Construction method and identification method of characteristic chromatogram of senecio scandens medicine and its decoction pieces, standard decoction and formula granules

Abstract

The application discloses a characteristic chromatogram construction method and identification method of Senecae Scandentae, its decoction pieces, standard decoction and formula granules, and relates to the technical field of traditional Chinese medicine identification. The characteristic chromatogram construction method comprises the following steps: methanol is used to extract the to-be-detected sample to obtain a test sample solution; methanol is used to dissolve 5-caffeoylquinic acid, chlorogenic acid, caffeic acid, 4-caffeoylquinic acid, rutin, hyperoside, isoquercitrin and 4,5-dicaffeoylquinic acid as a control sample solution; and a liquid chromatograph is used for determination. The chromatographic column of the liquid chromatograph uses phenyl-methyl polysiloxane as a stationary phase, acetonitrile as a mobile phase A and 0.05vol%-0.3vol% methanoic acid aqueous solution as a mobile phase B for gradient elution. The characteristic chromatogram constructed by the application is rich in information and can quickly and comprehensively reflect the quality of Senecae Scandentae, its decoction pieces, standard decoction and formula granules.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of traditional Chinese medicine identification, and particularly relates to a characteristic chromatogram construction method and identification method of Senecio scandens Buch. medicine and its decoction pieces, standard decoction and formula granules. BACKGROUND

[0002] Senecio scandens Buch. is collected in Chinese Pharmacopoeia, and is the dry aboveground part of Senecio scandens Buch. of Compositae. Senecio scandens Buch. is mainly distributed in East China, South Central China and Southwest China, and is mostly grown in hilly mountain forest edges, shrubbery grasslands and roadsides. Senecio scandens Buch. is collected in summer and autumn, and is one of the commonly used traditional Chinese medicines with a long application history. The dry whole plant is 60-100 cm long, or is cut into small pieces of 2-3 cm long. The stem is cylindrical, and the surface is brownish yellow. The leaves are mostly shriveled and broken, and are elliptical triangular or ovate lanceolate. The base is harpoon-shaped or truncated, the edges are irregularly notched, and the color is dark green or grayish brown. The leaves are brittle. Sometimes, the branch tips have dry yellow head-shaped inflorescences. Senecio scandens Buch. can be collected all year round, and is dried after removing impurities. Senecio scandens Buch. is cold in nature, and bitter in taste. Senecio scandens Buch. has the effects of clearing heat and resolving toxins, clearing the vision, and removing dampness. Senecio scandens Buch. is used for treating carbuncle and sore, common cold and fever, red eyes and swelling pain, diarrhea and dysentery, and skin eczema.

[0003] The chemical components of Senecio scandens Buch. medicine have been studied in the prior art. For example, in one prior art, acetonitrile-0.2% phosphoric acid is used as a mobile phase, and a characteristic chromatogram of Senecio scandens Buch. formula granules is established by isocratic elution. However, only six characteristic peaks are identified, and the chromatographic peak information is not rich enough, so that it is difficult to effectively realize comprehensive quality control of Senecio scandens Buch. medicine. SUMMARY

[0004] The technical problem to be solved by the present application is to provide a characteristic chromatogram construction method of Senecio scandens Buch. medicine and its decoction pieces, standard decoction and formula granules, which has rich characteristic peak information, and can comprehensively reflect the quality characteristics of Senecio scandens Buch. medicine or its decoction pieces, standard decoction and formula granules.

[0005] In order to solve the above technical problem, the present application provides a characteristic chromatogram construction method of Senecio scandens Buch. medicine and its decoction pieces, standard decoction and formula granules, which comprises the following steps:

[0006] extracting Senecio scandens Buch. medicine or its decoction pieces, standard decoction and formula granules by using an extraction solvent to obtain a test solution;

[0007] dissolving 5-caffeoylquinic acid, chlorogenic acid, caffeic acid, 4-caffeoylquinic acid, rutin, hyperoside, isoquercitrin and 4,5-dicaffeoylquinic acid reference substances by using the extraction solvent to obtain a reference solution;

[0008] The test product solution and the control product solution are determined by using a liquid chromatograph to obtain the characteristic chromatogram of the Senecio scandens Buch Ham medicinal material or its decoction pieces, standard decoction, and formula granules;

[0009] The extraction solvent is selected from methanol;

