing a characteristic spectrum of a hydrangea vine drug, and its characteristic spectrum and application in a quality control method of hydrangea vine formula granules, including obtaining a characteristic spectrum of hydrangea vine formula granules, the characteristic spectrum at least including chromatographic peaks corresponding to caffeic acid and ferulic acid, in the characteristic spectrum of hydrangea vine formula granules, taking characteristic peak No. 3 where caffeic acid is located as reference peak S1, the relative retention times of peak 1, peak 2, peak 4 and reference peak S1 are 0.77±0.08, 0.96±0.10, 1.14±0.11 respectively; taking characteristic peak No. 6 where ferulic acid is located as reference peak S2, the relative retention times of peak 5, peak 7, peak 8 and reference peak S2 are 0.91±0.09, 1.06±0.11, 1.34±0.13 respectively. Obtain the ratio of the peak area of ​​peak 3 to the total peak area of ​​8 characteristic peaks, which is not less than 0.064.

[0078] The method for obtaining the above quality control indicators is as follows:

[0079] 1. Confirmation of characteristic peaks and relative retention times in the quality control indicators of Hydrangea chinensis formula granules

[0080] First, 15 batches of standard decoctions prepared from hydrangea chinensis slices were obtained, and the standard decoctions were tested using the detection method of Example 1 to obtain the characteristic spectra of the standard decoctions of each batch and the retention time and relative retention time of each characteristic peak. The characteristic peak where caffeic acid is located is used as the reference peak S1, and the characteristic peak where ferulic acid is located is used as the reference peak S2. The relative retention times of peaks 1, 2, and 4 relative to peak 3, and the relative retention times of peaks 5, 7, and 8 relative to peak 6 are calculated, and the results are shown in Table 2.

[0081] Table 2 Actual retention time and relative retention time of 15 batches of standard decoction of Hydrangea chinensis dried paste powder

[0082]

[0083] From the characteristic spectra of 15 batches of standard decoction of Hydrangea chinensis, it can be seen that the RSD coefficients of the relative retention time of each characteristic peak are all within 2.0%, which meets the quality control requirements. Taking the characteristic peak No. 3 where caffeic acid is located as the reference peak S1, the relative retention times of peak 1, peak 2, peak 4 and reference peak S1 are calculated to be 0.77±0.08, 0.96±0.10, and 1.14±0.11, respectively; taking the characteristic peak No. 6 where ferulic acid is located as the reference peak S2, the relative retention times of peak 5, peak 7, peak 8 and reference peak S2 are 0.91±0.09, 1.06±0.11, and 1.34±0.13, respectively.

[0084] 2. Confirmation of the relative content of caffeic acid in hydrangea vine formula granules

[0085] In order to confirm the relative content of caffeic acid, the ratio of the peak area of ​​characteristic peak 3 to the total peak area of ​​8 characteristic peaks in each batch of standard decoction was calculated. The results are shown in Table 3 below.

[0086] Table 3 The ratio of peak area of ​​characteristic peak 3 to total peak area of ​​15 batches of standard decoction of Hydrangea chinensis

[0087] batch number ratio XQT-BT-01 0.0958 XQT-BT-02 0.0878 XQT-BT-03 0.0928 XQT-BT-04 0.0783 XQT-BT-05 0.0768 XQT-BT-06 0.0759 XQT-BT-07 0.0963 XQT-BT-08 0.2335 XQT-BT-09 0.0907 XQT-BT-10 0.0779 XQT-BT-11 0.0753 XQT-BT-12 0.0733 XQT-BT-13 0.0789 XQT-BT-14 0.1039 XQT-BT-15 0.0766 average value 0.0843 Average ± 30% 0.059-0.110 SD value 0.010 Mean ± 2SD range 0.064-0.104

[0088] The caffeic acid peak area ratio data of 15 batches of standard decoctions of Hydrangea chinensis contain a discrete value of 0.2335. After removing it, the actual measured range is 0.0753-0.1039, and the mean is 0.0843. The SD is 0.010, the mean ± 3SD range is 0.055-0.114, and the mean ± 30% range is 0.050-0.110. Although the mean ± 3SD range is wider than the mean ± 30% range, there are many batches of data far from the lower limit. The mean ± 2SD range is selected as the lower limit of the peak area ratio, that is, the ratio of the peak area of ​​characteristic peak 3 to the total peak area of ​​8 characteristic peaks should not be less than 0.064.

