Construction method of characteristic chromatogram of Yemingsha medicinal material, decoction piece, standard decoction and dispensing granule

Abstract

The present application relates to the technical field of analytical detection, in particular to a characteristic chromatogram construction method of Yemingsha medicinal materials, decoction pieces, standard decoction or formula granules, and provides a characteristic chromatogram construction method of Yemingsha medicinal materials, decoction pieces, standard decoction or formula granules, and a Yemingsha medicinal material, decoction piece, standard decoction or formula granule identification method.The present application performs single-species research on Yemingsha formula granules, scientifically identifies Yemingsha, has the characteristics of comprehensive detection and simple operation, can effectively identify Yemingsha formula granules, and can ensure the uniformity and stability of Yemingsha formula granule quality.The method is suitable for Yemingsha medicinal materials, decoction pieces, standard decoction or formula granules.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of analytical detection technology, in particular to the construction method of characteristic chromatogram of Yemingsha medicinal material, decoction piece, standard decoction or formula granule. BACKGROUND

[0002] Yemingsha has a long history of medicinal use, and was first recorded in Shennong Bencao Jing, and was listed as a medium grade. It was not until the Song Dynasty that it was referred to as Yemingsha. The medicinal Yemingsha is now the dried feces of bats of the Vespertilionidae family, such as Vespertilio superans Thomas and other bats. Yemingsha has been used by physicians of past dynasties to treat diseases. For example, Fangmai Zhengzong records: "for treating scrofula: Yemingsha 3 coins, white clam shell 5 coins (fire calcined). Grind into fine powder, make into pills with rice, like green beans. Take 2 coins every night, with white soup." Renzai Zhizhi Fang records: "for treating ulcer and pus discharge: Yemingsha 1 liang, cassia 0.5 liang, frankincense 1 fen. Make into powder, add 0.5 liang of dry granulated sugar, and apply with well water." Quanyou Xinjian records: "for treating all kinds of malnutrition and toxic: Yemingsha 5 coins, put into a pottery bottle, add 3 liang of lean pork, cut thinly, put into the bottle, cook with water until cooked, eat the meat with children, drink the juice, remove the toxic from the lower abdomen; next, use 4 liang of ginger, fry with skin, mix with 1 liang of Huanglian powder, make into pills with millet, take 3 times a day." Modern Ruan Xiande, editor of Zhonghua Yao Hai, records: "Yemingsha is bitter and cold, and can disperse and resolve, clear heat, and enter the liver meridian, and can remove blood heat from the liver meridian." Zhang Jimin reported that Yemingsha was used to treat central retinitis, vitreous opacity, corneal nebula, ametropia or non-emmetropia, and good results were achieved. Chen Yan used Yemingsha to treat 43 cases of herpes simplex keratitis combined with western medicine, and the total effective rate was 95.4%. Xu Huiguang found that Yemingsha was very effective in treating repeated bleeding in the white part of the eye. The research on the efficacy and clinical application of past dynasties shows that Yemingsha can clear heat and brighten the eyes, disperse blood and resolve accumulation. It is used for liver heat and red eyes, internal and external obstruction nebula, malnutrition, and blood stasis and pain. As one of the cold back small varieties, there is currently a small amount of demand in the market. At present, Yemingsha is traded in small quantities, and Yemingsha is sold in the market of Hehuachi, Bozhou, and Anguo. There is currently no research on content determination and characteristic chromatogram. SUMMARY

[0003] Therefore, the present application provides a construction method of characteristic chromatogram of Yemingsha medicinal material, decoction piece, standard decoction, and formula granule. The present application provides a construction method of characteristic chromatogram of Yemingsha medicinal material, decoction piece, standard decoction, and formula granule, and a method for identifying Yemingsha medicinal material, decoction piece, standard decoction, and formula granule. The present application studies Yemingsha formula granule as a single variety, scientifically identifies Yemingsha, has the characteristics of comprehensive detection and simple operation, can effectively identify Yemingsha formula granule, and can ensure the uniformity and stability of the quality of Yemingsha formula granule. The method is suitable for Yemingsha medicinal material, decoction piece, standard decoction, and formula granule.

[0004] To achieve the above-mentioned purposes of the application, the application provides the following technical solutions.

[0005] The application provides a construction method of characteristic chromatograms of Yemingsha medicinal materials, decoction pieces, standard decoctions and formula granules, comprising the following steps:

[0006] S1, dissolving and extracting the test sample raw material by using a solvent to obtain a test sample solution;

[0007] S2, determining the test sample solution of S1 by using liquid chromatography to obtain characteristic chromatograms of Yemingsha medicinal materials, decoction pieces, standard decoctions or formula granules;

[0008] The test sample raw material comprises Yemingsha medicinal materials, decoction pieces, standard decoctions or formula granules;

[0009] The chromatographic conditions of the liquid chromatography comprise the following: the chromatographic column is a C18 column; the mobile phase A is acetonitrile or methanol, and the mobile phase B is a 0.1% formic acid solution or a 0.1% phosphoric acid solution.

[0010] In some specific embodiments of the application, the mobile phase A is acetonitrile, and the mobile phase B is a 0.1% phosphoric acid solution.

[0011] In some specific embodiments of the application, the construction method further comprises the steps of preparing a reference solution of a control product and / or a reference solution of a control medicinal material;

[0012] The preparation method of the reference solution of the control product comprises the following steps: respectively taking guanine and uric acid, dissolving the guanine and the uric acid by using 0.5% ammonia water to obtain the reference solution of the control product;

[0013] The preparation method of the reference solution of the control medicinal material comprises the following steps: mixing Yemingsha control medicinal material and water, decocting, cooling, filtering, evaporating the filtrate to dryness, dissolving the residue by using 10% methanol, filtering, taking the filtrate to obtain the reference solution of the control medicinal material.

[0014] In some specific embodiments of the application, the decocting time is greater than or equal to 15 min.

[0015] In some specific embodiments of the application, the concentration of the reference solution of the control product comprises 10 μg / mL.

[0016] In some specific embodiments of the application, the solvent of S1 comprises 10% methanol and / or water.

[0017] In some specific embodiments of the application, the mass-volume ratio of (I) and (II) comprises (1:20) to (1:25) in g / mL.

[0018] (I), the Yemingsha medicinal materials, decoction pieces, standard decoctions or formula granules;

[0019] (II) the solvent.

[0020] In some embodiments of the present application, the extraction in S1 comprises decoction and / or ultrasonic extraction; the power of the ultrasonic extraction comprises 600 W, and the frequency comprises 40 kHz; the time of the extraction comprises 15-30 min.

