Quality detection method of Xinling oral liquid

Abstract

The invention belongs to the technical field of traditional Chinese medicine detection, and discloses a quality detection method of a Jinling oral liquid. According to the quality detection method, high performance liquid chromatography is adopted for detection, and the used chromatographic conditions are as follows: ZORBAX Eclipse Plus C18 is adopted as a filler of a chromatographic column; acetonitrile is used as a mobile phase A, a 0.04-0.06 mol / L ammonium acetate aqueous solution is used as a mobile phase B, and gradient elution is carried out. The detection method can be used for detecting 22 common chromatographic peaks in the Xianling oral liquid, identifying 6 chemical components and also belonging to 5 medicinal materials, has the characteristics of simplicity in operation, accuracy and stability, can be used for better controlling and managing the product quality of the Xianling oral liquid, and has a wide application prospect. And a more comprehensive scientific basis is provided for quality evaluation of the Xinling oral liquid.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine detection, and particularly relates to a quality detection method for Jingling oral liquid. Background Art

[0002] Jingling Oral Liquid is a Chinese patent medicine that nourishes Yin and suppresses Yang, calms the mind and improves intelligence. It is mainly used to treat ADHD in children, relieving clinical symptoms of kidney yin deficiency and liver yang excess, such as inattention, hyperactivity, hyperphagia, impulsiveness, learning difficulties, red tongue, and thin and rapid pulse. Over the past 30 years since it was launched on the market, clinical results have shown that the drug is highly safe and has significant efficacy, and has become a core Chinese patent medicine product for the treatment of ADHD in children.

[0003] It is known that the raw materials of Jingling oral liquid include Rehmannia root, yam, Poria cocos, peony bark, Alisma orientalis, Polygala tenuifolia, calcined dragon bone, Ligustrum lucidum, Phellodendron chinense, Anemarrhena asphodeloides, Schisandra chinensis, and Acorus calamus. As a compound Chinese patent medicine, its chemical composition is relatively complex and difficult to separate and detect. Currently, the quality standard of Jingling oral liquid is low, and the medicinal taste of Phellodendron chinense or berberine hydrochloride is often used as quality control indicators, which can hardly fully and effectively reflect the actual quality of the medicine.

[0004] In addition, although there are also documents that ultra-high performance liquid chromatography-mass spectrometry can be used to detect multiple components in Jingling oral liquid, this method has higher requirements for equipment and higher detection costs, and is difficult to be widely used in pharmaceutical manufacturers. Therefore, the present invention hopes to provide a method that can detect more components in Jingling oral liquid based on conventional high performance liquid chromatography to better control the quality of the drug. Summary of the invention

[0005] The present invention aims to solve at least one of the technical problems existing in the above-mentioned prior art. To this end, the present invention proposes a quality detection method for Jingling oral liquid, which can detect 22 common chromatographic peaks in Jingling oral liquid, identify 6 chemical components therein, and can also be attributed to 5 medicinal materials. It has the characteristics of simple operation, accuracy and stability, can better control and manage the product quality of Jingling oral liquid, and also provide a more comprehensive scientific basis for the quality evaluation of Jingling oral liquid.

[0006] The present invention provides a quality detection method for Jingling oral liquid, which adopts high performance liquid chromatography for detection, and the chromatographic conditions used are:

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

[0008] Acetonitrile was used as mobile phase A, and 0.04-0.06 mol / L ammonium acetate aqueous solution was used as mobile phase B for gradient elution;

[0009] The conditions of the gradient elution are:

[0010] 0-50min, 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] 80-83min, mobile phase A 40%→70%, mobile phase B 60%→40%;

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

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

[0016] In the above-mentioned gradient elution, the % shown in the mobile phase represents the volume percentage.

[0017] In the process of determining the mobile phase of high performance liquid chromatography, the present invention investigates a variety of mobile phases such as methanol water, acetonitrile water, acetonitrile phosphoric acid aqueous solution, acetonitrile-formic acid aqueous solution, acetonitrile-potassium dihydrogen phosphate aqueous solution, and acetonitrile-ammonium acetate aqueous solution, and the results show that the system chromatographic separation is significantly better when the acetonitrile-ammonium acetate aqueous solution mobile phase is selected. Since the chemical components in Jingling oral liquid are complex and the polarity difference is large, the present invention selects gradient elution to ensure that more chemical components can be effectively separated, which is conducive to qualitative identification of the main medicinal materials and components, thereby better controlling the quality of drug production.

