Construction method of compound moslae herba water fingerprint and application thereof

Abstract

The application discloses a method for constructing a compound elsholtzia ciliata water fingerprint and application thereof. The compound elsholtzia ciliata water fingerprint constructed by the application realizes the purpose of simultaneously detecting multiple volatile components in compound elsholtzia ciliata water, changes the technical means of quality control of single index component, can more comprehensively evaluate the volatile component quality of compound elsholtzia ciliata water, and is suitable for controlling the product quality of compound elsholtzia ciliata water.

Description

Technical Field

[0001] This invention belongs to the field of quality control technology of traditional Chinese medicine preparations, specifically relating to a method for constructing a water fingerprint spectrum of compound Elsholtzia ciliata and its application. Background Technology

[0002] Traditional Chinese medicine (TCM) fingerprinting refers to the chromatograms or spectra obtained by appropriate analysis of certain TCM materials or preparations after proper processing. These spectra comprehensively reflect the types and quantities of chemical components contained in the medicinal materials, effectively demonstrating the holistic and synergistic effects of TCM components. Due to its speed and accuracy, it has been widely used in TCM analysis, identification, and quality control. Gas chromatography fingerprinting is a hot topic in TCM quality control research and is suitable for quality monitoring of volatile components in TCM preparations.

[0003] Compound Elsholtzia Water is a traditional Chinese medicine preparation composed of nine herbs: Elsholtzia ciliata, Indigofera tinctoria, Pogostemon cablin, Perilla frutescens, Magnolia officinalis, Amomum villosum, Aucklandia lappa, Zingiber officinale, and Glycyrrhiza uralensis. It has the effects of relieving exterior syndromes and resolving dampness, invigorating the spleen and stomach, and is used for external wind-cold, internal summer-dampness, headache with chills and fever, abdominal distension and pain, nausea and vomiting, and diarrhea with borborygmus. Compound Elsholtzia ciliata water is listed in the National Medical Products Administration's drug standard YBZ00542022. The current standard only includes physicochemical identification, liquid chromatography identification of Magnolia officinalis, and thin-layer chromatography identification of Aucklandia lappa, etc., and the key medicinal ingredients in the prescription are not effectively monitored. Furthermore, seven medicinal ingredients in the prescription—Elsholtzia ciliata, Pogostemon cablin, Perilla frutescens, Amomum villosum, Aucklandia lappa, Magnolia officinalis, and Zingiber officinale—contain volatile components. These volatile ingredients account for a significant proportion (90.7% of the total medicinal materials in the prescription). The preparation process also involves volatile oil extraction from these medicinal ingredients and adding the volatile oil to the formulation. Therefore, for… To more scientifically and comprehensively characterize the intrinsic quality of Compound Elsholtzia Water, strictly control its quality stability, and improve the product's technical level, it is necessary to monitor the volatile components in Compound Elsholtzia Water (such as eucalyptol in cardamom, β-dehydroelushenone in Elsholtzia ciliata, perilla ketone in perilla leaf, baicalein in patchouli, dehydrocostus lactone in costus root, and magnolol and honokiol in magnolia bark) and establish a gas phase fingerprint spectrum for Compound Elsholtzia Water. This is of great significance for product quality control and can provide a comprehensive quality control method for Compound Elsholtzia Water. Summary of the Invention

[0004] To address the limitation of current quality evaluation systems for Compound Elsholtzia ciliata Water in comprehensively reflecting and controlling product quality, this invention aims to provide a method for constructing a fingerprint spectrum for Compound Elsholtzia ciliata Water. This method enables the simultaneous determination of multiple indicator components in Compound Elsholtzia ciliata Water, overcoming the shortcomings of existing standards that cannot comprehensively reflect the quality status of Compound Elsholtzia ciliata Water. This invention achieves its objective through the following technical solution:

[0005] This invention provides a method for constructing a water fingerprint spectrum of compound Elsholtzia ciliata, comprising the following steps:

[0006] 1) Preparation of test solution: Take the compound Elsholtzia water sample and extract it with an organic solvent to obtain the test solution;

[0007] 2) Preparation of reference solution: Take appropriate amounts of eucalyptol, perillaldehyde, baicalein, dehydrosaponin lactone, magnolol, and honokiol, dissolve them in organic solvents and dilute to volume to prepare reference solution.