[0021] The time of the decoction is ≥ 15 min.

[0022] In some embodiments of the present application, the preparation method of the test solution of the granules comprises: taking the granules, grinding, mixing with 10% methanol, ultrasonic treatment at a power of 600 W and a frequency of 40 kHz for 15 min, cooling, shaking, filtering, and taking the filtrate, to obtain the test solution of the granules.

[0023] In some embodiments of the present application, the preparation method of the test solution of the Yemingsha medicinal material comprises: taking the powder of the Yemingsha medicinal material, mixing with water, decocting for ≥ 15 min, cooling, filtering, evaporating the filtrate to dryness, dissolving the residue with 10% methanol, filtering, and taking the filtrate, to obtain the test solution of the Yemingsha medicinal material.

[0024] In some embodiments of the present application, the preparation method of the test solution of the decoction piece comprises: taking the powder of the decoction piece, mixing with water, decocting for ≥ 15 min, cooling, filtering, evaporating the filtrate to dryness, dissolving the residue with 10% methanol, filtering, and taking the filtrate, to obtain the test solution of the decoction piece.

[0025] In some embodiments of the present application, the preparation method of the test solution of the standard decoction comprises: taking the powder of the standard decoction, mixing with 10% methanol, ultrasonic treatment at a power of 600 W and a frequency of 40 kHz for 15 min, cooling, and taking the filtrate, to obtain the test solution of the standard decoction.

[0026] In some embodiments of the present application, the elution procedure of the liquid chromatography determination comprises:

[0027] 0-20 min, A phase 0.5-2%, B phase 99.5-98%;

[0028] 20-40 min, A phase 2-5%, B phase 98-95%;

[0029] 40-70 min, A phase 5%, B phase 95%.

[0030] In some embodiments of the present application, the sample loading amount of the liquid chromatography determination comprises 10 μL.

[0031] In some embodiments of the application, the flow rate for the liquid chromatography assay comprises 0.8 mL / min.

[0032] In some embodiments of the application, the detection wavelength for the liquid chromatography assay comprises 248 nm.

[0033] In some embodiments of the application, the column temperature for the liquid chromatography assay comprises 25 °C.

[0034] In some embodiments of the application, in the characteristic chromatogram of the Yumingzhu medicinal material, 2 peaks respectively correspond to the retention time of the peaks of the reference solution of the control substance, taking the peak corresponding to uric acid in the reference solution of the control substance as the S peak, calculating the relative retention time of the remaining characteristic peaks and the S peak, the relative retention time is within ±10% of the characteristic peak value, the characteristic peak values are respectively: 0.88 (peak 2), 1.07 (peak 4), 1.32 (peak 5), 3.68 (peak 6), 5.02 (peak 7), 5.55 (peak 8);

[0035] In the characteristic chromatogram of the decoction piece, 2 peaks respectively correspond to the retention time of the peaks of the reference solution of the control substance, taking the peak corresponding to uric acid in the reference solution of the control substance as the S peak, calculating the relative retention time of the remaining characteristic peaks and the S peak, the relative retention time is within ±10% of the characteristic peak value, the characteristic peak values are respectively: 0.87 (peak 2), 1.08 (peak 4), 1.32 (peak 5), 3.65 (peak 6), 4.98 (peak 7), 5.50 (peak 8);

[0036] In the characteristic chromatogram of the standard decoction, 2 peaks respectively correspond to the retention time of the peaks of the reference solution of the control substance, taking the peak corresponding to uric acid in the reference solution of the control substance as the S peak, calculating the relative retention time of the remaining characteristic peaks and the S peak, the relative retention time is within ±10% of the characteristic peak value, the characteristic peak values are respectively: 0.87 (peak 2), 1.09 (peak 4), 1.29 (peak 5), 3.55 (peak 6), 4.85 (peak 7), 5.37 (peak 8);

[0037] In the characteristic chromatogram of the formula granule, 2 peaks respectively correspond to the retention time of the peaks of the reference solution of the control substance, taking the peak corresponding to uric acid in the reference solution of the control substance as the S peak, calculating the relative retention time of the remaining characteristic peaks and the S peak, the relative retention time is within ±10% of the characteristic peak value, the characteristic peak values are respectively: 0.88 (peak 2), 1.10 (peak 4), 1.37 (peak 5), 3.76 (peak 6), 5.11 (peak 7), 5.65 (peak 8).

[0038] The application also provides a method for identifying Yemingsha medicinal materials, decoction pieces, standard decoctions or formula granules, taking a test sample, using the determination method to obtain a characteristic spectrum of the test sample, two peaks in the characteristic spectrum correspond to the retention time of the peaks of the reference solution of the control sample respectively, taking the peak corresponding to the uric acid as the S peak, calculating the relative retention time of each characteristic peak and the S peak, and identifying the Yemingsha medicinal materials, decoction pieces, standard decoctions or formula granules as the test sample when the relative retention time of each characteristic peak and the S peak in the characteristic spectrum of the test sample is within the range of the specified value ± 10 %;

[0039] identifying the non-Yemingsha medicinal materials, decoction pieces, standard decoctions or formula granules as the test sample when the relative retention time of each characteristic peak and the S peak in the characteristic spectrum of the test sample is outside the range of the specified value ± 10 %.

[0040] In the characteristic spectrum of the Yemingsha medicinal materials, the specified values are 0.88 (peak 2), 1.07 (peak 4), 1.32 (peak 5), 3.68 (peak 6), 5.02 (peak 7), 5.55 (peak 8) respectively;

[0041] In the characteristic spectrum of the decoction pieces, the specified values are 0.87 (peak 2), 1.08 (peak 4), 1.32 (peak 5), 3.65 (peak 6), 4.98 (peak 7), 5.50 (peak 8) respectively;

[0042] In the characteristic spectrum of the standard decoction, the specified values are 0.87 (peak 2), 1.09 (peak 4), 1.29 (peak 5), 3.55 (peak 6), 4.85 (peak 7), 5.37 (peak 8) respectively;

[0043] In the characteristic spectrum of the formula granule, the specified values are 0.88 (peak 2), 1.10 (peak 4), 1.37 (peak 5), 3.76 (peak 6), 5.11 (peak 7), 5.65 (peak 8) respectively.