[0010] The liquid chromatograph uses a chromatographic column with phenyl-bonded silica gel as a stationary phase, acetonitrile as a mobile phase A, and 0.05vol% to 0.3vol% of a methanol aqueous solution as a mobile phase B for gradient elution, and the gradient elution curve is as follows:

[0011] 0min to 15min, mobile phase A from 5% to 8%, and mobile phase B from 95% to 92%;

[0012] 15min to 22min, mobile phase A from 8% to 12%, and mobile phase B from 92% to 88%;

[0013] 22min to 38min, mobile phase A from 12% to 16%, and mobile phase B from 88% to 84%;

[0014] 38min to 50min, mobile phase A from 16% to 25%, and mobile phase B from 84% to 75%.

[0015] As an improvement of the above technical solution, the column length of the chromatographic column of the liquid chromatograph is 100mm to 150mm, the column diameter is 3mm to 5mm, and the particle size of the stationary phase is 2μm to 3μm.

[0016] As an improvement of the above technical solution, the chromatographic column is a Waters CORTECS Phenyl chromatographic column, the column length is 150mm, the column diameter is 4.6mm, and the particle size of the stationary phase is 2.7μm.

[0017] As an improvement of the above technical solution, the mobile phase B is a 0.2vol% methanol aqueous solution.

[0018] As an improvement of the above technical solution, the column temperature of the chromatographic column of the liquid chromatograph is 28℃ to 32℃, the flow rate is 0.7mL / min to 0.9mL / min, the injection amount of the test product solution is 1μL to 3μL, and the injection amount of the control product solution is 1μL to 3μL;

[0019] The detection wavelength of the liquid chromatograph is 300nm to 400nm.

[0020] As an improvement of the above technical solution, the column temperature of the chromatographic column of the liquid chromatograph is 30℃, the flow rate is 0.8mL / min, the injection amount of the test product solution is 2μL, and the injection amount of the control product solution is 2μL;

[0021] The detection wavelength of the liquid chromatograph is 360 nm.

[0022] As the improvement of the above technical solution, in the step of extracting the medicinal material or its decoction pieces, standard decoction or formula granules of Senecioninae Radix with the extraction solvent to obtain the test solution, the extraction solvent is 70vol% to 80vol% methanol, the extraction method is heating reflux extraction or ultrasonic extraction, and the extraction time is 20min to 90min.

[0023] As the improvement of the above technical solution, when the test solution is the medicinal material or the decoction pieces of Senecioninae Radix, the ratio of the weight of the test solution to the volume of the extraction solvent is (0.8-1.5)g:(20-30)mL, the extraction method is heating reflux extraction, and the extraction time is 40-80min.

[0024] When the test solution is the standard decoction or the formula granules of Senecioninae Radix, the ratio of the weight of the test solution to the volume of the extraction solvent is (0.8-1.5)g:(20-30)mL, the extraction method is ultrasonic extraction, the ultrasonic power is 200-400W, the frequency is 20-50kHz, and the extraction time is 20-50min.

[0025] As the improvement of the above technical solution, the characteristic spectrum includes 10 common peaks, wherein, peak 2 is 5-caffeoyl quinic acid, peak 3 is chlorogenic acid, peak 4 is caffeic acid, peak 5 is 4-caffeoyl quinic acid, peak 7 is rutin, peak 8 is hyperoside, peak 9 is isoquercitrin, and peak 10 is 4,5-dicaffeoyl quinic acid.

[0026] Correspondingly, the application also discloses a method for identifying Senecioninae Radix and its counterfeit products.

[0027] Providing a substance to be identified;

[0028] Constructing a characteristic spectrum of the substance to be identified by using the method for constructing the characteristic spectrum of Senecioninae Radix and its decoction pieces, standard decoction or formula granules;

[0029] If the characteristic spectrum only presents peaks 1-10, the substance to be identified includes or is Senecioninae Radix; otherwise, it is a counterfeit product.

[0030] The counterfeit product includes Senecioninae Radix.

