Testing methods and quality control methods for Tangut rhubarb products

Abstract

The present invention relates to the technical field of traditional Chinese medicine detection, and in particular to a detection method and quality control method for a Tangut rhubarb product. The detection method for the Tangut rhubarb product provided by the present invention uses octadecylsilane bonded silica gel as a filler, uses methanol-phosphoric acid aqueous solution as a mobile phase for gradient elution, selects a specific elution program, and obtains 18 common characteristic peaks, which not only significantly improves the number of characteristic peaks, but also achieves good separation of these common characteristic peaks, the elution program is simple, the obtained characteristic spectrum baseline is stable, each characteristic peak has a good peak shape, and the separation between each characteristic peak is high, so the detection method can effectively improve the accuracy of the characteristic spectrum of the Tangut rhubarb product, and can fully reflect the integrity and characteristic of the Tangut rhubarb product, which is conducive to more comprehensive monitoring of the quality of the Tangut rhubarb product.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine detection, and in particular to a detection method and a quality control method for a rhubarb tangut product. Background Art

[0002] The pharmacopoeia stipulates that rhubarb for human use should come from the dried roots and rhizomes of Rheum palmatum L., Rheum offcinale Baill., and Rheum tanguticum Maxim. ex Balf. It is dug up in late autumn when the stems and leaves wither or before germination in the next spring, the fine roots are removed, the outer skin is scraped off, the petals or segments are cut, and the dried in strings or directly dried.

[0003] Among them, Tangut rhubarb is bitter and cold in nature. It has the effects of clearing heat and clearing the intestines, cooling blood and detoxifying, clearing away heat and dampness, and removing blood stasis and dredging menstruation. It is used to treat constipation due to excess heat, abdominal pain due to accumulation, diarrhea, jaundice due to damp heat, red eyes, swollen throat, abdominal pain due to intestinal abscesses, carbuncles, furuncles, amenorrhea due to blood stasis, and external treatment of burns caused by water or fire, toothache caused by wind and fire, and injuries caused by falls.

[0004] Tangut rhubarb is an important source variety of medicinal rhubarb and has a wide range of uses. A detection method that can comprehensively and quickly detect it is needed to better control the quality of Tangut rhubarb products.

[0005] Characteristic spectrum detection method is a commonly used quality control method for traditional Chinese medicine. However, the existing characteristic spectrum detection method for Tangut rhubarb products can effectively separate fewer chemical components, and the separation effect of each component is poor. The number of characteristic peaks in the obtained characteristic spectrum is small, and it cannot comprehensively monitor the quality of Tangut rhubarb products. Summary of the invention

[0006] Therefore, the technical problem to be solved by the present invention is to overcome the defect that the relevant characteristic spectrum in the existing characteristic spectrum detection method of Tangut rhubarb products cannot comprehensively monitor the quality of Tangut rhubarb products due to the small number of characteristic peaks and low separation, thereby providing a detection method and quality control method for Tangut rhubarb products.

[0007] To this end, the present invention provides a method for detecting a Tangut rhubarb product, comprising the following steps:

[0008] Take the test solution and detect it by ultra-high performance liquid chromatography; wherein the chromatographic conditions of the ultra-high performance liquid chromatography include:

[0009] Octadecylsilane bonded silica gel is used as a filler, methanol is used as a mobile phase A, and a phosphoric acid aqueous solution, a formic acid aqueous solution or an acetic acid aqueous solution with a volume percentage of 0.03-0.20% is used as a mobile phase B for gradient elution. The gradient elution program includes: 0-1min, the volume percentage of methanol in the mobile phase is 5%→20%; 1-9min, the volume percentage of methanol in the mobile phase is 20%→35%; 9-17min, the volume percentage of methanol in the mobile phase is 35%→37%; 17-21min, the volume percentage of methanol in the mobile phase is 37%→48%; 21-24min, the volume percentage of methanol in the mobile phase is 48%→60%; 24-30min, the volume percentage of methanol in the mobile phase is 60%→100%; 30-35min, the volume percentage of methanol in the mobile phase is 100%.

[0010] Optionally, the chromatographic conditions of the ultra-high performance liquid chromatography method further include at least one of the following conditions:

[0011] 1) The detection wavelength is 260-270nm;

[0012] 2) Column temperature is 30-40°C;

[0013] 3) Flow rate is 0.20-0.30 mL / min;

[0014] 4) The injection volume is 0.5 to 2 μL;

[0015] 5) The chromatographic column is a CORTECS T3 chromatographic column with an inner diameter of 2.1 mm, a column length of 100 mm, and a particle size of 1.6 μm.

[0016] Optionally, the Rhubarb Tangut product includes at least one of Rhubarb Tangut medicinal materials, Rhubarb Tangut decoction pieces and Rhubarb Tangut preparations;

[0017] Optionally, the Tangut rhubarb decoction piece includes at least one of rhubarb (Tangut rhubarb), cooked rhubarb (Tangut rhubarb) and wine rhubarb (Tangut rhubarb);

[0018] Optionally, the Tangut rhubarb preparation includes at least one of rhubarb (Tangut rhubarb) formula granules, cooked rhubarb (Tangut rhubarb) formula granules and wine rhubarb (Tangut rhubarb) formula granules.

[0019] Optionally, when the tangut rhubarb product is a tangut rhubarb preparation, the preparation process of the test solution includes:

[0020] Taking Tangut rhubarb preparation powder, adding extraction solvent, extracting, solid-liquid separation, and taking liquid; wherein the extraction solvent is selected from any one of water, methanol or methanol aqueous solution, and the volume percentage of methanol in the methanol aqueous solution is not less than 60%;

[0021] Optionally, relative to 1g of the Tangut rhubarb preparation powder, the amount of the extraction solvent added is 50 to 150ml;

[0022] Optionally, the extraction is ultrasonic extraction, the ultrasonic power is 200W to 300W, and the ultrasonic time is 15 to 45 minutes.

[0023] Optionally, when the Rheum tanguticum product is Rheum tanguticum medicinal material and / or decoction pieces, the preparation process of the test solution includes:

[0024] Take the Tangut rhubarb medicinal material and / or the decoction pieces, add water, heat and reflux, separate the solid and liquid, and concentrate the obtained liquid to dryness to obtain a concentrated residue;

[0025] Adding an extraction solvent to the concentrated residue, extracting, separating the solid and the liquid, and taking the liquid; the extraction solvent is selected from any one of water, methanol or a methanol aqueous solution, and the volume percentage of methanol in the methanol aqueous solution is not less than 60%;

[0026] Optionally, the amount of water added is 25-75 ml relative to 1 g of the Tangut rhubarb medicinal material and / or decoction pieces;

[0027] Optionally, the heating reflux temperature is 80-100° C. and the time is 20-60 min;

[0028] Optionally, when adding the extraction solvent to the concentrated residue of the Rheum tanguticum medicinal material and / or the decoction pieces, the amount of the extraction solvent added is 25 to 75 ml relative to 1 g of the Rheum tanguticum medicinal material and / or the decoction pieces;

[0029] Optionally, the extraction is ultrasonic extraction, the ultrasonic power is 200W to 300W, and the ultrasonic time is 15min to 45min.

