Quality detection method for carpesium abrotanoides formula granules

Abstract

The invention discloses a quality detection method for carpesium abrotanoides formula granules, and belongs to the technical field of traditional Chinese medicine detection. The detection method comprises the following steps: testing the total content of the specific chromatogram, neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid, and testing the content of carabrone. The method for testing the specific chromatogram and the total content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid comprises the following steps: a, preparing a neochlorogenic acid reference substance solution, a chlorogenic acid reference substance solution and a cryptochlorogenic acid reference substance solution; b, preparing a test solution; and c, carrying out liquid chromatography detection, and calculating the total content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid. The carabrone content test comprises the following steps: A, preparing a carabrone reference substance solution; b, preparing a test solution; and C, carrying out liquid chromatography detection, and calculating the carabrone content. The quality of the carpesium abrotanoides formula granules is effectively controlled, and the medication effectiveness and safety of the carpesium abrotanoides formula granules are guaranteed.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine detection, and more specifically, to a method for detecting the quality of Tianmingjing formula granules. Background Art

[0002] Chinese medicine formula granules are made from Chinese medicine slices through extraction, concentration, drying, granulation and other steps. Compared with the raw materials, the formula granules have lost their original form, and the corresponding active ingredients have also changed accordingly. The quality inspection method for raw materials is not suitable for the quality inspection of formula granules.

[0003] Tianmingjing is a plant of the genus Tianmingjing in the Asteraceae family, which has the effects of clearing heat, resolving phlegm, detoxifying, killing insects, removing blood stasis, and stopping bleeding. However, there is currently no clear qualitative and quantitative quality testing method for Tianmingjing formula granules, making it difficult to comprehensively evaluate and control the process from Tianmingjing to the finished formula granules, and unable to meet the quality control requirements of traditional Chinese medicine formula granules, which limits the widespread use of Tianmingjing formula granules in clinical practice. Summary of the invention

[0004] The purpose of the present invention is to overcome the deficiencies of the above-mentioned prior art and provide a quality detection method for Tianmingjing formula granules, which can effectively control the quality of Tianmingjing formula granules and ensure their effectiveness and safety.

[0005] In order to achieve the above object, the present invention adopts the following technical solution:

[0006] A quality detection method for Tianmingjing formula granules, including characteristic spectrum and total content test of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid and content test of Tianmingjing lactone;

[0007] The characteristic spectrum and the total content test of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid include the following steps:

[0008] a. Prepare neochlorogenic acid reference solution, chlorogenic acid reference solution and cryptochlorogenic acid reference solution;

[0009] b. Take the Tianmingjing formula granule sample to prepare the test solution;

[0010] c. Perform liquid chromatography on the neochlorogenic acid reference solution, chlorogenic acid reference solution, cryptochlorogenic acid reference solution and test solution, respectively, and calculate the total content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid;

[0011] The test of the content of tianmingjing lactone comprises the following steps:

[0012] A. Prepare the reference solution of tianmingjing lactone;

[0013] B. Take the Tianmingjing formula granule sample to prepare the test solution;

[0014] C. Perform liquid chromatography on the tianmingjing lactone reference solution and the test solution respectively, and calculate the tianmingjing lactone content.

[0015] The present invention is further configured as follows: the Tianmingjing decoction pieces are decocted with water for multiple times, filtered, the filtrate is concentrated to a clear paste, the clear paste is dried to form a dry paste; the auxiliary material maltodextrin is added and mixed and granulated, or granulated directly without adding auxiliary materials to obtain the Tianmingjing formula granules.

[0016] The present invention is further configured that in the characteristic spectrum and the total content test of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid, the chromatographic conditions are:

[0017] Chromatographic column: CORTECS UPLC T3, 2.1mm×100mm, 1.6μm; detection wavelength: 330nm; flow rate: 0.3ml / min; column temperature: 35℃; injection volume: 1μl; mobile phase: methanol as mobile phase A, 0.1wt% phosphoric acid as mobile phase B; gradient elution program: during 0-8min, mobile phase A increased from 15% to 30%, and mobile phase B decreased from 85% to 70%; during 8-18min, mobile phase A increased from 30% to 36%, and mobile phase B decreased from 70% to 64%; during 18-25min, mobile phase A increased from 36% to 50%, and mobile phase B decreased from 64% to 50%; during 25-35min, mobile phase A increased from 50% to 70%, and mobile phase B decreased from 50% to 30%.