[0044] The application includes but is not limited to providing the following beneficial effects:

[0045] The application studies the single variety of Yemingsha formula granules, scientifically identifies Yemingsha, has the characteristics of comprehensive detection and simple operation, can effectively identify Yemingsha formula granules, and can ensure the uniformity and stability of the quality of Yemingsha formula granules. The method is suitable for Yemingsha medicinal materials, decoction pieces, standard decoctions and formula granules. BRIEF DESCRIPTION OF DRAWINGS

[0046] In order to more clearly illustrate the technical solutions in the embodiments of the application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or prior art description.

[0047] Figure 1 Investigation of the mobile phase under chromatographic conditions;

[0048] Figure 2 Identification of chromatographic peaks in the formulation of pyramine granules;

[0049] Figure 3 Different instrumental studies were conducted on the formulation and particle size of luminous sand.

[0050] Figure 4 Investigation of different chromatographic columns for the formulation of pyridine granules;

[0051] Figure 5 Verification of the particle size distribution of the luminous sand formula;

[0052] Figure 6 Verification of the medicinal material "Night-Shining Sand";

[0053] Figure 7 Verification of the use of night-shining cinnabar in medicinal slices;

[0054] Figure 8 Verification of the use of night-shining sand-based decoction;

[0055] Figure 9 The characteristic chromatogram of the formula granules of Yeming Sand is shown, in which peak 1: guanine; peak 3 (S): uric acid;

[0056] Figure 10 The characteristic chromatogram of the determination of the medicinal material of Night-shining Sand is shown, in which peak 1: guanine; peak 3: uric acid;

[0057] Figure 11 The characteristic chromatogram of the determination of the medicinal slices of Night-Brightening Sand is shown, in which peak 1: guanine; peak 3: uric acid;

[0058] Figure 12 The characteristic chromatogram of the standard decoction of Yemingsha is shown, in which peak 1: guanine; peak 3 (S): uric acid. Detailed Implementation

[0059] This invention discloses a method for constructing characteristic spectra of *Nyctaginus styracifolius* medicinal materials, processed slices, standard decoctions, and formulated granules. Those skilled in the art can refer to the content of this document and appropriately modify the process parameters to achieve the desired results. It is particularly important to note that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred embodiments. Those skilled in the art can clearly modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0060] Yeming sand has a long history of medicinal use, which is the dried feces of bats such as Vespertilio superans Thomas and many other bats. The product is the dried feces of bats such as Vespertilio superans Thomas and many other bats, which is processed into a formula granule according to the standard. In order to ensure the uniformity and stability of the quality of Yeming sand formula granules, quality control of Yeming sand medicinal materials, decoction pieces, standard decoction and formula granules must be carried out to ensure the stability of the quality. Therefore, HPLC method is used to control the quality of Yeming sand medicinal materials, decoction pieces, standard decoction and formula granules.

[0061] The present application provides a method for determining the characteristic spectrum of Yeming sand medicinal materials, decoction pieces, standard decoction and formula granules:

[0062] The chromatographic conditions and system suitability test use octadecylsilane bonded silica gel as the filler (the column length is 250 mm, the inner diameter is 4.6 mm, and the particle size is 5 μm); acetonitrile is used as the mobile phase A, and 0.1% phosphoric acid solution is used as the mobile phase B, which is gradient eluted according to the following table; the detection wavelength is 248 nm. The theoretical plate number should not be less than 5000 calculated according to the uric acid peak.

[0063]

[0064] The preparation of the reference solution takes Yeming sand control medicinal materials 2.5 g, adds water 50 mL, decocts for 15 minutes, cools, shakes uniformly, filters, evaporates the filtrate to dryness, adds 10% methanol 2 mL to dissolve, filters, takes the continuous filtrate as the reference solution of the control medicinal materials. Take the guanine control product in an appropriate amount, accurately weigh and add 0.5% ammonia solution to prepare a solution containing 10 μg per 1 mL as the control solution. Then take the uric acid control product in an appropriate amount, accurately weigh and add 0.5% ammonia to prepare a solution containing 10 μg per 1 mL as the control reference solution.

[0065] The preparation of the test solution takes Yeming sand formula granules in an appropriate amount, grinds them finely, takes about 1 g, puts it in a conical flask with a plug, adds 10% methanol 25 mL, ultrasonic treatment (power 600 W, frequency 40 kHz) for 15 minutes, cools, shakes uniformly, filters, takes the continuous filtrate, and gets it; or

[0066] Take Yeming sand medicinal material powder or Yeming sand decoction piece powder (pass through No. 3 sieve) 2.5 g, add water 50 mL, decoct for 15 minutes, cool, filter, evaporate the filtrate to dryness, add 10% methanol 2 mL to dissolve, filter, take the continuous filtrate, and get it; or

[0067] Take Yeming sand standard decoction powder about 1 g, put it in a conical flask, add 10% methanol 25 mL, ultrasonic treatment (power 600 W, frequency 40 kHz) for 15 minutes, cool, take the continuous filtrate, and get it.

[0068] Determination method respectively precision pipette control solution and test solution of each 10 μL, injection liquid chromatograph, determination, namely.

[0069] Test sample chromatography should present 8 characteristic peaks, and should be with the control drug reference material chromatography in the 8 characteristic peaks corresponding to the retention time, of which 2 peaks should be respectively corresponding to the retention time of the corresponding control reference peak; with the peak of uric acid control reference peak S peak, calculate the relative retention time of each characteristic peak and S peak, the relative retention time should be within the range of ± 10% of the specified value.

[0070] Among them, the characteristic spectrum determination method of formula granules is as follows:

[0071] Chromatographic conditions and system suitability test:

[0072] Octadecylsilane bonded silica gel as filler (column length of 250 mm, inner diameter of 4.6 mm, particle size of 5 μm); acetonitrile as mobile phase A, 0.1% phosphoric acid solution as mobile phase B, gradient elution according to the following table; detection wavelength is 248 nm. The theoretical plate number should not be less than 5000 according to the uric acid peak.

[0073]

[0074] The preparation of reference solution is as follows: 2.5 g of Yemingsha control drug is added to 50 mL of water and boiled for 15 minutes. After cooling, shaking and filtering, the filtrate is evaporated to dryness. The residue is dissolved in 2 mL of 10% methanol and filtered. The filtrate is taken as the control drug reference solution. An appropriate amount of guanine control is precisely weighed and dissolved in 0.5% ammonia solution to prepare a solution containing 10 μg per 1 mL as the control solution. Another appropriate amount of uric acid control is precisely weighed and dissolved in 0.5% ammonia to prepare a solution containing 10 μg per 1 mL as the control reference solution.