[0031] The application has the following beneficial effects:

[0032] The application constructs the characteristic spectrum of Senecionine medicinal materials and its decoction pieces, standard decoction and formula granules, the constructed characteristic spectrum information is rich, and good separation of different chromatographic components is realized, the characteristic spectrum has 10 common peaks, wherein 8 chromatographic peaks are identified, and they are: peak 2 is 5-caffeoyl quinic acid, peak 3 is chlorogenic acid, peak 4 is caffeic acid, peak 5 is 4-caffeoyl quinic acid, peak 7 is rutin, peak 8 is hyperoside, peak 9 is isoquercitrin, and peak 10 is 4, 5-di-caffeoyl quinic acid. Based on the characteristic spectrum, comprehensive quality control indexes can be provided for the quality and quantity transmission of Senecionine, and effective identification of Senecionine and common counterfeits can be realized. BRIEF DESCRIPTION OF DRAWINGS

[0033] Figure 1 is the characteristic spectrum of Senecionine formula granules in example 1 when different chromatographic columns are used;

[0034] Figure 2 is the characteristic spectrum of Senecionine formula granules in example 1 when different mobile phase B is used;

[0035] Figure 3 is the characteristic spectrum of Senecionine formula granules in example 1 when different mobile phase gradient is used;

[0036] Figure 4 is the investigation result graph of the specificity of the characteristic spectrum of Senecionine formula granules in example 1;

[0037] Figure 5 is the investigation result graph of the different temperature durability of the characteristic spectrum of Senecionine formula granules in example 1;

[0038] Figure 6 is the investigation result graph of the different flow rate durability of the characteristic spectrum of Senecionine formula granules in example 1;

[0039] Figure 7 is the superimposed graph of the characteristic spectrum of 13 batches of Senecionine medicinal materials in example 2;

[0040] Figure 8 is the superimposed graph of the characteristic spectrum of 13 batches of Senecionine decoction pieces in example 2;

[0041] Figure 9 is the superimposed graph of the characteristic spectrum of 3 batches of Senecionine formula granules in example 3;

[0042] Figure 10 is the characteristic spectrum of 2 batches of Senecionine and Senecionine medicinal materials in example 4;

[0043] Peak 2 is 5-caffeoylquinic acid, peak 3 is chlorogenic acid, peak 4 is caffeic acid, peak 5 is 4-caffeoylquinic acid, peak 7 is rutin, peak 8 is hyperoside, peak 9 is isoquercitrin, and peak 10 is 4,5-dicaffeoylquinic acid. DETAILED DESCRIPTION

[0044] In order to make the objects, technical solutions and advantages of the present application clearer, the present application will be further described in detail below with reference to the drawings and specific embodiments.

[0045] Example 1: Construction of characteristic chromatograms of Echinacea medicinal materials and their decoction pieces, standard decoction and formula granules

[0046] 1. Instruments and reagents

[0047] (1) Instruments: Waters H-Class Plus ultra-high performance liquid chromatograph (Waters Corporation); Thermo ultra-high performance liquid chromatograph (Vanquish, Thermo Fisher Scientific); Waters CORTECS Phenyl chromatographic column (4.6 mm x 150 mm, 2.7 μm); one-hundredth balance (ME204E, Mettler-Toledo); one-millionth balance (ABT220-5DM, Guangzhou Dexiang Technology Co., Ltd.); ten-millionth balance (XP26, Mettler-Toledo); electronic balance (JJ600, Changshu Shuangjie Testing Instrument Factory); numerical control ultrasonic cleaner (KQ500DE, Kunshan Ultrasonic Instrument Co., Ltd.); constant-temperature water bath (HWS28, Shanghai Yiheng Scientific Instruments); ultrapure water system (Villi-Q Direct, Merck); (2) Reagents: ethanol (Xilong Scientific Co., Ltd.), methanol (Xilong Scientific Co., Ltd.) analytical pure; phosphoric acid (Tianjin Kemio Chemical Reagent Co., Ltd.), methanol (Merck), acetonitrile (Merck) are chromatographic grade, and water is laboratory self-made ultrapure water.

[0048] (2) Reagents: ethanol (Xilong Scientific Co., Ltd.), methanol (Xilong Scientific Co., Ltd.) analytical pure; phosphoric acid (Tianjin Kemio Chemical Reagent Co., Ltd.), methanol (Merck), acetonitrile (Merck) are chromatographic grade, and water is laboratory self-made ultrapure water.