[0008] 3) Inject the above-mentioned test solution and reference solution into a gas chromatograph, measure and record the chromatograms. Process the chromatogram of the compound Xiangru water sample using the 2012 version of the "Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System" to obtain the fingerprint chromatogram of the compound Xiangru water. The gas chromatographic detection conditions are as follows:

[0009] The chromatographic column is an elastic quartz capillary column, with cross-linked 5% phenylmethyl polysiloxane or cyanopropyl polysiloxane as the stationary phase;

[0010] The column temperature is programmed, with an injection port temperature of 200–250°C, a detector temperature of 260–290°C, a gas flow rate of 1.0–2.0 mL / min, a split ratio of (5–15):1, and an injection volume of 0.3–1 μl. Preferably, the injection port temperature is 230°C, the detector temperature is 280°C, the gas flow rate is 1.0–2.0 mL / min, preferably 1.5 mL / min, the split ratio is 10:1, and the injection volume is 0.3–1 μl, preferably 1 μl.

[0011] In some embodiments, the heating program is as follows: the initial temperature is 80-120°C, held for 3-8 minutes, then heated to 160-200°C at a rate of 3-10°C per minute, and then heated to 220-260°C at a rate of 5-8°C per minute, and held for 5-15 minutes.

[0012] Preferably, the heating program is as follows: the initial temperature is 80-120°C, held for 3-8 minutes, then increased to 150-200°C at a rate of 3-10°C per minute, held for 0-2 minutes, and then increased to 220-260°C at a rate of 4-8°C per minute, held for 5-15 minutes.

[0013] More preferably, the heating program is as follows: the initial temperature is 100°C, held for 5 minutes, then heated to 160°C at a rate of 3°C per minute, and then heated to 240°C at a rate of 8°C per minute, and held for 10 minutes.

[0014] The temperature program can also be as follows: start at 100°C, hold for 8 minutes, increase to 150°C at a rate of 4°C per minute, hold for 2 minutes, then increase to 240°C at a rate of 4°C per minute, and hold for 15 minutes.

[0015] In some embodiments, the organic solvents described in steps 1) and 2) are each independently selected from diethyl ether and n-hexane, preferably n-hexane.

[0016] In some embodiments, the extraction method is liquid-liquid extraction.

[0017] In some embodiments, the chromatographic column is selected from Agilent HP-5 column (30m × 0.32mm, 0.25μm), DB-WAX column (30m × 0.32mm, 0.25μm), and Agilent DB-1701 column (30m × 0.25mm, 0.25μm).

[0018] In some embodiments, the detector used in the gas chromatography conditions is a flame ionization detector (FID).

[0019] In some embodiments, the present invention utilizes GC-MS technology to compare with reference standards and combines the retention time of the reference standards with information such as molecular ion peaks and fragmentation fragments from mass spectrometry to identify chromatographic peaks.

[0020] In some embodiments, the concentration of the reference solution (eucalyptol, perillaldehyde, baicalein, dehydroauric acid lactone, magnolol and honokiol) in step 2) is 80-120 μg / mL, preferably 100-120 μg / mL, and more preferably 100 μg / mL.

[0021] In some embodiments, the fingerprint spectrum contains 11 common peaks, wherein peak 1 is eucalyptol, peak 4 is perillaldehyde, peak 7 is baicalein, peak 9 is dehydroauric acid lactone, peak 10 is magnolol, and peak 11 is honokiol. Using the baicalein reference peak as a reference peak, the relative retention times of each chromatographic peak are: peak 1 0.1628, peak 2 0.3022, peak 3 0.3132, peak 4 0.3746, peak 5 0.4531, peak 6 0.6336, peak 7 1.0, peak 8 1.0429, peak 9 1.3375, peak 10 1.4646, and peak 11 1.5244. The deviation range of the relative retention times of each common peak is ±10%.

