Fingerprint spectrum construction method of rhizoma gastrodiae decoction pieces, fingerprint spectrum and application

Abstract

The invention discloses a gastrodia elata decoction piece fingerprint spectrum construction method, a fingerprint spectrum and application, the gastrodia elata decoction piece fingerprint spectrum meeting gastrodia elata formula granule quality requirements is constructed, and the fingerprint spectrum comprises six Mark peaks of gastrodin, p-hydroxybenzyl alcohol, parishin, parishin B, parishin C and parishin E; the control fingerprint spectrum generated in the common peak mode can be used for comparison to judge whether the fingerprint spectrum of the gastrodia elata decoction pieces meets requirements or not, and when the similarity of the fingerprint spectrum of the gastrodia elata decoction pieces is not lower than 0.76, the similarity of the fingerprint spectrum of the prepared gastrodia elata formula granules is not lower than 0.90. The problem that the quality of the gastrodia elata decoction pieces meeting the quality requirements of gastrodia elata formula granules is unstable is solved.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine formula granules, and specifically relates to a method for preparing Gastrodia elata slices that meets the quality requirements of Gastrodia elata formula granules, fingerprint spectrum construction and application thereof. Background Art

[0002] Gastrodia elata formula granules YBZ-PFKL-2021122 has been promulgated and implemented as a national drug standard. This standard uses water as the solvent, and the prescription and preparation amount is 4:1. It is stipulated that the content of the finished product is based on the peak area of ​​gastrodin, multiplied by the correction factor respectively, and the content of gastrodin, p-hydroxybenzyl alcohol, barisonoside E, barisonoside B, barisonoside C, and barisonoside should be 43.0-80.0 mg / g. The fingerprint spectrum stipulates that the fingerprint spectrum of the finished product should present chromatographic peaks with the same retention time as the chromatographic peak of the reference object, and Mark peak matching is adopted. According to the similarity evaluation system of traditional Chinese medicine chromatographic fingerprint spectrum, the similarity between the test sample fingerprint spectrum and the control fingerprint spectrum shall not be less than 0.90.

[0003] Gastrodia elata is the dried tuber of the perennial plant Gastrodia elata Bl. of the Orchidaceae family, and is included in the Chinese Pharmacopoeia. The Pharmacopoeia has requirements for the characteristic spectrum of Gastrodia elata raw medicinal materials, stipulating that the chromatogram of Gastrodia elata raw medicinal materials should show 6 characteristic peaks, and should correspond to the 6 characteristic peaks in the chromatogram of the reference medicinal material reference material. Among them, peak 1 and peak 2 should be consistent with the retention time of the reference material of gastrodin and p-hydroxybenzyl alcohol, and the substances corresponding to peaks 3, 4, 5, and 6 are barisonoside E, barisonoside B, barisonoside C, and barisonoside, respectively. The content index requirement is the total amount of gastrodin and p-hydroxybenzyl alcohol. There is no requirement for the characteristic spectrum of Gastrodia elata decoction pieces under the item of Gastrodia elata decoction pieces, and the content limit requirement of decoction pieces is consistent with that of the raw medicinal materials.

[0004] Gastrodia elata is a commonly used bulk Chinese herbal medicine, widely distributed in Anhui, Hubei, Shaanxi, Sichuan, Yunnan, Guizhou and other places, and has been included in the catalog of fresh processing varieties by the drug supervision departments of the above provinces. However, the processing of Gastrodia elata slices is quite particular. There has been a saying of "nine steaming and nine drying" since ancient times. The Chinese Pharmacopoeia requires a relatively simple processing process for Gastrodia elata slices. The original medicinal materials are washed, moistened or steamed, cut into thin slices, and dried. Modern research shows that the structure of barison glycosides is unstable and there is a mutual conversion relationship between them and gastrodin under different temperature conditions. Different processing methods, temperatures, and times will affect the quality of Gastrodia elata slices. As a fresh processing variety in the place of origin, Gastrodia elata has a large fluctuation in the quality of processed Gastrodia elata slices due to different processing methods and processes, which in turn affects the quality of the prepared Gastrodia elata formula granules. In particular, the similarity of the fingerprint spectrum of Gastrodia elata formula granules fluctuates greatly, and many unqualified cases occur.

[0005] In view of this, the present invention is proposed. Summary of the invention

[0006] The purpose of the present invention is to provide a method for preparing Gastrodia elata slices that meet the quality requirements of Gastrodia elata formula granules, and to construct a fingerprint spectrum of Gastrodia elata slices. By means of the preparation method, Gastrodia elata slices that meet the quality requirements of Gastrodia elata formula granules can be prepared. The constructed fingerprint spectrum and detection method can determine the similarity of the fingerprint spectrum of Gastrodia elata slices. When the similarity meets the requirements, the fingerprint spectrum of the prepared Gastrodia elata formula granules meets the requirements of national standards, thereby achieving accurate, objective and effective control of the quality of Gastrodia elata slices, and ensuring that the fingerprint spectrum of the prepared Gastrodia elata formula granules meets the requirements.

[0007] To achieve the above object, the technical solution adopted by the present invention is:

[0008] A method for constructing a fingerprint spectrum of Gastrodia elata decoction pieces comprises the following steps:

[0009] Step 1: Take the raw material of Gastrodia elata, wash it, steam it in a steamer for 5h to 10h, then cut it into thin slices, cool it at room temperature, dry it at 50℃ to 60℃, sieve out the fine powder, and obtain the Gastrodia elata slices;

[0010] Step 2, placing the prepared Gastrodia elata slices in a decoction machine, adding 8 times the amount of water of the slices, soaking for 30 minutes, boiling over high heat, decocting over low heat for 60 minutes, filtering while hot, and setting aside; adding 6 times the amount of water of the slices for the second time, boiling over high heat, decocting over low heat for 40 minutes, filtering while hot, and setting aside; combining the two extraction filtrates, concentrating under reduced pressure at 60°C to a material-liquid ratio of 1:1, and a relative density of 1.05-1.10 at 60°C, spray drying, adding auxiliary materials, mixing, and granulating to obtain the finished Gastrodia elata formula granules, each 1g of the formula granules is equivalent to 4g of the slices;

[0011] Step 3: Take the Tianma formula granules prepared in step 2, and detect the content, fingerprint, extract, properties, and thin layer identification indicators of the Tianma formula granules according to the requirements of the national drug standard YBZ-PFKL-2021122;

[0012] Step 4: According to the processing technology of the Gastrodia elata slices corresponding to the Gastrodia elata formula granules that have passed the test of all indicators in step 3, 15 different batches of Gastrodia elata slices are processed, and steps 2 and 3 are performed respectively;