[0049] (3) 4-caffeoylquinic acid (batch number: wkq18030107, content: 98.0%, Sichuan Weikeqi Biotechnology Co., Ltd.); 5-caffeoylquinic acid (batch number: DST210427-035, content: 94.9%, Chengdu Desite Biological Technology Co., Ltd.); chlorogenic acid (batch number: 110753-202018, content: 94.9%, China National Institute for Food and Drug Control); caffeic acid (batch number: 110885-201703, content: 99.7%, China National Institute for Food and Drug Control); rutin (batch number: 100080-202012, content: 91.6%, China National Institute for Food and Drug Control); hyperoside (batch number: 111521-201809, content: 94.9%, China National Institute for Food and Drug Control); isoquercitrin (batch number: 111809-201804, content: 99.4%, China National Institute for Food and Drug Control); 4,5-dicaffeoylquinic acid (batch number: 111894-202104, content: 95.1%, China National Institute for Food and Drug Control); Senecio control medicinal materials (batch number: 120964-201810, China National Institute for Food and Drug Control); Senecio formula granules (batch numbers: KL01, KL02, KL03; source: Guangdong Yifang Pharmaceutical Co., Ltd.).

[0050] 2. Preparation of test sample solution

[0051] Medicinal materials, decoction pieces: take the powder (pass through a No. 2 sieve) of the product, take about 1.0 g, accurately weigh, place in a conical flask with a stopper, accurately add 75% methanol 25 mL, weigh the weight, heat reflux for 1 hour, cool, make up the weight lost with 75% methanol, filter, take the supernatant, concentrate to an appropriate amount, make up to 5 mL, shake well, filter, take the filtrate, and obtain.

[0052] Standard decoction: take an appropriate amount of the product, grind finely, take about 1.0 g, accurately weigh, place in a conical flask with a stopper, accurately add 75% methanol 25 mL, weigh the weight, ultrasonic treatment (power 300 W, frequency 40 kHz) for 30 minutes, take out, cool, re-weigh, make up the weight lost with 75% methanol, shake well, filter, take the filtrate, and obtain.

[0053] Formula granules: take an appropriate amount of the product, grind finely, take about 1.0 g, accurately weigh, place in a conical flask with a stopper, accurately add 75% methanol 25 mL, weigh the weight, ultrasonic treatment (power 300 W, frequency 40 kHz) for 30 minutes, take out, cool, re-weigh, make up the weight lost with 75% methanol, shake well, filter, take the filtrate, and obtain.

[0054] 3. Preparation of control sample solution

[0055] Take 5-caffeoyl quinic acid reference substance, chlorogenic acid reference substance, caffeic acid reference substance, 4-caffeoyl quinic acid reference substance, rutin reference substance, hyperoside reference substance, isoquercitrin reference substance, 4, 5-dicaffeoyl quinic acid reference substance, and 75% methanol to prepare a mixed solution containing 25 μg per 1 mL as a reference solution.

[0056] 4. Chromatographic conditions

[0057] Chromatographic column: Waters CORTECS Phenyl chromatographic column (column length 150 mm, inner diameter 4.6 mm, particle size 2.7 μm); mobile phase: acetonitrile (A)-0.2% formic acid aqueous solution (B); elution mode: gradient elution; flow rate 0.8 mL / min, column temperature 30°C, detection wavelength 360 nm. The gradient elution program of the mobile phase is shown in Table 1.

[0058] Table 1 Elution program

[0059]

[0060] 5. Determination

[0061] Precisely take 2 μL of the reference solution and 2 μL of the test solution, respectively, inject into the liquid chromatograph, and determine, i.e. obtain.

[0062] 6. Methodology investigation

[0063] (1) Column investigation

[0064] This investigation examines the influence of different chromatographic columns on each characteristic peak.

[0065] Chromatographic conditions: Waters Cortecs T3 chromatographic column (2.1 mm x 100 mm, 1.6 μm); Thermo Accource C18 chromatographic column (2.1 mm x 100 mm, 1.5 μm); Waters CSH C18 chromatographic column (2.1 mm x 100 mm, 1.7 μm); Waters Cortecs Phenyl chromatographic column (4.6 mm x 150 mm, 2.7 μm); Waters Cortecs C18 chromatographic column (4.6 mm x 150 mm, 2.7 μm) were used as chromatographic columns; acetonitrile was used as mobile phase A, and 0.2% formic acid aqueous solution was used as mobile phase B; elution was performed according to the provisions of Table 2, the flow rate was 0.8 mL / min, the column temperature was 30°C, the detection wavelength was 360 nm, and the injection volume was 2 μL.

