A quality detection method of Jingling oral liquid

Abstract

The application belongs to the technical field of traditional Chinese medicine detection, and discloses a quality detection method of Jingling oral liquid. The quality detection method is detected by high performance liquid chromatography, and the chromatographic conditions are as follows: ZORBAX Eclipse Plus C18 is used as the filler of the chromatographic column; acetonitrile is used as the mobile phase A, and 0.04-0.06 mol / L ammonium acetate aqueous solution is used as the mobile phase B to perform gradient elution. The detection method can detect 22 common chromatographic peaks in the Jingling oral liquid, identify 6 chemical components, and attribute to 5 medicinal materials. The method has the characteristics of simple operation, accuracy and stability, can better control and manage the product quality of the Jingling oral liquid, and provides a more comprehensive scientific basis for the quality evaluation of the Jingling oral liquid.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine testing technology, specifically relating to a quality testing method for Jingling oral liquid. Background Technology

[0002] Jingling Oral Liquid is a traditional Chinese medicine that nourishes Yin and calms Yang, and is primarily used to treat ADHD in children. It alleviates clinical symptoms such as inattention, hyperactivity, talkativeness, impulsivity, learning difficulties, red tongue, and thready, rapid pulse, which are indicative of kidney Yin deficiency and liver Yang excess. With over 30 years on the market, clinical results have shown that this medicine is highly safe and effective, and it has become a core traditional Chinese medicine product for treating ADHD in children.

[0003] It is known that the raw materials of Jingling Oral Liquid include Rehmannia glutinosa, Dioscorea opposita, Poria cocos, Paeonia suffruticosa, Alisma plantago-aquatica, Polygala tenuifolia, calcined dragon bone, Ligustrum lucidum, Phellodendron chinense, Anemarrhena asphodeloides, Schisandra chinensis, and Acorus tatarinowii. As a compound traditional Chinese medicine, its chemical composition is relatively complex and difficult to separate and detect. At present, the quality standards of Jingling Oral Liquid are low, and Phellodendron chinense or berberine hydrochloride are often used as quality control indicators, which are difficult to comprehensively and effectively reflect the actual quality of the drug.

[0004] Furthermore, although some literature indicates that ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS / MS) can detect multiple components in Jingling oral liquid, this method has higher equipment requirements and higher detection costs, making it difficult to widely apply in pharmaceutical manufacturing companies. Therefore, this invention aims to provide a method for detecting more components in Jingling oral liquid based on conventional high performance liquid chromatography, thereby better controlling drug quality. Summary of the Invention

[0005] The present invention aims to at least solve one of the technical problems existing in the prior art. To this end, the present invention proposes a quality testing method for Jingling oral liquid. This method can detect 22 common chromatographic peaks in Jingling oral liquid, identify 6 chemical components, and attribute them to 5 medicinal herbs. It is characterized by its simple operation, accuracy, and stability, enabling better control and management of the product quality of Jingling oral liquid, and providing a more comprehensive scientific basis for the quality evaluation of Jingling oral liquid.

[0006] This invention provides a quality detection method for Jingling oral liquid, which uses high performance liquid chromatography (HPLC) for detection. The chromatographic conditions used are as follows:

[0007] ZORBAX Eclipse Plus C18 was used as the packing material for the chromatographic column;

[0008] Gradient elution was performed using acetonitrile as mobile phase A and 0.04-0.06 mol / L ammonium acetate aqueous solution as mobile phase B.

[0009] The conditions for gradient elution are as follows:

[0010] 0-50 min, mobile phase A 5%→28%, mobile phase B 95%→72%;

[0011] 50-65 min, mobile phase A 28%, mobile phase B 72%;

[0012] 65-80 min, mobile phase A 28%→40%, mobile phase B 72%→60%;

[0013] 80-83 min, mobile phase A 40%→70%, mobile phase B 60%→40%;

[0014] 83-85 min, mobile phase A 70%→20%, mobile phase B 40%→80%;

[0015] 85-90 min, mobile phase A 20% → 5%, mobile phase B 80% → 95%.