[0075] The preparation of test solution is as follows: an appropriate amount of Yemingsha granules is finely ground and about 1.0 g is taken in a conical flask with a plug. 25 mL of 10% methanol is added and ultrasonic treatment (power 600 W, frequency 40 kHz) is carried out for 15 minutes. After cooling, shaking and filtering, the filtrate is obtained.

[0076] Determination method respectively precision pipette reference solution and test solution of each 10 μL, injection liquid chromatograph, determination, namely.

[0077] Test sample chromatography should present 8 characteristic peaks( Figure 9), and should correspond to the retention time of 8 characteristic peaks in the reference chromatogram of the control medicinal material, wherein peak 1 and peak 3 should correspond to the retention time of the corresponding reference peaks of the control, and the peak corresponding to the reference peak of the uric acid control 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: 0.88 (peak 2), 1.10 (peak 4), 1.37 (peak 5), 3.76 (peak 6), 5.11 (peak 7), and 5.65 (peak 8).

[0078] The characteristic chromatogram determination method of the medicinal material is as follows:

[0079] The chromatographic conditions and system suitability test use octadecylsilane-bonded silica gel as the filler (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); acetonitrile as mobile phase A, 0.1% phosphoric acid solution as mobile phase B, gradient elution according to the following table, detection wavelength 248 nm. The theoretical plate number should not be less than 5000 calculated by the uric acid peak.

[0080]

[0081] The preparation of the reference solution takes 2.5 g of the control medicinal material of Yemingsha, adds 50 mL of water, decocts for 15 minutes, cools, filters, evaporates the filtrate to dryness, adds 2 mL of 10% methanol to the residue to dissolve, filters, and takes the subsequent filtrate as the control medicinal material reference solution. An appropriate amount of guanine control is precisely weighed, 0.5% ammonia solution is added to prepare a solution containing 10 μg per 1 mL, which is used as the control reference solution. Then take an appropriate amount of uric acid control, precisely weigh, add 0.5% ammonia to prepare a solution containing 10 μg per 1 mL, which is used as the control reference solution.

[0082] The preparation of the test solution takes 2.5 g of the test product powder (passed through a No. 3 sieve), adds 50 mL of water, decocts for 15 minutes, cools, filters, evaporates the filtrate to dryness, adds 2 mL of 10% methanol to dissolve, filters, and takes the subsequent filtrate.

[0083] The determination method precisely takes 10 μL of the reference solution and the test solution respectively, injects into the liquid chromatograph, and determines.

[0084] The test product chromatogram should present 8 characteristic peaks Figure 10) and should correspond to the retention time of 8 characteristic peaks in the reference chromatogram of the control medicinal material, of which 2 peaks should correspond to the retention time of the corresponding reference peaks of the control; the peak corresponding to the reference peak of the control of uric acid is the S peak, and the relative retention time of the other characteristic peaks to the S peak should be within ±10% of the specified value. The specified value is: 0.88 (peak 2), 1.07 (peak 4), 1.32 (peak 5), 3.68 (peak 6), 5.02 (peak 7), 5.55 (peak 8).

[0085] The characteristic chromatogram determination method of the decoction pieces is as follows:

[0086] The chromatographic conditions and system suitability test used octadecylsilane-bonded silica gel as the filler (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); acetonitrile as mobile phase A, 0.1% phosphoric acid solution as mobile phase B, gradient elution according to the following table; the detection wavelength was 248 nm. The theoretical plate number should not be less than 5000 calculated by the uric acid peak.

[0087]

[0088] The preparation of the reference solution took 2.5 g of the control medicinal material of Yemingsha, added 50 mL of water, decocted for 15 minutes, cooled, filtered, and the filtrate was evaporated to dryness. The residue was dissolved with 2 mL of 10% methanol by ultrasonic, filtered, and the filtrate was taken as the reference solution of the control medicinal material. An appropriate amount of the control of guanine was accurately weighed, dissolved in 0.5% ammonia solution to prepare a solution containing 10 μg per 1 mL, which was used as the reference solution of the control. An appropriate amount of the control of uric acid was accurately weighed, dissolved in 0.5% ammonia to prepare a solution containing 10 μg per 1 mL, which was used as the reference solution of the control.

[0089] The preparation of the test solution took 2.5 g of the powder of the product (passed through a No. 3 sieve), added 50 mL of water, decocted for 15 minutes, cooled, filtered, and the filtrate was evaporated to dryness. The residue was dissolved with 2 mL of 10% methanol, filtered, and the filtrate was taken as the test solution.

[0090] The determination method was to accurately pipette 10 μL of the reference solution and the test solution respectively, inject into the liquid chromatograph, and determine.

[0091] The test chromatogram should present 8 characteristic peaks ( Figure 11) and should correspond to the retention times of 8 characteristic peaks in the reference chromatogram of the control medicinal material, of which 2 peaks should correspond to the retention times of the corresponding reference peaks of the control; the peak corresponding to the reference peak of the control of uric acid is the S peak, and the relative retention times of the other characteristic peaks to the S peak should be within ±10% of the specified values, which are: 0.87 (peak 2), 1.08 (peak 4), 1.32 (peak 5), 3.65 (peak 6), 4.98 (peak 7), and 5.50 (peak 8).

[0092] The results of the standard decoction of 5 batches of Yemingsha are as follows:

[0093] The chromatographic conditions and system suitability test used octadecylsilane-bonded silica gel as the filler (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); acetonitrile as the mobile phase A and 0.1% phosphoric acid solution as the mobile phase B, gradient elution was performed according to the following table; the detection wavelength was 248 nm. The theoretical plate number should not be less than 5000 calculated by the uric acid peak.

[0094]

[0095] The preparation of the reference solution took 2.5 g of Yemingsha control medicinal material, added 50 mL of water, decocted for 15 minutes, cooled, filtered, and the filtrate was evaporated to dryness. The residue was dissolved with 2 mL of 10% methanol by ultrasonic treatment, filtered, and the filtrate was taken as the reference solution of the control medicinal material. An appropriate amount of guanine control was precisely weighed, dissolved in 0.5% ammonia solution to prepare a solution containing 10 μg per 1 mL, which was taken as the reference solution of the control. An appropriate amount of uric acid control was precisely weighed, dissolved in 0.5% ammonia to prepare a solution containing 10 μg per 1 mL, which was taken as the reference solution of the control.