[0022] This invention is the first to construct a gas chromatographic fingerprint of compound Elsholtzia ciliata water. Based on the structural characteristics and physicochemical properties of the active ingredients in compound Elsholtzia ciliata water, the analytical conditions, including sample processing methods, gas chromatographic conditions, detection wavelength, column temperature, and flow rate, were screened and optimized, and the systematic methodology was validated. The obtained fingerprint of compound Elsholtzia ciliata water exhibits high resolution, good peak shape, and good baseline separation for all characteristic chromatographic peaks. It demonstrates good stability and numerous characteristic peaks, enabling a comprehensive and accurate evaluation of the quality of volatile components in compound Elsholtzia ciliata water, and is suitable for controlling the quality of compound Elsholtzia ciliata water products. Attached Figure Description

[0023] Figure 1 The image shows the water chromatogram of the compound Elsholtzia ciliata from Example 1.1.

[0024] Figure 2 The image shows the water chromatogram of the compound Elsholtzia ciliata from Example 1.3.

[0025] Figure 3 The image shows the water chromatogram of the compound Elsholtzia ciliata from Example 1.4.

[0026] Figure 4 This is a chromatogram overlay for a precision test.

[0027] Figure 5 This is a chromatogram overlay for repeatability testing.

[0028] Figure 6 This is a chromatogram overlay for stability testing.

[0029] Figure 7 This is a superimposed chromatogram of 18 batches of samples of Compound Elsholtzia Water.

[0030] Figure 8 For comparison with fingerprint patterns.

[0031] Figure 9 This is a superimposed chromatogram of the reference standard and the test sample.

[0032] Figure 10 This is a superimposed image of the fingerprint spectra of each medicinal material and the test sample. Detailed Implementation

[0033] The present invention will be further described below with reference to specific embodiments and accompanying drawings, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.

[0034] Unless otherwise specified, all reagents and materials used in this invention are commercially available.

[0035] The instrument, reagent, and sample information are as follows:

[0036] 1. Instruments

[0037] Agilent 7890B gas chromatograph (FID detector), Agilent HP-5 column (30m × 0.32mm × 0.25μm), Agilent DB-1701 column (30m × 0.25mm × 0.25μm), electronic microbalance (Mettler-Toledo).

[0038] 2. Reagents

[0039] Reference standards: Eucalyptol (110788-202108, purity 99.4%, China National Institutes for Food and Drug Control), Perillaldehyde (F11IB207024, purity 98.5%, Shanghai Yuanye Biotechnology Co., Ltd.), Prunol (110772-201909, purity 100%, China National Institutes for Food and Drug Control), Dehydroausyl lactone (111525-201912, purity 99.5%, China National Institutes for Food and Drug Control); Magnolol (110729-202015, purity 99.0%, China National Institutes for Food and Drug Control), and Magnolol (110730-201313, purity 99.5%, China National Institutes for Food and Drug Control).

[0040] Diethyl ether was of analytical grade, and n-hexane was of chromatographic grade.

[0041] 3. Sample

[0042] The single medicinal herbs *Elsholtzia rugulosa*, *Elsholtzia ciliata*, *Pogostemon cablin*, *Perilla frutescens*, *Magnolia officinalis*, *Amomum villosum*, *Saussurea costus*, ginger, and licorice, as identified by the Quality Department of Guangzhou Yipinhong Pharmaceutical Co., Ltd., all meet the standards stipulated in the *Chinese Pharmacopoeia* (2020 edition). *Elsholtzia rugulosa* is the dried aerial part of *Elsholtzia rugulosa* hemsl. (Lamiaceae); *Elsholtzia ciliata* is the dried stem and leaves of *Piper boehmerifolium* (Miq.) C.DC. (Piperaceae); *Pogostemon cablin* is the dried aerial part of *Pogostemon cablin* (Blanco) Benth. (Lamiaceae); *Perilla frutescens* is the dried leaves (or with tender branches) of *Perilla frutescens* (L.) Britt. (Lamiaceae); and *Magnolia officinalis* is the *Magnolia officinalis* Rehd. et Wils. or *Magnolia officinalis* (Magnoliaceae). The dried bark, root bark, and branch bark of *Rehd. et Wils.* var. *biloba* Rehd. et Wils.; the dried mature fruit of *Amomum kravanh Pierre ex Gagnep.* or *Amomum compactum Soland ex Maton* (both belonging to the ginger family); the dried root of *Aucklandia lappa Decne.* (belonging to the Asteraceae family); the fresh rhizome of *Zingiber officinale Rosc.* (belonging to the ginger family); and the dried root and rhizome of *Glycyrrhiza uralensis* Fisch., *Glycyrrhiza inflata* Bat., or *Glycyrrhiza glabra* L. (belonging to the legume family).