[0030] Optionally, the detection method further comprises the steps of preparing a reference solution using a gallic acid reference substance, a catechin reference substance, a sennoside A reference substance, a rhein reference substance, a rhein reference substance, a chrysophanol reference substance, a physophanol methyl ether reference substance and an aloe-rhein reference substance, and the step of detecting the reference solution by ultra-high performance liquid chromatography according to any of the above detection methods to obtain a reference spectrum of the reference substances;

[0031] Optionally, the detection method further includes the step of preparing a control medicinal material solution using Rheum tanguticum as a control medicinal material, and the step of detecting the control medicinal material solution by ultra-high performance liquid chromatography according to any of the detection methods described above to obtain a control medicinal material reference atlas.

[0032] The present invention also provides the use of any of the above detection methods in the quality control of Rheum tanguticum products and / or the use of Rheum tanguticum products in the identification of Rheum tanguticum products.

[0033] The present invention also provides a quality control method for Tangut rhubarb products, comprising the steps of obtaining a characteristic spectrum of the Tangut rhubarb product to be tested according to any of the above detection methods, and comparing the characteristic spectrum with a control characteristic spectrum;

[0034] Among them, the control characteristic spectrum is obtained by fitting the characteristic spectrum obtained by using at least one batch of standard samples of Tangut rhubarb products according to any of the detection methods described above through the average value or median method.

[0035] Optionally, the control characteristic spectrum includes 18 characteristic peaks, peak 1 is a gallic acid peak, peak 2 is a catechin peak, peak 3 is an epicatechin-3-O-gallate peak, peak 4 is an iso-anectoside peak, peak 5 is anctoside peak, peak 6 is an aloe-emodin-8-O-β-D-glucoside peak, peak 7 is a resveratrol-4'-O-β-D-(6"-O-galloyl) glucoside peak, peak 8 is a rhein-8-O-β-D-glucoside peak, peak 9 is a sennoside A peak, peak 10 is a chrysophanol-1-O-β-D-glucoside peak, peak 11 is a chrysophanol-8-O-β-D-glucoside peak, peak 12 is a rhein-8-O-β-D-pyranoglucoside peak, peak 13 is a The peak 14 is the aloe-rhein peak, the peak 15 is the rhein peak, the peak 16 is the rhein peak, the peak 17 is the chrysophanol peak, and the peak 18 is the rhein peak. The rhein peak (peak 15) is the first reference peak. The relative retention time of each characteristic peak and the first reference peak is within ±10% of the specified value, and the specified value is: 0.29 (peak 3), 0.32 (peak 4), 0.33 (peak 5), 0.37 (peak 6), 0.44 (peak 7), 0.46 (peak 8), 0.81 (peak 10), 0.83 (peak 11), 0.86 (peak 12), and 0.93 (peak 13).

[0036] Optionally, in the control characteristic spectrum, the chrysophanol peak (peak 17) is used as the second reference peak, and the relative peak areas of peaks 5, 7, 8 and 9 and the second reference peak are within a specified range, and the specified range is: not less than 0.9 (peak 5), not less than 1.4 (peak 7), not less than 13.0 (peak 8) and not less than 1.7 (peak 9).

[0037] The technical solution of the present invention has the following advantages:

[0038] 1. The detection method of the Tangut rhubarb product provided by the present invention uses octadecylsilane bonded silica gel as a filler, methanol-phosphoric acid aqueous solution as a mobile phase for gradient elution, and selects a specific elution program to obtain 18 common characteristic peaks, which not only significantly increases the number of characteristic peaks, but also achieves good separation of these common characteristic peaks. The elution procedure is simple, the baseline of the obtained characteristic spectrum is stable, the peak shape of each characteristic peak is good, and the separation degree between each characteristic peak is high. Therefore, the detection method can effectively improve the accuracy of the characteristic spectrum of the Tangut rhubarb product, and can fully reflect the integrity and characteristic of the Tangut rhubarb product, which is conducive to more comprehensive monitoring of the quality of the Tangut rhubarb product.

[0039] 2. The detection method of the Tangut rhubarb product provided by the present invention contains 18 characteristic peaks in the characteristic spectrum obtained by detection, accurately locates the peak positions of aloe-rhein peak, rhein peak, rhein peak, chrysophanol peak and physophanol methyl ether peak, and identifies each of the remaining characteristic peaks, providing a basis for quality detection and control of the Tangut rhubarb product.

[0040] 3 The detection method of the Tangut rhubarb product provided by the present invention adopts a specific test solution preparation method, the preparation process is simple and rapid, and the chemical components in the Tangut rhubarb product can be extracted comprehensively and sufficiently, which can further improve the accuracy of the characteristic spectrum of the Tangut rhubarb product and can more comprehensively monitor the quality of the Tangut rhubarb product. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0042] Figure 1 It is a comparison chart of the chromatogram of the test sample, the chromatogram of the reference medicinal material and the chromatogram of the reference substance in Example 1 of the present invention;

[0043] Figure 2 It is a chromatogram of 21 batches of Tangut rhubarb products in Example 2 of the present invention;

[0044] Figure 3 It is a comparative characteristic spectrum of the Tangut rhubarb product in Example 2 of the present invention;

[0045] Figure 4 This is the negative control chromatogram for specificity verification in Example 3 of the present invention;

[0046] Figure 5This is the reference medicinal material chromatogram of rhubarb (tangut rhubarb) for specific property verification in Example 3 of the present invention;

[0047] Figure 6 This is a chromatogram of the specific property verification of rhubarb (tangut rhubarb) formula granules in Example 3 of the present invention;

[0048] Figure 7 It is the precision verification chromatogram in Example 3 of the present invention;

[0049] Figure 8 is the repeatability verification chromatogram in Example 3 of the present invention;

[0050] Fig. 9 It is the intermediate precision verification chromatogram in Example 3 of the present invention;

[0051] Fig.10 This is a chromatogram of stability study in Example 3 of the present invention;

[0052] Figure 11-13 This is a chromatogram obtained by investigating different column temperatures in Example 3 of the present invention;

[0053] Figure 14-16 This is a chromatogram of different flow rates in Example 3 of the present invention;

[0054] Figure 17-19 This is a chromatogram obtained by investigating different chromatographic columns in Example 3 of the present invention;

[0055] Figure 20-22 This is a chromatogram of the same type of chromatographic columns with different numbers in Example 3 of the present invention;

[0056] Figure 23-25 This is a chromatogram of the mobile phase system with different phosphoric acid concentrations in Example 3 of the present invention;