[0096] The preparation of the test solution took about 1 g of Yemingsha standard decoction powder, placed it in a conical flask, added 25 mL of 10% methanol, and treated it with ultrasonic waves (power 600 W, frequency 40 kHz) for 15 minutes, cooled, and took the filtrate, which was obtained.

[0097] The determination method precisely took 10 μL of the reference solution and the test solution, respectively, injected into the liquid chromatograph, and determined, which was obtained.

[0098] The test chromatogram should present 8 characteristic peaks ( Figure 12), and should correspond to the retention time of 8 characteristic peaks in the reference chromatogram of the control medicinal material, wherein 2 peaks should correspond to the retention time of the corresponding reference peaks of the control; the peak corresponding to the reference peak of the uric acid control is the S peak, and the relative retention time of the remaining characteristic peaks to the S peak is calculated, which should be within ±10% of the specified value. The specified value is: 0.87 (peak 2), 1.09 (peak 4), 1.29 (peak 5), 3.55 (peak 6), 4.85 (peak 7), and 5.37 (peak 8).

[0099] The experimental instruments and materials used in the present application are as follows:

[0100] Instrument 1: Agilent 1260 high performance liquid chromatograph;

[0101] Instrument 2: Waters e2695 high performance liquid chromatograph;

[0102] Instrument 3: Thermo Vanquish high performance liquid chromatograph

[0103] Electronic balance: ME204E / 02, MS205DM, XP26 (Mettler-Toledo Instruments Co., Ltd.);

[0104] Ultrapure water machine: CelluSafe 1810A (Shanghai Moore Scientific Instruments Co., Ltd.);

[0105] Ultrasonic cleaner: KQ5200DB (600W, 40KHz; Kunshan Ultrasonic Instrument Co., Ltd.);

[0106] Chromatographic column:

[0107] Chromatographic column 1: Agilent 5TC-C18, 4.6*250mm, 5μm,

[0108] Chromatographic column 2: Agilent ZORBAX SB-C18, 4.6*250mm, 5μm,

[0109] Chromatographic column 3: ZORBAX Eclipse Plus C18, 4.6*250mm, 5μm

[0110] Acetonitrile and phosphoric acid are chromatographically pure; water is ultrapure water, and other reagents are analytically pure.

[0111] Guanine control (China Food and Drug Inspection Research Institute, batch number: 140631-202008, content is 98.9%)

[0112] Uric acid control (Shanghai Shidand Standard Technology Service Co., Ltd., batch number: 9602, content is 98.0%);

[0113] YMS-YP-01, YMS-YP-02, YMS-YP-03, YMS-YP-04, YMS-YP-05.

[0114] YMS-YP-01, YMS-YP-02, YMS-YP-03, YMS-YP-04, YMS-YP-05.

[0115] YMS-YP-01, YMS-YP-02, YMS-YP-03, YMS-YP-04, YMS-YP-05.

[0116] YMS-YP-01, YMS-YP-02, YMS-YP-03, YMS-YP-04, YMS-YP-05.

[0117] YMS-YP-01, YMS-YP-02, YMS-YP-03, YMS-YP-04, YMS-YP-05.

[0118] The raw materials and reagents used in the determination method of the characteristic chromatogram of Yeshiming sand medicinal materials, decoction pieces, standard decoction or formula granules provided by the present application can be purchased from the market, unless otherwise specified.

[0119] The present application will be further described below in combination with examples:

[0120] Example characteristic methodology validation

[0121] 1. Investigation of mobile phase

[0122] Based on the above-mentioned experimental conditions, the separation effects of five different mobile phases were investigated, which were acetonitrile-water, acetonitrile-0.1% phosphoric acid solution, acetonitrile-0.1% formic acid solution, methanol-0.1% phosphoric acid solution and acetonitrile-0.1% phosphoric acid solution containing 0.1% triethylamine, as shown in Figure 1 .

[0123] As can be seen from the above figure, the baseline of the chromatographic peak under the gradient elution condition of acetonitrile-0.1% phosphoric acid solution and methanol-0.1% phosphoric acid solution is relatively stable, there are many chromatographic peaks, and the chromatographic peak shape and symmetry under the acetonitrile-0.1% phosphoric acid solution gradient elution condition are better than those under the methanol-0.1% phosphoric acid solution system, so the acetonitrile-0.1% phosphoric acid solution gradient elution is selected as the mobile phase system of the determination method of Yeshiming sand granule characteristic chromatogram.

[0124] 2. Chromatographic peak identification

[0125] Chromatographic conditions and system suitability test: octadecylsilane-bonded silica gel as the filling agent (column length 250 mm, inner diameter 4.5 mm, particle size 5 μm); acetonitrile as the mobile phase A, 0.1% phosphoric acid solution as the mobile phase B, gradient elution according to the provisions in the following table; detection wavelength 248 nm. The theoretical plate number should not be less than 5000 calculated by the peak of uric acid.

[0126]

[0127] Preparation of reference solution: take Yemingsha control medicinal material 2.5 g, add water 50 mL, decoct for 15 minutes, cool, shake up, filter, evaporate the filtrate to dryness, add 10% methanol 2 mL to dissolve, filter, take the filtrate as the reference solution of control medicinal material.

[0128] Preparation of test solution: take Yemingsha granules, grind finely, take about 1 g, put in a conical flask with a plug, add 10% methanol 25 mL, ultrasonic treatment (power 600 W, frequency 40 kHz) for 15 minutes, cool, shake up, filter, take the filtrate as the test solution of Yemingsha granules.

[0129] Preparation of control solution: take guanine control and uric acid control, accurately weigh, add 0.5% ammonia water to make a solution containing 10 μg per 1 mL, namely the control solution.

[0130] Preparation of negative control solution: according to the above proposed experimental conditions, prepare the negative control solution of Yemingsha granules. The preparation method is as follows: add 10% methanol 25 mL in a conical flask with a plug, ultrasonic treatment (power 600 W, frequency 40 kHz) for 15 minutes, cool, shake up, filter, take the filtrate as the negative control solution.

[0131] Determination method: accurately take 10 μL of reference solution, negative control solution, control solution and test solution respectively, inject into the liquid chromatograph, determine, and locate the peaks of Yemingsha formula granules characteristic spectrum as shown in the following table. Figure 2

[0132] The results show that peak 1 is guanine and peak 3 is uric acid. In the following methodology investigation, 8 characteristic peaks in the sample are investigated.

[0133] 3 Precision test

[0134] Take the test solution of Yemingsha formula granules (batch number: YMS-01), according to the proposed experimental method, continuously inject 6 times, 10 μL each time, calculate the retention time and peak area of each characteristic peak. As shown in table 1 and table 2.

