A method for constructing characteristic spectrum of Zanthoxylum bungeanum formula granules and its application

Abstract

The present invention provides a method for constructing a characteristic spectrum of a Zanthoxylum formula granule and its application, the method comprising the following steps: (1) mixing a sample with water, then mixing and extracting with an organic solvent, and taking an organic phase to obtain a test solution; (2) mixing a Zanthoxylum reference medicinal material with water and heating, then mixing and extracting with an organic solvent, and taking an organic phase to obtain a reference solution; (3) performing UPLC detection on the reference solution and the test solution respectively, screening out peaks with consistent retention time, good peak shape and high separation degree as characteristic peaks according to the detection results, and selecting S peaks with determined components and good peak shape from the characteristic peaks, calculating the relative retention times of other characteristic peaks relative to the S peak, and obtaining the characteristic spectrum of the Zanthoxylum formula granule. The characteristic spectrum of the Zanthoxylum formula granule obtained by the construction method provided by the present invention has high precision, good durability and high accuracy.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicine identification, and specifically relates to a method for constructing a characteristic spectrum of Zanthoxylum formula granules and an application thereof, and in particular to a method for constructing a characteristic spectrum of Zanthoxylum formula granules with high accuracy and an application thereof. Background Art

[0002] Zanthoxylum bungeanum, also known as pepper mortar, is the mature dry seed of Zanthoxylum bungeanum. It is nearly round, black, smooth and shiny, has the effect of diuresis and swelling, and is often used for phlegm and wheezing, edema and fullness; Zanthoxylum bungeanum is not included in the "Chinese Pharmacopoeia", but is included in the "Zhejiang Province Traditional Chinese Medicine Processing Specifications". There are few reports on the chemical composition and quality standards of Zanthoxylum bungeanum. Zanthoxylum bungeanum belongs to warming medicine, its medicinal properties are pungent and warm, and it belongs to the spleen, stomach, and kidney meridians. It has the effects of warming the middle and relieving pain, killing insects and relieving itching.

[0003] This medicine can be used to treat the following diseases clinically: 1. This medicine can be used to treat cold spleen and stomach syndrome. When used, it can be used with dried ginger, codonopsis, etc. for cold pain in the abdomen and abdomen caused by spleen and stomach deficiency and coldness, vomiting, etc. For abdominal pain, vomiting and diarrhea caused by cold and dampness in the middle, it can be used with Atractylodes lancea, Magnolia officinalis, etc. 2. This medicine can be used to treat ascaris abdominal pain, eczema itching, vaginal itching and other diseases. When used, it can be used with black plum, phellodendron, etc. for abdominal pain caused by worm accumulation, cold hands and feet, etc. For eczema itching, women's vaginal itching, etc., it can be used with Sophora flavescens, Phellodendron, etc. When using this medicine, it should be noted that this medicine is pungent, hot and dry, and the dosage should not be too large. Long-term use can also easily hurt yang and help fire. It is not suitable for patients with diseases such as real heat syndrome, yin deficiency and fire, and deficiency of body fluid and blood. It needs to be used with caution for pregnant patients.

[0004] At present, the research on Zanthoxylum bungeanum has found that its main components are unsaturated fatty acids, etc. Compared with the identification of the components of Zanthoxylum bungeanum, the identification of the components of Zanthoxylum bungeanum is relatively small, and there is a gap in the relevant research on its quality control methods. Therefore, how to provide a method for constructing a characteristic map of Zanthoxylum bungeanum to strengthen its specific identification and overall quality control has become an urgent problem to be solved. Summary of the invention

[0005] In view of the shortcomings of the prior art, the purpose of the present invention is to provide a method for constructing a characteristic spectrum of Zanthoxylum formula granules and its application, and in particular to provide a method for constructing a characteristic spectrum of Zanthoxylum formula granules with high accuracy and its application. The characteristic spectrum of Zanthoxylum formula granules obtained by the construction method provided by the present invention has high precision, good durability and high accuracy.