[0057] Fig.26 is the chromatogram when the column temperature is 30° C. in Example 4 of the present invention;

[0058] Fig. 27 is the chromatogram when the column temperature is 35° C. in Example 4 of the present invention;

[0059] Fig.28 is the chromatogram when the column temperature is 40° C. in Example 4 of the present invention;

[0060] Fig.29 This is a chromatogram when methanol-0.1% phosphoric acid is used as the mobile phase in Example 5 of the present invention;

[0061] Fig.30 This is a chromatogram when methanol-0.1% formic acid is used as the mobile phase in Example 5 of the present invention;

[0062] Fig.31This is a chromatogram when methanol-0.1% acetic acid is used as the mobile phase in Example 5 of the present invention;

[0063] Fig.32 This is a chromatogram when methanol-0.05% phosphoric acid is used as the mobile phase in Example 6 of the present invention;

[0064] Fig.33 This is a chromatogram when methanol-0.1% phosphoric acid is used as the mobile phase in Example 6 of the present invention;

[0065] Fig.34 This is a chromatogram when methanol-0.2% phosphoric acid is used as the mobile phase in Example 6 of the present invention;

[0066] Fig.35 is a chromatogram when water is used as the extraction solvent in Example 7 of the present invention;

[0067] Fig.36 is a chromatogram when methanol is used as the extraction solvent in Example 7 of the present invention;

[0068] Fig.37 is a chromatogram when ethanol is used as the extraction solvent in Example 7 of the present invention;

[0069] Fig.38 is a chromatogram when a 60% methanol solution is used as the extraction solvent in Example 7 of the present invention;

[0070] Fig.39 This is a chromatogram when 80% methanol solution is used as the extraction solvent in Example 7 of the present invention. DETAILED DESCRIPTION

[0071] The following examples are provided for a better understanding of the present invention, but are not intended to limit the best mode of implementation, nor to limit the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior arts shall fall within the protection scope of the present invention.

[0072] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in the field can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be obtained commercially.

[0073] The instruments, reagents and reagents involved in the embodiments of the present invention are as follows:

[0074] Instruments: The chromatograph is a Waters ACQUITY UPLC H-Class PLUS chromatography system, including a quaternary solvent manager (ACQ-QSM), an automatic sample injector (ACQ-FTN), an original imported chromatography column oven (ACQ-CM), a diode array UV detector (ACQ-PDA), and an Empower chromatography management system;

[0075] XS204, XS205, XSE205 one-tenth balance (Mettler, Switzerland), ME36S one-millionth balance (Mettler, Switzerland), ultrasonic instrument (Shanghai Kedao Ultrasonic Instrument Co., Ltd.), water bath;

[0076] Chromatographic columns: (1) YMC-Triart C18, 2.1 mm × 100 mm, 1.9 μm; (2) capcell pak C18, 2.0 mm × 100 mm, 2 μm; (3) CORTECS T3, 2.1 mm × 100 mm, 1.6 μm.

[0077] Reagents: Methanol was of chromatographic grade, water was ultrapure water; other reagents were of analytical grade.

[0078] Drug testing:

[0079] Rhubarb (Tangut rhubarb) formula granules (batch numbers: 2108001Y, 2108002Y, 2108003Y, source: China Resources Sanjiu Pharmaceutical Co., Ltd.);

[0080] Rhubarb (Rheum tanguticum) slices standard decoction (lyophilized powder) (Batch No.: 2005001Y, 2005002Y, 2005003Y, 2005004Y, 2005005Y, 2005006Y, 2005007Y, 2005008Y, 2005009Y, 2005010Y, 2005011Y, 2005012Y, 2005013Y, 2005014Y, 2005015Y, 210401Y, 210402Y, 210403Y, source: China Resources Sanjiu Pharmaceutical Co., Ltd.);

[0081] Gallic acid (batch number: 110831-201605, for content determination, calculated at 90.8%, purchased from China Food and Drug Inspection Institute), catechin (batch number: 110877-201604, for content determination, calculated at 99.2%, purchased from China Food and Drug Inspection Institute), aloe-emodin (batch number: 110795-202011, for content determination, calculated at 97.5%, purchased from China Food and Drug Inspection Institute), rhein (batch number: 110757-201607, for content determination, calculated at 99.3%, purchased from China Food and Drug Inspection Institute Food and Drug Inspection Institute), rhamnoside (batch number: 110756-201913, for content determination, calculated at 96.0%, purchased from China Food and Drug Inspection Institute), chrysophanol (batch number: 110796-201922, for content determination, calculated at 99.4%, purchased from China Food and Drug Inspection Institute), rhamnoside methyl ether (batch number: 110758-201616, for content determination, calculated at 99.0%, purchased from China Food and Drug Inspection Institute), sennoside A (batch number: 110824-202003, purchased from China Food and Drug Inspection Institute);

[0082] Rhubarb (Rheum tanguticum) control drug (batch number: 120902-201912, purchased from China Food and Drug Inspection Institute);

[0083] The preparation method of rhubarb (Tangut rhubarb) formula granules is as follows: take rhubarb (Tangut rhubarb) medicinal materials, remove impurities, wash, soak for half an hour, take out, steam until thoroughly moistened, cut into thick slices, and dry at 50°C. Processing obtains rhubarb (Tangut rhubarb) slices that meet the requirements. Take rhubarb (Tangut rhubarb) slices, add water and boil twice, add 9 times water for the first time, soak for 30 minutes, boil for 60 minutes, filter with 200-mesh filter cloth, add 7 times water for the second time, boil for 30 minutes, filter with 200-mesh filter cloth, combine the filtrate, concentrate to a fluid paste with a relative density of 1.06-1.12 g / mL (60-80°C), spray dry, add an appropriate amount of maltodextrin, mix well, dry granulate, and package to obtain;

[0084] The preparation method of the standard decoction (lyophilized powder) of rhubarb (Rheum tanguticum) slices is as follows: take about 100 g of the slices, put them in a casserole, add 900 mL of purified water and soak for 30 minutes, boil them over high heat (500 W), then simmer them over low heat (300 W) for 20 minutes, filter them through a 200-mesh filter cloth while they are hot, and quickly cool the filtrate to room temperature; add 700 mL of purified water to the residue, boil them over high heat (500 W), then simmer them over low heat (300 W) for 15 minutes, filter them through a 200-mesh filter cloth while they are hot, and quickly cool the filtrate to room temperature, combine the two filtrates, and weigh the filtrate.