[0016] In the gradient elution described above, the percentage (%) in the mobile phase represents a volume percentage.

[0017] In determining the mobile phase for high-performance liquid chromatography (HPLC), this invention investigated various mobile phases, including methanol-water, acetonitrile-water, acetonitrile-phosphate aqueous solution, acetonitrile-formic acid aqueous solution, acetonitrile-potassium dihydrogen phosphate aqueous solution, and acetonitrile-ammonium acetate aqueous solution. The results showed that the system achieved significantly better chromatographic separation when using acetonitrile-ammonium acetate aqueous solution as the mobile phase. Because the chemical components of Jingling oral liquid are complex and exhibit significant polarity differences, this invention employs gradient elution to ensure the effective separation of more chemical components. This facilitates the qualitative identification of the main medicinal materials and components, thereby better controlling the quality of drug production.

[0018] The above-mentioned quality testing method was used to test Jingling oral liquid. According to the obtained fingerprint spectrum, there were 22 common peaks, which could be attributed to five medicinal herbs: Phellodendron bark, Polygala root, Ligustrum lucidum, Paeonia suffruticosa root bark, and Schisandra fruit. Six chemical components, namely berberine hydrochloride, berberine hydrochloride, schisandra methanol, paeoniflorin, paeonol, and ligustrol, could be identified. This allows for better control and management of the product quality of Jingling oral liquid.

[0019] Preferably, the quality inspection method includes the following steps:

[0020] Weigh out berberine hydrochloride, schisandra methanol, paeoniflorin, paeonol, and ligustrum lucidum glycoside, respectively, and dissolve them in solvent to prepare solutions of each reference standard.

[0021] Weigh out Jingling oral liquid, dissolve it in a solvent, and prepare the test solution;

[0022] Based on the 12 medicinal materials of Jingling Oral Liquid, a single herb was removed, and the remaining herbs were mixed in equal mass, and water was added for extraction to obtain a negative sample lacking a single herb; the negative sample lacking a single herb was taken, evaporated to dryness, and dissolved in a solvent to obtain a negative sample solution lacking a single herb.

[0023] The reference solution, the test solution, and the negative sample solution lacking a single herb were injected into a high-performance liquid chromatograph for detection, and chromatograms of the reference solution, the test solution, and the negative sample solution lacking a single herb were obtained. The chromatograms were then analyzed.

[0024] In the preparation of the reference solution, the test solution, and the negative sample solution lacking a single herb, the solvent is methanol or a methanol solution.

[0025] Experiments show that samples obtained by evaporation followed by dissolution with methanol solution have a larger response value.

[0026] The concentration of the reference solution is 30-120 μg / mL.

[0027] The single herb mentioned is selected from any one of the following: Rehmannia glutinosa (processed), Dioscorea opposita, Poria cocos, Paeonia suffruticosa, Alisma plantago-aquatica, Polygala tenuifolia, calcined dragon bone, Ligustrum lucidum, Phellodendron chinense, Anemarrhena asphodeloides, Schisandra chinensis, and Acorus tatarinowii. For example, when Phellodendron chinense is selected as the single herb, a corresponding negative sample solution lacking Phellodendron chinense is prepared.

[0028] Preferably, the process of analyzing the chromatogram includes: performing drug component attribution analysis based on the chromatograms of the test sample and the negative sample lacking a single herb; identifying the drug components based on the chromatograms of the reference standard and the test sample; or establishing a liquid phase fingerprint of Jingling oral liquid based on the chromatogram of the test sample.

[0029] Preferably, the quality detection method employs multi-wavelength detection. Because the active ingredients in Jingling oral liquid are complex, and the maximum absorption wavelengths of each ingredient differ significantly, multi-wavelength detection is employed in this invention to consider detection sensitivity.

[0030] More preferably, the wavelengths of the multi-wavelength detection include 220nm, 237nm, 293nm and 320nm.