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

[0007] In one aspect, the present invention provides a method for constructing a characteristic spectrum of Zanthoxylum bungeanum formula particles, the construction method comprising the following steps:

[0008] (1) Mix the sample with water, then mix and extract with an organic solvent, and take the organic phase to obtain the test solution;

[0009] (2) mixing the Zanthoxylum order reference medicinal material with water and heating it, then mixing and extracting it with an organic solvent, and taking the organic phase to obtain a reference substance solution;

[0010] (3) The reference solution and the test solution are respectively subjected to UPLC detection, and according to the detection results, the peaks with consistent retention time, good peak shape and high separation are screened as characteristic peaks, and the peaks with determined components and good peak shape are selected as S peaks among the characteristic peaks, and the relative retention times of other characteristic peaks relative to the S peak are calculated, so as to obtain the characteristic spectrum of the Zanthoxylum bungeanum formula granules.

[0011] There is no order distinction between steps (1) and (2).

[0012] The above method is extracted through Zanthoxylum formula granules, compared with Zanthoxylum control medicinal materials, and jointly constructs a characteristic spectrum. The obtained characteristic spectrum can not only effectively identify the Zanthoxylum formula granules; but also the characteristic spectrum obtained by UPLC detection using a specific method has high accuracy, high precision and good durability.

[0013] Preferably, the liquid-to-solid ratio of the sample to water in step (1) is 1:(40-60) g / mL, for example 1:40 g / mL, 1:45 g / mL, 1:50 g / mL, 1:55 g / mL or 1:60 g / mL, but is not limited to the above-listed values. Other values ​​not listed within the above numerical range are also applicable.

[0014] Preferably, in step (2), the solid-liquid ratio of the Zanthoxylum order reference medicinal material to water is 1:(50-70) g / mL.

[0015] Preferably, the heating temperature in step (2) is 90-100° C. and the heating time is 20-40 min.

[0016] Among them, the material-liquid ratio can be 1:50g / mL, 1:55g / mL, 1:60g / mL, 1:65g / mL or 1:70g / mL, etc., the temperature can be 90°C, 91°C, 92°C, 93°C, 94°C, 95°C, 96°C, 97°C, 98°C, 99°C or 100°C, etc., and the time can be 20min, 25min, 30min, 35min or 40min, etc., but are not limited to the values ​​listed above, and other values ​​not listed within the above numerical range are also applicable.

[0017] Preferably, the organic solvent comprises ethyl acetate.

[0018] Preferably, the filler of the chromatographic column for UPLC detection in step (3) is octadecylsilane bonded silica gel.

[0019] Preferably, the column temperature of the UPLC detection in step (3) is 33-37°C.

[0020] Preferably, the flow rate of the UPLC detection in step (3) is 0.28-0.32 mL / min.

[0021] The column temperature can be 33°C, 34°C, 35°C, 36°C or 37°C, etc., and the flow rate can be 0.28mL / min, 0.29mL / min, 0.3mL / min, 0.31mL / min or 0.32mL / min, etc., but are not limited to the values ​​listed above. Other values ​​not listed within the above numerical range are also applicable.

[0022] Preferably, the mobile phase for UPLC detection in step (3) comprises mobile phase A and mobile phase B, wherein mobile phase A is acetonitrile, and mobile phase B is an acid aqueous solution, wherein the mass fraction of the acid aqueous solution is 0.08-1%, for example, 0.08%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1%, etc., but are not limited to the values ​​listed above, and other values ​​not listed within the above numerical range are equally applicable.

[0023] Preferably, the acid comprises any one of formic acid, acetic acid or phosphoric acid or a combination of at least two thereof, preferably acetic acid.

[0024] Preferably, the elution procedure of the UPLC detection in step (3) is gradient elution, and the specific process of the gradient elution is as follows:

[0025] In the first 0-2 minutes, the volume fraction of mobile phase A was changed from 5% to 10% at a constant speed, and the volume fraction of mobile phase B was changed from 95% to 90% at a constant speed;

[0026] From 2 to 5 minutes, the volume fraction of mobile phase A was 10%, and the volume fraction of mobile phase B was 90%;

[0027] From 5 to 10 minutes, the volume fraction of mobile phase A changes from 10% to 36% at a constant rate, and the volume fraction of mobile phase B changes from 90% to 64% at a constant rate;