[0089] The ratio of the peak area of ​​characteristic peak 3 to the total peak area of ​​8 characteristic peaks was also calculated for the medicinal pieces, and the mean-2SD range was selected as the lower limit of the peak area ratio, and the lower limit of the peak area ratio was 0.061. In order to be consistent with the range of the standard decoction, the quality control index was set as: the ratio of the peak area of ​​characteristic peak 3 to the total peak area of ​​8 characteristic peaks should not be less than 0.064.

[0090] In this example, three batches of Hydrangea chinensis slices were used to prepare the formula granules using the preparation method of the formula granules in the above-mentioned Example 2 to verify the use of the Hydrangea chinensis formula granules. After detection using the same method as in Example 1, the retention time and relative retention time of the characteristic spectra, as well as the peak area ratios of the three batches of Hydrangea chinensis formula granules are shown in Tables 4 and 5.

[0091] Table 4 Retention time and relative retention time of three batches of hydrangea vine formula granules

[0092]

[0093] Table 5 Relative peak area ratios of three batches of hydrangea vine formula granules

[0094]

[0095] It can be seen from Tables 4 and 5 that the quality control indicators prepared by the method for preparing the formula granules of the present invention are all within the specified range values, further proving the consistency of the composition and content of the formula granules with the standard decoction.

[0096] Example 4

[0097] This embodiment provides a method for establishing a characteristic spectrum of a hydrangea chinensis medicament, and its characteristic spectrum and application, including confirmation of the ferulic acid content in the quality control index of hydrangea chinensis granules.

[0098] The content range and yield of ferulic acid in different batches of standard decoctions of Hydrangea chinensis obtained by the method of Examples 1-2 were converted according to the prepared amount (i.e., one gram of Hydrangea chinensis formula granules is equivalent to 8.3 grams of decoction pieces) to obtain the content of ferulic acid in the formula granules (mg / g). The results are shown in Table 6.

[0099] Conversion calculation formula: Ferulic acid content in formula granules = paste yield * Ferulic acid content in standard decoction * 8.3

[0100] Table 6 Calculation results of ferulic acid content in 15 batches of Hydrangea chinensis granules

[0101] Sample batch number Cream yield / % Standard soup dry paste powder content mg / g Converted into formula particle content mg / g XQT-BT-01 10.92 0.352 0.32 XQT-BT-02 9.99 0.216 0.18 XQT-BT-03 9.96 0.398 0.33 XQT-BT-04 8.45 0.440 0.31 XQT-BT-05 9.25 0.519 0.40 XQT-BT-06 9.04 0.372 0.28 XQT-BT-07 9.73 0.321 0.26 XQT-BT-08 8.85 0.407 0.30 XQT-BT-09 8.86 0.528 0.39 XQT-BT-10 8.72 0.303 0.22 XQT-BT-11 9.37 0.474 0.37 XQT-BT-12 8.16 0.235 0.16 XQT-BT-13 8.45 0.469 0.33 XQT-BT-14 8.84 0.475 0.35 XQT-BT-15 9.54 0.327 0.26

[0102] According to the results of Table 6, the content of ferulic acid in the standard decoction of Hydrangea chinensis is in the range of 0.22mg / g-0.53mg / g; according to the amount of preparation, the content is converted (i.e., each gram of Hydrangea chinensis formula granules is equivalent to 8.3 grams of crude drug), and the content of ferulic acid in the formula granules of Hydrangea chinensis is in the range of 0.18mg / g-0.40mg / g. According to the results of the standard decoction, the measured range is 8.16%-10.92%, the mean is 9.21%, the mean ±30% range is 6.45%-11.97%, and the mean ±3SD range is 7.03%-11.39%. The cream yields of 15 batches of standard decoctions are within the mean ±30% and ±3SD of the cream yield, and no outliers appear. By comparison, the cream yield of Chuanmutong is relatively low, and the range of ±30% cream yield is relatively wide, which is more conducive to the control of product quality, so ±30% is used as its cream yield range. The yield of the standard decoction of Caulis Atractylodes lancea was determined to be in the range of 6.5%-12.0%, i.e. the preparation amount = 1 / 8.3.

[0103] According to the formula: the content of ferulic acid in the formula granules = the paste yield * the content of ferulic acid in the standard decoction * 8.3, the average content of 15 batches of dry paste powder of Chuanmutong standard decoction converted into Chuanmutong formula granules is 0.30 mg / g, and the range of the average value ± 3SD is 0.08-0.51 mg / g.