[0018] The present invention is further configured such that in step a, the neochlorogenic acid reference solution, the chlorogenic acid reference solution and the cryptochlorogenic acid reference solution are all prepared using methanol as a solvent, and the concentration of each reference solution is 30 μg / ml.

[0019] The present invention is further configured that step b specifically comprises: taking 0.5 g of Tianmingjing formula granule sample powder, adding 25 ml of 50 wt% methanol, ultrasonically treating, cooling, weighing again, supplementing the lost weight with 50 wt% methanol, shaking well, filtering, and taking the filtrate to obtain.

[0020] The present invention is further configured that, in the test of the content of tianmingjing lactone, the chromatographic conditions are:

[0021] Chromatographic column: CORTECS UPLC T3, 2.1mm×100mm, 1.6μm; detection wavelength: 210nm; flow rate: 0.15ml / min; column temperature: 30℃; injection volume: 1μl; mobile phase: acetonitrile as mobile phase A, 0.1wt% phosphoric acid as mobile phase B; gradient elution program: during 0-5min, mobile phase A was maintained at 35%, and mobile phase B was maintained at 65%; during 5-7.5min, mobile phase A increased from 35% to 40%, and mobile phase B decreased from 65% to 60%; during 7.5-8.8min, mobile phase A increased from 40% to 95%, and mobile phase B decreased from 60% to 5%; during 8.8-10min, mobile phase A was maintained at 95%, and mobile phase B was maintained at 5%; during 10-12min, mobile phase A decreased from 95% to 35%, and mobile phase B increased from 5% to 65%.

[0022] The present invention is further configured as follows: in step A, the tianmingjing lactone reference substance solution is prepared using methanol as a solvent, and the concentration of the tianmingjing lactone reference substance solution is 30 μg / ml.

[0023] The present invention is further configured that step B specifically comprises: taking 0.5 g of Tianmingjing formula granule powder, adding 25 ml of 50 wt% methanol solution, heating to reflux, cooling, weighing again, supplementing the lost weight with 50 wt% methanol, shaking well, filtering, and taking the filtrate to obtain.

[0024] In summary, the present invention has the following beneficial effects:

[0025] The quality of Tianmingjing formula granules is evaluated by constructing a characteristic map and determining the contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, and Tianmingjing lactone ketone. Further, a feasible quality standard for Tianmingjing formula granules is established to achieve effective control of the quality of Tianmingjing formula granules and ensure the effectiveness and safety of clinical use. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 The chromatogram of the new chlorogenic acid reference substance solution was obtained to verify the characteristic spectrum of Tianmingjing formula granules and the positioning and system applicability of the determination method of the total content of new chlorogenic acid, chlorogenic acid and cryptochlorogenic acid;

[0027] Figure 2 The chromatogram of the chlorogenic acid reference solution was obtained to verify the characteristic spectrum of Tianmingjing Formula Granules and the positioning and system applicability of the determination method of the total content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid;

[0028] Figure 3 The chromatogram of the cryptochlorogenic acid reference solution was obtained to verify the characteristic spectrum of Tianmingjing formula granules and the positioning and system applicability of the determination method of the total content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid;

[0029] Figure 4 The chromatogram of the test sample solution was obtained to verify the characteristic spectrum of Tianmingjing formula granules and the positioning and system applicability of the determination method of the total content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid;

[0030] Figure 5 To verify the characteristic spectrum of Tianmingjing formula granules and the chromatogram obtained by the determination method of the total content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid at 35°C column temperature durability;

[0031] Figure 6 To verify the characteristic spectrum of Tianmingjing formula granules and the chromatogram obtained by the determination method of the total content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid at 30°C column temperature durability;

[0032] Figure 7 To verify the characteristic spectrum of Tianmingjing formula granules and the chromatogram obtained by the determination method of the total content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid at 40°C column temperature durability;