[0018] The above-mentioned quality inspection method was used to detect Jingling oral liquid. According to the obtained fingerprint spectrum, there were 22 common peaks, which can be attributed to five medicinal flavors: Phellodendron chinense, Polygala tenuifolia, Ligustrum lucidum, Paeonia suffruticosa, and Schisandra chinensis. Six chemical components, including phellodendronine hydrochloride, berberine hydrochloride, schisandra chinensis methanol, paeoniflorin, paeonol, and Ligustrum lucidum glycosides, can also be identified, which can better control and manage the product quality of Jingling oral liquid.

[0019] Preferably, the quality detection method comprises the following steps:

[0020] Weigh phellodendronine hydrochloride, berberine hydrochloride, schisandra methanol, paeoniflorin, paeonol, and ligustri lucidifolia glycosides respectively, add solvent to dissolve, and prepare each reference substance solution;

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

[0022] On the basis of the 12 medicinal materials of Jingling oral liquid, a single medicinal material is removed, and the remaining medicinal materials are mixed in equal mass, and water is added for extraction to obtain a negative sample lacking a single medicinal material; the negative sample lacking a single medicinal material is taken, evaporated to dryness, and dissolved in a solvent to obtain a negative sample solution lacking a single medicinal material;

[0023] The reference substance solution, the test substance solution and the single-drug-deficient negative sample solution are injected into a high performance liquid chromatograph for detection, chromatograms of the reference substance, the test substance and the single-drug-deficient negative sample are obtained, and the chromatograms are analyzed.

[0024] In the preparation of the reference substance solution, the test substance solution and the single-drug-deficient negative sample solution, the solvent is methanol or a methanol solution.

[0025] The experiment showed that the response value was greater when the sample was evaporated to dryness and then dissolved in methanol solution.

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

[0027] The single herb is selected from any one of Radix Rehmanniae Preparata, Rhizoma Dioscoreae, Poria, Cortex Moutan, Rhizoma Alismatis, Radix Polygalae, Radix Achyranthis Bidentatae, Fructus Ligustri Lucidi, Cortex Phellodendri, Rhizoma Anemarrhenae, Fructus Schisandrae Chinensis, and Rhizoma Acorus Calamus. For example, when the single herb is Cortex Phellodendri, a corresponding Cortex Phellodendri negative sample solution 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 drug, identifying the drug components based on the chromatograms of the reference sample and the test sample, or establishing a liquid fingerprint of Jingling oral liquid from the chromatogram of the test sample.

[0029] Preferably, the quality detection method adopts multi-wavelength detection. Since the effective components contained in Jingling oral liquid are complex and the maximum absorption wavelengths of each component are significantly different, the present invention adopts multi-wavelength detection in view of detection sensitivity.

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

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

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

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

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

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

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

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

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

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

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

[0041] Preferably, the specification of the chromatographic column is 250×4.6 mm, and the filler particle size of the chromatographic column is 5 μm.

[0042] Preferably, the chromatographic conditions further include: an injection volume of 5-15 μL. Further 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 tert-lucidin is a relatively stable component in Jingling oral liquid with a relatively high content, has a small loss during the extraction process, has a moderate retention time in liquid chromatography, a high response value and a good separation degree, and can be used as an index component for content determination in Jingling oral liquid, the quality detection method of the present invention selects tert-lucidin as a reference peak. The theoretical plate number is calculated based on the reference tert-lucidin, and should not be less than 5000.

[0047] The quality detection method provided by the present invention is used for detection. There are 22 common chromatographic peaks in the liquid fingerprint of Jingling oral liquid, among which Peak 12 is the chromatographic peak with the same peak retention time as the reference substance, terebinthrin, and is defined as the reference peak. Compared with the reference peak retention time (counted as 1), the relative retention time of each chromatographic peak is: 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 (Huangbai alkaloid hydrochloride) 0.628±10%, Peak 10 (peoniflorin) 0.737±10%, Peak 11 0.762±10%. ±10%, Peak 12 (special privet 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 (schisandrin A) 2.247±10%.

[0048] In the liquid 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; peaks 2, 3, 4, 5, 6, 7, 8, 9, and 17 are derived from Phellodendron chinense.