[0104] This example also obtained the main indicators of standard decoctions and formula granules prepared from 15 batches of medicinal pieces. The results are shown in Table 7.

[0105] Table: Main indicators of standard decoctions and formula granules prepared from 15 batches of decoction pieces

[0106] project Standard decoction Formula Granules Cream yield 6.5％-12.0％ 7.0％→11.5％ content 0.10mg / g-0.70mg / g 0.08mg / g-0.51mg / g Transfer rate 22.6％-60.5％ 22.6％-60.5％ leachate Not less than 18.0% Not less than 14.0%

[0107] The above index range limits are compared to show that the medicinal substances of the formula granules prepared by the present invention are consistent with those of the standard decoction.

[0108] Example 5

[0109] This example verifies the repeatability, intermediate precision, stability and linearity of the characteristic spectrum and ferulic acid content determination obtained by the detection method in Example 1, as follows:

[0110] 1. Repeatability

[0111] According to the detection method in Example 1, 6 test sample solutions were prepared from the same hydrangea vine formula granules to obtain the characteristic spectrum of the test sample, and caffeic acid and ferulic acid were used as reference solutions.

[0112] Taking characteristic peak No. 3 where caffeic acid is located as reference peak S1, the relative retention times of peaks 1, 2, and 4 with respect to reference peak S1 were calculated; taking characteristic peak No. 6 where ferulic acid is located as reference peak S2, the relative retention times of peaks 5, 7, and 8 were calculated. The RSD of the retention time of each characteristic peak was in the range of 0.02-0.42%, and the RSD of the relative peak area was in the range of 0.52-1.61%, indicating that the characteristic spectrum had good repeatability.

[0113] At the same time, the ferulic acid content was determined, and the average ferulic acid content was 0.165 mg / g, with an RSD of 1.80%, which met the repeatability requirements of the methodology validation.

[0114] 2. Accuracy

[0115] Take an appropriate amount of ferulic acid reference substance, accurately weigh it, and add 70% methanol solution to make a solution containing 53.12μg of ferulic acid per 1ml. Take the formula granules with known content under the repeatability item, grind them finely, take about 0.3g, accurately weigh them, and add 9 portions in total. Add 1ml, 2ml, and 3ml of ferulic acid reference substance solution to each 3 portions respectively, and then add 70% methanol to 20ml respectively, and operate according to Example 1. The results show that the recovery rate measured ranges from 100.24% to 104.57%, the average recovery rate is 102.74%, and the RSD value is 1.58%, which meets the recovery rate requirements of the methodology verification, indicating that the results measured by this method are accurate and suitable for the content determination of this product.

[0116] 3. Intermediate precision

[0117] Different analysts used Waters, PDA detector detector at different times, took the test solution, and measured it according to the method in Example 1 to obtain its characteristic spectrum. The characteristic peak No. 3 where caffeic acid is located was used as the reference peak S1, and the relative retention times of peaks 1, 2, and 4 with respect to the reference peak S1 were calculated; the characteristic peak No. 6 where ferulic acid is located was used as the reference peak S2, and the relative retention times of peaks 5, 7, and 8 were calculated, and the RSD% was calculated, and the ferulic acid content was determined at the same time.

[0118] Results: The relative retention time RSD of each characteristic peak in the characteristic spectrum of Hydrangea chinensis formula granules was determined by Waters PDA detector, and the RSD of the relative peak area was in the range of 0.1%-0.84%, and 0.7%-1.3%, indicating that the relative retention time of the characteristic spectrum between different instruments meets the analysis requirements.

[0119] The average content of ferulic acid was 0.167 mg / g, the RSD value between different instruments was 1.60%, and the RSD of the test results of the repeatability test was 1.80%, which met the precision requirements of the method validation.

[0120] 4. Stability

[0121] Take the same test solution, and determine the characteristic spectrum according to the method of Example 1 at 0, 2, 4, 6, 8, 12, and 24 hours, respectively. Take the characteristic peak No. 3 where caffeic acid is located as the reference peak S1, and calculate the relative retention time of peak 1, peak 2, peak 4 and the reference peak S1; take the characteristic peak No. 6 where ferulic acid is located as the reference peak S2, calculate the relative retention time of peak 5, peak 7, and peak 8, calculate the RSD%, and determine the ferulic acid content at the same time. As a result, the relative retention time RSD of each characteristic peak is 0%, and the relative peak area ratio of each characteristic peak is between 0.06% and 0.75% of the specified value, indicating that the stability of the characteristic spectrum of the hydrangea vine test solution is good at 24 hours.