[0085] The filtrate is concentrated under reduced pressure (50°C to 65°C) to a concentrated extract with a relative density of 1.06g / ml to 1.12g / ml (60°C±5°C), the concentrated extract is placed in a freeze dryer, freeze-dried (-80°C, 0Mpa) to a dry state, taken out, weighed, crushed, and packaged in vials to obtain a standard decoction (lyophilized powder) of rhubarb (tangut rhubarb) slices;

[0086] The preparation method of the negative sample of rhubarb (Tangut rhubarb) formula granules is as follows: take maltodextrin as auxiliary material, dry granulate, and the granulation parameters are roller speed: 4-7rpm; extrusion pressure: 5-9Mpa; feeding speed: 20-30rpm.

[0087] Example 1

[0088] The detection method of rhubarb (Tanggut rhubarb) formula granules (batch numbers: 2108001Y, 2108002Y, 2108003Y) includes:

[0089] (1) Preparation of test solution: Take an appropriate amount of the product, grind it into powder, take about 0.5 g, place it in a stoppered conical flask, add 50 ml of 80% methanol, and ultrasonically treat (power 250 W, frequency 40 kHz) for 30 minutes. Cool, shake well, filter, and take the filtrate to obtain the solution;

[0090] (2) Preparation of reference solution: Take appropriate amount of gallic acid, catechin, sennoside A, rhein, emodin, chrysophanol, physophanol methyl ether, and aloe-emodin reference substances, accurately weigh them, and add methanol solution to prepare solutions containing 140 μg of gallic acid, 20 μg of catechin, 20 μg of sennoside A, 16 μg of rhein, 16 μg of emodin, 16 μg of chrysophanol, 16 μg of physophanol methyl ether, and 8 μg of aloe-emodin per 1 mL, to obtain reference solution;

[0091] (3) Preparation of reference medicinal material solution: Take 2 g of rhubarb (Rheum tanguticum) as reference medicinal material, place it in a stoppered conical flask, add 50 ml of water, heat and reflux for 1 hour, filter, evaporate the filtrate to dryness, add 50 ml of 80% methanol to the residue, ultrasonically treat (power 250 W, frequency 40 kHz) for 30 minutes, cool, shake well, filter, and take the filtrate to obtain the reference medicinal material solution.

[0092] (4) Ultra-high performance liquid chromatography detection:

[0093] The chromatographic column is a CORTECS T3 chromatographic column (2.1 mm×100 mm, 1.6 μm); methanol is used as mobile phase A, 0.05% by volume phosphoric acid aqueous solution is used as mobile phase B, and gradient elution is performed according to Table 1; the flow rate is 0.25 mL / min; the column temperature is 35° C.; the detection wavelength is 265 nm; the injection volume is 1 μL. The theoretical plate number should be no less than 2000 calculated based on the chromatographic peak of rhein.

[0094] Accurately pipette 1 μL of each reference solution, reference medicinal material solution and test solution, inject into ultra-high performance liquid chromatography, and measure to obtain the result.

[0095] Table 1 Gradient elution table

[0096]

[0097] Test results such as Figure 1 As shown, Figure 1 It is a spectrum comparison diagram of the test sample chromatogram, the reference medicinal material chromatogram and the reference substance chromatogram, wherein S1 is the test sample chromatogram, S2 is the reference medicinal material chromatogram, and S3 is the reference substance chromatogram. The obtained test sample chromatogram presents 18 characteristic peaks, and the retention times of the 18 characteristic peaks in the reference medicinal material chromatogram correspond to each other, wherein peak 1, peak 2, peak 9, peak 14, peak 15, peak 16, peak 17, and peak 18 should correspond to the reference peak retention times of the corresponding gallic acid, catechin, sennoside A, aloe-emodin, rhein, rhein, chrysophanol, and physophanol methyl ether reference substances, respectively. Taking the rhein peak (peak 15) as the first reference peak, the relative retention time of each characteristic peak and the first reference peak is within ±10% of the specified value, and the specified value is: 0.29 (peak 3), 0.32 (peak 4), 0.33 (peak 5), 0.37 (peak 6), 0.44 (peak 7), 0.46 (peak 8), 0.81 (peak 10), 0.83 (peak 11), 0.86 (peak 12), and 0.93 (peak 13); taking the chrysophanol peak (peak 17) as the second reference peak, the relative peak areas of Peak 5, Peak 7, Peak 8 and Peak 9 and the second reference peak are within the specified range, and the specified range is: not less than 0.9 (peak 5), not less than 1.4 (peak 7), not less than 13.0 (peak 8) and not less than 1.7 (peak 9).

[0098] Example 2

[0099] This example is used to illustrate the establishment process of the comparative characteristic spectrum of Tangut rhubarb products:

[0100] Take the above 3 batches of rhubarb (Rheum tanguticum) formula granules and the above 18 batches of rhubarb (Rheum tanguticum) decoction piece standard decoction (lyophilized powder), and prepare the test solution according to the method of step (1) in Example 1.

[0101] The 21 batches of test sample solutions obtained above were subjected to ultra-high performance liquid chromatography according to the method of step (4) in Example 1 to obtain liquid phase spectra, as shown in FIG. Figure 2 shown. Figure 2In the figure, S1-S3 are liquid phase spectra of 3 batches of rhubarb (Tangut rhubarb) formula granules, S4-S21 are liquid phase spectra of 18 batches of rhubarb (Tangut rhubarb) decoction piece standard decoction (lyophilized powder), and the corresponding product batch numbers are: S1: 2108001Y, S2: 2108002Y, S3: 2108003Y, S4: 2005001Y, S5: 2005002Y, S6: 2005003Y, S7: 2005004Y, S8: 2005005Y, S9: 2005006Y, S10: 2005007Y, S11: 2005008Y, S12: 2005009Y, S13: 2005010Y, S14: 2005011Y, S15: 2005012Y, S16: 2005013Y, S17: 2005014Y, S18: 2005015Y, S19: 210401Y, S20: 210402Y, S21: 210403Y.

[0102] Will Figure 2 The chromatograms of 21 batches of Tangut rhubarb products were imported into the fingerprint similarity evaluation software "Chinese Herbal Chromatographic Fingerprint Similarity Evaluation System 2012 Edition" compiled by the Pharmacopoeia Committee, and the "multi-point correction, MARK peak matching" mode was used for fitting to generate a reference characteristic spectrum. The reference characteristic spectrum of Tangut rhubarb products showed 18 common characteristic peaks, such as Figure 3 shown.

[0103] The 18 characteristic peaks were identified and assigned by HPLC and LC / MS / MS using reference substances. Peak 1 was determined to be gallic acid, peak 2 was catechin, peak 3 was epicatechin-3-O-gallate, peak 4 was iso-anecarboxylic acid, peak 5 was anecarboxylic acid, peak 6 was aloe-emodin-8-O-β-D-glucoside, peak 7 was resveratrol-4'-O-β-D-(6"-O-galloyl) glucoside, peak 8 was rhein-8 -O-β-D-glucoside, peak 9 is sennoside A, peak 10 is chrysophanol-1-O-β-D-glucoside, peak 11 is chrysophanol-8-O-β-D-glucoside, peak 12 is rhein-8-O-β-D-pyranoglucoside, peak 13 is physophanol methyl ether-8-O-β-D-glucoside, peak 14 is aloe-emodin, peak 15 is rhein, peak 16 is rhein, peak 17 is chrysophanol, and peak 18 is physophanol methyl ether.