[0028] From 10 to 20 minutes, the volume fraction of mobile phase A changes from 36% to 50% at a constant rate, and the volume fraction of mobile phase B changes from 64% to 50% at a constant rate;

[0029] From 20 to 30 minutes, the volume fraction of mobile phase A changes from 50% to 66% at a constant rate, and the volume fraction of mobile phase B changes from 50% to 34% at a constant rate;

[0030] From 30 to 38 minutes, the volume fraction of mobile phase A changes from 66% to 78% at a constant rate, and the volume fraction of mobile phase B changes from 34% to 32% at a constant rate;

[0031] From 38 to 43 minutes, the volume fraction of mobile phase A changed from 78% to 100% at a constant speed, and the volume fraction of mobile phase B changed from 22% to 0 at a constant speed.

[0032] Preferably, in the characteristic spectrum of the Zanthoxylum bungeanum formula particles in step (3), the number of characteristic peaks is 14, and the S peak includes S1 peak and S2 peak.

[0033] Sorted by retention time from small to large, with peak 2 as S1 peak, the relative retention times of peak 1, peak 3, peak 4, peak 5, peak 6, peak 7, peak 8, peak 9, peak 10 and S1 peak were 0.29±10%, 1.06±10%, 1.10±10%, 1.51±10%, 1.54±10%, 1.66±10%, 1.69±10%, 1.71±10%, 1.76±10%, respectively; with peak 11 as S2 peak, the relative retention times of peak 12, peak 13, peak 14 and S2 peak were 1.05±10%, 1.10±10%, 1.21±10%, respectively.

[0034] The component of the peak 2 is hydroxy-α-sanshool, the component of the peak 3 is hydroxy-β-sanshool, the component of the peak 11 is α-linolenic acid, the component of the peak 13 is linoleic acid, and the component of the peak 14 is oleic acid.

[0035] In a second aspect, the present invention also provides the application of the construction method as described above in the quality control of Zanthoxylum bungeanum formula granules.

[0036] Compared with the prior art, the present invention has the following beneficial effects:

[0037] The present invention provides a method for constructing a characteristic spectrum of Zanthoxylum formula granules. Extraction is performed through the Zanthoxylum formula granules, and the mixture is compared with a Zanthoxylum control medicinal material to jointly construct a characteristic spectrum. The obtained characteristic spectrum can effectively identify the Zanthoxylum formula granules; and the characteristic spectrum obtained by UPLC detection using a specific method has high accuracy, high precision and good durability. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 It is the characteristic spectrum result when the detection wavelength is 284nm in Example 1;

[0039] Figure 2 It is the characteristic spectrum result when the detection wavelength is 203nm in Example 1;

[0040] Figure 3It is the liquid phase result when the detection wavelength is 284nm in Example 2;

[0041] Figure 4 It is the liquid phase result when the detection wavelength is 203nm in Example 2;

[0042] Figure 5 It is the liquid phase result when the detection wavelength is 284nm in Example 3;

[0043] Figure 6 It is the liquid phase result when the detection wavelength is 203nm in Example 3;

[0044] Figure 7 It is the liquid phase result when the detection wavelength is 284nm in Example 4;

[0045] Figure 8 It is the liquid phase result when the detection wavelength is 203nm in Example 4;

[0046] Fig. 9 It is the liquid phase result when the detection wavelength is 284nm in Example 5;

[0047] Fig.10 It is the liquid phase result when the detection wavelength is 203nm in Example 5;

[0048] Fig.11 It is the liquid phase result when the detection wavelength is 284nm in Example 6;

[0049] Fig.12 It is the liquid phase result when the detection wavelength is 203nm in Example 6;

[0050] Fig.13 It is the liquid phase result when the detection wavelength is 284nm in Example 7;

[0051] Fig.14 This is the liquid phase result in Comparative Example 1. DETAILED DESCRIPTION

[0052] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0053] Example 1

[0054] This embodiment provides a method for constructing a characteristic spectrum of Zanthoxylum bungeanum formula particles, which is as follows:

[0055] 1. Test instruments and test drugs

[0056] Instruments: analytical balance (Sartorius, SQP QUINTIX224-1CN, Germany), Waters high performance liquid chromatograph (equipped with TUV detector, temperature-controlled column oven, temperature-controlled autosampler, USA), high-speed desktop centrifuge (SIGMA, 1-14, Germany), ultrasonic cleaner (Tianjin Aotesense, AS20500BT, China).