[0013] Step 5: Take the Gastrodia elata decoction pieces corresponding to the Gastrodia elata formula granules with all the qualified indicators in step 4 as samples, and carry out the following steps:

[0014] A) Take the Gastrodia elata slices, crush them, pass them through a No. 4 pharmacopoeia sieve, and take the sieve contents as the test sample;

[0015] B) taking a sample of the test substance, dissolving it in a solvent, extracting it, and obtaining a test substance solution;

[0016] C) using ultra-high performance liquid chromatography to measure the test solution to obtain UPLC characteristic spectra of each batch of Gastrodia elata slices;

[0017] D) Importing the UPLC characteristic spectrum corresponding to each batch of medicinal materials into the Chinese medicine chromatographic fingerprint similarity evaluation system, using Mark peak matching to generate a common peak pattern spectrum, and constructing a reference fingerprint spectrum of Gastrodia elata decoction pieces;

[0018] Step 6: The Gastrodia elata slices with different steaming times are respectively subjected to the characteristic spectrum detection according to the method described in step 5, and the Mark peaks are matched with the constructed fingerprint spectrum of the Gastrodia elata slices, and the similarity between the fingerprint spectrum and the control fingerprint spectrum is calculated;

[0019] Step seven, with the offset of the fingerprint spectra of the Gastrodia elata slices at different steaming times in step six as the horizontal coordinate and the similarity of the fingerprint spectra of the corresponding Gastrodia elata formula granules as the vertical coordinate, a linear regression equation is fitted. According to the fitted linear regression equation, if the similarity of the fingerprint spectra of the Gastrodia elata formula granules is not less than 0.90, the similarity of the fingerprint spectra of the corresponding Gastrodia elata slices shall not be less than 0.76.

[0020] In the present invention, the steaming time in step 1 is 5h, 6h, 7h, 8h, 9h, 10h, and the low-temperature drying temperature is 55°C.

[0021] In the present invention, the steaming time range of the 15 different batches of Gastrodia elata slices during preparation in step 4 is 5h-10h.

[0022] In the present invention, the sample weight of the crushed and sieved material of the Gastrodia elata decoction pieces in step 5 is 1.0 g.

[0023] In the present invention, the method for preparing the test solution in step 5 is specifically as follows:

[0024] Take 1.0g of Gastrodia elata powder, accurately weigh it, put it in a stoppered conical flask, accurately add 50ml of water, heat and reflux for 1 hour, cool, filter, accurately measure 10ml of the filtrate, evaporate to dryness, add 30% methanol to dissolve and make up to 5ml, ultrasonically treat (power 250W, frequency 40kHz) for 30 minutes, cool and filter with a 0.22μm syringe filter to obtain.

[0025] The chromatographic conditions of the ultra-high performance liquid chromatography described in step 5 are: octadecyl bonded silica gel as the filler, a column length of 150 mm, a column inner diameter of 2.1 mm, a particle size of 1.6 μm, and a column temperature of 30°C; a flow rate of 0.30 mL per minute, and a detection wavelength of 220 nm; an injection volume of 1 μL, and a theoretical plate number calculated based on gastrodin should not be less than 3000; acetonitrile is used as the mobile phase A, and 0.05% phosphoric acid solution is used as the mobile phase B, and elution is performed according to the following gradient elution program:

[0026]

[0027] In the present invention, the C18 column has a size of 2.1×150 mm, 1.6 μm, and a column temperature of 30° C.

[0028] In the present invention, the flow rate is 0.30 mL per minute, the detection wavelength is 220 nm, the injection volume is 1 μL, and the theoretical plate number should be no less than 3000 calculated based on gastrodin.

[0029] In the present invention, the 0.05% phosphoric acid solution is the mobile phase B in volume fraction.

[0030] The present invention also includes preparing a reference substance solution, specifically: taking gastrodin, p-hydroxybenzyl alcohol, barisonoside, barisonoside B, barisonoside C, and barisonoside E reference substances, accurately weighing them, placing them in a volumetric flask, adding methanol to dissolve and mix them, and preparing solutions containing 200 μg, 5 μg, 200 μg, 150 μg, 50 μg, and 200 μg per 1 mL, respectively, as reference substance solutions.

[0031] The Mark peak matching described in the present invention refers to the six characteristic peaks of gastrodin, p-hydroxybenzyl alcohol, barisonoside, barisonoside B, barisonoside C and barisonoside E, which are injected under the same chromatographic conditions respectively, and the gastrodin, p-hydroxybenzyl alcohol, barisonoside, barisonoside B, barisonoside C and barisonoside E reference substances are used to locate and identify the chromatographic peaks.

[0032] The Gastrodia elata slices with different steaming times described in step six are used as comparisons. In addition to the steaming time described in step one, in order to comprehensively study the effects of different steaming times on the characteristic spectra of Gastrodia elata slices, Gastrodia elata slices steamed for 2h, 3h, and 4h are also included. These Gastrodia elata slices are also prepared by the methods described in steps two, three, and five, and the Gastrodia elata formula granules are tested for various indicators of the formula granules.

[0033] In step 7, the similarity of the fingerprint spectrum of the Gastrodia elata decoction pieces is used as the abscissa (X), and the similarity of the fingerprint spectrum of the Gastrodia elata formula granules corresponding to the Gastrodia elata decoction pieces is used as the ordinate (Y) to perform linear regression equation fitting, and the fitted linear regression equation is:

[0034] Y = 0.4714X + 0.5544, correlation coefficient R 2 =0.9768.

[0035] In the present invention, according to the fitted linear regression equation, if the fingerprint similarity of the Tianma formula granules is not less than 0.90, the corresponding fingerprint similarity of the Tianma decoction pieces should not be less than 0.733. Taking into account the errors in the detection and processing process, combined with multiple batches of data, it is corrected that the fingerprint similarity of the Tianma decoction pieces should not be less than 0.76.

[0036] Gastrodia elata is a variety that is processed while fresh at the place of origin. Different processing methods and processing techniques make the quality of Gastrodia elata slices vary greatly. In order to meet the special requirements of the national standard for Gastrodia elata formula granules, it is necessary to establish quality judgment standards for Gastrodia elata slices, especially the fingerprint spectrum similarity requirements; if the original Gastrodia elata is purchased and processed by oneself, the specific processing technique and the quality requirements of the slices after processing need to be clarified. Based on this, the technical solution of the present invention has the following advantages:

[0037] 1. The present invention combines the preparation process of Gastrodia elata formula granules and conducts in-depth research on the key process of Gastrodia elata decoction pieces processing, namely the steaming process in step one. The content, characteristic spectrum and other indicators of the prepared Gastrodia elata formula granules meet the standard requirements as the result orientation, and establishes an industrial processing method for Gastrodia elata decoction pieces that meets the quality requirements of Gastrodia elata formula granules.