[0043] 18 batches of Compound Elsholtzia Water (specification: 10mL / bottle) were produced by Yipinhong Biopharmaceutical Co., Ltd. The serial numbers and production batch numbers are shown in Table 1.

[0044] Table 1 Sample Information Table

[0045]

[0046]

[0047] Example 1: Investigation of Gas Chromatographic Conditions for Compound Elsholtzia ciliata Water

[0048] 1.1 Investigation of chromatographic temperature program conditions

[0049] 1. Preparation of test solution: Take 20 mL of this product (batch number 10622010), place it in a 50 mL centrifuge tube, extract it three times with n-hexane, 5 mL each time, combine the n-hexane solutions, blow nitrogen down to 5 mL, shake well, and the solution is obtained.

[0050] 2. Chromatographic conditions: The column was an Agilent HP-5 capillary column (30 m in length, 0.32 mm in inner diameter, and 0.25 μm in membrane thickness); the temperature program was as follows: initial temperature 100 °C, held for 5 minutes, increased to 160 °C at a rate of 3 °C / min, then increased to 240 °C at a rate of 8 °C / min, and held for 10 minutes. The injection port temperature was 230 °C, the detector temperature was 280 °C, the gas flow rate was 2 mL / min, and the split ratio was 10:1.

[0051] 3. Determination: Accurately pipette 1 μl of the test solution and inject it into the gas chromatograph for determination. Record the chromatogram. Results are shown below. Figure 1 The results showed that the main chromatographic peaks appeared within 40 minutes, with good peak resolution and shape, and the response values ​​of the main chromatographic peaks were suitable.

[0052] 1.2 Investigation of chromatographic temperature program conditions

[0053] 1. Preparation of test solution: Take 20 mL of this product (batch number 10622010), place it in a 50 mL centrifuge tube, extract it three times with 5 mL of ether each time, combine the ether solutions, blow nitrogen down to 5 mL, shake well, and the solution is ready.

[0054] 2. Chromatographic conditions: The column was an Agilent HP-5 capillary column (30 m in length, 0.32 mm in inner diameter, and 0.25 μm in membrane thickness); the temperature program was as follows: initial temperature 120 °C, held for 6 minutes, increased to 200 °C at a rate of 10 °C / min, then increased to 260 °C at a rate of 5 °C / min, and held for 15 minutes. The injection port temperature was 250 °C, the detector temperature was 270 °C, the gas flow rate was 1.5 mL / min, and the split ratio was 15:1.

[0055] 3. Determination: Accurately pipette 1 μl of the test solution and inject it into the gas chromatograph for determination, then record the chromatogram. The results show that this method has good peak resolution and excellent peak shape.

[0056] 1.3 Investigation of chromatographic temperature program conditions

[0057] 1. Preparation of test solution: Take 20 mL of this product (batch number 10621010), place it in a 50 mL centrifuge tube, extract it three times with n-hexane, 5 mL each time, combine the n-hexane solutions, blow nitrogen down to 5 mL, shake well, and the solution is obtained.

[0058] 2. Chromatographic conditions: The column was an Agilent DB-1701 capillary column (30 m in length, 0.25 mm in inner diameter, and 0.25 μm in membrane thickness); the temperature program was as follows: initial temperature 100 °C, held for 8 minutes, increased to 150 °C at a rate of 4 °C per minute, held for 2 minutes, then increased to 240 °C at a rate of 4 °C per minute, and held for 15 minutes. The injection port temperature was 230 °C, the detector temperature was 260 °C, the gas flow rate was 1.0 mL / min, and the split ratio was 10:1.