[0057] Test drug: Zanthoxylum bungeanum formula granules (batch numbers 20040571, 19044601, 19044621), prepared by Beijing Kangrentang Pharmaceutical Co., Ltd.;

[0058] Hydroxy-α-sanshool (Batch No.: AF21032006, Chengdu Aifa Co., Ltd.).

[0059] Reagents: acetonitrile (Tianjin Bohai Chemical Reagent Co., Ltd., 20210220, chromatographic grade), methanol (Tianjin Bohai Chemical Reagent Co., Ltd., 20210301, analytical grade), formic acid (Tokyo Chemical Industry Co., Ltd., 3C7ON-NJ, chromatographic grade).

[0060] 2. Test solution

[0061] Take an appropriate amount of Zanthoxylum bungeanum granules, grind them into powder, weigh 0.5g, accurately weigh, place in a stoppered conical flask, add 25mL of water, shake well, extract with 75mL of ethyl acetate, take the ethyl acetate solution, evaporate to dryness, add 70% methanol to dissolve the residue and transfer it to a 5mL volumetric flask, dilute to the scale with 70% methanol, shake well, filter, and take the filtrate to obtain.

[0062] 3. Comparative medicinal material reference solution

[0063] Take 1.0 g of Zanthoxylum bungeanum reference medicinal material, add 30 mL of water, boil at 100°C for 30 min, cool, shake, filter, extract with 75 mL of ethyl acetate, take the ethyl acetate solution, evaporate to dryness, add 70% methanol to dissolve the residue and transfer to a 5 mL volumetric flask, dilute to the scale with 70% methanol, shake, filter, and take the filtrate as the reference solution of the reference medicinal material.

[0064] 4. Chromatographic conditions

[0065] Octadecylsilane bonded silica gel is used as filler (column length 100 mm, inner diameter 2.1 mm, particle size 1.7 μm); acetonitrile is used as mobile phase A, 0.1% acetic acid solution is used as mobile phase B, and gradient elution is performed according to the provisions in the following table; the flow rate is 0.3 mL per minute; the column temperature is 35°C; the detection wavelength is 284 nm (0-30 min) and 203 nm (30 min-43 min). The theoretical plate number calculated based on the hydroxy-α-sanshool peak should not be less than 5000.

[0066]

[0067] The above-mentioned reference medicinal material solutions and test sample solutions were subjected to UPLC detection, and the results were summarized to obtain characteristic peaks and characteristic spectra (see Figure 1-2 , the numbers in the figure represent the corresponding characteristic peaks), they are sorted from small to large by retention time, with peak 2 as the S1 peak, and the relative retention times of peak 1, peak 3, peak 4, peak 5, peak 6, peak 7, peak 8, peak 9, peak 10 and S1 peak are 0.29, 1.06, 1.10, 1.51, 1.54, 1.66, 1.69, 1.71, 1.76 respectively; with peak 11 as the S2 peak, the relative retention times of peak 12, peak 13, peak 14 and S2 peak are 1.05, 1.10, 1.21 respectively.

[0068] After comparison with the standard reference substance, it was confirmed that the component of peak 2 was hydroxy-α-sanshool, the component of peak 3 was hydroxy-β-sanshool, the component of peak 11 was α-linolenic acid, the component of peak 13 was linoleic acid, and the component of peak 14 was oleic acid.

[0069] 5. Methodological Validation

[0070] 5.1 Precision

[0071] Take an appropriate amount of Zanthoxylum bungeanum formula granules (batch number: 20040571), measure according to the above chromatographic conditions, use hydroxy-α-sanshool peak and α-linolenic acid as reference peaks, calculate the relative retention time and relative peak area, calculate the RSD value, and the results are as follows.