[0038] 2. The present invention establishes a fingerprint detection method for Gastrodia elata slices for the first time. Through this method, 15 batches of Gastrodia elata slices that meet the quality requirements of Gastrodia elata formula granules are used to construct a control fingerprint of Gastrodia elata slices, and a linear regression equation for the similarity of the fingerprint of Gastrodia elata slices and the fingerprint of Gastrodia elata formula granules is established. Through this detection method, the constructed fingerprint and the linear regression equation, the fingerprint of Gastrodia elata slices can be measured, thereby controlling the quality of Gastrodia elata slices and improving the product qualification rate of Gastrodia elata formula granules. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 This is the chromatogram of the gastrodin reference substance solution;

[0040] Figure 2 This is the chromatogram of p-hydroxybenzyl alcohol reference solution;

[0041] Figure 3 This is the chromatogram of the barisenoside E reference substance solution;

[0042] Figure 4 This is the chromatogram of the barisenoside B reference substance solution;

[0043] Figure 5 This is the chromatogram of the barisenoside C reference solution;

[0044] Figure 6 This is the chromatogram of the barisin glycoside reference solution;

[0045] Figure 7 Chromatograms of 15 batches of Gastrodia elata slices;

[0046] Figure 8 It is the fingerprint of Gastrodia elata decoction pieces;

[0047] Fig. 9 It is a linear relationship diagram between the fingerprint similarity of Gastrodia elata slices and the fingerprint similarity of Gastrodia elata formula granules;

[0048] Fig.10 To investigate the precision of fingerprint spectrum of Gastrodia elata decoction pieces;

[0049] Fig.11 To investigate the stability of fingerprint spectrum of Gastrodia elata decoction pieces;

[0050] Fig.12 This study investigates the repeatability of fingerprints of Gastrodia elata decoction pieces. DETAILED DESCRIPTION

[0051] The present application is further explained below in conjunction with specific implementation methods, but the embodiments do not limit the present application in any form.

[0052] instrument

[0053] Agilent 1290 series ultra-high performance liquid chromatograph (Agilent Technologies, Inc.); chromatographic column: CORTECST3 C18 column (2.1×150 mm, 1.6 μm); SQP electronic analytical balance (Sartorius, Germany); CNC ultrasonic cleaner (Shanghai Kedao Ultrasonic Instrument Co., Ltd.); PURELAB classic UV ultrapure water meter (ELGA, UK); digital display constant temperature water bath (Changzhou Guohua Electric Co., Ltd.); HYDJ20 micro-pressure circulation decoction machine (Tianjin Huayanyuan Machinery Co., Ltd.); R-1020 rotary evaporator (Zhengzhou Great Wall Science and Technology Industry and Trade Co., Ltd.); OPD-8 spray dryer (Wuxi Heming Drying Equipment Co., Ltd.); PSJ-1000 jet cleaning machine (Jiangsu Baowei Machinery Technology Co., Ltd.); CT-C-II hot air circulation oven (Hubei Hengfeng Medical Pharmaceutical Equipment Co., Ltd.); QWZL-300D linear reciprocating medicine cutting machine (Taizhou Boda Pharmaceutical Machinery Technology Co., Ltd.); LG100 dry granulator (Nanjing Canaan Pharmaceutical Equipment Co., Ltd.).

[0054] Drug testing

[0055] Gastrodin reference substance (batch number: 110807-202010, purity 95.5%, China Food and Drug Inspection Institutes).

[0056] p-Hydroxybenzyl alcohol reference substance (batch number: 111970-201702, purity 99.4%, China Food and Drug Inspection Institutes).

[0057] Balisong glycoside (batch number: 020141-202208, purity 98.0%, Shanghai Hongyong Biotechnology Co., Ltd.).

[0058] Balisonoside B (batch number: 020144-202205, purity 98.0%, Shanghai Hongyong Biotechnology Co., Ltd.). Balisonoside C (batch number: 020154-202211, purity 98.0%, Shanghai Hongyong Biotechnology Co., Ltd.).

[0059] Balisonoside E (batch number: 020162-202112, purity 98.0%, Shanghai Hongyong Biotechnology Co., Ltd.).

[0060] Acetonitrile was chromatographically pure and was purchased from Fisher, USA.

[0061] Phosphoric acid was chromatographically pure and was purchased from Sinopharm Chemical Reagent Co., Ltd.

[0062] Water was ultrapure water, and other reagents were of analytical grade.

[0063] Table 1 Information on 15 batches of Gastrodia elata medicinal materials and formula granules

[0064]

[0065]

[0066] Example 1

[0067] A method for preparing Gastrodia elata decoction pieces that meets the quality requirements of Gastrodia elata formula granules comprises the following steps:

[0068] Step 1: Take the raw material of Gastrodia elata (medicinal material number T1), wash it, steam it in a steamer for 5 hours, cut it into thin slices using a linear reciprocating medicine cutter, cool it at room temperature, and then place it in a hot air circulation oven to dry it at 50°C, sieve out the fine powder, and you will get the Gastrodia elata slices.

[0069] Step 2. Take the Gastrodia elata slices prepared in step 1, place them in a decoction machine, add 8 times the amount of water as the slices, soak for 30 minutes, boil over high heat, simmer for 60 minutes, filter while hot, and set aside; add 6 times the amount of water as the slices for the second time, boil over high heat, simmer for 40 minutes, filter while hot, and set aside; combine the two extraction filtrates, concentrate under reduced pressure at 60°C to a relative density of 1.08 (60°C), spray dry, add an appropriate amount of dextrin, mix well, and granulate to obtain the finished Gastrodia elata formula granules (each 1g of formula granules is equivalent to 4g of slices).

[0070] Step 3. Take the Gastrodia elata formula granules prepared in step 2, and detect the content, fingerprint, extract, properties, thin layer identification and other indicators of the formula granules according to the method under the content determination and fingerprint of Gastrodia elata formula granules YBZ-PFKL-2021122.

[0071] Example 2

[0072] A method for preparing Gastrodia elata decoction pieces that meets the quality requirements of Gastrodia elata formula granules comprises the following steps:

[0073] Step 1: Take the raw material of Gastrodia elata (medicinal material number T1), wash it, steam it in a steamer for 6 hours, cut it into thin slices using a linear reciprocating medicine cutter, cool it at room temperature, and then place it in a hot air circulation oven to dry it at 55°C, sieve out the fine powder, and you will get the Gastrodia elata slices.