[0066] Table 2 Elution program

[0067]

[0068] The results are shown inFigure 1 From the figure, it can be seen that different brand chromatographic columns have a greater impact on the separation of each characteristic peak. When using Waters CORTECS Phenyl (4.6 mm x 150 mm, 2.7 μm) chromatographic column, the separation effect of each characteristic peak is better, so Waters CORTECS Phenyl (4.6 mm x 150 mm, 2.7 μm) chromatographic column is selected.

[0069] (2) Investigation of mobile phase concentration and type

[0070] This investigation studies the influence of different mobile phase B on each characteristic peak in the characteristic spectrum.

[0071] Chromatographic conditions: Waters Cortecs Phenyl chromatographic column (4.6 mm x 150 mm, 2.7 μm); acetonitrile as mobile phase A, 0.01% formic acid, 0.05% formic acid, 0.2% formic acid, 0.2% phosphoric acid, 0.2% acetic acid as mobile phase B; eluted according to the provisions of Table 3, the flow rate is 0.8 mL per minute; the column temperature is 30°C; the detection wavelength is 360 nm; the injection volume is 2 μL.

[0072] Table 3 Elution program

[0073]

[0074]

[0075] The characteristic spectrum corresponding to different mobile phase B is shown in Figure 2 From the figure, it can be seen that when 0.05% formic acid and 0.2% formic acid are used as mobile phase B, the separation effect of each peak is better. When 0.2% formic acid is used as mobile phase B, the separation effect of each peak is the best.

[0076] (2) Investigation of mobile phase gradient

[0077] This investigation studies the influence of different gradient elution curves on each characteristic peak in the characteristic spectrum.

[0078] Chromatographic conditions: Waters Cortecs Phenyl chromatographic column (4.6 mm x 150 mm, 2.7 μm); acetonitrile as mobile phase A, 0.2% formic acid as mobile phase B; eluted according to the provisions of Table 4, the flow rate is 0.8 mL per minute; the column temperature is 30°C; the detection wavelength is 360 nm; the injection volume is 2 μL.

[0079] Table 4 Gradient elution table

[0080]

[0081] The results are shown in Figure 3As can be seen from the figure, when using gradient 3 for program elution, the separation degree between each chromatographic peak is good.

[0082] 7. Methodology validation

[0083] (1) Specificity

[0084] Take the above test solution, control solution and blank solvent (75% methanol) as appropriate, and test under the characteristic chromatogram chromatographic conditions. The results show that the blank solvent has no interference Figure 4 ).

[0085] (2) Durability

[0086] (2.1) Investigation of different column temperatures

[0087] Take the Senecio vulgaris formula granules (KL01) sample, prepare the test solution according to the test solution preparation method determined in item "2", except that the column temperature is 28°C, 30°C and 32°C respectively, and the other chromatographic conditions are the same as specified in item "4", sample analysis, take peak 8 as reference peak S, calculate the relative retention time and relative peak area of each characteristic peak to S peak, and calculate the RSD value. The results are shown in Figure 5 . The results show that the RSD values of relative retention time and relative peak area are all less than 5.0%, indicating that the method has good durability at different column temperatures.

[0088] (2.2) Investigation of different flow rates

[0089] Take the Senecio vulgaris formula granules (KL01) sample, prepare the test solution according to the test solution preparation method determined in item "2", except that the flow rate is 0.7mL / min, 0.8mL / min and 0.9mL / min respectively, and the other chromatographic conditions are the same as specified in item "4", sample analysis, take peak 8 as reference peak S, calculate the relative retention time and relative peak area of each characteristic peak to S peak, and calculate the RSD value. The results are shown in Figure 6 , the RSD values of relative retention time and relative peak area are all less than 5.0%, indicating that the method has good durability at different column temperatures.

[0090] (3) Stability investigation

[0091] Take the same batch of Senecio vulgaris formula granules (KL01) sample, prepare the test solution according to the method in item "2", and sample analysis under the chromatographic conditions in item "4" at 0h, 6h, 8h, 12h, 16h, 20h, 24h, take peak 8 as reference peak S, record the relative retention time and relative peak area of each characteristic peak to S peak, and calculate the RSD value, the results show that the RSD values of relative retention time and relative peak area of each characteristic peak are all less than 5%, indicating that the test solution has good stability within 24 hours.