[0072]

[0073] Chromatographic peak number 11(S2) 12 13 14 11(S2) Precision 1 1.000 1.051 1.097 1.207 1.000 Precision 2 1.000 1.051 1.096 1.207 1.000 Precision 3 1.000 1.051 1.096 1.207 1.000 Precision 4 1.000 1.051 1.097 1.208 1.000 Precision 5 1.000 1.051 1.097 1.208 1.000 Precision 6 1.000 1.051 1.097 1.209 1.000 RSD / % 0.00 0.02 0.03 0.05 0.00

[0074] The results showed that the RSD of the relative retention time and relative peak area of ​​the characteristic peaks were both less than 3%, indicating good precision.

[0075] 5.2 Intermediate precision

[0076] Take an appropriate amount of Zanthoxylum bungeanum formula granules (batch number: 20040571), measure according to the above chromatographic conditions, use hydroxy-α-sanshool peak and α-linolenic acid as reference peaks, calculate the relative retention time and relative peak area, calculate the RSD value, and the results are as follows.

[0077]

[0078] Chromatographic peak number 11(S2) 12 13 14 11(S2) Intermediate precision 1 1.000 1.051 1.097 1.208 1.000 Intermediate precision 2 1.000 1.051 1.097 1.208 1.000 Intermediate precision 3 1.000 1.051 1.097 1.208 1.000 Intermediate precision 4 1.000 1.051 1.097 1.208 1.000 Intermediate precision 5 1.000 1.051 1.097 1.208 1.000 Intermediate precision 6 1.000 1.051 1.097 1.208 1.000 RSD / % 0.00 0.02 0.02 0.02 0.00

[0079] The results showed that the RSD of the relative retention time and relative peak area of ​​the characteristic peaks were both less than 3%, indicating good repeatability.

[0080] 5.3 Durability

[0081] 5.3.1 Stability

[0082] Take the test solution and inject it at 0, 2, 4, 8, 12, and 24 hours according to the above chromatographic conditions, record the retention time and peak area of ​​the identified peak, and the results are as follows. Take the hydroxy-α-sanshool peak and α-linolenic acid as reference peaks, calculate the relative retention time of other characteristic peaks and S peak, and the results are as follows.

[0083]

[0084] Chromatographic peak number 11(S2) 12 13 14 11(S2) 0h 1.000 1.050 1.095 1.200 0h 2h 1.000 1.050 1.095 1.200 2h 4h 1.000 1.050 1.095 1.200 4h 8h 1.000 1.050 1.095 1.200 8h 12h 1.000 1.050 1.095 1.200 12h 24h 1.000 1.050 1.095 1.200 24h RSD / % 0.00 0.00 0.00 0.00 RSD / %

[0085] The results showed that the RSD of the relative retention time and relative peak area of ​​the characteristic peaks were both less than 1%, indicating that the test sample had good stability within 24 hours.

[0086] 5.3.2 Investigation of different column temperatures

[0087] Take an appropriate amount of Zanthoxylum bungeanum particles, grind them into fine powder, and prepare the test solution according to the preparation method of the test solution mentioned above. Use different column temperatures (33℃, 35℃ and 37℃) to measure according to the method in the text to investigate the durability of the experimental method for column temperature. The results are shown in the following table.

[0088]

[0089]

[0090] The results of the investigation at different column temperatures showed that the relative retention time RSD of 14 identification peaks in the samples at different temperatures was in the range of 0.0% to 1.58%, indicating that this method has good durability for column temperature.

[0091] 5.3.3 Investigation of different flow rates

[0092] Take an appropriate amount of Zanthoxylum bungeanum particles, grind them into fine powder, and prepare the test solution according to the preparation method of the test solution in the text. Use different flow rates (0.28mL / min, 0.30mL / min, 0.32mL / min) to measure according to the above method to examine the durability of the experimental method for column temperature. The results are shown in the following table.

[0093]

[0094]

[0095] The results of the investigation at different flow rates showed that when different flow rates (0.28ml / min, 0.30ml / min, 0.32ml / min) were investigated respectively, the relative retention time RSD of 14 identification peaks in the sample was in the range of 0.00% to 3.61%, indicating that this method has good durability to flow rate.

[0096] Summary: Judging from the durability test results at different column temperatures and flow rates, this method has good durability.