[0074] Step 2. Take the Gastrodia elata slices prepared in step 1, place them in a decoction machine, add 8 times the amount of water as the slices, soak for 30 minutes, boil over high heat, simmer for 60 minutes, filter while hot, and set aside; add 6 times the amount of water as the slices for the second time, boil over high heat, simmer for 40 minutes, filter while hot, and set aside; combine the two extraction filtrates, concentrate under reduced pressure at 60°C to a relative density of 1.09 (60°C), spray dry, add an appropriate amount of dextrin, mix well, and granulate to obtain the finished Gastrodia elata formula granules (each 1g of formula granules is equivalent to 4g of slices).

[0075] Step 3. Take the Gastrodia elata formula granules prepared in step 2, and detect the content, fingerprint, extract, properties, thin layer identification and other indicators of the formula granules according to the method under the content determination and fingerprint of Gastrodia elata formula granules YBZ-PFKL-2021122.

[0076] Example 3

[0077] A method for preparing Gastrodia elata decoction pieces that meets the quality requirements of Gastrodia elata formula granules comprises the following steps:

[0078] Step 1: Take the raw material of Gastrodia elata (medicinal material number T1), wash it, steam it in a steamer for 7 hours, cut it into thin slices using a linear reciprocating medicine cutter, cool it at room temperature, and then place it in a hot air circulation oven to dry it at 60°C, sieve out the fine powder, and you will get the Gastrodia elata slices.

[0079] Step 2. Take the Gastrodia elata slices prepared in step 1, place them in a decoction machine, add 8 times the amount of water as the slices, soak for 30 minutes, boil over high heat, simmer for 60 minutes, filter while hot, and set aside; add 6 times the amount of water as the slices for the second time, boil over high heat, simmer for 40 minutes, filter while hot, and set aside; combine the two extraction filtrates, concentrate under reduced pressure at 60°C to a relative density of 1.05 (60°C), spray dry, add an appropriate amount of dextrin, mix well, and granulate to obtain the finished Gastrodia elata formula granules (each 1g of formula granules is equivalent to 4g of slices).

[0080] Step 3. Take the Gastrodia elata formula granules prepared in step 2, and detect the content, fingerprint, extract, properties, thin layer identification and other indicators of the formula granules according to the method under the content determination and fingerprint of Gastrodia elata formula granules YBZ-PFKL-2021122.

[0081] Example 4

[0082] A method for preparing Gastrodia elata decoction pieces that meets the quality requirements of Gastrodia elata formula granules comprises the following steps:

[0083] Step 1: Take the raw material of Gastrodia elata (medicinal material number T1), wash it, steam it in a steamer for 8 hours, cut it into thin slices using a linear reciprocating medicine cutter, cool it at room temperature, and then place it in a hot air circulation oven to dry it at 50°C, sieve out the fine powder, and you will get the Gastrodia elata slices.

[0084] Step 2. Take the Gastrodia elata slices prepared in step 1, place them in a decoction machine, add 8 times the amount of water as the slices, soak for 30 minutes, boil over high heat, simmer for 60 minutes, filter while hot, and set aside; add 6 times the amount of water as the slices for the second time, boil over high heat, simmer for 40 minutes, filter while hot, and set aside; combine the two extraction filtrates, concentrate under reduced pressure at 60°C to a relative density of 1.05 (60°C), spray dry, add an appropriate amount of dextrin, mix well, and granulate to obtain the finished Gastrodia elata formula granules (each 1g of formula granules is equivalent to 4g of slices).

[0085] Step 3. Take the Gastrodia elata formula granules prepared in step 2, and detect the content, fingerprint, extract, properties, thin layer identification and other indicators of the formula granules according to the method under the content determination and fingerprint of Gastrodia elata formula granules YBZ-PFKL-2021122.

[0086] Example 5

[0087] A method for preparing Gastrodia elata decoction pieces that meets the quality requirements of Gastrodia elata formula granules comprises the following steps:

[0088] Step 1: Take the raw material of Gastrodia elata (medicinal material number T1), wash it, steam it in a steamer for 9 hours, cut it into thin slices using a linear reciprocating medicine cutter, cool it at room temperature, and then place it in a hot air circulation oven to dry it at 55°C, sieve out the fine powder, and you will get the Gastrodia elata slices.

[0089] Step 2. Take the Gastrodia elata slices prepared in step 1, place them in a decoction machine, add 8 times the amount of water as the slices, soak for 30 minutes, boil over high heat, simmer for 60 minutes, filter while hot, and set aside; add 6 times the amount of water as the slices for the second time, boil over high heat, simmer for 40 minutes, filter while hot, and set aside; combine the two extraction filtrates, concentrate under reduced pressure at 60°C to a relative density of 1.08 (60°C), spray dry, add an appropriate amount of dextrin, mix well, and granulate to obtain the finished Gastrodia elata formula granules (each 1g of formula granules is equivalent to 4g of slices).

[0090] Step 3. Take the Gastrodia elata formula granules prepared in step 2, and detect the content, fingerprint, extract, properties, thin layer identification and other indicators of the formula granules according to the method under the content determination and fingerprint of Gastrodia elata formula granules YBZ-PFKL-2021122.

[0091] Example 6

[0092] A method for preparing Gastrodia elata decoction pieces that meets the quality requirements of Gastrodia elata formula granules comprises the following steps:

[0093] Step 1: Take the raw material of Gastrodia elata (medicinal material number T1), wash it, steam it in a steamer for 10 hours, cut it into thin slices using a linear reciprocating medicine cutter, cool it at room temperature, and then place it in a hot air circulation oven to dry it at 60°C, sieve out the fine powder, and you will get the Gastrodia elata slices.

[0094] Step 2. Take the Gastrodia elata slices prepared in step 1, place them in a decoction machine, add 8 times the amount of water as the slices, soak for 30 minutes, boil over high heat, simmer for 60 minutes, filter while hot, and set aside; add 6 times the amount of water as the slices for the second time, boil over high heat, simmer for 40 minutes, filter while hot, and set aside; combine the two extraction filtrates, concentrate under reduced pressure at 60°C to a relative density of 1.10 (60°C), spray dry, add an appropriate amount of dextrin, mix well, and granulate to obtain the finished Gastrodia elata formula granules (each 1g of formula granules is equivalent to 4g of slices).

[0095] Step 3. Take the Gastrodia elata formula granules prepared in step 2, and detect the content, fingerprint, extract, properties, thin layer identification and other indicators of the formula granules according to the method under the content determination and fingerprint of Gastrodia elata formula granules YBZ-PFKL-2021122.

[0096] Comparative Example 1

[0097] A method for preparing Gastrodia elata decoction pieces that meets the quality requirements of Gastrodia elata formula granules comprises the following steps:

[0098] Step 1: Take the raw material of Gastrodia elata (medicinal material number T1), wash it, steam it in a steamer for 2 hours, cut it into thin slices using a linear reciprocating medicine cutter, cool it at room temperature, and then place it in a hot air circulation oven to dry it at 50°C, sieve out the fine powder, and you will get the Gastrodia elata slices.