[0031] Specifically, the following wavelength switching methods can be used for detection:

[0032] 0-21.25 min, wavelength 320 nm;

[0033] 21.25-21.5 min, wavelength from 320 to 293 nm;

[0034] 21.5-25.5 min, wavelength 293 nm;

[0035] 25.5-26 min, wavelength 293→220 nm;

[0036] 26-48 minutes, wavelength 220nm;

[0037] 48-49 min, wavelength 220→320 nm;

[0038] 49-70 minutes, wavelength 320nm;

[0039] 70-71 min, wavelength from 320 to 237 nm;

[0040] 71-90 min, wavelength 237 nm.

[0041] Preferably, the chromatographic column has a size of 250×4.6mm and the packing material has a particle size of 5μm.

[0042] Preferably, the chromatographic conditions further include an injection volume of 5-15 μL. More preferably, the injection volume is 10 μL.

[0043] Preferably, the chromatographic conditions further include a flow rate of 0.7-0.9 mL / min.

[0044] Preferably, the chromatographic conditions further include a column temperature of 28-32°C.

[0045] Preferably, the chromatographic conditions further include: the detector is a diode array detector (DAD detector).

[0046] Since ligustrol is a relatively abundant and stable component in Jingling oral liquid, it suffers minimal loss during extraction, exhibits moderate retention time, high response value, and good separation in liquid chromatography, and can serve as an indicator component for content determination in Jingling oral liquid, ligustrol is selected as the reference peak in the quality detection method of this invention. The theoretical plate number, calculated based on the reference ligustrol, should not be less than 5000.

[0047] Using the quality detection method provided by this invention, the liquid chromatographic fingerprint of Jingling oral liquid showed 22 common chromatographic peaks. Peak 12, with the same retention time as the reference peak (specifically, ligustrazine), was designated as the reference peak. Compared to the retention time of the reference peak (calculated as 1), the relative retention times of each chromatographic peak were as follows: Peak 1 0.133±10%, Peak 2 0.190±10%, Peak 3 0.227±10%, Peak 4 0.268±10%, Peak 5 0.340±10%, Peak 6 0.373±10%, Peak 7 0.447±10%, Peak 8 0.518±10%, Peak 9 (berberine hydrochloride) 0.628±10%, Peak 10 (paeoniflorin) 0.737±10%, and Peak 11 0.762±10%. Peak 12 (Ligustrum lucidum glycoside) 1.000±10%, Peak 13 1.209±10%, Peak 14 1.339±10%, Peak 15 1.363±10%, Peak 16 1.408±10%, Peak 17 (Berberine hydrochloride) 1.428±10%, Peak 18 1.542±10%, Peak 19 1.583±10%, Peak 20 1.631±10%, Peak 21 (Paeonol) 2.174±10%, Peak 22 (Schisandrol A) 2.247±10%.

[0048] In the liquid phase fingerprint of Jingling oral liquid: peaks 1 and 22 are derived from Schisandra chinensis; peaks 11 and 12 are derived from Ligustrum lucidum; peaks 10 and 21 are derived from Paeonia suffruticosa; peaks 13, 14, 15, 16, 18, 19, and 20 are derived from Polygala tenuifolia; and peaks 2, 3, 4, 5, 6, 7, 8, 9, and 17 are derived from Phellodendron chinense.

[0049] This invention uses the Chinese medicine chromatographic fingerprint similarity evaluation system provided by the National Pharmacopoeia Commission as the fingerprint similarity calculation software. After multiple experiments and studies, and by comparing it with the method of calculating relative retention time and relative peak area, the evaluation conclusions obtained are consistent and there is no significant difference, indicating that the quality detection method of Jingling oral liquid is effective, stable and reliable.