[0097] 6. Data processing

[0098] According to the requirements of traditional Chinese medicine fingerprint technology, the above method was used to analyze the characteristic spectra of three batches of Zanthoxylum bungeanum granules. The "Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System" (2004.A version) was used to calculate the similarity between the characteristic spectra of the three batches of Zanthoxylum bungeanum granules and the characteristic spectra in Section 4. The results are shown in the table below.

[0099]

[0100] Summary: The similarities between the characteristic spectra of the three batches of Zanthoxylum granules and the control characteristic spectra were all greater than 0.95, indicating that there was basically no difference in the spectra, indicating that this method has high accuracy.

[0101] Example 2-3

[0102] Examples 2-3 respectively provide a method for constructing a characteristic spectrum of Zanthoxylum bungeanum formula particles, which are consistent with Example 1 except that the concentration of the acetic acid solution in the mobile phase is 0.5% and 1% respectively.

[0103] The results are as follows Figure 3-6 shown.

[0104] It can be seen from the spectrum that the above acid concentration has little effect on the characteristic peaks of the characteristic spectrum, indicating that the above acid concentration is suitable for this characteristic method.

[0105] Embodiment 4-5

[0106] Examples 4-5 respectively provide a method for constructing a characteristic spectrum of Zanthoxylum bungeanum formula particles, which is consistent with Example 1 except that the acetic acid solution in the mobile phase is replaced by formic acid solution and phosphoric acid solution of equal concentrations.

[0107] The results are as follows Figure 7-10 shown.

[0108] It can be seen from the spectrum that the above acid types have little effect on the characteristic peaks of the characteristic spectrum, indicating that the above acid types are suitable for this characteristic method.

[0109] Example 6

[0110] Example 6 provides a method for constructing a characteristic spectrum of Zanthoxylum bungeanum formula particles, which is consistent with Example 1 except that the elution procedure is as follows.

[0111]

[0112] The results are as follows Fig.11 shown.

[0113] It can be seen from the spectrum that the peak shapes of the characteristic peaks in this elution procedure are poor.

[0114] Example 7

[0115] Example 7 provides a method for constructing a characteristic spectrum of Zanthoxylum bungeanum formula particles, which is consistent with Example 1 except that the elution procedure is as follows.

[0116]

[0117] The results are as follows Fig.12 shown.

[0118] It can be seen from the spectrum that the peak shapes of the characteristic peaks in this elution procedure are poor.

[0119] Comparative Example 1

[0120] Comparative Example 1 provides a method for constructing a characteristic spectrum of Zanthoxylum formula particles, and the specific chromatographic conditions are as follows:

[0121] Octadecylsilane bonded silica gel is used as filler; methanol is used as mobile phase A and water is used as mobile phase B, and gradient elution is performed according to the provisions in the following table; the detection wavelength is 254nm. The theoretical plate number calculated based on the peak of zanthoxylum bungeanum should not be less than 10,000.

[0122]

[0123] Preparation of test solution:

[0124] Take an appropriate amount of the sample to be tested, grind it into powder, weigh 1.0g, accurately weigh it, put it in a stoppered conical flask, accurately add 25mL of 70% methanol solution, stopper it, weigh it, and treat it with ultrasound (power 300w, frequency 40kHz) for 30 minutes. Let it cool, weigh it again, make up the lost weight with 70% methanol, shake it well, filter it, and take the filtrate to obtain it.

[0125] The results are as follows Fig.14 As shown. The above method refers to the detection of pepper formula granules in the Shanghai Traditional Chinese Medicine Formula Granules Quality Standard (Fourth Batch). It can be seen from the figure that most of the peaks cannot be effectively detected. Although pepper is the seed coat of the Zanthoxylum order, the pepper method is not suitable for the detection of Zanthoxylum order varieties.

