Characteristic chromatogram of heart-stabilizing traditional Chinese medicine composition and construction method thereof

Abstract

The invention provides a characteristic chromatogram of a heart-stabilizing traditional Chinese medicine composition and a construction method thereof, the construction method adopts high performance liquid chromatography, takes acetonitrile-water as a mobile phase, screens out reasonable chromatographic conditions, detects a reference substance solution and a test solution, realizes complete separation of a plurality of effective components, and improves the accuracy of the characteristic chromatogram of the heart-stabilizing traditional Chinese medicine composition. Five common peaks are identified and identified, and a characteristic spectrum is established; an original method for detecting chemical components in one or two of codonopsis pilosula, pseudo-ginseng and rhizoma nardostachyos is improved into a method capable of detecting the chemical components in the three components at the same time, so that the quality control of the heart-stabilizing traditional Chinese medicine composition and the preparation thereof is more comprehensive.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of quality control and detection methods of traditional Chinese medicine, in particular to a characteristic chromatogram of Wenxin traditional Chinese medicine composition, a chromatogram construction method and their applications. BACKGROUND

[0002] Wenxin traditional Chinese medicine composition has Wenxin tablets, Wenxin capsules and Wenxin granules on the market, which is composed of five medicinal materials such as Dangshen, Huangjing, Sanqi, Succus and Gansong, and has the effects of benefiting qi and activating blood, and stabilizing palpitation and restoring pulse.

[0003] The characteristic chromatogram of traditional Chinese medicine preparation refers to the chromatogram that is established by studying the chromatogram of traditional Chinese medicine preparation samples after appropriate treatment, using appropriate analysis methods, which can reflect the information of multiple components and embody the quality characteristics. The characteristic chromatogram of traditional Chinese medicine preparation is of great significance for identifying the key quality attributes of traditional Chinese medicine preparation, studying the quantity and quality transmission, evaluating the uniformity and stability of traditional Chinese medicine preparation quality, and improving the overall quality control level of traditional Chinese medicine preparation.

[0004] Wenxin preparation contains five kinds of medicinal materials, indicating that multiple chemical components exist in the preparation at the same time. However, the existing technology only establishes a quality control method for the content detection of the main components of Dangshen and Sanqi (two medicinal materials) or the qualitative or quantitative determination of volatile components in Gansong (one medicinal material), which cannot simultaneously evaluate the situation of three (such as Dangshen, Sanqi and Gansong) or more medicinal materials in Wenxin preparation, and has a big defect for the overall evaluation and control of Wenxin product quality. SUMMARY

[0005] The present application aims to provide a characteristic chromatogram of Wenxin traditional Chinese medicine composition and a construction method thereof. The construction method can simultaneously separate multiple effective components in Wenxin traditional Chinese medicine composition, and the obtained chromatogram has good peak shape and separation effect. The method can be used for constructing a characteristic chromatogram. The characteristic chromatogram constructed by the aforementioned construction method can comprehensively reflect the chemical component information of the whole compound of Wenxin traditional Chinese medicine composition, and can be used for quality control of Wenxin traditional Chinese medicine composition.

[0006] Specifically, in a first aspect, the present application provides a characteristic chromatogram of Wenxin traditional Chinese medicine composition, which presents five characteristic peaks. The peak corresponding to the reference substance ginsenoside Rg1 is the reference peak (i.e. S peak), and the relative retention time of each characteristic peak to the reference peak is within ±10% of the specified value. The specified values of peak 1-peak 5 are 0.78, 0.85, 0.88, 1.00 and 1.97, respectively.

[0007] In some embodiments, the aforementioned characteristic chromatogram of Wenxin traditional Chinese medicine composition has characteristic peaks comprising substantially as shown in Figure 4 .