[0033] Figure 8 To verify the chromatogram obtained by the characteristic spectrum of Tianmingjing formula granules and the flow rate durability of the determination method of the total content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid at a flow rate of 0.27ml / min;

[0034] Fig. 9 To verify the chromatogram obtained by the characteristic spectrum of Tianmingjing formula granules and the flow rate durability of the determination method of the total content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid at a flow rate of 0.33ml / min;

[0035] Fig.10 To verify the characteristic spectrum of Tianmingjing formula granules and the chromatogram obtained by the determination method of the total content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid at a wavelength of 325nm;

[0036] Fig.11 To verify the characteristic spectrum of Tianmingjing formula granules and the chromatogram obtained by the determination method of the total content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid at a wavelength of 335nm;

[0037] Fig.12 The chromatogram of the Tianmingjing lactone reference substance solution obtained to verify the positioning of the determination method and system suitability of the Tianmingjing lactone content in Tianmingjing formula granules;

[0038] Fig.13 The chromatogram of the test sample solution obtained to verify the positioning of the determination method and system suitability of the content of Tianmingjing lactone in Tianmingjing formula granules;

[0039] Fig.14 The chromatogram obtained to verify the column temperature durability at 30°C of the method for determining the content of Tianmingjing lactone in Tianmingjing formula granules;

[0040] Fig.15 The chromatogram obtained to verify the column temperature durability at 25°C of the method for determining the content of Tianmingjing lactone in Tianmingjing formula granules;

[0041] Fig.16 The chromatogram obtained to verify the column temperature durability of the determination method of Tianmingjing lactone content in Tianmingjing formula granules at 35°C;

[0042] Fig.17 The chromatogram obtained to verify the flow rate durability of the method for determining the content of Tianmingjing lactone in Tianmingjing formula granules at a flow rate of 0.13ml / min;

[0043] Fig.18 The chromatogram obtained to verify the flow rate durability of the method for determining the content of Tianmingjing lactone in Tianmingjing formula granules at a flow rate of 0.17ml / min;

[0044] Fig.19 The chromatogram obtained to verify the wavelength durability of the determination method of Tianmingjing lactone content in Tianmingjing formula granules at a wavelength of 205nm;

[0045] Fig. 20 This chromatogram was obtained to verify the wavelength durability of the determination method of Tianmingjing lactone content in Tianmingjing formula granules at a wavelength of 215nm. DETAILED DESCRIPTION

[0046] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0047] 1. Preparation of Tianmingjing Formula Granules

[0048] Take Tianmingjing slices, add water and boil for several times, filter, concentrate the filtrate to a clear paste, dry the clear paste into a dry paste (the yield of the clear paste to the dry paste is 15-40%), add the auxiliary material maltodextrin, mix and granulate, or directly granulate without adding auxiliary materials to obtain Tianmingjing formula granules.

[0049] Take 3 portions of Tianmingjing decoction pieces and prepare Tianmingjing formula granules according to the above method and the parameters shown in Table 1.

[0050] Table 1

[0051]

[0052] 2. Quality inspection of Tianmingjing formula granules

[0053] (1) Characteristic spectrum and total content test of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid

[0054] 1.1 Test Method

[0055] 1.1.1 Preparation of reference solution: Take new chlorogenic acid (purchased from Shanghai Shidande Standard Technical Service Co., Ltd., batch number: ST06230120, content 98.0%), chlorogenic acid (purchased from China Food and Drug Inspection Institute, batch number: 110753-202119, content 96.3%), and cryptochlorogenic acid (purchased from Shanghai Shidande Standard Technical Service Co., Ltd., batch number: ST07850120, content 98.0%) reference substances, accurately weigh them, and add methanol to prepare new chlorogenic acid reference solution, chlorogenic acid reference solution and cryptochlorogenic acid reference solution with a concentration of 30μg / ml respectively.

[0056] 1.1.2 Preparation of test solution: Grind 0.5 g of Tianmingjing granule sample into powder, accurately weigh it, put it in a stoppered conical flask, accurately add 25 ml of 50 wt% methanol, weigh it, ultrasonically treat it (power 250 W, frequency 50 kHz) for 15 minutes, cool it, weigh it again, make up the lost weight with 50 wt% methanol, shake it well, filter it, and take the filtrate to obtain the solution.