[0099] Step 2. Take the Gastrodia elata slices prepared in step 1, place them in a decoction machine, add 8 times the amount of water as the slices, soak for 30 minutes, boil over high heat, simmer for 60 minutes, filter while hot, and set aside; add 6 times the amount of water as the slices for the second time, boil over high heat, simmer for 40 minutes, filter while hot, and set aside; combine the two extraction filtrates, concentrate under reduced pressure at 60°C to a relative density of 1.08 (60°C), spray dry, add an appropriate amount of dextrin, mix well, and granulate to obtain the finished Gastrodia elata formula granules (each 1g of formula granules is equivalent to 4g of slices).

[0100] Step 3. Take the Gastrodia elata formula granules prepared in step 2, and detect the content, fingerprint, extract, properties, thin layer identification and other indicators of the formula granules according to the method under the content determination and fingerprint of Gastrodia elata formula granules YBZ-PFKL-2021122.

[0101] Comparative Example 2

[0102] A method for preparing Gastrodia elata decoction pieces that meets the quality requirements of Gastrodia elata formula granules comprises the following steps:

[0103] Step 1: Take the raw material of Gastrodia elata (medicinal material number T1), wash it, steam it in a steamer for 3 hours, cut it into thin slices using a linear reciprocating medicine cutter, cool it at room temperature, and then place it in a hot air circulation oven to dry it at 55°C, sieve out the fine powder, and you will get the Gastrodia elata slices.

[0104] Step 2. Take the Gastrodia elata slices prepared in step 1, place them in a decoction machine, add 8 times the amount of water as the slices, soak for 30 minutes, boil over high heat, simmer for 60 minutes, filter while hot, and set aside; add 6 times the amount of water as the slices for the second time, boil over high heat, simmer for 40 minutes, filter while hot, and set aside; combine the two extraction filtrates, concentrate under reduced pressure at 60°C to a relative density of 1.08 (60°C), spray dry, add an appropriate amount of dextrin, mix well, and granulate to obtain the finished Gastrodia elata formula granules (each 1g of formula granules is equivalent to 4g of slices).

[0105] Step 3. Take the Gastrodia elata formula granules prepared in step 2, and detect the content, fingerprint, extract, properties, thin layer identification and other indicators of the formula granules according to the method under the content determination and fingerprint of Gastrodia elata formula granules YBZ-PFKL-2021122.

[0106] Comparative Example 3

[0107] A method for preparing Gastrodia elata decoction pieces that meets the quality requirements of Gastrodia elata formula granules comprises the following steps:

[0108] Step 1: Take the raw material of Gastrodia elata (medicinal material number T1), wash it, steam it in a steamer for 4 hours, cut it into thin slices using a linear reciprocating medicine cutter, cool it at room temperature, and then place it in a hot air circulation oven to dry it at 60°C, sieve out the fine powder, and you will get the Gastrodia elata slices.

[0109] Step 2. Take the Gastrodia elata slices prepared in step 1, place them in a decoction machine, add 8 times the amount of water as the slices, soak for 30 minutes, boil over high heat, simmer for 60 minutes, filter while hot, and set aside; add 6 times the amount of water as the slices for the second time, boil over high heat, simmer for 40 minutes, filter while hot, and set aside; combine the two extraction filtrates, concentrate under reduced pressure at 60°C to a relative density of 1.07 (60°C), spray dry, add an appropriate amount of dextrin, mix well, and granulate to obtain the finished Gastrodia elata formula granules (each 1g of formula granules is equivalent to 4g of slices).

[0110] Step 3. Take the Gastrodia elata formula granules prepared in step 2, and detect the content, fingerprint, extract, properties, thin layer identification and other indicators of the formula granules according to the method under the content determination and fingerprint of Gastrodia elata formula granules YBZ-PFKL-2021122.

[0111] Table 2 Comparison of preparation process of each embodiment and comparative example

[0112]

[0113] It can be seen from Table 2 that all indicators of the Gastrodia elata formula granules prepared in Examples 1, 2, 3, 4, 5, and 6 meet the requirements of national standards, and the content, extract, properties, thin layer identification, and paste yield of the Gastrodia elata formula granules in Comparative Examples 1, 2, and 3 meet the requirements of national standards, but the fingerprint similarities are all lower than 0.90, which does not meet the national standard requirements for Gastrodia elata formula granules.

[0114] The similarity of the fingerprint spectra of the formula granules in each embodiment is greater than 0.90, and with the extension of the steaming time of the medicinal pieces, the similarity of the fingerprint spectra of the prepared Gastrodia elata formula granules shows an upward trend, but the content of the formula granules shows a downward trend with the extension of the steaming time of the medicinal pieces, which may be related to the mutual transformation of gastrodin and other ingredients under high temperature conditions.

[0115] Combined with the test results of various indicators, the processing technology of Gastrodia elata slices was determined as follows: take the original Gastrodia elata medicinal material, wash it, steam it in a steamer for 5h to 10h, cut it into thin slices, cool it at room temperature, and then place it in a hot air circulation oven at 50℃ to 60℃ for low-temperature drying, sieve out the fine powder, and you can get Gastrodia elata slices.

[0116] Example 7

[0117] The invention discloses a construction of a fingerprint of Gastrodia elata slices that meets the quality requirements of Gastrodia elata formula granules, comprising: using 15 batches of Gastrodia elata raw medicinal materials (medicinal material numbers T1-T15) of different batches and origins as raw materials, preparing them into Gastrodia elata slices according to the above-mentioned processing technology, selecting Gastrodia elata slices with all indicators of the prepared Gastrodia elata formula granules qualified as test samples and preparing test sample solutions, using ultra-high performance liquid chromatography to obtain characteristic spectra of each test sample solution, and then importing each characteristic spectrum into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system, using Mark peak matching to generate a common peak pattern spectrum, and constructing a fingerprint of Gastrodia elata slices.

[0118] Specifically, the 15 batches of Gastrodia elata raw medicinal materials numbered T1-T15 in this embodiment are processed according to the processing process determined above, the processed medicinal pieces are prepared into formula granules according to the Gastrodia elata formula granule preparation method, and the various indicators of the formula granules are tested according to the quality standard requirements of Gastrodia elata formula granules YBZ-PFKL-2021122.

[0119] Table 3 Test results of various indicators of Tianma formula granules prepared from 15 batches of Tianma decoction pieces

[0120]

[0121] As can be seen from Table 3, 15 batches of Gastrodia elata raw medicinal materials of different batches and origins were processed into Gastrodia elata decoction pieces according to the processing technology determined above. The content, fingerprint, extract, properties, thin layer identification, paste yield and other indicators of the prepared Gastrodia elata formula granules all met the national standard requirements. The 15 batches of Gastrodia elata decoction pieces all met the quality requirements of the formula granules and could be used as test samples for the construction of the fingerprint of Gastrodia elata decoction pieces.