[0050] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0051] (1) The fingerprint spectrum of Jingling oral liquid established by the present invention by high performance liquid chromatography identified a total of 22 common chromatographic peaks and identified 6 index chemical components, namely berberine hydrochloride, berberine hydrochloride, schisandra methanol, paeoniflorin, paeonol, and privetin; and can be attributed to 5 medicinal materials, namely Phellodendron bark, Polygala tenuifolia, Ligustrum lucidum, Paeonia suffruticosa, and Schisandra chinensis, thus realizing the qualitative identification of the above-mentioned medicinal materials.

[0052] (2) Through precision tests, stability tests, and repeatability tests, the quality detection method of this invention has been confirmed to have good detection effect, high reproducibility, and good reliability, and can be used for qualitative quality control of Jingling oral liquid. Furthermore, the quality detection method of this invention was used to test multiple batches of samples, and 22 common peaks were detected in all samples. Six chemical components were identified and attributed to five medicinal herbs, and all test results were qualified. This further demonstrates that the quality detection method is simple to operate, accurate, and stable, enabling better control and management of the product quality of Jingling oral liquid, and providing a more comprehensive scientific basis for the quality evaluation of Jingling oral liquid. Attached Figure Description

[0053] Figure 1 The chromatogram (293 nm) shows a comparison of the test solution, the negative sample solution lacking Phellodendron bark, and the reference solution of Phellodendron hydrochloride in Example 1.

[0054] Figure 2 The chromatogram (320 nm) shows the test solution in Example 1 compared with the negative sample solution lacking Phellodendron bark and the berberine hydrochloride reference solution.

[0055] Figure 3 The chromatogram (219 nm) shows a comparison of the test solution, the negative sample solution lacking peony bark, and the paeoniflorin reference solution in Example 1.

[0056] Figure 4 The chromatogram (237 nm) shows the comparison of the test solution, the negative sample solution lacking peony bark, and the paeonol reference solution in Example 1.

[0057] Figure 5 The chromatogram (342 nm) shows the comparison between the test solution in Example 1 and the negative sample solution lacking privet fruit, and the purslane.

[0058] Figure 6 The chromatograms (237 nm) of the test sample solution in Example 1 are compared with those of the negative sample solution lacking Schisandra chinensis and the methanol chromatogram of Schisandra chinensis.

[0059] Figure 7 The chromatogram (342 nm) shows the comparison between the test solution and the negative sample solution lacking Polygala tenuifolia in Example 1, and the polygala tenuifolia sago ketone III.

[0060] Figure 8 The chromatogram (342 nm) shows the comparison between the test solution and the negative sample solution of Acorus tatarinowii in Example 1.

[0061] Figure 9 The chromatogram (342 nm) shows a comparison between the test solution and the negative sample solution lacking Rehmannia glutinosa in Example 1.

[0062] Figure 10The chromatogram (275 nm) shows the comparison between the test solution and the negative sample solution lacking Anemarrhena asphodeloides in Example 1, and Anemarrhena saponin BⅡ.

[0063] Figure 11 The chromatogram (342nm) shows a comparison of the test solution in Example 1 with the negative sample solution lacking yam and the negative sample solution lacking poria.

[0064] Figure 12 The chromatogram (342 nm) shows a comparison between the test solution and the negative sample solution lacking Alisma plantago-aquatica in Example 1.

[0065] Figure 13 This is the chromatogram of the negative sample lacking a single herb in Example 1;

[0066] Figure 14 This is a chromatogram of the identified components of the reference standard in Example 1. Detailed Implementation

[0067] To enable those skilled in the art to more clearly understand the technical solutions described in this invention, the following embodiments are provided for illustration. It should be noted that the following embodiments do not constitute a limitation on the scope of protection claimed by this invention.

[0068] The instruments, reagents, and reference standards used in the following examples, as well as the specific operating procedures and protocols, are shown below. Unless otherwise specified, all other raw materials, reagents, or devices can be obtained from conventional commercial sources or through existing known methods.