[0008] In a second aspect, the present application provides a method for constructing a characteristic map of a heart-stabilizing traditional Chinese medicine composition, which comprises the following steps:

[0009] 1) Preparation of a control solution: take a proper amount of a control substance, dissolve it in 80% methanol, and shake well to obtain the control solution; the control substance is Panax Notoginseng Saponins R1 control substance, Panax Ginseng Saponins Rg1 control substance, Panax Ginseng Saponins Rb1 control substance, and Codonopsis Glycoside control substance;

[0010] 2) Preparation of a test solution: take a proper amount of a test substance, extract it with 80% methanol, perform ultrasonic treatment, cool, weigh, supplement with 80% methanol, shake well, filter, and take the filtrate to obtain the test solution;

[0011] 3) High-performance liquid chromatography: use a DiamonsiL PLus C18 column with a specification of 5um, 250x4.6mm; use acetonitrile as mobile phase A and water as mobile phase B for gradient elution; the gradient elution program is as follows:

[0012] 0-12min, 16%-19%A, 84%-81%B;

[0013] 12-30min, 19%-22%A, 81%-78%B;

[0014] 30-35min, 22%A, 78%B;

[0015] 35-42min, 22%-28%A, 78%→72%B;

[0016] 42-65min, 28%-31%A, 72%-69%B;

[0017] 65-75min, 31%-43%A, 69%-57%B.

[0018] In some embodiments, the detection wavelength used in the chromatographic conditions in step 3) is 203nm.

[0019] In some embodiments, the injection amount used in the chromatographic conditions in step 3) is 10uL.

[0020] In some embodiments, the column temperature used in the chromatographic conditions in step 3) is 20-25℃, and / or the flow rate is 0.8-1.0mL / min, the detection wavelength is 203nm, and the injection amount is 10uL; preferably, the column temperature is 25℃, and / or the flow rate is 1.0mL / min.

[0021] The peaks in the obtained characteristic spectrum have suitable peak time, good resolution and stable baseline by using the column temperature and flow rate provided by the embodiment of the present application; the characteristic spectrum of the heart-stabilizing traditional Chinese medicine composition and the construction method thereof can be applied to quality detection of various heart-stabilizing products (i.e. the heart-stabilizing traditional Chinese medicine composition) such as heart-stabilizing granules, heart-stabilizing tablets and heart-stabilizing capsules.

[0022] In a third aspect, the present application further provides a detection method of the heart-stabilizing traditional Chinese medicine composition, comprising the following steps:

[0023] 1) Preparation of the solution of the sample to be detected: an appropriate amount of the sample to be detected of the heart-stabilizing traditional Chinese medicine composition is taken, extracted with 80% methanol, ultrasonically treated, cooled, weighed, supplemented with 80% methanol, shaken, filtered, and the filtrate is taken, thereby obtaining the solution of the sample to be detected;

[0024] 2) The solution of the sample to be detected is detected by high performance liquid chromatography, and the obtained chromatogram of the solution of the sample to be detected is compared with the characteristic spectrum constructed by the aforementioned construction method;

[0025] The chromatographic conditions are as follows: the chromatographic column is DiamonsiL PLus C18 with a specification of 5um and 250x4.6mm; acetonitrile is used as the mobile phase A and water is used as the mobile phase B for gradient elution, and the gradient elution program is as follows:

[0026] 0-12min, 16%-19%A, 84%-81%B;

[0027] 12-30min, 19%-22%A, 81%-78%B;

[0028] 30-35min, 22%A, 78%B;

[0029] 35-42min, 22%-28%A, 78%→72%B;

[0030] 42-65min, 28%-31%A, 72%-69%B;

[0031] 65-75min, 31%-43%A, 69%-57%B.

[0032] In some embodiments, the column temperature in step 3) is 20-25℃, and / or the flow rate is 0.8-1.0mL / min; preferably, the column temperature is 25℃, and / or the flow rate is 1.0mL / min.

[0033] In some embodiments, the detection wavelength in step 3) is 203nm.

[0034] In some embodiments, the injection amount in step 3) is 10uL.