[0135] Table 1 Precision investigation-retention time

[0136] ​

[0137] Table 2 Precision Investigation - Peak Area

[0138]

[0139] The results show that the instrument precision is good.

[0140] 4. Reproducibility Investigation

[0141] Take Yeming Sand Formula Granules (Batch No.: YMS-01) 6 parts, prepare and determine according to the proposed experimental method. Calculate the relative retention time of each characteristic peak, as shown in Table 3.

[0142] Table 3 Reproducibility Investigation - Relative Retention Time

[0143]

[0144]

[0145] The results show that the method is reproducible.

[0146] 5. Intermediate Precision Investigation

[0147] 5.1 Different Instrument Investigation

[0148] On the basis of the above proposed experimental conditions, take Yeming Sand Formula Granules (Batch No.: YMS-01) to prepare the test solution, and determine on Instrument 1: Agilent 1260 type high performance liquid chromatograph, Instrument 2: Waters e2695 type high performance liquid chromatograph, Instrument 3: Thermo Vanquish type high performance liquid chromatograph. Calculate the relative retention time of each characteristic peak, as shown in Table 4, Figure 3 .

[0149] Table 4 Instrument Durability Investigation

[0150]

[0151] The results show that the different instrument durability is good.

[0152] 5.2 Different Personnel and Time Investigation

[0153] On the basis of the above proposed experimental conditions, take Yeming Sand Formula Granules (Batch No.: YMS-01) by different personnel (A, B) at different times (T1, T2) each two parts, prepare the test sample, and determine. As shown in Table 5.

[0154] Table 5 Personnel and Time Investigation - Relative Retention Time Ratio

[0155]

[0156] 6 Durability investigation

[0157] 6.1 Durability investigation of chromatographic column

[0158] Based on the above experimental conditions, the following investigations were carried out on chromatographic column 1: Agilent 5TC-C18, 4.6*250mm, 5μm, chromatographic column 2: Agilent ZORBAX SB-C18, 4.6*250mm, 5μm, chromatographic column 3: ZORBAX Eclipse Plus C18, 4.6*250mm, 5μm. The results are shown in Table 6. Figure 4

[0159] Table 6 Durability investigation of chromatographic column

[0160]

[0161] The results show that the above three chromatographic columns are stable for detecting the sample.

[0162] 6.2 Stability investigation

[0163] Based on the above experimental conditions, the same sample solution was taken and measured at 0h, 2h, 6h, 8h, 12h, 24h. The results are shown in Table 7.

[0164] Table 7 Stability investigation

[0165]

[0166] The results show that the sample solution is relatively stable within 24 hours.

[0167] In summary, the RSD of the relative retention time of each characteristic peak meets the requirements in the above investigations, and the method is good. The above eight characteristic peaks were included in the subsequent investigation.

[0168] Example 13 Verification results of 8 batches of Yimingsha formula granules

[0169] Chromatographic conditions and system suitability test:

[0170] Octadecylsilane-bonded silica gel was used as the filler (column length 250mm, inner diameter 4.6mm, particle size 5μm); acetonitrile was used as mobile phase A, and 0.1% phosphoric acid solution was used as mobile phase B, which was gradient eluted according to the following table; the detection wavelength was 248nm. The theoretical plate number should not be less than 5000 calculated by the uric acid peak.

[0171]

[0172] ​Reference solution preparation: 2.5 g of Yemingsha control drug was added to 50 mL of water, decocted for 15 minutes, cooled, shaken, filtered, and the filtrate was evaporated to dryness. The residue was dissolved in 2 mL of 10% methanol, filtered, and the filtrate was used as the control drug reference solution. An appropriate amount of guanine control was precisely weighed, dissolved in 0.5% ammonia water to prepare a solution containing 10 μg per 1 mL, and used as the control solution. An appropriate amount of uric acid control was precisely weighed, dissolved in 0.5% ammonia water to prepare a solution containing 10 μg per 1 mL, and used as the control reference solution.

[0173] Test solution preparation: an appropriate amount of Yemingsha granules was finely ground, about 1.0 g was taken, 25 mL of 10% methanol was added, and ultrasonic treatment (power 600 W, frequency 40 kHz) was performed for 15 minutes. The mixture was cooled, shaken, filtered, and the filtrate was obtained.

[0174] Determination: 10 μL of the reference solution and the test solution were precisely taken and injected into the liquid chromatograph, and determination was performed.

[0175] The above proposed method was used to determine the characteristic chromatograms of 3 batches of samples. As shown in Table 8, Figure 5

[0176] Table 8 Yemingsha granules verification

[0177]

[0178] The results showed that the method could effectively detect Yemingsha granules.

[0179] According to the principle of stable relative retention time and the detection of each batch of samples with relatively high peaks, a total of 8 peaks with good reproducibility were selected as characteristic peaks. Peak 1 and peak 3 should correspond to the retention time of the corresponding control reference peaks, respectively. The peak corresponding to the uric acid control reference peak is S peak, and the relative retention time of each characteristic peak to S peak should be within ±10% of the specified value. The specified value is: 0.88 (peak 2), 1.10 (peak 4), 1.37 (peak 5), 3.76 (peak 6), 5.11 (peak 7), and 5.65 (peak 8).

[0180] The results of each investigation item and verification of the method are shown in Table 9:

[0181] Table 9 RSD% of each item of the method

[0182]

[0183] ​From the above table, each characteristic peak relative retention time is stable, and in the average value ± 10% range, the reproducibility and applicability of the method are good, so the relative retention time of each characteristic peak is temporarily set as

[0184] ±10%.

[0185] Effect example 25 batch of cinnabar medicinal material verification results

[0186] Chromatographic conditions and system suitability test With octadecylsilane bonded silica gel as the filler (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); with acetonitrile as mobile phase A, and 0.1% phosphoric acid solution as mobile phase B, gradient elution according to the provisions in the table below; the detection wavelength is 248 nm. The theoretical plate number should not be less than 5000 calculated by the uric acid peak.

[0187]

[0188] Preparation of reference solution Take 2.5 g of cinnabar reference drug, add 50 mL of water, decoct for 15 minutes, cool, filter, evaporate the filtrate to dryness, add 2 mL of 10% methanol to the residue and ultrasonic to dissolve, filter, take the filtrate as the reference solution of the reference drug. Take an appropriate amount of guanine reference substance, accurately weigh, add 0.5% ammonia solution to prepare a solution containing 10 μg per 1 mL as the reference substance reference solution. Take an appropriate amount of uric acid reference substance, accurately weigh, add 0.5% ammonia to prepare a solution containing 10 μg per 1 mL as the reference substance reference solution.