[0069] 1. Instruments and equipment:

[0070] High-performance liquid chromatograph (HPLC), digitally controlled ultrasonic cleaner, constant temperature and humidity chamber, chromatographic column, electronic balance (0.0001 g), electric thermostatic water bath, etc.

[0071] 2. Reagents and reference standards:

[0072] Reagents: Acetonitrile, methanol, formic acid, phosphoric acid, potassium dihydrogen phosphate, ammonium acetate, ultrapure water, etc.

[0073] Reference standards: rehmannia glycoside D, rhodioloside, berberine hydrochloride, paeoniflorin, ligustrum lucidum glycoside, berberine hydrochloride, and paeonol, etc.

[0074] 3. Specific operating procedures and plans:

[0075] The determination was performed according to the high performance liquid chromatography method (General Chapter 0512, Part IV, Chinese Pharmacopoeia 2020 Edition).

[0076] Example 1

[0077] This embodiment provides a quality testing method for Jingling oral liquid, including the following steps:

[0078] Preparation of the test solution: Take 10 mL of Jingling oral liquid, evaporate it to dryness in an evaporating dish, dissolve it in 5 mL of 70% methanol solution, take the supernatant, filter it through a 0.22 μm organic microporous membrane, and take the filtrate.

[0079] Preparation of reference solutions: Take berberine hydrochloride, schisandra syrup, paeoniflorin, paeonol, and ligustrum lucidum glycoside, respectively, and dissolve them in methanol to obtain the corresponding reference solutions.

[0080] Preparation of negative sample solution for single herb deficiency: (1) Based on the 12 herbs in Jingling oral liquid, remove the single herb, mix the remaining herbs in equal mass, add water equal to 8 times the mass of the total herbs, boil and extract for 3.5 hours to obtain negative sample for single herb deficiency; (2) Take the negative sample for single herb deficiency, evaporate to dryness, add methanol to dissolve, filter with a 0.22μm organic microporous membrane, take the filtrate, and thus obtain the corresponding negative sample solution for single herb deficiency.

[0081] The following chromatographic conditions were used for detection:

[0082] The chromatographic column used was a ZORBAX Eclipse Plus C18 (250×4.6mm, 5μm).

[0083] Acetonitrile was used as mobile phase A, and 0.05 mol / L ammonium acetate aqueous solution was used as mobile phase B. Gradient elution was performed under the conditions shown in Table 1.

[0084] Table 1

[0085] Time (min) Liquidity A (%) Liquidity B (%) -15 to 0 5 95 0-50 5→28 95→72 50-65 28 72 65-80 28→40 72→60 80-83 40→70 60→30 83-85 70→20 30→80 85-90 20→5 80→95

[0086] Injection volume: 10 μL; Column temperature: 30℃; Flow rate: 0.8 mL / min; Detector: DAD detector. The time from -15 to 0 min is the column equilibration time.

[0087] Based on the above chromatographic conditions, the components were first screened using multiple wavelengths, and the peak localization of the fingerprint spectrum of Jingling oral liquid was studied.

[0088] Accurately pipette 10 μL each of the test solution, reference solution, and negative sample solution lacking a single herb, inject them into the liquid chromatograph, perform qualitative analysis, and record the chromatograms.

[0089] The components were screened using multiple wavelengths, and the results were obtained from... Figure 1-12 As shown, the chromatograms of the test sample, the reference sample, and the negative sample lacking a single herb are classified as the following herbs: Phellodendron bark, Polygala root, Ligustrum lucidum, Paeonia suffruticosa root bark, and Schisandra fruit. The chemical components identified are berberine hydrochloride, berberine hydrochloride, schisandra methanol, paeoniflorin, paeonol, and ligustrum lucidum glycoside.

[0090] Based on the results of the peak localization study above, this embodiment sets the detection wavelengths to 220nm, 320nm, 237nm, and 293nm, and simultaneously enables full-wavelength scanning as the final confirmed wavelength detection method. The wavelength switching methods are shown in Table 2.