[0092] (4) Precision examination

[0093] Take the same batch of radix sophorae flavescentis formula granules (KL01), respectively prepare the test solution according to the method in item (2) and continuously inject 6 times under the chromatographic conditions in item (4), take peak 8 as the reference peak S, record the relative retention time and relative peak area of each characteristic peak to S peak, and calculate the RSD value, the RSD value of the relative retention time and relative peak area of each characteristic peak is less than 5%, indicating that the instrument precision is good.

[0094] (5) Reproducibility examination

[0095] Take 6 portions of the same batch of radix sophorae flavescentis formula granules (KL01) sample powder, prepare 6 test solution in parallel according to the method in item (2), and inject for determination under the chromatographic conditions in item (4), take peak 8 as the reference peak S, record the relative retention time and relative peak area of each characteristic peak to S peak, and calculate the RSD value, the RSD value of the relative retention time and relative peak area of each characteristic peak is less than 5%, indicating that the method has good reproducibility.

[0096] 8. Identification of common peaks

[0097] Take radix sophorae flavescentis formula granules (KL01) sample, prepare the test solution according to the test solution preparation method in item (2), and inject for analysis under the chromatographic conditions specified in item (4), determine that peak 2 is 5-caffeoyl quinic acid, peak 3 is chlorogenic acid, peak 4 is caffeic acid, peak 5 is 4-caffeoyl quinic acid, peak 7 is rutin, peak 8 is hyperoside, peak 9 is isoquercitrin, and peak 10 is 4, 5-dicaffeoyl quinic acid.

[0098] Example 2 Determination of radix sophorae flavescentis medicinal material and decoction piece samples

[0099] Take 13 batches of radix sophorae flavescentis medicinal materials and decoction pieces, prepare the test solution according to the test solution preparation method in item (2) in Example 1, inject for determination under the chromatographic conditions in item (4) in Example 1, and use the "Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation Software" to identify the common peaks of the characteristic chromatograms of the 13 batches of radix sophorae flavescentis medicinal materials and decoction pieces. The results show that the relative retention time RSD values of the 13 batches of radix sophorae flavescentis medicinal materials are in the range of 0.03% to 0.22%, and the relative peak area RSD values are in the range of 40.86% to 95.81%; the relative retention time RSD values of the 13 batches of radix sophorae flavescentis decoction pieces are in the range of 0.03% to 0.33%, and the relative peak area RSD values are in the range of 40.20% to 118.96%; see Figure 7 , Figure 8 .

[0100] Example 3 Determination of radix sophorae flavescentis formula granule samples

[0101] Take three batches of Senecionis Scandentis Herba Dispensing Granules (batch number: KL01, KL02, KL03), prepare the test solution according to the test solution preparation method in item 2 of Example 1; inject for determination according to the chromatographic conditions in item 4 of Example 1; use the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation Software to identify the common peaks of the characteristic chromatograms of the three batches of Senecionis Scandentis Herba Dispensing Granules, and the results are shown in Table 5-Table 6, Figure 9 .

[0102] Table 5 Characteristic chromatogram of Senecionis Scandentis Herba Dispensing Granules Sample Determination (Relative Retention Time)

[0103]

[0104] Table 6 Characteristic chromatogram of Senecionis Scandentis Herba Dispensing Granules Sample Determination (Relative Peak Area)

[0105]

[0106] The above is the preferred embodiment of the application. It should be noted that for those skilled in the art, without departing from the principles of the application, several improvements and refinements can be made, which are also considered within the scope of protection of the application.

[0107] Example 4 Application of Senecionis Scandentis Herba and Its Adulterants

[0108] Take Senecionis Scandentis Herba reference materials and Senecionis Scandentis Herba, respectively, and prepare the test solution according to the test solution preparation method in item 2 of Example 1; inject for determination according to the chromatographic conditions in item 4 of Example 1; use the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation Software to study the specificity of the characteristic chromatograms of Senecionis Scandentis Herba and its adulterants, and the results show that Senecionis Scandentis Herba lacks peaks 1 and 6, and has characteristic peaks at retention time 38min-45min, indicating that the characteristic chromatogram method can effectively distinguish Senecionis Scandentis Herba and its adulterants, and the specific results are shown in Figure 10 .