[0035] In the present application, substantially as Figure 4 refers to the characteristic map of the Wenxin traditional Chinese medicine composition contains 5 characteristic peaks, with peak 4 as the reference peak (S peak, relative retention time is 1.00), the relative retention time of the other four characteristic peaks is 1 peak (0.78), 2 peak (0.85), 3 peak (0.88), 5 peak (1.97), respectively. The relative retention time of each characteristic peak is within ±10% of the specified value. In addition to the above 5 identified characteristic peaks, a plurality of common peaks can also be optionally included, such as the attached Figure 4 .

[0036] The present application has the following beneficial effects:

[0037] (1) The present application provides a method for constructing the characteristic map of the Wenxin traditional Chinese medicine composition. Through a large number of experiments, a specific chromatographic column and a specific elution program are selected to achieve the complete separation of multiple chemical components of ginseng, sanchi and gansong in the Wenxin traditional Chinese medicine composition. The peak shape of each characteristic peak in the characteristic map obtained by the construction method is good, and the baseline is stable. Each component measured can be completely separated. The construction method provides a basis for quality detection and control of the Wenxin traditional Chinese medicine composition and its preparations, and can fully reflect the integrity and characteristics of the Wenxin traditional Chinese medicine composition and its preparations. The method is simple to operate and suitable for quality control of various Wenxin preparations.

[0038] (2) The present application uses high performance liquid chromatography to construct the characteristic map of the Wenxin traditional Chinese medicine composition. Through the identification of reference substances, peak 4 is ginsenoside Rg1, which has good separation degree and appropriate area. The other characteristic peaks are determined by reference to it, and the accuracy of the quality control of the Wenxin traditional Chinese medicine composition can be achieved by qualitative method. The five characteristic peaks in the characteristic map determined by the method are the stability common peaks of 15 batches of Wenxin granules test samples. The method has good precision, repeatability, durability and specificity, so it can well guarantee the accuracy and stability of the method when applied to the quality control of the Wenxin traditional Chinese medicine composition and its preparations.

[0039] (3) Compared with the prior art, the detection method provided by the present application realizes the simultaneous separation and qualitative identification of more main effective components. Through the construction method of the present application, the problem of adhesion between the chemical components of sanchi saponin R1, ginsenoside Rb1, ginsenoside Rg1 and gansong and the nearby impurity peaks, or the problem of impure chromatographic peaks are solved, thereby effectively solving the technical problem of interference verification between chromatographic peaks caused by the large number of impurities in the Wenxin traditional Chinese medicine composition due to the large number of medicinal materials. The simultaneous detection of the chemical components of ginseng, sanchi and gansong and the stable control of the intermediate or preparation product quality are realized. BRIEF DESCRIPTION OF DRAWINGS

[0040] Figure 1 The chromatogram obtained by the detection method of examples 1-4.

[0041] Figure 2 Characteristics of the Example 4 stable heart granule intermediate.

[0042] Figure 3 Comparison chart of the software-generated control characteristics and the characteristics of 15 batches of stable heart granule intermediates.

[0043] Figure 4 Software-generated control characteristics.

[0044] Figure 5 Comparison chart of the software-generated control characteristics and the reference chart of the control.

[0045] Figure 6 Specificity investigation results. DETAILED DESCRIPTION

[0046] The following content exemplarily illustrates the technical solutions of the present application, which is a further full description of the present application, facilitating the understanding of the implementation conditions and means of the present application, and should not be considered as a limitation of the present application.

[0047] Instrument

[0048] High performance liquid chromatograph (AgiLent 1260Infinity II, each component: pump G7111B, sample injector G7129A, column oven G7116A, detector G7115A), one-millionth scale (Ohaus), ten-millionth scale (Swiss Mettler), ultrasonic instrument (model: KQ-500DE).