[0122] Specifically, the chromatographic conditions of the ultra-high performance liquid chromatography in this embodiment are: octadecyl bonded silica gel as the filler, column length of 150 mm, column inner diameter of 2.1 mm, particle size of 1.6 μm, column temperature of 30°C; flow rate of 0.30 mL per minute, detection wavelength of 220 nm; injection volume of 1 μL, theoretical plate number calculated based on gastrodin should not be less than 3000; acetonitrile as mobile phase A, 0.05% phosphoric acid solution as mobile phase B, elution according to the following gradient elution program:

[0123]

[0124] Preparation of reference substance solutions: Take appropriate amounts of gastrodin, p-hydroxybenzyl alcohol, barisonoside, barisonoside B, barisonoside C, and barisonoside E, accurately weigh them, place them in a volumetric flask, add methanol to dissolve and mix them, and prepare solutions containing 200μg, 5μg, 200μg, 150μg, 50μg, and 200μg per 1mL, respectively, as reference substance solutions.

[0125] Preparation of test solution: Take 1g of Gastrodia elata powder, accurately weigh it, put it in a stoppered conical flask, accurately add 50ml of water, heat and reflux for 1 hour, cool, filter, accurately measure 10ml of the filtrate, evaporate to dryness, add 30% methanol to dissolve and make up to 5ml, ultrasonically treat (power 250W, frequency 40kHz) for 30 minutes, cool and filter with a 0.22μm syringe filter to obtain the solution.

[0126] Determination method: Accurately pipette 1 μL of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0127] Specifically, in this embodiment, there are 6 characteristic peaks in the characteristic spectrum of 15 batches of Gastrodia elata slices, which are located by 6 reference substances respectively. In order of peak appearance time, they are gastrodin, p-hydroxybenzyl alcohol, barisonoside E, barisonoside B, barisonoside C, and barisonoside. The chromatograms of the reference substance solutions are shown in Figure 1-6 .

[0128] In this embodiment, in order to construct the reference fingerprint of Gastrodia elata slices, 15 batches of characteristic spectrum detection chromatograms of Gastrodia elata slices of different batches and origins were introduced into the Chinese medicine chromatographic fingerprint similarity evaluation system (2012.130723 version), and the similarity of 15 batches of Gastrodia elata slices samples was evaluated. The evaluation results showed that the similarity of the slices between each batch was above 0.95, see Figure 7 , as shown in Table 4.

[0129] The six characteristic peaks of gastrodin, p-hydroxybenzyl alcohol, barisonoside E, barisonoside B, barisonoside C and barisonoside were selected as Mark peaks, and the common peak pattern spectrum was generated by Mark peak matching to obtain the reference fingerprint spectrum of Gastrodia elata decoction pieces, as shown in Figure 8 shown.

[0130]

[0131] Example 8

[0132] An application of fingerprint spectrum of Gastrodia elata decoction pieces that meets the quality requirements of Gastrodia elata formula granules includes using an established fingerprint spectrum detection method for Gastrodia elata decoction pieces and a constructed fingerprint spectrum of Gastrodia elata decoction pieces to perform fingerprint spectrum determination on Gastrodia elata decoction pieces with different steaming times, and establishing a linear relationship between the similarity of fingerprint spectrum of Gastrodia elata decoction pieces and the similarity of fingerprint spectrum of Gastrodia elata formula granules, so as to evaluate whether the quality of Gastrodia elata decoction pieces meets the requirements of Gastrodia elata formula granules.

[0133] Specifically, in this embodiment, the Gastrodia elata slices prepared with the above-mentioned different steaming times (2h, 3h, 4h, 5h, 6h, 7h, 8h, 9h, 10h) were subjected to fingerprint determination according to the above-mentioned fingerprint detection method, and Mark peak matching was used and introduced into the Chinese medicine chromatographic fingerprint similarity evaluation system to detect the similarity between each slice and the control fingerprint.

[0134] Specifically, in this embodiment, the similarity of the fingerprints of Gastrodia elata slices at different steaming times is used as the horizontal coordinate (X), and the similarity of the fingerprints of the corresponding Gastrodia elata formula granules is used as the vertical coordinate (Y), and a linear relationship between the two is established. The results show that the similarity of the fingerprints of Gastrodia elata slices and the fingerprints of Gastrodia elata formula granules show a typical positive correlation. The linear regression equation is: Y = 0.4714X + 0.5544, and the correlation coefficient R 2 =0.9768, such as Fig. 9 shown.

[0135] According to the linear regression equation, if the similarity of the fingerprint spectrum of Tianma formula granules is not less than 0.90, the similarity of Tianma decoction pieces should not be less than 0.733. Taking into account the errors in the detection and processing process, combined with multiple batches of data, it is corrected to the similarity of the fingerprint spectrum of Tianma decoction pieces should not be less than 0.76.

[0136] Table 5 Similarity test results of fingerprint spectra of Gastrodia elata slices and Gastrodia elata granules with different steaming times

[0137]

[0138] In particular, in order to investigate the rationality of the application of the constructed fingerprint, the following three comparative examples were conducted.

[0139] Comparative Example 4

[0140] An application of fingerprint spectrum of Gastrodia elata decoction pieces that meets the quality requirements of Gastrodia elata formula granules comprises the following steps:

[0141] Step 1: Take the raw material of Gastrodia elata (No. T7), wash it, steam it in a steamer for 1.5 hours, cut it into thin slices using a linear reciprocating medicine cutter, cool it at room temperature, and then place it in a hot air circulation oven to dry it at 55°C, sieve out the fine powder, and obtain the Gastrodia elata slices.

[0142] Step 2. Take the Gastrodia elata slices prepared in step 1, place them in a decoction machine, add 8 times the amount of water as the slices, soak for 30 minutes, boil over high heat, simmer for 60 minutes, filter while hot, and set aside; add 6 times the amount of water as the slices for the second time, boil over high heat, simmer for 40 minutes, filter while hot, and set aside; combine the two extraction filtrates, concentrate under reduced pressure at 60°C to a relative density of 1.09 (60°C), spray dry, add an appropriate amount of dextrin, mix well, and granulate to obtain the finished Gastrodia elata formula granules (each 1g of formula granules is equivalent to 4g of slices).

[0143] Step 3. Take the Gastrodia elata formula granules prepared in step 2, and detect the content, fingerprint, extract, properties, thin layer identification and other indicators of the formula granules according to the method under the content determination and fingerprint of Gastrodia elata formula granules YBZ-PFKL-2021122.