[0057] 1.1.3 Determination: Accurately pipette 1 μl of each of the neochlorogenic acid reference solution, chlorogenic acid reference solution, cryptochlorogenic acid reference solution, and test solution, and inject them into the liquid chromatograph for testing. The chromatographic conditions are:

[0058] Chromatographic column: CORTECS UPLC T3, 2.1mm×100mm, 1.6μm; detection wavelength: 330nm; flow rate: 0.3ml / min; column temperature: 35℃; injection volume: 1μl; mobile phase: methanol as mobile phase A, 0.1wt% phosphoric acid as mobile phase B, and elution according to the gradient specified in Table 2.

[0059] Table 2

[0060]

[0061] 1.2 Methodological validation

[0062] 1.2.1 Positioning and system applicability

[0063] Take appropriate amounts of new chlorogenic acid, chlorogenic acid, and cryptochlorogenic acid reference substances, accurately weigh them, and add methanol to prepare a new chlorogenic acid reference solution containing 44.69 μg of new chlorogenic acid per 1 ml, a chlorogenic acid reference solution containing 49.38 μg of chlorogenic acid per 1 ml, and a cryptochlorogenic acid reference solution containing 53.94 μg of cryptochlorogenic acid per 1 ml.

[0064] Accurately weigh 0.5 g of Tianmingjing formula granule sample (batch number: TMJ-F-241025-01), place in a stoppered conical flask, accurately add 25 ml of 50 wt% methanol, weigh the weight, ultrasonically treat (power 250 W, frequency 50 kHz) for 15 minutes, cool, weigh again, make up the lost weight with 50 wt% methanol, shake well, filter, and take the filtrate as the test solution.

[0065] Accurately pipette 1 μl of each of the neochlorogenic acid reference solution, chlorogenic acid reference solution, cryptochlorogenic acid reference solution and test solution, inject into the high performance liquid chromatograph for determination, and record the chromatogram. The relevant chromatogram is shown in Figure 1-Figure 4 It can be seen that in the chromatogram of the new chlorogenic acid reference solution, the retention time of new chlorogenic acid is 2.443min (see Figure 1 Peak 1), the theoretical plate number is 8077, and the tailing factor is 1.25; in the chromatogram of the chlorogenic acid reference solution, the retention time of chlorogenic acid is 5.270min (see Figure 2 Peak 1), the theoretical plate number is 16706, and the tailing factor is 1.10; in the chromatogram of the chlorogenic acid reference solution, the retention time of chlorogenic acid is 5.737 min (see Figure 3 Peak 1), the theoretical plate number is 34133, and the tailing factor is 1.10. The retention time of neochlorogenic acid in the chromatogram of the test solution is 2.490min (see Figure 4 Peak 1), the theoretical plate number is 9119, the tailing factor is 1.17; the retention time of chlorogenic acid is 5.357min (see Figure 4 Peak 2), the theoretical plate number is 30807, the tailing factor is 1.13; the retention time of cryptochlorogenic acid is 5.817min (see Figure 4 Peak 4), the theoretical plate number is 35874, and the tailing factor is 1.10. It can be seen that the characteristic spectrum and the system applicability of the test method for the total content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid are good.

[0066] 1.2.2 Durability test

[0067] 1.2.2.1 Column temperature durability

[0068] According to the specified chromatographic conditions, change the column temperature by ±5°C, take the Tianmingjing Formula Granule sample (batch number: TMJ-F-241025-01), prepare the test solution according to the method under "1.1.2 Preparation of Test Solution", accurately pipette 1μl of the test solution, and inject it into the high performance liquid chromatograph for testing. Record the retention time of the common peak, take the peak corresponding to the chlorogenic acid reference peak as the S peak, calculate the relative retention time and RSD value of peak 3, peak 5, peak 6, peak 7, peak 8, peak 9, peak 10, peak 11, peak 12, peak 13, peak 14 and S peak, the results are shown in Table 3, and the relevant chromatograms are shown in Figure 5-Figure 7; The test results of the total contents of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid are shown in Table 4.