[0049] Reagents and medicinal materials

[0050] Acetonitrile and methanol are chromatographically pure, water is ultrapure water, and all other reagents are analytically pure; Panax notoginseng saponin R1 (purchased from China Food and Drug Inspection Research Institute, batch number 110745-202322); ginsenoside Rg1 (purchased from China Food and Drug Inspection Research Institute, batch number 110703-202235); ginsenoside Rb1 (purchased from China Food and Drug Inspection Research Institute, batch number 110704-202331); codonopsin (purchased from Chengdu Glipl Biological Technology Co., Ltd., batch number 23030902); stable heart granule test sample (self-made stable heart granule intermediate or finished product); 80% methanol in the present application refers to 80% methanol by volume; 80% ethanol refers to 80% ethanol by volume;

[0051] Preparation example

[0052] 1. Preparation of self-made stable heart granule intermediate

[0053] Prescription: Codonopsis 300g, Polygonatum 400g, Panax notoginseng 60g, Amber 40g, Gamsong 200g

[0054] Process: The above five flavors, amber powder into fine powder, volatile oil extraction, extraction of water solution collected separately; three seven powder into coarse powder, reflux extraction with 80% ethanol twice, 2 hours each time, filter, filter combined, concentrated under reduced pressure to the relative density of 1.13~1.15 (60 ℃) clear paste, the residue plus water decoction two times, the first time 2 hours, the second time 1.5 hours, decoction combined: decoction with the above two times, filter; filter concentrate to the relative density of 1.22~1.24 (60 ℃) clear paste, add ethanol to make the alcohol content of 65%, stirring, standing for 24 hours, filter, filter concentrate to 1.30~1.33 (60 ℃) thick paste, with three seven clear paste, mix evenly, get stable heart granules intermediate.

[0055] 2, the preparation of self-made stable heart granules

[0056] Amber fine powder 40g Aspartame 1.37g Dextrin 88g Soluble starch 264g

[0057] Take the above stable heart granules intermediate, add the above prescription amount of amber fine powder, aspartame dextrin and soluble starch, mix evenly, granulation, drying, spray into volatile oil of glycosmis, mix evenly, made into granules 556g (no sucrose), packed, get stable heart granules finished product.

[0058] Example 1

[0059] Preparation of reference solution: take ginsenoside R1 reference, ginsenoside Rg1 reference, ginsenoside Rb1 reference, party ginseng glycoside reference, accurately weighed, add 80% methanol to make 1mL containing ginsenoside R1 0.04mg, ginsenoside Rg10.15mg, ginsenoside Rb1 0.1mg, party ginseng glycoside 0.04mg mixed solution, shake well, get.

[0060] Preparation of test solution: take 0.5g of stable heart granules intermediate of preparation example 1, accurately weighed, put into a conical flask with plug, accurately add 25mL of 80% methanol, seal, weigh, ultrasonic treatment (power 500W, frequency 40kHz) 45 minutes, take out, cool, weigh again, make up the weight loss with 80% methanol, shake well, filter, take the filtrate, get.

[0061] Determination by high performance liquid chromatography: high performance liquid chromatograph AgiLent 1260Infinity II, using chromatographic column Luna C18(2)(5μm, 250×4.6mm); with acetonitrile as mobile phase A, water as mobile phase B, gradient elution according to the following table 1; flow rate is 1.0mL / min, column temperature is 25℃, detection wavelength is 203nm, injection volume is 10uL, the determination results are as followsFigure 1 S2(3) is shown in the middle.

[0062] Table 1: Mobile phase elution gradient 1

[0063]

[0064] Example 2

[0065] Preparation of the control solution: Take a proper amount of ginsenoside R1 control, ginsenoside Rg1 control, ginsenoside Rb1 control, and loxogrisein control, and accurately weigh, add 80% methanol to prepare a mixed solution containing 0.04 mg of ginsenoside R1, 0.15 mg of ginsenoside Rg1, 0.1 mg of ginsenoside Rb1, and 0.04 mg of loxogrisein per 1 mL, shake well, and obtain.