[0104] Peaks 1 to 18 were analyzed by LC / MS / MS, and it was confirmed that their molecular structures were consistent with those of gallic acid, catechin, epicatechin-3-O-gallate, isoanecarboxylic acid, anthein, aloe-emodin-8-O-β-D-glucoside, resveratrol-4'-O-β-D-(6"-O-galloyl) glucoside, rhein-8-O-β-D-glucoside, sennoside A, chrysophanol-1-O-β-D-glucoside, chrysophanol-8-O-β-D-glucopyranoside, physcion-8-O-β-D-glucoside, aloe-emodin, rhein, rhein, chrysophanol, and physcion reference substances, respectively.

[0105] Among the 18 characteristic peaks identified above, only gallic acid, catechins, sennoside A, aloe-emodin, rhein, rhein, chrysophanol, and physophanol methyl ether can be obtained through the legal institution China Food and Drug Inspection Institute, among which aloe-emodin, rhein, rhein, chrysophanol, and physophanol methyl ether are index components for content determination. Therefore, five anthraquinone components are used as reference peaks to better evaluate the relative retention time and relative peak area of ​​each characteristic peak. Among them, the response value of rhein is higher and the retention time is relatively moderate. Therefore, the rhein peak (peak 15) is used as the first reference peak as S1 peak, and the relative retention time of each characteristic peak is calculated. In combination with the detection results of multiple batches of samples, the differentiation study of different original samples and the differentiation study of different processed products, chrysophanol (peak 17) is used as the second reference peak S2 peak, and the relative peak area ratios of peak 5 (nelumboside), peak 7 (resveratrol-4'-O-β-D-(6"-O-galloyl) glucoside), peak 8 (rhein-8-O-β-D-glucoside) and peak 9 (sennoside A) are calculated for the differentiation of different original samples and to improve the control level of formula granules.

[0106] The relative retention time of the characteristic spectrum is determined according to the research results: it is determined that the characteristic spectrum of rhubarb (Tangut rhubarb) formula granules has a total of 18 characteristic peaks, and should correspond to the 18 characteristic peaks in the chromatogram of the reference medicinal material reference material, among which peak 1, peak 2, peak 9, peak 14, peak 15, peak 16, peak 17, and peak 18 should correspond to the corresponding gallic acid, catechin, sennoside A, aloe-emodin, rhein, rhein, chrysophanol, and physophanol reference material peak retention times, respectively; considering that among the known ingredients, aloe-emodin, rhein, rhein, chrysophanol, and physophanol are content determination indicator components, among which the content level and peak area of ​​rhein account for a relatively high proportion, and the retention time position of rhein in the characteristic peak is relatively moderate, without interference from impurity peaks, selecting it as the reference peak can better evaluate the relative retention time and relative peak area of ​​other characteristic peaks. Therefore, the peak corresponding to the rhein reference peak (peak 15) is selected as the S1 peak, and the relative retention time of each characteristic peak and S peak 1 is calculated as follows: 0.29 (peak 3), 0.32 (peak 4), 0.33 (peak 5), 0.37 (peak 6), 0.44 (peak 7), 0.46 (peak 8), 0.81 (peak 10), 0.83 (peak 11), 0.86 (peak 12), 0.93 (peak 13). In combination with the verification results of the characteristic spectrum method of Tanggut rhubarb product, the relative retention time of the characteristic peak is specified to be within the range of ±10% of the specified value. The chrysophanol peak (peak 17) is selected as the second reference peak S2, and the relative peak areas of peaks 5, 7, 8 and 9 and the second reference peak are calculated to be within the specified range, which is: not less than 0.9 (peak 5), not less than 1.4 (peak 7), not less than 13.0 (peak 8) and not less than 1.7 (peak 9).

[0107] Example 3

[0108] This example is used to verify the method of the present invention:

[0109] (1) Specificity verification

[0110] Take rhubarb (Tanggut rhubarb) formula granules (batch number: 2108001Y) and the above rhubarb (Tanggut rhubarb) formula granules negative sample, prepare the test solution and the negative control solution according to the method of step (1) in Example 1, and perform ultra-high performance liquid chromatography detection according to the method of step (4) in Example 1. The detection results are as follows: Figure 4-Figure 6 As shown, Figure 4 is the negative control chromatogram. Figure 5 This is the reference medicinal material chromatogram of rhubarb (Rheum tanguticum). Figure 6 This is a chromatogram of rhubarb (Tangut rhubarb) formula granules. The result shows that the negative control solution has no interference, indicating that the system adaptability and specificity of the method of the present invention are good, and it can be used as a detection method for the characteristic spectrum of Tangut rhubarb products.

[0111] (2) Precision verification

[0112] Take the same portion of rhubarb (tangut rhubarb) formula granules (batch number: 2108001Y), prepare the test solution according to the method of step (1) in Example 1, and perform ultra-high performance liquid chromatography detection according to the method of step (4) in Example 1, repeat the injection 6 times, and obtain the characteristic spectrum of each detection, such as Figure 7 shown.

[0113] The relative retention time of each characteristic peak and the first reference peak was calculated by taking the rhein peak (peak 15) as the first reference peak S1, and the relative peak area of ​​each characteristic peak and the second reference peak was calculated by taking the chrysophanol peak (peak 17) as the second reference peak S2, and the RSD value was calculated. The calculation results are shown in Tables 2 and 3.

[0114] Table 2 Precision verification retention time and relative retention time table

[0115]

[0116]

[0117] Table 3 Precision verification peak area and relative peak area

[0118]

[0119]

[0120] It can be seen from Table 2 and Table 3 that the method of the present invention has good precision.

[0121] (3) Repeatability verification

[0122] The same batch of rhubarb (tangut rhubarb) formula granules (batch number: 2108001Y) was taken, and 6 test solutions were repeatedly prepared according to the method of step (1) in Example 1. Ultra-high performance liquid chromatography was performed according to the method of step (4) in Example 1 to obtain characteristic spectra of the 6 test samples, such as Figure 8 shown.

[0123] Taking the rhein peak (peak 15) as the first reference peak S1, the relative retention time of each characteristic peak and the first reference peak was calculated. Taking the chrysophanol peak (peak 17) as the second reference peak S2, the relative peak area of ​​each characteristic peak and the second reference peak was calculated, and the RSD value was calculated. The calculation results are shown in Tables 4 and 5.

[0124] Table 4 Repeatability verification retention time and relative retention schedule

[0125]

[0126] Table 5 Repeatability verification peak area and relative peak area

[0127]

[0128]

[0129] It can be seen from Table 4 and Table 5 that the method of the present invention has good repeatability.