[0091] Table 2

[0092]

[0093]

[0094] Accurately pipette 10 μL each of the test solution, reference solution, and negative sample solution lacking a single herb, and inject them into the liquid chromatograph. Perform qualitative analysis using the above-confirmed detection method and record the chromatograms.

[0095] according to Figure 13-14 The liquid chromatogram of Jingling Oral Solution shows 22 common chromatographic peaks. Peak 12 has the same retention time as the reference peak (specifically, ligustrazine) and is designated as the reference peak. Compared with the retention time of the reference peak (calculated as 1), the relative retention times of each peak are as follows: Peak 1 0.133±10%, Peak 2 0.190±10%, Peak 3 0.227±10%, Peak 4 0.268±10%, Peak 5 0.340±10%, Peak 6 0.373±10%, Peak 7 0.447±10%, Peak 8 0.518±10%, Peak 9 (berberine hydrochloride) 0.628±10%, Peak 10 (paeoniflorin) 0.737±10%, Peak 11 0.762±10%. Peak 12 (Ligustrum lucidum glycoside) 1.000±10%, Peak 13 1.209±10%, Peak 14 1.339±10%, Peak 15 1.363±10%, Peak 16 1.408±10%, Peak 17 (Berberine hydrochloride) 1.428±10%, Peak 18 1.542±10%, Peak 19 1.583±10%, Peak 20 1.631±10%, Peak 21 (Paeonol) 2.174±10%, Peak 22 (Schisandrol A) 2.247±10%.

[0096] The classification and chemical composition of each medicinal ingredient are shown in Table 3.

[0097] Table 3

[0098]

[0099]

[0100] Table 3 shows that peaks 1 and 22 are derived from Schisandra chinensis; peaks 11 and 12 from Ligustrum lucidum; peaks 10 and 21 from Paeonia suffruticosa; peaks 13, 14, 15, 16, 18, 19, and 20 from Polygala tenuifolia; and peaks 2, 3, 4, 5, 6, 7, 8, 9, and 17 from Phellodendron chinense. The medicinal herbs associated with these peaks are Polygala tenuifolia, Phellodendron chinense, Ligustrum lucidum, Paeonia suffruticosa, and Schisandra chinensis. The identified chemical components are berberine hydrochloride, berberine hydrochloride, paeonol, paeoniflorin, schisandrin A, and ligustrazine.

[0101] Example 2

[0102] Precision test: Take Jingling oral liquid product and operate according to the preparation and quality detection method of the test solution in Example 1. Inject the sample 6 times consecutively, calculate the relative retention time and similarity, and the specific test results are shown in Table 4-5.

[0103] Table 4. Calculation Results of Relative Retention Time for Fingerprint Precision

[0104]

[0105]

[0106] Table 5. Precision Examination Similarity Evaluation Results

[0107] Similarity R S2 S3 S4 S5 S6 S7 R 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S2 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S3 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S4 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S5 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S6 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S7 1.000 1.000 1.000 1.000 1.000 1.000 1.000

[0108] Wherein, R: reference fingerprint spectrum, S2: precision 1, S3: precision 2, S4: precision 3, S5: precision 4, S6: precision 5, S7: precision 6.

[0109] Table 4-5 shows that the fingerprint chromatogram of the test sample exhibits chromatographic peaks with the same retention times as the reference chromatographic peaks, and presents 22 major chromatographic peaks. The relative retention time RSD is less than 2.0%. Comparison with the generated control fingerprint chromatogram shows a similarity greater than 0.90, indicating good instrument precision.

[0110] Example 3

[0111] Stability test: Take Jingling oral liquid product and operate according to the preparation and quality testing method of the test solution in Example 1. Samples were injected and measured after 0, 3, 6, 9, 12 and 24 hours after preparation. The relative retention time and similarity were calculated. The specific test results are shown in Table 6-7.

[0112] Table 6. Calculation results of relative retention time for fingerprint spectrum stability.