[0189] Preparation of test solution Take 2.5 g of cinnabar medicinal material powder (pass through No. 3 sieve), add 50 mL of water, decoct for 15 minutes, cool, filter, evaporate the filtrate to dryness, dissolve the residue with 2 mL of 10% methanol, filter, and take the filtrate.

[0190] Determination method: precisely take 10 μL of reference solution and test solution respectively, inject into the liquid chromatograph, and determine.

[0191] The above proposed method was used to determine the characteristic spectrum of 5 batches of samples, and the relative retention time was calculated. As shown in Figure 6 , Table 10.

[0192] Table 105 batches of cinnabar medicinal material relative retention time

[0193]

[0194] The results showed that the chromatogram of the test sample contained eight characteristic peaks, which should correspond to the retention times of the eight characteristic peaks in the chromatogram of the reference medicinal material. Two of these peaks should correspond to the retention times of the corresponding reference standard peaks. The peak corresponding to the uric acid reference standard peak was the S peak. The relative retention times of the remaining characteristic peaks and the S peak were calculated, and their relative retention times should be within ±10% of the specified values. The specified values ​​were: 0.88 (peak 2), 1.07 (peak 4), 1.32 (peak 5), 3.68 (peak 6), 5.02 (peak 7), and 5.55 (peak 8). This method can effectively detect the eight characteristic peaks of five batches of *Vaccaria segetalis* medicinal material. Furthermore, the relative retention times of each characteristic peak and the S peak were within ±10% of the specified values.

[0195] Results of 35 batches of Night-Shining Sand Decoction Pieces

[0196] Chromatographic conditions and system suitability tests were performed using octadecylsilane-bonded silica gel as the packing material (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); acetonitrile was used as mobile phase A, and 0.1% phosphoric acid solution was used as mobile phase B, with gradient elution performed according to the specifications in the table below; the detection wavelength was 248 nm. The theoretical plate number, calculated based on the uric acid peak, should not be less than 5000.

[0197]

[0198]

[0199] Preparation of reference solutions: Take 2.5g of *Vaccaria segetalis* reference material, add 50mL of water, decoct for 15 minutes, cool, filter, evaporate the filtrate to dryness, add 2mL of 10% methanol to the residue, sonicate to dissolve, filter, and take the filtrate as the reference solution. Take an appropriate amount of guanine reference standard, accurately weigh it, and add 0.5% ammonia water to prepare a solution containing 10μg per 1mL as the reference solution. Take an appropriate amount of uric acid reference standard, accurately weigh it, and add 0.5% ammonia water to prepare a solution containing 10μg per 1mL as the reference solution.

[0200] Preparation of the test solution: Take 2.5g of powdered night-shining sand (passed through a No. 3 sieve), add 50mL of water, decoct for 15 minutes, cool, filter, evaporate the filtrate to dryness, dissolve the residue in 2mL of 10% methanol, filter, and take the filtrate to obtain the test solution.

[0201] The assay involves precisely pipetting 10 μL each of the reference solution and the test solution into a liquid chromatograph and measuring the results.

[0202] The aforementioned proposed method was used to determine the characteristic spectra of five batches of this product, and the relative retention times were calculated. For example... Figure 7 As shown in Table 11.

[0203] Table 115 Yeshiming sand pieces verification

[0204]

[0205] The results show that the test product chromatography presents 8 characteristic peaks, which should correspond to the retention time of 8 characteristic peaks in the reference chromatogram of the control medicinal material. Among them, 2 peaks should correspond to the retention time of the corresponding reference peak of the control product, respectively. The peak corresponding to the reference peak of the uric acid control product is the S peak. The relative retention time of each characteristic peak to the S peak is calculated, which should be within ±10% of the specified value. The specified value is: 0.87 (peak 2), 1.08 (peak 4), 1.32 (peak 5), 3.65 (peak 6), 4.98 (peak 7), 5.50 (peak 8). This method can effectively detect 8 characteristic peaks of 5 batches of Yeshiming sand pieces. And the relative retention time of each characteristic peak to the S peak is within ±10% of the specified value. Effect example 45 batches of Yeshiming sand standard decoction certificate results

[0206] Chromatographic conditions and system suitability test With octadecylsilane bonded silica gel as the filler (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); with acetonitrile as mobile phase A, and 0.1% phosphoric acid solution as mobile phase B, gradient elution according to the following table; the detection wavelength is 248 nm. The theoretical plate number should not be less than 5000 calculated by the uric acid peak.

[0207]

[0208] Preparation of reference solution Take Yeshiming sand control medicinal material 2.5 g, add water 50 mL, decoct for 15 minutes, cool, filter, evaporate the filtrate to dryness, add 10% methanol 2 mL to the residue to dissolve, filter, take the filtrate as the control medicinal material reference solution. Take guanine control product in appropriate amount, accurately weigh, add 0.5% ammonia solution to prepare a solution containing 10 μg per 1 mL as the control product reference solution. Take uric acid control product in appropriate amount, accurately weigh, add 0.5% ammonia to prepare a solution containing 10 μg per 1 mL as the control product reference solution.

[0209] Preparation of test solution Take Yeshiming sand standard decoction powder about 1 g, put it in a conical flask, add 10% methanol 25 mL, ultrasonic treatment (power 600 W, frequency 40 kHz) for 15 minutes, cool, take the filtrate, and get it.

[0210] Determination method: accurately pipette 10 μL of reference solution and test solution respectively, inject into the liquid chromatograph, and determine.

[0211] The proposed method was used to determine the characteristic spectrum of 5 batches of samples, and the relative retention time was calculated. As shown in Figure 8 , Table 12.

[0212] Table 125 batch Yiming sand standard decoction verification

[0213]

[0214]

[0215] The results show that 8 characteristic peaks are presented in the chromatogram of the test sample, and should correspond to the retention time of 8 characteristic peaks in the reference chromatogram of the control medicinal material, wherein 2 peaks should correspond to the retention time of the corresponding reference peaks of the control sample respectively; the peak corresponding to the reference peak of the uric acid control sample is the S peak, and the relative retention time of each characteristic peak to the S peak is calculated, which should be within ±10% of the specified value. The specified value is: 0.87 (peak 2), 1.09 (peak 4), 1.29 (peak 5), 3.55 (peak 6), 4.85 (peak 7), 5.37 (peak 8). The method can effectively detect 8 characteristic peaks of 5 batches of Yiming sand standard decoction. And the relative retention time of each characteristic peak to the S peak is within ±10% of the specified value.