[0069] Table 3

[0070]

[0071] Table 4

[0072]

[0073] 1.2.2.2 Flow rate durability

[0074] According to the specified chromatographic conditions, change the flow rate by ±0.03ml / min, take the Tianmingjing formula granule sample (batch number: TMJ-F-241025-01), prepare the test solution according to the method under "1.1.2 Preparation of test solution", accurately pipette 1μl of the test solution, and inject it into the high performance liquid chromatograph for testing. Record the retention time of the common peak, take the peak corresponding to the chlorogenic acid reference peak as the S peak, calculate the relative retention time and RSD value of peak 3, peak 5, peak 6, peak 7, peak 8, peak 9, peak 10, peak 11, peak 12, peak 13, peak 14 and S peak, the results are shown in Table 5, and the relevant chromatograms are shown in Figure 8-Figure 9 ; The test results of the total contents of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid are shown in Table 6.

[0075] Table 5

[0076]

[0077] Table 6

[0078]

[0079] 1.2.2.3 Wavelength durability

[0080] According to the specified chromatographic conditions, change the wavelength ± 5nm, take the Tianmingjing formula granule sample (batch number: TMJ-F-241025-01), prepare the test solution according to the method under "1.1.2 Preparation of test solution", accurately pipette 1μl of the test solution, and inject it into the high performance liquid chromatograph for testing. Record the retention time of the common peak, take the peak corresponding to the chlorogenic acid reference peak as the S peak, calculate the relative retention time and RSD value of peak 3, peak 5, peak 6, peak 7, peak 8, peak 9, peak 10, peak 11, peak 12, peak 13, peak 14 and S peak, the results are shown in Table 7, and the relevant chromatograms are shown in Figure 10-11 ; The test results of the total contents of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid are shown in Table 8.

[0081] Table 7

[0082]

[0083] Table 8

[0084]

[0085] The results of the above durability tests showed that the durability of the column temperature, flow rate, and wavelength of this method met the requirements of the characteristic spectrum and the determination of the total content of neochlorogenic acid, chlorogenic acid, and cryptochlorogenic acid.

[0086] 1.3 Sample testing

[0087] Three batches of Tianmingjing formula granules (batch numbers: TMJ-K-241125-01, TMJ-K-241125-02, TMJ-K-241125-03) were tested according to "1.1 Test Method". The relative retention time of the characteristic spectrum is shown in Table 9; the test results of the total content of neochlorogenic acid, chlorogenic acid, and cryptochlorogenic acid are shown in Table 10.

[0088] Table 9

[0089]

[0090] Table 10

[0091]

[0092] 2-day test of sperm lactone content

[0093] 2.1. Test methods

[0094] 2.1.1 Preparation of reference solution: Take an appropriate amount of Tianmingjing lactone reference standard (purchased from Shanghai Shidande Standard Technology Service Co., Ltd., batch number: 18364, content 99.8%), accurately weigh it, and add methanol to make a solution containing 30μg per 1ml.

[0095] 2.1.2 Preparation of test solution: Grind 0.5 g of Tianmingjing formula granules into fine powder, weigh accurately, put it in a stoppered conical flask, add 25 ml of 50 wt% methanol solution accurately, weigh the weight, heat and reflux for 30 minutes, cool, weigh again, make up the lost weight with 50 wt% methanol, shake well, filter, and take the filtrate to obtain the solution.

[0096] 2.1.3 Determination: Accurately pipette 1 μl of the reference solution and the test solution, respectively, and inject them into the liquid chromatograph for determination. Calculate the content of tianmingjing lactone according to the obtained chromatogram. The chromatographic conditions are:

[0097] Chromatographic column: CORTECS UPLC T3, 2.1mm×100mm, 1.6μm; detection wavelength: 210nm; flow rate: 0.15ml / min; column temperature: 30℃; injection volume: 1μl; mobile phase: acetonitrile as mobile phase A, 0.1wt% phosphoric acid as mobile phase B, and elution according to the gradient specified in Table 11.