[0059] 3. Determination: Accurately pipette 1 μl of the test solution and inject it into the gas chromatograph for determination. Record the chromatogram. Results are shown below. Figure 2 The results showed that the main chromatographic peaks appeared within 52 minutes, the chromatographic peaks were well separated and had good peak shapes, and the response values ​​of the main chromatographic peaks were suitable.

[0060] 1.4 Investigation of Chromatographic Temperature Program Conditions (Comparative Example)

[0061] 1. Preparation of test solution: Take 20 mL of this product (batch number 10622010), place it in a 50 mL centrifuge tube, extract it three times with n-hexane, 5 mL each time, combine the n-hexane solutions, blow nitrogen down to 5 mL, shake well, and the solution is obtained.

[0062] 2. Chromatographic conditions: The column was an Agilent HP-5 capillary column (30 m in length, 0.32 mm in inner diameter, and 0.25 μm in membrane thickness); the temperature program was as follows: initial temperature 40 °C, held for 3 minutes, increased to 120 °C at a rate of 3 °C per minute, and then increased to 280 °C at a rate of 10 °C per minute. The injection port temperature was 230 °C, and the detector temperature was 280 °C; the gas flow rate was 1.5 mL / min; and the split ratio was 10:1.

[0063] 3. Determination: Accurately pipette 1 μl of the test solution and inject it into the gas chromatograph for determination. Record the chromatogram. Results are shown below. Figure 3 The results showed that this method had poor peak separation and shape, and high baseline noise.

[0064] As can be seen, this invention has screened and optimized the processing method and gas chromatography conditions of the test sample based on the structural characteristics and physicochemical properties of the effective components contained in the compound Elsholtzia ciliata water, resulting in chromatograms with high resolution and good peak shape.

[0065] Example 2 Methodological Investigation

[0066] 2.1 Precision Test

[0067] Samples from batch number 10622010 were taken, and test solutions were prepared according to the method in Example 1.1. The samples were injected six times consecutively under the chromatographic conditions in Example 1.1. The relative retention time, relative peak area, and similarity were used as evaluation indicators. The results showed that the RSD of the relative retention time of the common peaks was less than 1.0%, the RSD of the relative peak area was less than 2.0%, and the similarity was greater than 0.95, indicating that the instrument has good precision.

[0068] chromatogram overlay Figure 4 .

[0069] 2.2 Repeatability Test

[0070] Six samples from batch number 10622010 were taken and test solutions were prepared according to the method in Example 1.1. The samples were injected six times consecutively under the chromatographic conditions in Example 1.1. The relative retention time, relative peak area, and similarity were used as evaluation indicators. The results showed that the RSD of the relative retention time of the common peaks was less than 2.0%, the RSD of the relative peak area was less than 3.0%, and the similarity was greater than 0.96, indicating that the method had good repeatability.

[0071] chromatogram overlay Figure 5 .

[0072] 2.3 Stability Test

[0073] Samples from batch 10622010 of the precision test were injected at 0, 6, 12, 24, 32, 48, and 60 hours according to the chromatographic conditions in Example 1.1. The relative retention time, relative peak area, and similarity were used as evaluation indicators. The results showed that the RSD of the relative retention time of the common peaks was less than 3%, the RSD of the relative peak area was less than 2.5%, and the similarity was greater than 0.95, indicating that the samples had good stability within 60 hours.

[0074] chromatogram overlay Figure 6 .

[0075] Example 3: Construction of aqueous vapor phase fingerprint of compound Elsholtzia ciliata

[0076] 1) Following the test solution preparation method and gas chromatography detection conditions described in Example 1.1, 18 batches of Compound Elsholtzia ciliata water samples were taken and analyzed. The superimposed chromatograms of the 18 batches of Compound Elsholtzia ciliata water samples are shown below. Figure 7 .