[0066] Preparation of the test solution: Take 0.5 g of the intermediate of the Wenxin granules of Preparation Example 1, accurately weigh, place in a conical flask with a plug, accurately add 25 mL of 80% methanol, tightly plug, weigh, ultrasonic treat (power 500 W, frequency 40 kHz) for 45 minutes, take out, cool, re-weigh, make up the weight loss with 80% methanol, shake well, filter, and take the filtrate, and obtain.

[0067] High performance liquid chromatography determination: AgiLent 1260Infinity II high performance liquid chromatograph, using a DiamonsiL PLus C18 column (5 μm, 250 x 4.6 mm); acetonitrile as mobile phase A, water as mobile phase B, gradient elution according to the provisions of Table 1 in Example 1; flow rate 1.0 mL / min, column temperature 25℃, detection wavelength 203 nm, injection volume 10 uL, and the determination results are as follows Figure 1 S3(5) is shown in the middle.

[0068] Example 3

[0069] Preparation of the control solution: Take a proper amount of ginsenoside R1 control, ginsenoside Rg1 control, ginsenoside Rb1 control, and loxogrisein control, and accurately weigh, add 80% methanol to prepare a mixed solution containing 0.04 mg of ginsenoside R1, 0.15 mg of ginsenoside Rg1, 0.1 mg of ginsenoside Rb1, and 0.04 mg of loxogrisein per 1 mL, shake well, and obtain.

[0070] Preparation of the test solution: Take 0.5 g of the intermediate of the Wenxin granules of Preparation Example 1, accurately weigh, place in a conical flask with a plug, accurately add 25 mL of 80% methanol, tightly plug, weigh, ultrasonic treat (power 500 W, frequency 40 kHz) for 45 minutes, take out, cool, re-weigh, make up the weight loss with 80% methanol, shake well, filter, and take the filtrate, and obtain.

[0071] High performance liquid chromatography: AgiLent 1260 Infinity II high performance liquid chromatograph, using a chromatographic column DiamonsiL PLus C18 (5 μm, 250 x 4.6 mm); acetonitrile as mobile phase A, water as mobile phase B, gradient elution according to the following Table 2; flow rate of 1.0 mL / min, column temperature 25 ℃, detection wavelength of 203 nm, injection volume of 10 uL, and the determination results are as follows Figure 1 S4(5) is shown in the middle.

[0072] Table 2: mobile phase elution gradient 2

[0073]

[0074] Example 4

[0075] Preparation of the reference solution: Take notoginsenoside R1 reference, ginsenoside Rg1 reference, ginsenoside Rb1 reference, and loxogrisein reference, accurately weigh, add 80% methanol to prepare a mixed solution containing 0.04 mg of notoginsenoside R1, 0.15 mg of ginsenoside Rg1, 0.1 mg of ginsenoside Rb1, and 0.04 mg of loxogrisein per 1 mL, shake well, and obtain.

[0076] Preparation of the test solution: Take 0.5 g of the intermediate of Wenxin granules prepared in Preparation Example 1, accurately weigh, place in a conical flask with a plug, accurately add 25 mL of 80% methanol, tightly plug, weigh, ultrasonic treatment (power 500 W, frequency 40 kHz) for 45 minutes, take out, cool, re-weigh, make up the weight loss with 80% methanol, shake well, filter, and take the filtrate, which is the test solution.

[0077] High performance liquid chromatography: AgiLent 1260 Infinity II high performance liquid chromatograph, using a chromatographic column DiamonsiL PLus C18 (5 μm, 250 x 4.6 mm); acetonitrile as mobile phase A, water as mobile phase B, gradient elution according to the following Table 3; flow rate of 1.0 mL / min, column temperature 25 ℃, detection wavelength of 203 nm, injection volume of 10 uL. The determination results are as follows Figure 1 S1(5) and Figure 2 S4(5) are shown in the middle.