[0130] (4) Intermediate precision (different operators) verification

[0131] The same batch of rhubarb (Tanggut rhubarb) formula granules (batch number: 2108001Y) was taken, and three inspectors prepared the test solution according to the method of step (1) in Example 1 at different times, and performed ultra-high performance liquid chromatography detection according to the method of step (4) in Example 1 to obtain a characteristic spectrum, such as Fig. 9 shown.

[0132] Taking the rhein peak (peak 15) as the first reference peak S1, the relative retention time of each characteristic peak and the first reference peak was calculated. Taking the chrysophanol peak (peak 17) as the second reference peak S2, the relative peak area of ​​each characteristic peak and the second reference peak was calculated, and the RSD value was calculated. The calculation results are shown in Tables 6 and 7.

[0133] Table 6 Intermediate precision verification retention time and relative retention time table

[0134]

[0135] Table 7 Intermediate precision verification peak area and relative peak area

[0136]

[0137] It can be seen from Tables 6 and 7 that the intermediate precision of the method of the present invention is good.

[0138] (5) Stability test

[0139] Take rhubarb (tangut rhubarb) formula granules (batch number: 2108001Y), prepare a test solution according to the method of step (1) in Example 1, and perform ultra-high performance liquid chromatography detection according to the method of step (4) in Example 1 at time points such as 0, 4, 8, 12, 16, and 24 hours after the preparation of the test solution to obtain a characteristic spectrum, such as Fig.10 shown.

[0140] Taking the rhein peak (peak 15) as the first reference peak S1, the relative retention time of each characteristic peak and the first reference peak was calculated. Taking the chrysophanol peak (peak 17) as the second reference peak S2, the relative peak area of ​​each characteristic peak and the second reference peak was calculated, and the RSD value was calculated. The calculation results are shown in Tables 8 and 9.

[0141] Table 8 Retention time and relative retention time table of stability study

[0142]

[0143]

[0144] Table 9 Stability test peak area and relative peak area

[0145]

[0146] It can be seen from Tables 8 and 9 that the test solution used in the method of the present invention is stable within 24 hours and meets the measurement requirements.

[0147] (6) Investigation of different column temperatures

[0148] Take rhubarb (tangut rhubarb) formula granules (batch number: 2108001Y), prepare the test solution according to the method of step (1) in Example 1, and then set the column temperature to 33°C, 35°C, and 37°C respectively, and perform ultra-high performance liquid chromatography detection according to the method of step (4) in Example 1 to obtain a characteristic spectrum, such as Figure 11-13 As shown, Fig.11 This is the chromatogram when the column temperature is 33°C. Fig.12 This is the chromatogram when the column temperature is 35°C. Fig.13 The chromatogram is shown when the column temperature is 37°C.

[0149] Taking the rhein peak (peak 15) as the first reference peak S1, the relative retention time of each characteristic peak and the first reference peak was calculated. Taking the chrysophanol peak (peak 17) as the second reference peak S2, the relative peak area of ​​each characteristic peak and the second reference peak was calculated, and the RSD value was calculated. The calculation results are shown in Tables 10 and 11.

[0150] Table 10 Retention time and relative retention time table at different column temperatures

[0151]

[0152]

[0153] Table 11 Peak area and relative peak area at different column temperatures

[0154]

[0155] It can be seen from Tables 10 and 11 that when the column temperature changes, the retention time of each characteristic chromatographic peak will change to a certain extent, indicating that the retention time of each characteristic chromatographic peak is affected by the column temperature, but the degree of influence is within an acceptable range. Considering the separation degree and peak shape of individual peaks, the optimal column temperature of the method of the present invention is 35°C.

[0156] (7) Investigation of different flow rates

[0157] Take rhubarb (tangut rhubarb) formula granules (batch number: 2108001Y), prepare the test solution according to the method of step (1) in Example 1, and then set the flow rate to 0.20ml / min, 0.25ml / min, and 0.30ml / min respectively, and perform ultra-high performance liquid chromatography detection according to the method of step (4) in Example 1 to obtain a characteristic spectrum, such as Figure 14-16 As shown, Fig.14 This is the chromatogram at a flow rate of 0.20 ml / min. Fig.15 This is the chromatogram at a flow rate of 0.25 ml / min. Fig.16 This is the chromatogram at a flow rate of 0.30 ml / min.

[0158] Taking the rhein peak (peak 15) as the first reference peak S1, the relative retention time of each characteristic peak and the first reference peak was calculated. Taking the chrysophanol peak (peak 17) as the second reference peak S2, the relative peak area of ​​each characteristic peak and the second reference peak was calculated, and the RSD value was calculated. The calculation results are shown in Tables 12 and 13.

[0159] Table 12 Retention time and relative retention time table at different flow rates

[0160]

[0161]

[0162] Table 13 Peak area and relative peak area at different flow rates

[0163]

[0164] It can be seen from Tables 12 and 13 that the deviation of the relative peak areas of Peaks 4 and 9 is large when the flow rate changes; Figure 14-16 It can be seen that different flow rates will affect the retention time and peak area of ​​each peak, as well as the separation of each characteristic peak and the surrounding small peaks, but in the range of 0.20-0.30ml / min, the degree of influence is within the allowable range; combined with the investigation results of different flow rates and column pressures, the optimal flow rate of the method of the present invention is 0.25ml / min.

[0165] (8) Investigation of different chromatographic columns

[0166] Take rhubarb (tanggut rhubarb) formula granules (batch number: 2108001Y), prepare a test solution according to the method of step (1) in Example 1, and then use chromatographic columns ①YMC-Triart C18 (2.1mm×100mm, 1.9μm), ②capcell pak C18 (2.0mm×100mm, 2μm), and ③CORTECS T3 (2.1mm×100mm, 1.6μm) to perform ultra-high performance liquid chromatography detection according to the method of step (4) in Example 1 to obtain a characteristic spectrum, such as Figure 17-19 As shown, Fig.17 FIG18 is a chromatogram when using the chromatographic column ①YMC-Triart C18 (2.1 mm × 100 mm, 1.9 μm), and FIG19 is a chromatogram when using the chromatographic column ②capcell pak C18 (2.0 mm × 100 mm, 2 μm). Fig.19 This is the chromatogram when using chromatographic column ③ CORTECS T3 (2.1 mm×100 mm, 1.6 μm).

[0167] Taking the rhein peak (peak 15) as the first reference peak S1, the relative retention time of each characteristic peak and the first reference peak was calculated. Taking the chrysophanol peak (peak 17) as the second reference peak S2, the relative peak area of ​​each characteristic peak and the second reference peak was calculated, and the RSD value was calculated. The calculation results are shown in Tables 14 and 15.