[0077] 2) The main chromatographic peaks obtained from the above 18 batches of Compound Elsholtzia ciliata water all appeared within 40 minutes. The obtained fingerprint chromatograms were imported into the "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine" (2012 edition) of the National Pharmacopoeia Commission. The chromatographic peaks before 2 minutes were clipped. Using the median method and a time window width set to 0.1, a multi-point correction method was applied to perform Mark peak matching on the chromatograms to generate a control fingerprint chromatogram. Eleven common peaks were identified. (See attached image) Figure 8 .

[0078] It is easy to see that the compound Elsholtzia water fingerprint spectrum detected by this method has many chemical components, moderate peak heights, stable baseline, and good separation and peak shape.

[0079] 3) Take appropriate amounts of eucalyptol, perillaldehyde, baicalein, dehydroauric acid lactone, magnolol, and honokiol reference standards, respectively, and prepare solutions with a concentration of 100 μg / mL. Inject each reference standard solution sequentially according to the chromatographic conditions in Example 1.1, and record the chromatograms of the reference standards. We used GC-MS technology to compare with the reference standards to assign chromatographic peaks. Combining the retention time of the reference standards with information such as molecular ion peaks and fragmentation fragments from the mass spectrometry, we identified six chromatographic peaks: peak 1 was eucalyptol, peak 4 was perillaldehyde, peak 7 was baicalein, peak 9 was dehydroauric acid lactone, peak 10 was honokiol, and peak 11 was honokiol. The chromatograms of the above reference standards and the test sample were obtained, as shown in [reference needed]. Figure 9 .

[0080] Experiment Example 4: Assignment of Common Peaks

[0081] (1) Preparation of the test solution of medicinal materials

[0082] 1) Weigh out 10g of Elsholtzia ciliata, 5g of Pogostemon cablin, 5g of Perilla frutescens, 1g of Magnolia officinalis, 1.5g of Amomum villosum, 1.0g of Zingiber officinale, and 0.4g of Glycyrrhiza uralensis. Grind them into coarse powder, add 1000mL of water to each powder, and extract for 2 hours. Collect the volatile oil, filter, and reserve the filtrate. Boil the residue twice more, adding 1000mL of water each time for 2 hours each time, filter, combine the above filtrates, concentrate under reduced pressure, add ethanol solution, stir well, and adjust the total volume to 100ml to obtain the extract of Elsholtzia ciliata, Pogostemon cablin, Perilla frutescens, Magnolia officinalis, Amomum villosum, Zingiber officinale, and Glycyrrhiza uralensis.

[0083] 2) Weigh 1.0g of Costus root and 1.0g of Indigofera tinctoria, grind them into coarse powder, moisten with 60% ethanol, soak for 48 hours, then begin percolation. Collect 6.5ml of the initial percolate and set aside. Continue collecting the subsequent percolate until 1000ml is obtained, then stop percolation. Concentrate the percolate under reduced pressure, add ethanol solution to adjust the total volume to 1000ml, and obtain the Costus root and Indigofera tinctoria extract.

[0084] Take 20 ml of each of the above-mentioned herbal extracts and place them in a 50 mL centrifuge tube. Extract with n-hexane three times, 5 ml each time. Combine the n-hexane solutions, blow with nitrogen to 5 ml, and shake well to obtain the test solutions of each herbal material.

[0085] (2) Shared peak attribution

[0086] Take the above-mentioned medicinal material test solutions and compound Elsholtzia water test solutions (prepared according to Example 1.1), respectively, and inject them according to the chromatographic conditions in Example 1.1, and record the chromatograms. Through chromatographic peak matching and analysis, the common peaks were assigned, as shown in Table 2 and... Figure 10 The results showed that peaks 1 and 2 belonged to cardamom; peaks 3 and 5 belonged to Elsholtzia ciliata; peaks 4, 6, and 8 belonged to Perilla frutescens; peak 7 belonged to patchouli; peak 9 belonged to Saussurea costus; and peaks 10 and 11 belonged to Magnolia officinalis.