[0078] Table 3: mobile phase elution gradient 3

[0079]

[0080] In order to completely separate and detect the five components in the test sample by high performance liquid chromatography, the applicant carried out screening experiments (Examples 1-4), the specific process is as follows:

[0081] The detection results showed that peak 4 was seriously adhered to the impurity peak when eluted with gradient 1 using Luna C18(2) (5 μm, 250 x 4.6 mm) as the chromatographic column, resulting in poor separation of peak 4 (see Figure 2S3 of Example 2); Figure 1

[0082] To solve the problem of separation of peak 4, the chromatographic column was replaced with DiamonsiL Plus C18 (5 μm, 250 x 4.6 mm), and the detection results showed that peak 4 was completely separated from the adjacent peaks, but peaks 2 and 3 were still slightly adhered to the impurity peaks, and peak 2 was detected by the diode array detector as a non-pure peak (see Figure 2S3 (5) of Example 2); Figure 1

[0083] It can be seen that although the separation problem of peak 4 has been solved by changing the type of chromatographic column, peaks 2 and 3 still cannot be separated, so on the basis of the chromatographic column being DiamonsiL Plus C18 (5 μm, 250 x 4.6 mm), the elution program was further changed to gradient 2 for detection, and the results showed that peaks 3 and 4 were completely separated from the adjacent peaks, but peak 2 was still slightly adhered to the impurity peaks (see Figure 2S4 (5) of Example 2); Figure 1

[0084] Further optimization of the elution gradient, the chromatographic column remained unchanged, and elution was carried out with gradient 3, and the detection results showed that peaks 1, 2, 3, 4 and 5 were completely separated, and the peak shapes were good (see Figure 2S1 (5) of Example 2). Figure 1

[0085] In summary, the high performance liquid chromatography method of Example 4 can effectively separate the five components in the test sample at the same time.

[0086] Confirmation of characteristic peaks of the characteristic spectrum of Example 5

[0087] Take 15 batches of stable heart granule intermediates prepared in Preparation Example 1, respectively, according to the method of Example 4 to prepare the test sample solution, and determine under the chromatographic conditions of Example 4, obtain the chromatograms of 15 batches of stable heart granule intermediates, see Figure 3 (wherein R(5) is the control characteristic spectrum, S1(5) ~ S15(5) are the chromatograms of 15 batches of stable heart granule intermediates, respectively); the results of the chromatograms of 15 batches of stable heart granule intermediates are fitted, and the traditional Chinese medicine chromatographic fingerprint similarity evaluation system (2012 version) software is used to generate the test sample control characteristic spectrum (see Figure 4 ) by the average method, calculate the similarity of each characteristic peak, all greater than 0.9, see Table 4 below:

[0088] Table 4: Similarity calculation results of 15 batches of stable heart granule test samples

[0089] ​​​​

[0090] Take the appropriate amount of notoginseng saponin Rl reference substance, ginsenoside Rg1 reference substance, ginsenoside Rb1 reference substance, and codonopsin reference substance, respectively, and accurately weigh, add 80% methanol to prepare a solution containing 0.04 mg of notoginseng saponin Rl, 0.15 mg of ginsenoside Rg1, 0.10 mg of ginsenoside Rb1, and 0.04 mg of codonopsin per 1 mL, respectively, and shake well to obtain four kinds of reference substance reference solution. Under the chromatographic conditions of Example 4, the reference substance reference solution is determined to obtain the reference substance reference spectrum. Compare the test product control characteristic spectrum with the reference substance reference spectrum, see Figure 5 (wherein S1(5) is the control characteristic spectrum, and S2-S5 are the reference substances).

[0091] Conclusion: The characteristic spectrum of the test product presents five characteristic peaks, which correspond to the retention times of the four characteristic peaks in the reference substance reference solution. Peak 4 is ginsenoside Rg1, and this peak is labeled as the S peak. The relative retention times of the other characteristic peaks to the S peak are calculated to be the specified values, which are: 0.78 (peak 1), 0.85 (peak 2), 0.88 (peak 3), 1.00 (peak 4), and 1.97 (peak 5). The relative retention times of each characteristic peak to the S peak are within ±10% of the specified values.