Claims

1. A method for constructing a characteristic chromatogram of Senecae Radix and its decoction pieces, standard decoction and formula granules, characterized in that, The method comprises the following steps: The medicinal material of Senecionis Scandentis or its decoction pieces, standard decoction and formula granules are extracted by using an extraction solvent to obtain a test solution; The 5-caffeoylquinic acid control sample, chlorogenic acid control sample, caffeic acid control sample, 4-caffeoylquinic acid control sample, rutin control sample, hyperoside control sample, isoquercitrin control sample and 4, 5-dicaffeoylquinic acid control sample are dissolved by using the extraction solvent to obtain a control solution; The test solution and the control solution are determined by using a liquid chromatograph to obtain a characteristic spectrum of the medicinal material of Senecionis Scandentis or its decoction pieces, standard decoction and formula granules; The extraction solvent is methanol; The column of the liquid chromatograph is a Waters CORTECS Phenyl column, the column length is 150 mm, the column diameter is 4.6 mm, the particle size of the stationary phase is 2.7 μm, acetonitrile is used as the mobile phase A, and 0.05 vol%~0.2 vol% methanol aqueous solution is used as the mobile phase B for gradient elution, and the gradient elution curve is as follows: 0 min~15 min, the mobile phase A is from 5%→8%, and the mobile phase B is from 95%→92%; 15 min~22 min, the mobile phase A is from 8%→12%, and the mobile phase B is from 92%→88%; 22 min~38 min, the mobile phase A is from 12%→16%, and the mobile phase B is from 88%→84%; 38 min~50 min, the mobile phase A is from 16%→25%, and the mobile phase B is from 84%→75%; The detection wavelength of the liquid chromatograph is 300 nm~400 nm.

2. The method for constructing the characteristic chromatogram of Senecio scandens Buch Ham and its decoction pieces, standard decoction and formula granules according to claim 1, characterized in that, The mobile phase B is 0.2 vol% methanol aqueous solution.

3. The method for constructing the characteristic chromatogram of Senecio scandens Buch Ham and its decoction pieces, standard decoction and formula granules according to claim 1, characterized in that, The column temperature of the column of the liquid chromatograph is 28℃~32℃, the flow rate is 0.7 mL / min~0.9 mL / min, the injection amount of the test solution is 1 μL~3 μL, and the injection amount of the control solution is 1 μL~3 μL.

4. The method for constructing the characteristic chromatogram of Senecae Radix and its slices, standard decoction, and formula granules according to claim 1 or 3, characterized in that, The column temperature of the column of the liquid chromatograph is 30℃, the flow rate is 0.8 mL / min, the injection amount of the test solution is 2 μL, and the injection amount of the control solution is 2 μL; The detection wavelength of the liquid chromatograph is 360 nm.

5. The method for constructing the characteristic chromatogram of Senecio scandens Buch Ham and its decoction pieces, standard decoction and formula granules according to claim 1, characterized in that, In the step of extracting the medicinal material of Senecionis Scandentis or its decoction pieces, standard decoction and formula granules by using the extraction solvent to obtain the test solution, the extraction solvent is 70 vol%~80 vol% methanol, the extraction mode is heating reflux extraction or ultrasonic extraction, and the extraction time is 20 min~90 min.

6. The method for constructing the characteristic chromatogram of Senecio as the medicinal material and its decoction pieces, standard decoction and formula granules according to claim 1 or 5, characterized in that, When the test sample is the medicinal material of Senecionis Scandentis or the decoction piece of Senecionis Scandentis, the ratio of the weight of the test sample to the volume of the extraction solvent is (0.8~1.5) g:(20~30) mL, the extraction mode is heating reflux extraction, and the extraction time is 40~80 min; When the test sample is the standard decoction of Senecionis Scandentis or the formula granule of Senecionis Scandentis, the ratio of the weight of the test sample to the volume of the extraction solvent is (0.8~1.5) g:(20~30) mL, the extraction mode is ultrasonic extraction, the ultrasonic power is 200~400 W, the frequency is 20~50 kHz, and the extraction time is 20~50 min.

7. The method for constructing the characteristic chromatogram of the medicinal material of Senecionis Scandentis Herba and its slices, standard decoction, and formula granules according to claim 1, characterized in that, The characteristic spectrum comprises 10 common peaks, wherein peak 2 is 5-caffeoylquinic acid, peak 3 is chlorogenic acid, peak 4 is caffeic acid, peak 5 is 4-caffeoylquinic acid, peak 7 is rutin, peak 8 is hyperoside, peak 9 is isoquercitrin, and peak 10 is 4,5-dicaffeoylquinic acid.

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