[0216] The above only describes the preferred embodiments of the present application, and it should be noted that for those skilled in the art, without departing from the principles of the present application, several improvements and refinements can be made, and these improvements and refinements should also be considered within the scope of protection of the present application.

Claims

1. A method for constructing the characteristic chromatogram of Yemingsha medicinal material, decoction piece, standard decoction or formula granule, characterized in that, Comprise: S1, the test sample raw material is dissolved by solvent, extraction, to obtain the test sample solution; the solvent comprises 10% methanol and / or water; the extraction comprises decoction and / or ultrasonic extraction; S2, the test sample solution of S1 is determined by liquid chromatography, to obtain the characteristic spectrum of Yemingsha medicinal material, decoction piece, standard decoction or formula granule; The test sample raw material comprises Yemingsha medicinal material, decoction piece, standard decoction or formula granule; The chromatographic conditions of the liquid chromatography comprise: the chromatographic column is C18 column; the mobile phase A is acetonitrile or methanol, and the mobile phase B is 0.1% formic acid solution or 0.1% phosphoric acid solution; The elution program of the liquid chromatography determination comprises: 0~20min, A phase 0.5~2%, B phase 99.5~98%; 20~40min, A phase 2~5%, B phase 98~95%; 40~70min, A phase 5%, B phase 95%; The detection wavelength of the liquid chromatography determination comprises 248nm.

2. The construction method of claim 1, wherein, The construction method further comprises the steps of preparing a control reference solution and / or a control medicinal material reference solution; The preparation method of the control reference solution comprises: taking guanine and uric acid respectively, dissolving with 0.5% ammonia water to obtain the control reference solution; The preparation method of the control medicinal material reference solution comprises: mixing Yemingsha control medicinal material and water, decocting, cooling, filtering, evaporating the filtrate to dryness, dissolving the residue with 10% methanol, filtering, taking the filtrate to obtain the control medicinal material reference solution.

3. The construction method of claim 1, wherein, The mass-volume ratio of (I) and (II) comprises (1:20)~(1:50) in g / mL: (I), the Yemingsha medicinal material, decoction piece, standard decoction or formula granule; (II), the solvent.

4. The construction method of claim 1, wherein, In S1, the power of the ultrasonic extraction comprises 600W, and the frequency comprises 40kHz; the extraction time comprises 15~30min; The decoction time is greater than or equal to 15min.

5. The construction method of claim 1 wherein, The sample amount of the liquid chromatography determination comprises 10μL.

6. The construction method of claim 1 wherein, The theoretical plate number of the liquid chromatography determination is not less than 5000 according to the uric acid peak.

7. The construction method of claim 1 wherein, In the characteristic spectrum of the Yemingsha medicinal material, two peaks respectively correspond to the retention time of the peaks of the control reference solution, taking the peak corresponding to uric acid as S peak, calculating the relative retention time of each characteristic peak and the S peak, the relative retention time is within ±10% of the characteristic peak value, and the characteristic peak values are respectively: 0.88 (peak 2), 1.07 (peak 4), 1.32 (peak 5), 3.68 (peak 6), 5.02 (peak 7), 5.55 (peak 8); In the characteristic spectrum of the decoction piece, two peaks respectively correspond to the retention time of the peaks of the control reference solution, taking the peak corresponding to uric acid in the control reference solution as S peak, calculating the relative retention time of each characteristic peak and the S peak, the relative retention time is within ±10% of the characteristic peak value, and the characteristic peak values are respectively: 0.87 (peak 2), 1.08 (peak 4), 1.32 (peak 5), 3.65 (peak 6), 4.98 (peak 7), 5.50 (peak 8); In the characteristic spectrum of the standard decoction, two peaks respectively correspond to the retention time of the peaks of the reference solution of the control sample, and the peak corresponding to uric acid in the reference solution of the control sample is S peak. The relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time is within ±10% of the characteristic peak value. The characteristic peak values are 0.87 (peak 2), 1.09 (peak 4), 1.29 (peak 5), 3.55 (peak 6), 4.85 (peak 7), and 5.37 (peak 8). In the characteristic spectrum of the formula granules, two peaks respectively correspond to the retention time of the peaks of the reference solution of the control sample, and the peak corresponding to uric acid in the reference solution of the control sample is S peak. The relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time is within ±10% of the characteristic peak value. The characteristic peak values are 0.88 (peak 2), 1.10 (peak 4), 1.37 (peak 5), 3.76 (peak 6), 5.11 (peak 7), and 5.65 (peak 8).

8. A method for identifying Yemingsha medicinal material, decoction piece, standard decoction or formula granule, characterized in that, The characteristic spectrum of the test sample is obtained by using the construction method according to any one of claims 1 to 7. In the characteristic spectrum, two peaks respectively correspond to the retention time of the peaks of the reference solution of the control sample, and the peak corresponding to uric acid in the reference solution of the control sample is S peak. The relative retention time of each characteristic peak and the S peak is calculated, and the relative retention time of each characteristic peak and the S peak in the characteristic spectrum of the test sample is within ±10% of the specified value. It is identified as Yimingsha medicinal material, decoction piece, standard decoction or formula granules. If the relative retention time of each characteristic peak and the S peak in the characteristic spectrum of the test sample is outside the ±10% range of the specified value, it is identified as non-Yimingsha medicinal material, decoction piece, standard decoction or formula granules. In the characteristic spectrum of the Yimingsha medicinal material, the specified values are 0.88 (peak 2), 1.07 (peak 4), 1.32 (peak 5), 3.68 (peak 6), 5.02 (peak 7), and 5.55 (peak 8). In the characteristic spectrum of the decoction piece, the specified values are 0.87 (peak 2), 1.08 (peak 4), 1.32 (peak 5), 3.65 (peak 6), 4.98 (peak 7), and 5.50 (peak 8). In the characteristic spectrum of the standard decoction, the specified values are 0.87 (peak 2), 1.09 (peak 4), 1.29 (peak 5), 3.55 (peak 6), 4.85 (peak 7), and 5.37 (peak 8). In the characteristic spectrum of the formula granules, the specified values are 0.88 (peak 2), 1.10 (peak 4), 1.37 (peak 5), 3.76 (peak 6), 5.11 (peak 7), and 5.65 (peak 8).

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