[0098] Table 11

[0099]

[0100] Methodological validation

[0101] 2.2.1 Positioning and system applicability

[0102] Take an appropriate amount of tianmingjing lactone reference substance, weigh it accurately, and add methanol to prepare a solution containing 36.29 μg of tianmingjing lactone per 1 ml, which is used as the reference substance solution.

[0103] Accurately weigh 0.5 g of Tianmingjing formula granule sample (batch number: TMJ-F-241025-01), place in a stoppered conical flask, accurately add 25 ml of 50 wt% methanol solution, weigh the weight, heat under reflux for 30 minutes, cool, weigh again, make up the lost weight with 50 wt% methanol, shake well, filter, and take the filtrate as the test solution.

[0104] Accurately pipette 1 μl of the reference solution and the test solution respectively, inject into the high performance liquid chromatograph for determination, and record the chromatogram. Figure 12-13 As can be seen from the figure, the retention time of tianmingjing lactone in the chromatogram of the reference solution is 8.610min, the theoretical plate number is 15456, and the tailing factor is 1.05; the retention time of tianmingjing lactone in the chromatogram of the test solution is 8.657min, the theoretical plate number is 17854, and the tailing factor is 1.05. Therefore, the system applicability of the tianmingjing lactone content test method is good.

[0105] 2.2.2 Durability test

[0106] 2.2.2.1 Column temperature durability

[0107] According to the specified chromatographic conditions, the column temperature was changed by ±5°C, and the Tianmingjing formula granule sample (batch number: TMJ-F-241025-01) was taken, and the test solution was prepared according to the method under "2.1.2 Preparation of test solution" to determine the content of Tianmingjing lactone. The results are shown in Table 12 and the relevant chromatograms are shown in Figure 14-16 (Peak 1 in each figure is asparagine lactone).

[0108] Table 12

[0109]

[0110] 2.2.2.2 Flow rate durability

[0111] According to the specified chromatographic conditions, the flow rate was changed by ±0.02 ml / min, and the Tianmingjing formula granule sample (batch number: TMJ-F-241025-01) was taken, and the test solution was prepared according to the method under "2.1.2 Preparation of test solution" to determine the content of Tianmingjing lactone. The results are shown in Table 13 and the relevant chromatograms are shown in Figure 17-Figure 18 (Peak 1 in each figure is asparagine lactone).

[0112] Table 13

[0113]

[0114] 2.2.2.3 Wavelength durability

[0115] According to the specified chromatographic conditions, the wavelength was changed by ±5nm, and the Tianmingjing formula granule sample (batch number: TMJ-F-241025-01) was taken. The test solution was prepared according to the method under "2.1.2 Preparation of test solution" and the content of Tianmingjing lactone was determined. The results are shown in Table 14 and the relevant chromatograms are shown in Figure 19-20 (Peak 1 in each figure is asparagine lactone).

[0116] Table 14

[0117]

[0118] The results of the above durability tests indicate that the durability of the column temperature, flow rate and wavelength of this method all meet the requirements for the determination of the content of tianmingjing lactone.

[0119] 2.3 Sample determination

[0120] Three batches of Tianmingjing formula granule samples (batch numbers: TMJ-K-241125-01, TMJ-K-241125-02, TMJ-K-241125-03) were taken and the content of Tianmingjing lactone was determined according to “2.1 Test Method”. The results are shown in Table 15.

[0121] Table 15

[0122]

[0123] The above is only a preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above embodiments. All technical solutions under the concept of the present invention belong to the protection scope of the present invention. It should be pointed out that for ordinary technicians in this technical field, some improvements and modifications without departing from the principle of the present invention should also be regarded as the protection scope of the present invention.

Claims

1. A method for testing the quality of Tianmingjing granules, characterized in that: Including characteristic spectra and total content test of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid and tianmingjing lactone content test; The characteristic spectrum and the total content test of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid include the following steps: a. Prepare neochlorogenic acid reference solution, chlorogenic acid reference solution and cryptochlorogenic acid reference solution; b. Take the Tianmingjing formula granule sample to prepare the test solution; c. Perform liquid chromatography on the neochlorogenic acid reference solution, chlorogenic acid reference solution, cryptochlorogenic acid reference solution and test solution, respectively, and calculate the total content of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid; The test of the content of tianmingjing lactone comprises the following steps: A. Prepare the reference solution of tianmingjing lactone; B. Take the Tianmingjing formula granule sample to prepare the test solution; C. Perform liquid chromatography on the tianmingjing lactone reference solution and the test solution respectively, and calculate the tianmingjing lactone content.