[0122] At the same time, the peak area of ​​ferulic acid was measured. Within 24 hours, the RSD value of the peak area of ​​ferulic acid was 0.52%, indicating that the sample was stable within 24 hours and met the requirements for content determination.

[0123] 5. Linear

[0124] Accurately weigh 6.68 mg (99.4%) of the reference substance ferulic acid, dissolve it in 70% methanol and dilute to 25 mL, shake well, and the concentration of the ferulic acid reference substance mother solution is 265.60 mg / L, which is used as the reference substance solution stock solution. Take 0.5 ml, 1.0 ml, 2.0 ml, 3.0 ml, 4.0 ml, and 5.0 ml of the mother solution to a 25 ml volumetric flask in turn and dilute to the mark with 70% methanol to obtain 6 reference substance solutions of different concentration levels, namely DZ-1, DZ-2, DZ-3, DZ-4, DZ-5, and DZ-6.

[0125] Accurately pipette 2 μl of the above filtrate and inject it into the ultra-high performance liquid chromatograph. Determine the peak area of ​​the ferulic acid chromatographic peak according to the method in Example 1. Draw a standard curve with the peak area of ​​the ferulic acid chromatographic peak as the ordinate and the concentration of ferulic acid as the abscissa. Figure 4 As shown, the regression equation of ferulic acid was obtained as y=36.406x-26.285, R=0.9993, and its linear range was 5.31 μg / ml-53.12 μg / ml.

[0126] Example 6

[0127] This example uses a method for detecting hydrangea chinensis and formula granules to identify the origins of hydrangea chinensis, akebia schoenensis and standard decoctions, as follows:

[0128] The characteristic spectrum obtained by the method of Example 1 was used to identify the chromatographic peaks of caffeic acid and ferulic acid. The peak area ratios of caffeic acid / ferulic acid in Hydrangea chinensis, Akebia dasyphylla and standard decoctions are shown in Tables 8 and 9, and the two base source comparison spectra are shown in Spectrum 5. Discrete values ​​2.670 and 2.418 were removed from Hydrangea chinensis decoction pieces, the relative peak area ratio average value was 1.621, the average value ± 30% range was 1.135-2.108, and the average value ± 2SD range was 1.210-1.621, so the relative peak area ratio of the decoction pieces was 1.1-2.1. A discrete value 1.741 was removed from the corresponding standard decoction, and the relative peak area ratio of 14 batches of standard decoctions was 0.842, the average value ± 30% range was 0.589-1.094, and the average value ± 2SD range was 0.621-1.063, so the relative peak area ratio of the standard decoction was 0.59-1.1. Based on the comprehensive results, when the peak area ratio of caffeic acid / ferulic acid is 0.59-2.1, the base source corresponding to the medicinal pieces and standard decoction is Hydrangea chinensis.

[0129] The discrete value 40.90 was removed from the Xiaoketong decoction pieces, the relative peak area ratio average was 13.280, the average ± 30% range was 9.296-17.264, and the average ± 2SD range was 7.277-19.283, so the relative peak area ratio of the decoction pieces was 7.3-19.3. The corresponding standard decoctions removed a discrete value of 39.73 and 99.60, and the relative peak area ratio of 14 batches of standard decoctions was 11.579, the average ± 30% range was 8.105-15.052, and the average ± 2SD range was 11.177-11.981, so the relative peak area ratio of the standard decoction was 8.1-15.1. Taking into account the discrete values ​​in the Xiaoketong decoction pieces and the standard decoction, it was comprehensively obtained that when the peak area ratio of caffeic acid / ferulic acid was ≥7.3, the base source corresponding to the decoction pieces and the standard decoction was Xiaoketong.