[0168] Table 14 Retention time and relative retention time table of different chromatographic columns

[0169]

[0170]

[0171] Table 15 Peak area and relative peak area of ​​different chromatographic columns

[0172]

[0173] It can be seen from Table 14 and Table 15 that the chromatographic column of the method of the present invention has good durability. Combined with the separation effects of various chromatographic columns, the best chromatographic column of the method of the present invention is CORTECS T3 (2.1 mm×100 mm, 1.6 μm).

[0174] Three CORTECS T3 (2.1 mm × 100 mm, 1.6 μm) chromatographic columns with different numbers (column 1 number 1213123715304, column 2 number 1213123715308, column 3 number 1133924115409) were used to perform ultra-high performance liquid chromatography according to the method of step (4) in Example 1 to obtain characteristic spectra, such as Figure 20-22 As shown, Fig. 20 This is the chromatogram when column 1 is used. Fig.21 This is the chromatogram when column 2 is used. Fig. 22 This is the chromatogram when column 3 is used.

[0175] Taking the rhein peak (peak 15) as the first reference peak S1, the relative retention time of each characteristic peak and the first reference peak was calculated. Taking the chrysophanol peak (peak 17) as the second reference peak S2, the relative peak area of ​​each characteristic peak and the second reference peak was calculated, and the RSD value was calculated. The calculation results are shown in Tables 16 and 17.

[0176] Table 16 Retention time and relative retention time of different CORTECS T3 columns

[0177]

[0178]

[0179] Table 17 Peak area and relative peak area of ​​different CORTECS T3 columns

[0180]

[0181] It can be seen from Tables 16 and 17 that when CORTECS T3 chromatographic columns with different numbers were used, the retention time and relative peak area of ​​each characteristic chromatographic peak did not change much.

[0182] (9) Investigation of mobile phase systems with different phosphoric acid concentrations

[0183] Take rhubarb (Tanggut rhubarb) formula granules (batch number: 2108001Y), prepare the test solution according to the method of step (1) in Example 1, and then use methanol-0.03% phosphoric acid solution, methanol-0.05% phosphoric acid solution, and methanol-0.07% phosphoric acid solution as mobile phases, respectively, and perform ultra-high performance liquid chromatography detection according to the method of step (4) in Example 1 to obtain a characteristic spectrum, such as Figure 23-25 As shown, Fig.23 This is the chromatogram when using methanol-0.03% phosphoric acid solution as the mobile phase. Fig.24 This is the chromatogram when the mobile phase is methanol-0.05% phosphoric acid solution. Fig.25 The chromatogram is shown when a mobile phase of methanol-0.07% phosphoric acid solution is used.

[0184] Taking the rhein peak (peak 15) as the first reference peak S1, the relative retention time of each characteristic peak and the first reference peak was calculated. Taking the chrysophanol peak (peak 17) as the second reference peak S2, the relative peak area of ​​each characteristic peak and the second reference peak was calculated, and the RSD value was calculated. The calculation results are shown in Tables 18 and 19.

[0185] Table 18 Retention time and relative retention time table of different phosphoric acid concentration mobile phases

[0186]

[0187] Table 19 Peak area and relative peak area of ​​different phosphoric acid concentration mobile phases

[0188]

[0189] It can be seen from Tables 18 and 19 that when the concentration of the phosphoric acid solution in the mobile phase changes, the relative retention time of all chromatographic peaks is slightly affected. Combined with the investigation results, it is determined that the best mobile phase for the method of the present invention is methanol-0.05% phosphoric acid solution.

[0190] Example 4

[0191] The characteristic spectrum of the rhubarb (Tanggut rhubarb) formula granules was detected according to the method of Example 1, except that the column temperature was set at 30°C, 35°C, and 40°C in this example. The obtained chromatogram is as follows: Figure 26-28 As shown, Fig.26 This is the chromatogram when the column temperature is 30°C. Fig. 27 This is the chromatogram when the column temperature is 35°C. Fig.28 This is the chromatogram when the column temperature is 40°C.

[0192] Depend on Figure 26-28 It can be seen that within the range of column temperature of 30-40°C, the method of the present invention can achieve good detection effect.

[0193] Example 5

[0194] The characteristic spectrum of the rhubarb (Tanggut rhubarb) formula granules was detected according to the method of Example 1, except that methanol-0.1% phosphoric acid, methanol-0.1% formic acid and methanol-0.1% acetic acid were used as mobile phases for detection in this example. The obtained chromatogram is shown in FIG. Figure 29-31 As shown, Fig.29 This is the chromatogram when methanol-0.1% phosphoric acid is used as the mobile phase. Fig.30 This is the chromatogram when methanol-0.1% formic acid is used as the mobile phase. Fig.31 This is the chromatogram when methanol-0.1% acetic acid is used as the mobile phase.

[0195] Depend on Figure 29-31 It can be seen that when three different mobile phases are used, the method of the present invention can achieve better detection effects.

[0196] Example 6

[0197] The characteristic spectrum of the rhubarb (Tanggut rhubarb) formula granules was detected according to the method of Example 1, except that methanol-0.05% phosphoric acid, methanol-0.1% phosphoric acid and methanol-0.2% phosphoric acid were used as mobile phases for detection in this example. The obtained chromatogram is shown in FIG. Figure 32-34 As shown, Fig.32 This is the chromatogram when methanol-0.05% phosphoric acid is used as the mobile phase. Fig.33 This is the chromatogram when methanol-0.1% phosphoric acid is used as the mobile phase. Fig.34 This is the chromatogram when methanol-0.2% phosphoric acid is used as the mobile phase.

[0198] Depend on Figure 32-34 It can be seen that when the phosphoric acid concentration of the mobile phase B is in the range of 0.05%-0.2%, the method of the present invention can achieve good detection effect.

[0199] Example 7

[0200] The characteristic spectrum of the rhubarb (Tanggut rhubarb) formula granules was detected according to the method of Example 1, except that water, methanol, ethanol, 60% methanol solution, and 80% methanol solution were used as the extraction solvents of the test solution in this embodiment. The detection results are as follows: Figure 35-39 As shown, Fig.35 This is the chromatogram when water is used as the extraction solvent. Fig.36 This is the chromatogram when methanol is used as the extraction solvent. Fig.37 This is the chromatogram when ethanol is used as the extraction solvent. Fig.38 This is the chromatogram when 60% methanol solution is used as the extraction solvent. Fig.39 This is the chromatogram when 80% methanol solution is used as the extraction solvent.

[0201] Depend on Figure 35-39 It can be seen that when methanol, water, 60% methanol solution and 80% methanol solution are used as extraction solvents, there is no obvious difference in the detection results, and the method of the present invention can achieve good detection effects; when ethanol is used as the extraction solvent, the response value of the chromatogram peak is lower.