[0092] Methodology verification of Example 6

[0093] 1. Stability

[0094] Take the appropriate amount of test product, prepare the test product solution according to the method of Example 1, inject 10 μL into the liquid chromatograph, and continuously inject 6 times to obtain the corresponding peak area and retention time, and calculate the RSD value. The results are shown in Tables 5 and 6 below. According to the results, the relative retention time and relative peak area of each chromatographic peak in the stability test meet the requirements (RSD ≤ 3%), indicating that the method has good stability.

[0095] Table 5: Stability test (relative retention time)

[0096]

[0097] Table 6: Stability test (relative peak area)

[0098]

[0099] 2. Reproducibility

[0100] Take the test product, parallel to take 6, according to the method of example 1 preparation of test solution, injection volume is 10 μL, inject liquid chromatograph, record peak area and retention time, calculate RSD value, the results are shown in table 7 and table 8. From the results, the repeatability of each chromatographic peak relative retention time and relative peak area RSD are less than 3.0%, indicating that the method has good repeatability.

[0101] Table 7: repeatability test (relative peak area)

[0102]

[0103] Table 8: repeatability test (relative retention time)

[0104]

[0105] 3, durability

[0106] Take the preparation of example 1 of the intermediate of Wenxin granules, according to the method of example 1 preparation of test solution, to investigate the influence of the determination conditions under the specific range of fluctuation on the determination results. The test mainly investigates the durability of the experimental method under different column temperature (20 ℃ ~ 25 ℃), different flow rate (0.8 mL / min ~ 1.0 mL / min) variation factors, and the results show that when the column temperature is 20 ~ 25 ℃ and the flow rate is 0.8 ~ 1.0 mL / min, the chromatographic peak separation degree is better.

[0107] 4, specificity

[0108] Take the preparation of example 1 of the intermediate of Wenxin granules, negative sample, according to the method of example 1 preparation of test solution, according to the following method to prepare the negative sample solution, and detect the characteristic map, to investigate the specificity of the method, the results are shown in Figure 6 (S1 ~ S5 is negative sample, S6 is test product, S7 ~ S10 is control product).

[0109] Preparation of negative sample solution: take the negative sample 0.5 g, accurately weighed, put into the conical flask with plug, accurately add 80% methanol 25 mL, tightly plug, weigh, ultrasonic treatment (power 500 W, frequency 40 kHz) 45 minutes, take out, cool, weigh again, make up the weight loss with 80% methanol, shake evenly, filter, take the filtrate, get it.

[0110] The results show that the total saponins of panax notoginseng (panax notoginseng saponin R1, ginsenoside Rg1, ginsenoside Rb1) are from panax notoginseng decoction pieces, party a glycoside is from party a decoction pieces, 3 peak is from gansong decoction pieces, and the negative sample has no obvious chromatographic peak at the corresponding retention time of the test product chromatogram, indicating that the method has good specificity.

[0111] The above embodiments are only preferred specific embodiments, and do not represent the actual protection scope. Those skilled in the art can make various improvements and changes to the specific embodiments of the present application without departing from the spirit of the present application. If only simple changes, equivalent replacements or simple modifications are made to the above content, it can be considered that the improved or changed technical solutions still belong to the protection scope of the present application.

Claims

1. A characteristic chromatogram of a traditional Chinese medicine composition for stabilizing heart, characterized in that, The characteristic spectrum presents five characteristic peaks, and the peak corresponding to the reference substance ginsenoside Rg1 is the reference peak, the relative retention time of each characteristic peak to the reference peak is within ±10% of the specified value, and the specified values of peaks 1-5 are 0.78, 0.85, 0.88, 1.00, and 1.97, respectively.