[0113]

[0114]

[0115] Table 7. Similarity Evaluation Results of Stability Assessment

[0116] Similarity R S2 S3 S4 S5 S6 S7 R 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S2 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S3 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S4 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S5 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S6 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S7 1.000 1.000 1.000 1.000 1.000 1.000 1.000

[0117] Wherein, R: reference fingerprint spectrum, S2: stability 0h, S3: stability 3h, S4: stability 6h, S5: stability 9h, S6: stability 12h, S7: stability 24h.

[0118] Table 6-7 shows that the fingerprint chromatogram of the test sample exhibits chromatographic peaks with the same retention times as the reference chromatographic peaks, and presents 22 major chromatographic peaks. Its relative retention time RSD is less than 2.0%. Compared with the generated control fingerprint chromatogram, the similarity is greater than 0.90, indicating that the test sample solution has good stability within 24 hours.

[0119] Example 4

[0120] Repeatability test: Take 6 portions of Jingling oral liquid product, 10 mL each, and operate according to the preparation and quality testing method of the test solution in Example 1. Calculate the relative retention time and similarity. The specific test results are shown in Table 8-9.

[0121] Table 8. Calculation results of relative retention time for fingerprint repeatability.

[0122]

[0123] Table 9. Results of Similarity Evaluation for Repeatability Tests

[0124] Similarity R S2 S3 S4 S5 S6 S7 R 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S2 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S3 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S4 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S5 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S6 1.000 1.000 1.000 1.000 1.000 1.000 1.000 S7 1.000 1.000 1.000 1.000 1.000 1.000 1.000

[0125] Wherein, R: reference fingerprint spectrum, S2: repeatability 1, S3: repeatability 2, S4: repeatability 3, S5: repeatability 4, S6: repeatability 5, S7: repeatability 6.

[0126] Table 8-9 shows that the fingerprint chromatogram of the test sample exhibits chromatographic peaks with the same retention times as the reference chromatographic peaks, and presents 22 major chromatographic peaks. The relative retention time (RSD) is less than 2.0%. Compared with the generated control fingerprint chromatogram, the similarity is greater than 0.90, indicating that the repeatability of this quality detection method is good.

[0127] Example 5

[0128] Durability test: Based on the quality test method in Example 1, this example further investigates the durability of different flow rates (0.7 ml / min, 0.8 ml / min, 0.9 ml / min) and different column temperatures (28℃, 30℃, 32℃) on chromatographic conditions.

[0129] The results showed that the measurements were basically consistent under different flow rates and column temperatures. The common peaks in the chromatograms were sharp, symmetrical, and well-resolved. This indicates that the method is robust under different flow rates and column temperatures.

[0130] Example 6

[0131] Multiple batches of samples were tested: Jingling Oral Liquid is an exclusive product of Liaoning Dongfangren Pharmaceutical Co., Ltd. (National Drug Approval Number Z10910056). In this experiment, 10 different batches of Jingling Oral Liquid samples were tested using the quality detection method in Example 1. The results showed that 22 common peaks could be detected in all 10 batches of samples, and 6 chemical components (berberine hydrochloride, schisandra methanol, paeoniflorin, paeonol, and privetin) could be identified and attributed to 5 medicinal materials (Phellodendron bark, Polygala root, Ligustrum lucidum, Paeonia suffruticosa root bark, and Schisandra chinensis). The test results were all qualified, further indicating that the quality detection method has a good identification effect and can be used for the qualitative detection of Jingling Oral Liquid.

[0132] Comparative Example 1

[0133] The study investigated the detection results under different mobile phase conditions: Preliminary exploratory experiments showed that using mobile phase systems such as methanol-water, acetonitrile-water, acetonitrile-phosphoric acid aqueous solution, acetonitrile-formic acid aqueous solution, and acetonitrile-potassium dihydrogen phosphate aqueous solution all resulted in low peak response values, low resolution, and fewer identified chemical components or medicinal flavors. After continuous screening and optimization of chromatographic conditions, acetonitrile-ammonium acetate aqueous solution was finally selected as the mobile phase system. This mobile phase system was found to have excellent performance in the detection of Jingling oral liquid and could meet the actual detection requirements.