[0202] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A method for detecting a product of Rhubarb tanguticus, characterized in that: The steps include: Take the test solution and detect it by ultra-high performance liquid chromatography; wherein the chromatographic conditions of the ultra-high performance liquid chromatography include: Octadecylsilane bonded silica gel is used as a filler, methanol is used as a mobile phase A, and a phosphoric acid aqueous solution with a volume percentage of 0.03-0.20% is used as a mobile phase B for gradient elution, and the gradient elution program includes: 0-1min, the volume percentage of methanol in the mobile phase is 5%→20%; 1-9min, the volume percentage of methanol in the mobile phase is 20%→35%; 9-17min, the volume percentage of methanol in the mobile phase is 35%→37%; 17-21min, the volume percentage of methanol in the mobile phase is 37%→48%; 21-24min, the volume percentage of methanol in the mobile phase is 48%→60%; 24-30min, the volume percentage of methanol in the mobile phase is 60%→100%; 30-35min, the volume percentage of methanol in the mobile phase is 100%; The chromatographic column was a CORTECS T3 column with an inner diameter of 2.1 mm, a column length of 100 mm, and a particle size of 1.6 μm; In the case where the tangut rhubarb product is a tangut rhubarb preparation, the preparation process of the test solution includes: Taking Tangut rhubarb preparation powder, adding extraction solvent, extracting, solid-liquid separation, and taking liquid; wherein the extraction solvent is selected from any one of water, methanol or methanol aqueous solution, and the volume percentage of methanol in the methanol aqueous solution is not less than 60%; In the case where the Rheum tanguticum product is a Rheum tanguticum medicinal material and / or a decoction piece, the preparation process of the test solution includes: Take the Tangut rhubarb medicinal material and / or the decoction pieces, add water, heat and reflux, separate the solid and liquid, and concentrate the obtained liquid to dryness to obtain a concentrated residue; Adding an extraction solvent to the concentrated residue, extracting, separating the solid and the liquid, and taking the liquid; the extraction solvent is selected from any one of water, methanol or a methanol aqueous solution, and the volume percentage of methanol in the methanol aqueous solution is not less than 60%; The detection method also includes the steps of preparing a reference substance solution by using gallic acid reference substance, catechin reference substance, sennoside A reference substance, rhein reference substance, rhein reference substance, chrysophanol reference substance, physophanol methyl ether reference substance and aloe-rhein reference substance, and the step of detecting the reference substance solution by using the ultra-high performance liquid chromatography method to obtain a reference spectrum of the reference substances.

2. The detection method according to claim 1, characterized in that: The chromatographic conditions of the ultra-high performance liquid chromatography method also include at least one of the following conditions: 1) The detection wavelength is 260-270nm; 2) Column temperature is 30-40°C; 3) Flow rate is 0.20-0.30 mL / min; 4) The injection volume is 0.5 to 2 μL.

3. The detection method according to claim 1, characterized in that: The tangut rhubarb product includes at least one of tangut rhubarb medicinal materials, tangut rhubarb decoction pieces and tangut rhubarb preparations.

4. The detection method according to claim 3, characterized in that: The amount of the extraction solvent added is 50 to 150 ml relative to 1 g of the tangutic rhubarb preparation powder.

5. The detection method according to claim 4, characterized in that: The extraction is ultrasonic extraction, the ultrasonic power is 200W-300W, and the ultrasonic time is 15-45min.

6. The detection method according to claim 3, characterized in that: Relative to 1g of the Tangut rhubarb medicinal material and / or decoction pieces, the amount of water added is 25-75ml; The heating reflux temperature is 80-100° C. and the time is 20-60 min.

7. The detection method according to claim 6, characterized in that: When the extraction solvent is added to the concentrated residue of the Rheum tanguticum medicinal material and / or the decoction pieces, the amount of the extraction solvent added is 25 to 75 ml relative to 1 g of the Rheum tanguticum medicinal material and / or the decoction pieces.

8. The detection method according to claim 6, characterized in that: The extraction is ultrasonic extraction, the ultrasonic power is 200W to 300W, and the ultrasonic time is 15min to 45min.

9. The detection method according to claim 1, characterized in that: The detection method also includes the step of using Tangut rhubarb as a reference medicinal material to prepare a reference medicinal material solution, and the step of using the ultra-high performance liquid chromatography method to detect the reference medicinal material solution to obtain a reference medicinal material atlas.

10. Use of the detection method according to any one of claims 1 to 9 in quality control of Rheum tanguticum products and / or in identification of Rheum tanguticum products.

11. A quality control method for Tangut rhubarb products, characterized in that: The method comprises the steps of obtaining a characteristic spectrum of the tangut rhubarb product to be tested according to the detection method of any one of claims 1 to 9, and comparing the characteristic spectrum with a control characteristic spectrum; The control characteristic spectrum is obtained by fitting the characteristic spectrum obtained by the detection method described in any one of claims 1 to 9 using at least one batch of standard products of Tangut rhubarb products through the average value or median method.

12. The quality control method according to claim 11, characterized in that: The control characteristic spectrum includes 18 characteristic peaks, peak 1 is gallic acid peak, peak 2 is catechin peak, peak 3 is epicatechin-3-O-gallate peak, peak 4 is iso-anecarboxylic acid peak, peak 5 is anecarboxylic acid peak, peak 6 is aloe-emodin-8-O-β-D-glucoside peak, peak 7 is resveratrol-4'-O-β-D-(6"-O-galloyl) glucoside peak, peak 8 is rhein-8-O-β-D-glucoside peak, peak 9 is sennoside A peak, peak 10 is chrysophanol-1-O-β-D-glucoside peak, peak 11 is chrysophanol-8-O-β-D-glucoside peak, peak 12 is rhein-8-O-β-D-pyranoglucoside peak, peak 13 is rhubarb The peak of physophanol-8-O-β-D-glucoside is as follows: peak 14 is aloe-rhein peak, peak 15 is rhein peak, peak 16 is rhein peak, peak 17 is chrysophanol peak, peak 18 is physophanol peak, and the rhein peak is taken as the first reference peak. The relative retention time of each characteristic peak and the first reference peak is within ±10% of the specified value, the specified value of peak 3 is 0.29, the specified value of peak 4 is 0.32, the specified value of peak 5 is 0.33, the specified value of peak 6 is 0.37, the specified value of peak 7 is 0.44, the specified value of peak 8 is 0.46, the specified value of peak 10 is 0.81, the specified value of peak 11 is 0.83, the specified value of peak 12 is 0.86, and the specified value of peak 13 is 0.

93.

13. The quality control method according to claim 12, characterized in that: Taking the chrysophanol peak as the second reference peak, the relative peak areas of Peak 5, Peak 7, Peak 8 and Peak 9 to the second reference peak are within the specified range, the specified range for Peak 5 is not less than 0.9, the specified range for Peak 7 is not less than 1.4, the specified range for Peak 8 is not less than 13.0 and the specified range for Peak 9 is not less than 1.7.

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