2. The characteristic chromatogram of the heart-stabilizing traditional Chinese medicine composition according to claim 1, characterized in that, has characteristic peaks substantially as shown in Figure 4.

3. A method for constructing a characteristic map of a traditional Chinese medicine composition for stabilizing heart, characterized in that, The method comprises the following steps: 1) Preparation of the control solution: take the control substance, dissolve in 80% methanol, shake well, and obtain; The control substance is notoginsenoside R1 control substance, ginsenoside Rg1 control substance, ginsenoside Rb1 control substance, and codonopsin control substance. 2) Preparation of the test solution: take the test substance, extract with 80% methanol, ultrasonic treatment, cool, weigh, supplement with 80% methanol, shake well, filter, and take the filtrate, and obtain; 3) Detect the control solution and the test solution by high performance liquid chromatography, and the chromatographic conditions are as follows: The chromatographic column is DiamonsiL PLus C18, with a specification of 5um, 250x4.6mm; Use acetonitrile as mobile phase A and water as mobile phase B for gradient elution, and the gradient elution program is as follows: 0-12min, 16%-19%A, 84%-81%B; 12-30min, 19%-22%A, 81%-78%B; 30-35min, 22%A, 78%B; 35-42min, 22%-28%A, 78%→72%B; 42-65min, 28%-31%A, 72%-69%B; 65-75min, 31%-43%A, 69%-57%B.

4. The method of constructing a feature map according to claim 3, wherein, The column temperature for the chromatographic conditions in step 3) is 20-25℃, and / or the flow rate is 0.8-1.0mL / min.

5. The method of constructing a feature map according to claim 3 or 4, wherein, The column temperature for the chromatographic conditions in step 3) is 25℃, and / or the flow rate is 1mL / min.

6. The method of constructing a feature map according to any one of claims 3 to 5, wherein, The detection wavelength for the chromatographic conditions in step 3) is 203nm.

7. The method of constructing a feature map according to any one of claims 3 to 6, wherein, The characteristic spectrum presents five characteristic peaks, and the peak corresponding to the reference substance ginsenoside Rg1 is the reference peak, the relative retention time of each characteristic peak to the reference peak is within ±10% of the specified value, and the specified values of peaks 1-5 are 0.78, 0.85, 0.88, 1.00, and 1.97, respectively.

8. The characteristic spectrum of any one of claims 1-2 or the construction method of any one of claims 3-7 or the application in the quality detection of the stabilizing heart traditional Chinese medicine composition.

9. A method for detecting a Chinese medicine composition for stabilizing heart, characterized in that, The method comprises the following steps: 1) Preparation of the test solution: take the test substance, extract with 80% methanol, ultrasonic treatment, cool, weigh, supplement with 80% methanol, shake well, filter, and take the filtrate, and obtain; 2) Detect the test solution by high performance liquid chromatography, and compare the color spectrum of the obtained test solution with the characteristic spectrum constructed by the construction method of any one of claims 1-7; The chromatographic conditions are as follows: The chromatographic column is DiamonsiL PLus C18, with a specification of 5um, 250x4.6mm; The gradient elution program is as follows: 0-12 min, 16%-19% A, 84%-81% B; 12-30 min, 19%-22% A, 81%-78% B; 30-35 min, 22% A, 78% B; 35-42 min, 22%-28% A, 78%→72% B; 42-65 min, 28%-31% A, 72%-69% B; 65-75 min, 31%-43% A, 69%-57% B.

10. The detection method according to claim 9, characterized in that, The column temperature for the chromatography condition in step 2) is 20-25 ℃, and / or the flow rate is 0.8-1.0 mL / min.

11. The detection method according to claim 9 or 10, characterized in that, The column temperature for the chromatography condition in step 2) is 25 ℃, and / or the flow rate is 1 mL / min.

12. The assay method according to any one of claims 9 to 11, characterized in that, The detection wavelength for the chromatography condition in step 2) is 203 nm.

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