[0144] Comparative Example 5

[0145] An application of fingerprint spectrum of Gastrodia elata decoction pieces that meets the quality requirements of Gastrodia elata formula granules comprises the following steps:

[0146] Step 1: Take the Gastrodia elata slices steamed for 1.5 hours in Example 1, place them in a steamer and steam them for another 5 hours. After steaming, cool them at room temperature, place them in a hot air circulation oven and dry them at 55°C, sieve out the fine powder, and obtain the Gastrodia elata slices.

[0147] Step 2. Take the Gastrodia elata slices prepared in step 1, place them in a decoction machine, add 8 times the amount of water as the slices, soak for 30 minutes, boil over high heat, simmer for 60 minutes, filter while hot, and set aside; add 6 times the amount of water as the slices for the second time, boil over high heat, simmer for 40 minutes, filter while hot, and set aside; combine the two extraction filtrates, concentrate under reduced pressure at 60°C to a relative density of 1.08 (60°C), spray dry, add an appropriate amount of dextrin, mix well, and granulate to obtain the finished Gastrodia elata formula granules (each 1g of formula granules is equivalent to 4g of slices).

[0148] Step 3. Take the Gastrodia elata formula granules prepared in step 2, and detect the content, fingerprint, extract, properties, thin layer identification and other indicators of the formula granules according to the method under the content determination and fingerprint of Gastrodia elata formula granules YBZ-PFKL-2021122.

[0149] Comparative Example 6

[0150] An application of fingerprint spectrum of Gastrodia elata decoction pieces that meets the quality requirements of Gastrodia elata formula granules comprises the following steps:

[0151] Step 1: Take the raw material of Gastrodia elata (No. T7), wash it, place it in a pool, add appropriate amount of water (completely cover the material) and moisten it for 10 hours, take it out, filter out the water, cut it into thin slices using a linear reciprocating medicine cutter, and then place it in a hot air circulation oven at 55°C to dry it, sieve out the fine powder, and you will get the Gastrodia elata slices.

[0152] Step 2. Take the Gastrodia elata slices prepared in step 1, place them in a decoction machine, add 8 times the amount of water as the slices, soak for 30 minutes, boil over high heat, simmer for 60 minutes, filter while hot, and set aside; add 6 times the amount of water as the slices for the second time, boil over high heat, simmer for 40 minutes, filter while hot, and set aside; combine the two extraction filtrates, concentrate under reduced pressure at 60°C to a relative density of 1.08 (60°C), spray dry, add an appropriate amount of dextrin, mix well, and granulate to obtain the finished Gastrodia elata formula granules (each 1g of formula granules is equivalent to 4g of slices).

[0153] Step 3. Take the Gastrodia elata formula granules prepared in step 2, and detect the content, fingerprint, extract, properties, thin layer identification and other indicators of the formula granules according to the method under the content determination and fingerprint of Gastrodia elata formula granules YBZ-PFKL-2021122.

[0154] Table 6 Experimental results of comparative examples 4, 5 and 6

[0155]

[0156] It can be seen from the experimental results of the comparative examples that when the steaming time of the Gastrodia elata slices does not meet the processing technology requirements, the similarity of the fingerprints of the processed Gastrodia elata slices is only 0.514, and the similarity of the fingerprints of the prepared formula granules is 0.793, which is lower than the national standard requirements (Comparative Example 4); when the steaming time of the slices is extended to meet the processing technology requirements, the similarity of the fingerprints of the Gastrodia elata slices increases from 0.514 to 0.783, and the similarity of the fingerprints of the formula granules also increases from 0.793 to 0.934, which meets the national standard requirements (Comparative Example 5); when the moistening process is adopted, the similarity of the fingerprints of the processed Gastrodia elata slices is 0.503, and the similarity of the fingerprints of the prepared formula granules is 0.772, which does not meet the national standard requirements (Comparative Example 6), indicating that the processing technology of the present invention and the constructed fingerprint have good applicability.

[0157] Example 9

[0158] This example conducts a methodological investigation on the fingerprint detection method in Example 7, including chromatographic peak identification, precision test, stability test, and repeatability test, as follows:

[0159] 1. Chromatographic peak identification

[0160] Preparation of reference substance solutions: Take appropriate amounts of gastrodin, p-hydroxybenzyl alcohol, barisonoside, barisonoside B, barisonoside C, and barisonoside E, accurately weigh them, place them in a volumetric flask, add methanol to dissolve and mix them, and prepare solutions containing 200μg, 5μg, 200μg, 150μg, 50μg, and 200μg per 1mL, respectively, as reference substance solutions.

[0161] Preparation of test solution: Take 1g of Gastrodia elata powder, accurately weigh it, put it in a stoppered conical flask, accurately add 50ml of water, heat and reflux for 1 hour, cool, filter, accurately measure 10ml of the filtrate, evaporate to dryness, add 30% methanol to dissolve and make up to 5ml, ultrasonically treat (power 250W, frequency 40kHz) for 30 minutes, cool and filter with a 0.22μm syringe filter to obtain the solution.

[0162] According to the retention time and full-wavelength absorption curve of each reference substance, the characteristic spectral peaks of Gastrodia elata slices were identified, and peak No. 1 was identified as gastrodin, peak No. 2 was identified as p-hydroxybenzyl alcohol, peak No. 3 was identified as balisonoside E, peak No. 4 was identified as balisonoside B, peak No. 5 was identified as balisonoside C, and peak No. 6 was identified as balisonoside.

[0163] 2. Precision test

[0164] Take the same batch of Gastrodia elata slices, prepare 1 sample solution according to the sample solution preparation method in Example 7, and inject 6 times continuously according to the chromatographic conditions. The results show that the similarity between the 6 injections is greater than 0.9; taking gastrodin as the reference peak, the RSD of the relative retention time of each common peak is 0.19%, and the RSD of the relative peak area is 0.16%, indicating that the instrument has good precision. For related test results, see Fig.10 .

[0165] Table 7 Precision test similarity determination results

[0166]

[0167] Table 8 Relative retention time and relative peak area of ​​gastrodin

[0168]

[0169] 3. Stability test

[0170] Take the same batch of Gastrodia elata slices, prepare 1 sample solution according to the sample solution preparation method in Example 7, and inject and test at 0, 2, 4, 6, 8, and 12 hours according to the chromatographic conditions. The results show that the similarity between the 6 injections is greater than 0.9; taking gastrodin as the reference peak, the RSD of the relative retention time of each common peak is 0.85%, and the RSD of the relative peak area is 0.31%, indicating that the sample solution has good stability within 12 hours. For related test results, see Fig.11 .