2. According to the method for testing the quality of Tianmingjing granules according to claim 1, it is characterized in that: The Tianmingjing decoction pieces are boiled in water for several times, filtered, and the filtrate is concentrated to a clear paste, and the clear paste is dried to form a dry paste; the auxiliary material maltodextrin is added and mixed to granulate, or granulate directly without adding auxiliary materials to obtain the Tianmingjing formula granules.

3. A method for detecting the quality of Tianmingjing granules according to claim 1, characterized in that: In the characteristic spectrum and the total content test of neochlorogenic acid, chlorogenic acid and cryptochlorogenic acid, the chromatographic conditions are: Chromatographic column: CORTECS UPLC T3, 2.1mm×100mm, 1.6μm; detection wavelength: 330nm; flow rate: 0.3ml / min; column temperature: 35℃; injection volume: 1μl; mobile phase: methanol as mobile phase A, 0.1wt% phosphoric acid as mobile phase B; gradient elution program: during 0-8min, mobile phase A increased from 15% to 30%, and mobile phase B decreased from 85% to 70%; during 8-18min, mobile phase A increased from 30% to 36%, and mobile phase B decreased from 70% to 64%; during 18-25min, mobile phase A increased from 36% to 50%, and mobile phase B decreased from 64% to 50%; during 25-35min, mobile phase A increased from 50% to 70%, and mobile phase B decreased from 50% to 30%.

4. A method for detecting the quality of Tianmingjing granules according to claim 1, characterized in that: In step a, the neochlorogenic acid reference solution, the chlorogenic acid reference solution and the cryptochlorogenic acid reference solution were all prepared using methanol as solvent, and the concentration of each reference solution was 30 μg / ml.

5. A method for detecting the quality of Tianmingjing granules according to claim 1, characterized in that: Step b is specifically as follows: take 0.5 g of Tianmingjing formula granule sample powder, add 25 ml of 50 wt% methanol, ultrasonically treat, cool, weigh again, make up the lost weight with 50 wt% methanol, shake well, filter, and take the filtrate to obtain.

6. A method for detecting the quality of Tianmingjing granules according to claim 1, characterized in that: In the test of the content of tianmingjing lactone, the chromatographic conditions are: Chromatographic column: CORTECS UPLC T3, 2.1mm×100mm, 1.6μm; detection wavelength: 210nm; flow rate: 0.15ml / min; column temperature: 30℃; injection volume: 1μl; mobile phase: acetonitrile as mobile phase A, 0.1wt% phosphoric acid as mobile phase B; gradient elution program: during 0-5min, mobile phase A was maintained at 35%, and mobile phase B was maintained at 65%; during 5-7.5min, mobile phase A increased from 35% to 40%, and mobile phase B decreased from 65% to 60%; during 7.5-8.8min, mobile phase A increased from 40% to 95%, and mobile phase B decreased from 60% to 5%; during 8.8-10min, mobile phase A was maintained at 95%, and mobile phase B was maintained at 5%; during 10-12min, mobile phase A decreased from 95% to 35%, and mobile phase B increased from 5% to 65%.

7. A method for detecting the quality of Tianmingjing granules according to claim 1, characterized in that: In step A, the tianmingjing lactone reference solution is prepared using methanol as a solvent, and the concentration of the tianmingjing lactone reference solution is 30 μg / ml.

8. A method for testing the quality of Tianmingjing granules according to claim 1, characterized in that: Step B is specifically as follows: take 0.5 g of Tianmingjing formula granule powder, add 25 ml of 50 wt% methanol solution, heat to reflux, cool, weigh again, make up the lost weight with 50 wt% methanol, shake well, filter, and take the filtrate to obtain.

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