[0130] Table 8 Peak area ratios of caffeic acid / ferulic acid in 15 batches of standard decoctions of Hydrangea chinensis

[0131]

[0132] Table 9 Peak area ratio results of caffeic acid / ferulic acid in 15 batches of standard decoction of Akebia trifoliata

[0133]

[0134] Example 7

[0135] This embodiment provides a method for screening hydrangea chinensis slices, and uses a method for detecting hydrangea chinensis and hydrangea chinensis formula granules to screen the slices, as follows:

[0136] The characteristic spectrum of the hydrangea vine slices was obtained by the method of Example 1, and the slices with a ratio of the peak area of ​​peak 3 to the total peak area of ​​8 characteristic peaks of ≤0.2 were screened out. The yield of the standard decoction prepared from the corresponding batch of slices and the content of ferulic acid were obtained for verification. As shown in Table 10, the slices screened by the screening method had a higher content of ferulic acid and the quality of each batch was more stable.

[0137] Table 10 Quality comparison of different batches It can be seen from the above table that when the peak 3 / total peak area ratio in the decoction piece is not more than 0.20, the ferulic acid content in the standard decoction ranges from 0.398mg / g to 0.528mg / g, the paste yield ranges from 8.45% to 9.96%, and the extract ranges from 24.65% to 29.45%; the determination method of the extract refers to the hot soaking method under (General Rules 2201 of the 2020 edition of the Chinese Pharmacopoeia). 1g of hydrangea vine formula granules is equivalent to 8.3g of crude drug, and the preparation amount is 12%. When the peak 3 / total peak area ratio of the decoction piece is greater than 0.20, the ferulic acid content in the standard decoction ranges from 0.216mg / g to 0.372mg / g, which is relatively low; the paste yield ranges from 8.16% to 10.92%, with a slightly larger range fluctuation, and the extract ranges from 21.43% to 25.22%, which is relatively low. Therefore, when the ratio of the peak area of ​​the 3rd peak of the medicinal material to the total peak area of ​​the 8 characteristic peaks is stipulated by the characteristic spectrum to be no more than 0.20, the quality of the prepared hydrangea vine formula granules is stable and high.

[0138] In order to prove its effect, three batches of samples were used for verification in this embodiment. Different processing techniques were used for the same batch of medicinal materials, so that the peak 3 / total peak area ratio of the hydrangea vine formula granules before feeding was greater than 0.20 and less than 0.20, respectively. The same production process was used for verification, and the obtained product results are shown in Table 11 below.

[0139] Table 11 Comparison of validation batch quality

[0140]

[0141] Through data comparison, it can be seen that when the ratio of the area of ​​the decoction piece peak 3 / total peak is no more than 0.20, the ferulic acid content of the obtained formula granules is higher and the paste yield range is more stable and moderate, which meets the quality control of the product, and the quality of the formula granules is more stable and reliable.

[0142] Example 8

[0143] The difference between this embodiment and embodiment 1 is that the detection wavelength is different, as follows:

[0144] A diode array detector was used to scan the test solution of the dry paste powder of the standard decoction of Hydrangea chinensis at a wavelength of 190nm-400nm, and the analysis was performed through the isobsorption line diagram. The results showed that when the detection wavelength was around 331nm, the response values ​​of each chromatographic peak were moderate and the separation was good. After comprehensive consideration, 331nm was finally determined as the detection wavelength.

[0145] Take the same test solution and measure it according to the method of Example 1, change the detection wavelength to 294.2nm and 320nm, and obtain its characteristic spectrum. Figure 6 and Figure 7As shown. Taking characteristic peak No. 3 where caffeic acid is located as reference peak S1, the relative retention times of peaks 1, 2, and 4 with reference peak S1 are calculated; taking characteristic peak No. 6 where ferulic acid is located as reference peak S2, the relative retention times of peaks 5, 7, and 8 are calculated, and the retention time RSD of each characteristic peak is in the range of 0.02-0.42%.

[0146] At the same time, the changes in the content of ferulic acid at different wavelengths were recorded, and the RSD value of the ferulic acid content was 0.7%, indicating that the method had good durability at different wavelengths and met the analysis requirements.

[0147] Example 9

[0148] The difference between this embodiment and Example 1 is that the detection flow rate is different, as follows: take the same test solution, measure according to the method of Example 1, only change the flow rate to 0.30ml / min, 0.20ml / min, obtain the difference between this embodiment and Example 1 is that the detection flow rate is different, as follows: take the same test solution, measure according to the method of Example 1, only change the flow rate to 0.30ml / min, 0.20ml / min, obtain the control characteristic spectrum of the test sample at 0.30ml / min, 0.20ml / min, such as Figure 8 As shown. Taking the characteristic peak No. 3 where caffeic acid is located as the reference peak S1, the relative retention times of peaks 1, 2, and 4 with reference peak S1 are calculated; taking the characteristic peak No. 6 where ferulic acid is located as the reference peak S2, the relative retention times of peaks 5, 7, and 8 are calculated. The results show that the relative retention time of each characteristic peak is between 0.11% and 0.76% of the specified value, indicating that the characteristic spectrum method has good durability for different flow rates.