[0171] Table 9 Stability test similarity determination results

[0172]

[0173]

[0174] Table 10 Relative retention time and relative peak area of ​​gastrodin

[0175]

[0176] 4. Repeatability test

[0177] Take the same batch of Gastrodia elata slices, prepare 6 test solutions according to the test solution preparation method in Example 7, and measure according to the chromatographic conditions. The results show that the similarity between the 6 test solutions is greater than 0.9; taking gastrodin as the reference peak, the RSD of the relative retention time of each common peak is 0.30%, and the results show that the method has good repeatability. For related test results, see Fig.12 .

[0178] Table 11 Results of similarity determination of repeatability test

[0179]

[0180] Table 12 Relative retention time of gastrodin

[0181]

[0182] The above is only a preferred embodiment of the present invention, not a limitation of the embodiment. For ordinary technicians in the field, several forms of changes or modifications can be made based on the above description. It is not necessary and impossible to describe all the embodiments here, and these changes or modifications are still within the protection scope of the present invention.

Claims

1. A fingerprint spectrum construction and detection method for Gastrodia elata decoction pieces, characterized in that: The following steps are involved: Step 1: Take the raw material of Gastrodia elata, wash it, steam it in a steamer for 5h to 10h, then cut it into thin slices, cool it at room temperature, dry it at 50℃ to 60℃, sieve out the fine powder, and obtain the Gastrodia elata slices; Step 2, placing the prepared Gastrodia elata slices in a decoction machine, adding 8 times the amount of water of the slices, soaking for 30 minutes, boiling over high heat, decocting over low heat for 60 minutes, filtering while hot, and setting aside; adding 6 times the amount of water of the slices for the second time, boiling over high heat, decocting over low heat for 40 minutes, filtering while hot, and setting aside; combining the two extraction filtrates, concentrating under reduced pressure at 60°C to a material-liquid ratio of 1:1, and a relative density of 1.05-1.10 at 60°C, spray drying, adding auxiliary materials, mixing, and granulating to obtain the finished Gastrodia elata formula granules, each 1g of the formula granules is equivalent to 4g of the slices; Step 3: Take the Tianma formula granules prepared in step 2, and detect the content, fingerprint, extract, properties, and thin layer identification indicators of the Tianma formula granules according to the requirements of the national drug standard YBZ-PFKL-2021122; Step 4: According to the processing technology of the Gastrodia elata slices corresponding to the Gastrodia elata formula granules that have passed the test of all indicators in step 3, 15 different batches of Gastrodia elata slices are processed, and steps 2 and 3 are performed respectively; Step 5: Take the Gastrodia elata decoction pieces corresponding to the Gastrodia elata formula granules with all the qualified indicators in step 4 as samples, and carry out the following steps: A) Take the Gastrodia elata slices, crush them, pass them through a No. 4 pharmacopoeia sieve, and take the sieve contents as the test sample; B) taking a sample of the test substance, dissolving it in a solvent, extracting it, and obtaining a test substance solution; C) using ultra-high performance liquid chromatography to measure the test solution to obtain UPLC characteristic spectra of each batch of Gastrodia elata slices; D) Importing the UPLC characteristic spectrum corresponding to each batch of medicinal materials into the Chinese medicine chromatographic fingerprint similarity evaluation system, using Mark peak matching to generate a common peak pattern spectrum, and constructing a reference fingerprint spectrum of Gastrodia elata decoction pieces; Step 6: The Gastrodia elata slices with different steaming times are respectively subjected to the characteristic spectrum detection according to the method described in step 5, and the Mark peaks are matched with the constructed fingerprint spectrum of the Gastrodia elata slices, and the similarity between the fingerprint spectrum and the control fingerprint spectrum is calculated; Step seven, with the offset of the fingerprint spectra of the Gastrodia elata slices at different steaming times in step six as the horizontal coordinate and the similarity of the fingerprint spectra of the corresponding Gastrodia elata formula granules as the vertical coordinate, a linear regression equation is fitted. According to the fitted linear regression equation, if the similarity of the fingerprint spectra of the Gastrodia elata formula granules is not less than 0.90, the similarity of the fingerprint spectra of the corresponding Gastrodia elata slices shall not be less than 0.

76.

2. The method according to claim 1, characterized in that In step 1, the steaming time is 5h, 6h, 7h, 8h, 9h, and 10h, and the low-temperature drying temperature is 55°C.

3. The method according to claim 1, characterized in that The steaming time range of the 15 different batches of Gastrodia elata slices during preparation in step 4 is 5h-10h.

4. The method according to claim 1, characterized in that The method for preparing the test solution in step 5 is specifically as follows: Take 1.0g of Gastrodia elata powder, accurately weigh it, put it in a stoppered conical flask, accurately add 50ml of water, heat and reflux for 1 hour, cool, filter, accurately measure 10ml of the filtrate, evaporate to dryness, add 30% methanol to dissolve and make up to 5ml, ultrasonically treat (power 250W, frequency 40kHz) for 30 minutes, cool and filter with a 0.22μm syringe filter to obtain.

5. The method according to claim 1, characterized in that The chromatographic conditions for obtaining the fingerprint of Gastrodia elata slices are: Chromatographic conditions: octadecyl bonded silica gel as filler, column length of 150 mm, column inner diameter of 2.1 mm, particle size of 1.6 μm, column temperature of 30°C; flow rate of 0.30 mL per minute, detection wavelength of 220 nm; injection volume of 1 μL, theoretical plate number calculated based on gastrodin should not be less than 3000; acetonitrile as mobile phase A, 0.05% phosphoric acid solution as mobile phase B, elution according to the following gradient elution program:

6. The method according to claim 1, characterized in that The method also includes preparing a reference substance solution, specifically: taking gastrodin, p-hydroxybenzyl alcohol, barisonoside, barisonoside B, barisonoside C, and barisonoside E reference substances, accurately weighing them, placing them in a volumetric flask, adding methanol to dissolve and mix them, and making solutions containing 200 μg, 5 μg, 200 μg, 150 μg, 50 μg, and 200 μg per 1 mL, respectively, as reference substance solutions.

7. The method according to claim 1, characterized in that Mark peaks refer to the six characteristic peaks of gastrodin, p-hydroxybenzyl alcohol, barisonoside, barisonoside B, barisonoside C and barisonoside E, which were injected under the same chromatographic conditions, and gastrodin, p-hydroxybenzyl alcohol, barisonoside, barisonoside B, barisonoside C and barisonoside E reference substances were used to locate and identify the chromatographic peaks.

8. The method according to claim 1, characterized in that In step 7, the similarity of the fingerprint spectrum of the Gastrodia elata decoction pieces is used as the abscissa (X), and the similarity of the fingerprint spectrum of the Gastrodia elata formula granules corresponding to the Gastrodia elata decoction pieces is used as the ordinate (Y) to perform linear regression equation fitting, and the fitted linear regression equation is: Y = 0.4714X + 0.5544, correlation coefficient R 2 =0.9768.

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