[0087] In addition, using peak 6 (Baiqiu Lichun) as a reference peak, the average relative retention times (peak numbers) of the other 10 common peaks in the 18 batches of samples were calculated as follows: 0.1628(1), 0.3022(2), 0.3132(3), 0.3746(4), 0.4531(5), 0.6336(6), 1.0429(8), 1.3375(9), 1.4646(10), and 1.5244(11). The relative deviations were all within 2%.

[0088] Table 2. Assignment of common peaks in fingerprint spectral data.

[0089]

[0090] Example 5 Similarity Evaluation

[0091] The similarity evaluation system for chromatographic fingerprints of traditional Chinese medicine (2012 edition) published by the National Pharmacopoeia Commission was used to analyze the similarity between the fingerprint data of 18 batches of samples and the generated control chromatograms. The similarity of the 18 batches of samples is shown in Table 3. The data in the table show that the similarity of the 18 batches of samples is high, all greater than 0.9, indicating that the quality stability between sample batches is good. This fingerprint is suitable for quality control in the production process of compound Xiangru water.

[0092] Table 3 Similarity of 18 batches of samples

[0093]

[0094]

[0095] Obviously, the above embodiments are merely illustrative examples for clarity and are not intended to limit the implementation. Those skilled in the art will recognize that various variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations. However, any obvious variations or modifications derived therefrom remain within the scope of this invention.

Claims

1. A method for constructing a compound Shangye water fingerprint, comprising the following steps: 1) preparing a test solution: taking a compound Shangye water sample, extracting with an organic solvent to obtain a test solution; 2) preparing a control solution: taking eucalyptol, perilene, baiquloli, dehydroeudesmanol, magnolol and honokiol in proper amounts, dissolving and constant volume in an organic solvent respectively to prepare a control solution; 3) injecting the test solution and the control solution into a gas chromatograph, determining and recording the chromatogram, and processing the chromatogram of the compound Shangye water sample by using the Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System 2012 version to obtain the fingerprint of the compound Shangye water, wherein the conditions for detection by the gas chromatograph are as follows: the chromatographic column is an Agilent HP-5 chromatographic column or a DB-1701 chromatographic column, and the stationary phase is cross-linked 5% phenylmethyl polysiloxane or cyano propyl polysiloxane; the column temperature is programmed, the injection port temperature is 200-250℃, the detector temperature is 260-290℃, the gas flow rate is 1.0-2.0 mL / min, the split ratio is (5-15) : 1, and the injection volume is 0.3-1 μl; the temperature program is as follows: the initial temperature is 80-120℃, maintaining for 3-8 minutes, increasing at a rate of 3-10℃ per minute to 150-200℃, maintaining for 0-2 minutes, and then increasing at a rate of 4-8℃ per minute to 220-260℃, maintaining for 5-15 minutes; the organic solvent in step 1) and step 2) is independently selected from diethyl ether and n-hexane. 2.The method according to claim 1, wherein the extraction in step 1) is liquid-liquid extraction. 3.The method according to claim 1 or 2, wherein the detector used in the gas chromatograph is a flame ionization detector. 4.The method according to any one of claims 1-3, wherein the concentration of the control solution is 80-120 μg / ml. 5.The method according to any one of claims 1-4, wherein the temperature program is as follows: the initial temperature is 100℃, maintaining for 5 minutes, increasing at a rate of 3℃ per minute to 160℃, and then increasing at a rate of 8℃ per minute to 240℃, maintaining for 10 minutes. 6.The method according to any one of claims 1-5, wherein the temperature program is as follows: the initial temperature is 100℃, maintaining for8 minutes, increasing at a rate of 4℃ per minute to 150℃, maintaining for 2 minutes, and then increasing at a rate of 4℃ per minute to 240℃, maintaining for 15 minutes. 7.The fingerprint obtained by the method according to any one of claims 1-6. 8.The use of the fingerprint obtained by the method according to any one of claims 1-7 in the quality control of compound Shangye water.

2. The construction method of claim 1, wherein, ​ 3. The method of claim 1, wherein, ​ 4. The method of claim 1, wherein, ​ 5. The method of claim 1, wherein, ​ 6. The method of claim 1, wherein, ​ 7. The method of claim 1, wherein, ​ ​

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