[0134] The embodiments of this application have been described in detail above with reference to the accompanying drawings. However, this application is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of this application.

Claims

1. A quality testing method for Jingling oral liquid, characterized in that, Includes the following steps: Weigh out berberine hydrochloride, schisandra syrup, paeoniflorin, paeonol, and ligustrum lucidum glycoside, respectively, and dissolve them in methanol or methanol solution to prepare solutions of each reference standard. Weigh out Jingling oral liquid and dissolve it in methanol or a methanol solution to prepare the test solution; Fingerprint chromatograms were detected using high-performance liquid chromatography (HPLC), and the chromatographic conditions used were as follows: ZORBAX Eclipse Plus C18 was used as the packing material for the chromatographic column; Gradient elution was performed using acetonitrile as mobile phase A and 0.04-0.06 mol / L ammonium acetate aqueous solution as mobile phase B. The conditions for gradient elution are as follows: From 0 to 50 min, mobile phase A increased from 5% to 28%, and mobile phase B increased from 95% to 72%. 50-65 min, mobile phase A 28%, mobile phase B 72%; 65-80 min, mobile phase A 28%→40%, mobile phase B 72%→60%; 80-83 min, mobile phase A 40%→70%, mobile phase B 60%→40%; 83-85 min, mobile phase A 70%→20%, mobile phase B 40%→80%; 85-90 min, mobile phase A 20%→5%, mobile phase B 80%→95%; The following wavelength switching method is used for detection: 0-21.25 min, wavelength 320 nm; 21.25-21.5 min, wavelength from 320 to 293 nm; 21.5-25.5 min, wavelength 293 nm; 25.5-26 min, wavelength 293→220 nm; 26-48 minutes, wavelength 220nm; 48-49 min, wavelength 220→320 nm; 49-70 minutes, wavelength 320nm; 70-71 min, wavelength from 320 to 237 nm; 71-90 min, wavelength 237 nm.

2. The quality inspection method according to claim 1, characterized in that, It also includes the following steps: Based on the 12 medicinal materials of Jingling Oral Liquid, a single herb was removed, and the remaining herbs were mixed in equal mass, and water was added for extraction to obtain a negative sample lacking a single herb; the negative sample lacking a single herb was taken, evaporated to dryness, and dissolved in a solvent to obtain a negative sample solution lacking a single herb. The reference solution, the test solution, and the negative sample solution lacking a single herb were injected into a high-performance liquid chromatograph for detection, and chromatograms of the reference solution, the test solution, and the negative sample solution lacking a single herb were obtained. The chromatograms were then analyzed.

3. The quality inspection method according to claim 2, characterized in that, The process of analyzing the chromatograms includes: performing drug component attribution analysis based on the chromatograms of the test sample and negative samples lacking a single herb; identifying drug components based on the chromatograms of the reference standard and the test sample; or establishing a liquid phase fingerprint of Jingling oral liquid based on the chromatogram of the test sample.

4. The quality inspection method according to claim 2, characterized in that, In preparing the negative sample solution for the absence of a single herb, the solvent is methanol or a methanol solution.

5. The quality inspection method according to any one of claims 1-3, characterized in that, The chromatographic column has a specification of 250×4.6mm and the packing material has a particle size of 5μm.

6. The quality inspection method according to any one of claims 1-3, characterized in that, The chromatographic conditions also include: injection volume of 5-15 μL.

7. The quality inspection method according to any one of claims 1-3, characterized in that, The chromatographic conditions also include a flow rate of 0.7-0.9 mL / min.

8. The quality inspection method according to any one of claims 1-3, characterized in that, The chromatographic conditions also include: column temperature 28-32℃.

9. The quality inspection method according to any one of claims 1-3, characterized in that, The chromatographic conditions also include: the detector is a diode array detector.

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