[0126] The applicant declares that the present invention illustrates the construction method and application of the characteristic spectrum of the Zanthoxylum formula granules of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0127] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0128] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A method for constructing a characteristic spectrum of Zanthoxylum bungeanum formula particles, characterized in that: The construction method comprises the following steps: (1) Mixing the sample with water, then mixing with an organic solvent for extraction, and taking the organic phase to obtain a test solution; the organic solvent is ethyl acetate; (2) mixing the Zanthoxylum bungeanum reference medicinal material with water and heating it, then mixing it with an organic solvent for extraction, and taking the organic phase to obtain a reference medicinal material solution; (3) The control medicinal material solution and the test solution are respectively subjected to UPLC detection, and the peaks with consistent retention time, good peak shape and high separation are screened out as characteristic peaks according to the detection results, and the peak with a certain component and good peak shape is selected as the S peak among the characteristic peaks, and the relative retention time of other characteristic peaks relative to the S peak is calculated, so as to obtain the characteristic spectrum of the Zanthoxylum bungeanum formula granules; the flow rate of the UPLC detection is 0.28-0.32 mL / min; The filler of the chromatographic column detected by UPLC is octadecylsilane bonded silica gel; The mobile phase for UPLC detection includes mobile phase A and mobile phase B, wherein mobile phase A is acetonitrile, and mobile phase B is an aqueous acid solution, wherein the mass fraction of the aqueous acid solution is 0.08-1%; and the acid includes any one of formic acid, acetic acid or phosphoric acid, or a combination of at least two thereof; The detection wavelengths of the UPLC detection are 0-30min, 284nm and 30min-43min, 203nm; The elution procedure of the UPLC detection is gradient elution, and the specific process of the gradient elution is as follows: In the first 0-2 minutes, the volume fraction of mobile phase A was changed from 5% to 10% at a constant speed, and the volume fraction of mobile phase B was changed from 95% to 90% at a constant speed; From 2 to 5 min, the volume fraction of mobile phase A was 10%, and the volume fraction of mobile phase B was 90%; From 5 to 10 minutes, the volume fraction of mobile phase A changes at a constant rate from 10% to 36%, and the volume fraction of mobile phase B changes at a constant rate from 90% to 64%; From 10 to 20 minutes, the volume fraction of mobile phase A was changed from 36% to 50% at a constant rate, and the volume fraction of mobile phase B was changed from 64% to 50% at a constant rate; From 20 to 30 minutes, the volume fraction of mobile phase A changed from 50% to 66% at a constant rate, and the volume fraction of mobile phase B changed from 50% to 34% at a constant rate; From 30 to 38 minutes, the volume fraction of mobile phase A changed from 66% to 78% at a constant rate, and the volume fraction of mobile phase B changed from 34% to 32% at a constant rate; From 38 to 43 minutes, the volume fraction of mobile phase A changed from 78% to 100% at a constant rate, and the volume fraction of mobile phase B changed from 22% to 0 at a constant rate; In the characteristic spectrum of the Zanthoxylum formula granules, the number of characteristic peaks is 14, and the S peak includes the S1 peak and the S2 peak; Sorted by retention time from small to large, with peak 2 as S1 peak, the relative retention times of peak 1, peak 3, peak 4, peak 5, peak 6, peak 7, peak 8, peak 9, peak 10 and S1 peak were 0.29±10%, 1.06±10%, 1.10±10%, 1.51±10%, 1.54±10%, 1.66±10%, 1.69±10%, 1.71±10%, 1.76±10%, respectively; with peak 11 as S2 peak, the relative retention times of peak 12, peak 13, peak 14 and S2 peak were 1.05±10%, 1.10±10%, 1.21±10%; The component of peak 2 is hydroxy-α-sanshool, and the component of peak 11 is α-linolenic acid; There is no order in which steps (1) and (2) are performed.

2. The construction method according to claim 1, characterized in that: The solid-liquid ratio of the sample to water in step (1) is 1:(40-60) g / mL.

3. The construction method according to claim 1, characterized in that: The solid-liquid ratio of the Zanthoxylum order reference medicinal material to water in step (2) is 1:(50-70) g / mL.

4. The construction method according to claim 1, characterized in that: The heating temperature in step (2) is 90-100°C and the heating time is 20-40 min.

5. The construction method according to claim 1, characterized in that: The column temperature of the UPLC detection in step (3) is 33-37°C.

6. The construction method according to claim 1, characterized in that: The acid includes acetic acid.

7. An application of the construction method according to any one of claims 1 to 6 in the quality control of Zanthoxylum bungeanum formula granules.

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