[0149] At the same time, the change of ferulic acid content under different flow rates was determined. Under different flow rates, the RSD value of ferulic acid content was 1.2%, indicating that the method has good durability for different flow rates and meets the analysis requirements.

[0150] Example 10

[0151] The difference between this embodiment and embodiment 1 is that the detection column temperature is different, as follows:

[0152] Take the same test solution and measure it according to the method of Example 1 above, only changing the column temperature to 25°C and 35°C, and obtain the control characteristic spectrum of the test solution at 25°C and 35°C, as shown in Fig. 9 As shown. Taking characteristic peak No. 3 where caffeic acid is located as reference peak S1, the relative retention times of peaks 1, 2, and 4 with reference peak S1 are calculated; taking characteristic peak No. 6 where ferulic acid is located as reference peak S2, the relative retention times of peaks 5, 7, and 8 are calculated, and the RSD is calculated.

[0153] Results The relative retention time of each characteristic peak was between 0.41% and 0.87% of the specified value, indicating that the characteristic spectrum method had good durability for different column temperatures.

[0154] At the same time, the content change of ferulic acid at different column temperatures was determined. The RSD value of the content of ferulic acid at different column temperatures was 1.3%, indicating that the method had good durability for different column temperatures and met the analysis requirements.

[0155] Embodiment 11

[0156] The difference between this embodiment and embodiment 1 is that the chromatographic columns are different, as follows:

[0157] Take the same test solution and measure it according to the method of Example 1 above. According to the determined chromatographic conditions, two different chromatographic columns (① Waters T3 (2.1×100mm, 1.6μm; ②AgilentZORBAX Eclipse Plus C18 (2.1mm×100mm, 1.8μm); ③Waters ACQUITY BEH C18 (2.1 mm × 100 mm, 1.7 μm); The results are shown in Table 12. Figure 10-12 .

[0158] Table 12 Relative retention time tables of different chromatographic columns

[0159]

[0160] However, peak 2 in column ② obviously overlaps, and peak 8 collapses; peak 1 in column ③ overlaps, and the baseline is not flat from 13 minutes to 18 minutes. Column 1 has the best separation effect. This shows that this method is more suitable for column ①, and it is necessary to use a chromatographic column Waters T3 (2.1×100 mm, 1.6 μm).

[0161] Example 12

[0162] This embodiment provides another method for preparing hydrangea vine formula granules, which is different from the method for preparing hydrangea vine formula granules disclosed in Example 2 in that the same batch of medicinal pieces is selected, and the amount of water added is changed twice, namely, Comparative Example 1 (10 times, 8 times), Comparative Example 2 (10 times, 10 times), and Comparative Example 3 (12 times, 12 times), and the filtration mesh is 300 mesh; the detection method of Example 1 is used for determination; spray drying is used; the auxiliary material is micro-powder silica gel, and the results are shown in Table 13.

[0163] Table 13 Investigation of different water addition amounts

[0164] experiment Sample weight (g) Water addition amount (times) Cream yield (%) Ferulic acid content mg / g Comparative Example 1 100.15 First fry 10; second fry 8 8.34 0.344 Comparative Example 2 100.13 First fry 10; second fry 10 9.42 0.431 Comparative Example 3 100.23 12 for one fry; 12 for two fry 10.56 0.498 Example 2 100.19 12 for one fry; 10 for two fry 10.54 0.502

[0165] By comparison, the paste yield and ferulic acid content of the formula granules prepared by the method in this Example 2 are higher than those of the three comparative examples. Therefore, the method in Example 2 is the most suitable preparation method.

[0166] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

[0167] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for establishing a characteristic map of hydrangea chinensis medicine, characterized in that: include: Samples were taken to prepare a hydrangea vine test solution, and ultra-high performance liquid chromatography was used to obtain the characteristic spectrum of the test solution and the reference peak of the reference solution. The chromatographic conditions of the ultra-high performance liquid chromatography were as follows: Chromatographic column: chromatographic column with octadecylsilane bonded silica gel as filler; Detection wavelength: 321-331nm; Mobile phase A: acetonitrile, mobile phase B: 0.05-0.2% phosphoric acid solution; Column temperature: 25-35°C; Flow rate: 0.2-0.3ml / min; The number of theoretical plates calculated based on the ferulic acid peak is not less than 10,000; Reference solution: Ferulic acid concentration 10μg / ml, caffeic acid concentration 20μg / ml Gradient elution.

2. The method for establishing the characteristic map of the hydrangea chinensis medicine according to claim 1, characterized in that: The gradient elution method is: 0-6min, the volume ratio of mobile phase A to mobile phase B was 3-9%:97-91%; 6-9min, the volume ratio of mobile phase A to mobile phase B is 9-10%:91-90%; 9-11min, the volume ratio of mobile phase A to mobile phase B is 10-15%:90-85%; 11-15min, the volume ratio of mobile phase A to mobile phase B is 15-25%:85-75%; 15-20min, the volume ratio of mobile phase A to mobile phase B is 25-30%:75-70%; 20-25min, the volume ratio of mobile phase A to mobile phase B is 30-35%:70-65%.

3. The method for establishing the characteristic spectrum of the hydrangea chinensis medicine according to claim 1, characterized in that: The test sample is at least one of hydrangea chinensis medicinal materials, hydrangea chinensis decoction pieces, hydrangea chinensis water extract and hydrangea chinensis formula granules.

4. The method for establishing the characteristic map of the hydrangea chinensis medicine according to claim 3, characterized in that: When the test sample is hydrangea vine medicinal material or hydrangea vine decoction pieces, the preparation method of the test sample includes: taking the test sample powder, adding water to heat and reflux, cooling, shaking, filtering and taking the filtrate to obtain; when the test sample is hydrangea vine water extract or hydrangea vine formula granules, the preparation method of the test sample includes: taking the test sample powder, adding solvent, weighing quantitatively, ultrasonic treatment, cooling and weighing again, making up the lost weight with solvent, shaking, filtering and taking the filtrate to obtain.

5. The method for establishing the characteristic map of hydrangea chinensis medicine according to claim 1, characterized in that: The method also includes the preparation of reference substance solution, injecting reference substance solutions of different concentrations by ultra-high performance liquid chromatography, and drawing a standard curve of the reference substance with the injection concentration as the horizontal axis and the peak area as the vertical axis.

6. A characteristic spectrum of a hydrangea vine drug, characterized in that: The characteristic spectrum is established by the establishment method described in any one of claims 1 to 4, wherein the characteristic spectrum includes 8 characteristic peaks, with peak 3 where caffeic acid is located as the reference peak S1, and the relative retention times of peak 1, peak 2, peak 4 and reference peak S1 are 0.77±0.08, 0.96±0.10, and 1.14±0.11, respectively; with peak 6 where ferulic acid is located as the reference peak S2, the relative retention times of peak 5, peak 7, peak 8 and reference peak S2 are 0.91±0.09, 1.06±0.11, and 1.34±0.13, respectively.

7. The use of the characteristic map of the hydrangea chinensis medicine as claimed in claim 5 in the quality control method of the hydrangea chinensis formula granules, characterized in that: This includes obtaining a characteristic spectrum of hydrangea vine formula particles, in which the substance content of peak No. 6 in the characteristic spectrum ranges from 0.08 mg / g to 0.51 mg / g.

8. The use of the characteristic map of the hydrangea chinensis medicine as claimed in claim 7 in the quality control method of the hydrangea chinensis formula granules, characterized in that: The ratio of the area of ​​peak No. 3 to the total area of ​​the eight characteristic peaks is not less than 0.

064.

9. The use of the characteristic map of the hydrangea chinensis medicine as claimed in claim 5 in the identification of the origins of hydrangea chinensis and Akebia scabra, characterized in that: Including obtaining the characteristic spectra of the corresponding products respectively, calculating the area ratio of Peak 3 to Peak 6, when the area ratio of Peak 3 / Peak 6 is 0.59-2.1, the base source of the corresponding product is Hydrangea chinensis, and when the area ratio of Peak 3 / Peak 6 is ≥7.3, the base source of the corresponding product is Akebia trifoliata.

10. The use of the characteristic map of the hydrangea chinensis medicine according to claim 5 in the screening of hydrangea chinensis decoction pieces, characterized in that: The ratio of the area of ​​peak No. 3 to the total area of ​​the eight characteristic peaks is ≤0.2.

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