[0049] The present invention adopts the Chinese medicine chromatographic fingerprint similarity evaluation system provided by the State Pharmacopoeia Committee as the fingerprint similarity calculation software. After multiple experiments and studies, and by comparing with the method of calculating relative retention time and relative peak area, the evaluation conclusions obtained remain consistent without obvious differences, 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 of Jingling oral liquid established by the present invention through high performance liquid chromatography determined a total of 22 common chromatographic peaks, and identified and confirmed 6 index chemical components, namely phellodendronine hydrochloride, berberine hydrochloride, schisandra methanol, paeoniflorin, paeonol, and privet glycosides; and can be attributed to the five medicinal flavors of phellodendron, polygala, privet fruit, moutan bark, and schisandra, thus achieving the qualitative identification of the above medicinal materials.

[0052] (2) Through precision tests, stability tests, repeatability tests and other tests, it was confirmed that the quality detection method of the present invention has good detection effect, high reproducibility and good reliability, and can be used for qualitative quality control of Jingling oral liquid. In addition, the quality detection method of the present invention was used to detect multiple batches of samples, and 22 common peaks were detected, and 6 chemical components were identified and attributed to 5 medicinal materials. The test results were all qualified, which further showed that the quality detection method has the characteristics of simple operation, accuracy and stability, and can better control and manage the product quality of Jingling oral liquid, and also provide a more comprehensive scientific basis for the quality evaluation of Jingling oral liquid. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Figure 1 It is a chromatogram comparison chart (293nm) of the test solution, the negative sample solution lacking Phellodendron chinense, and the reference solution of Phellodendron chinense hydrochloride in Example 1;

[0054] Figure 2 The chromatograms of the test solution, the negative sample solution lacking Phellodendron chinense, and the berberine hydrochloride reference solution in Example 1 are compared (320 nm);

[0055] Figure 3 It is a chromatogram comparison diagram (219 nm) of the test solution, the negative sample solution lacking Paeonia suffruticosa Andrews, and the paeoniflorin reference solution in Example 1;

[0056] Figure 4 It is a chromatogram comparison diagram (237nm) of the test solution, the negative sample solution lacking Paeonia suffruticosa Andrews, and the paeonol reference solution in Example 1;

[0057] Figure 5 It is a chromatogram comparison chart (342nm) of the test solution, the negative sample solution lacking Ligustrum lucidum, and the Ligustrum lucidum in Example 1;

[0058] Figure 6 It is a comparison chart of the test solution, the negative sample solution lacking Schisandra chinensis, and the Schisandra chinensis methanol chromatogram in Example 1 (237 nm);

[0059] Figure 7 It is a chromatogram comparison of the test solution, the negative sample solution lacking Polygala tenuifolia, and Polygala tenuifolia yamkoningone III in Example 1 (342 nm);

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

[0061] Fig. 9 The chromatogram comparison diagram (342 nm) of the test solution and the negative sample solution lacking Rehmannia glutinosa in Example 1;

[0062] Fig.10The chromatograms of the test solution, the negative sample solution lacking Anemarrhena asphodeloides, and Anemarrhena asphodeloside BⅡ in Example 1 are compared (275 nm);

[0063] Fig.11 It is a chromatogram comparison diagram (342nm) of the test solution, the negative sample solution lacking Chinese yam, and the negative sample solution lacking Poria in Example 1;

[0064] Fig.12 It is a chromatogram comparison diagram (342nm) of the test solution and the negative sample solution lacking Alisma orientalis in Example 1;

[0065] Fig.13 The chromatogram is the attribution chromatogram of the negative sample lacking a single herb in Example 1;

[0066] Fig.14 The chromatogram is the identified component of the reference substance in Example 1. DETAILED DESCRIPTION

[0067] In order to make the technical scheme of the present invention more clearly understood by those skilled in the art, the following embodiments are listed for illustration. It should be pointed out that the following embodiments do not limit the protection scope of the present invention.

[0068] The instruments, reagents and reference substances used in the following examples, as well as the specific operation procedures and operation schemes are as follows. The remaining raw materials, reagents or devices, unless otherwise specified, can be obtained from conventional commercial channels or can be obtained by existing known methods.

[0069] 1. Instruments and equipment:

[0070] High performance liquid chromatography (HPLC), CNC ultrasonic cleaner, constant temperature and humidity chamber, chromatographic column, electronic balance (one ten-thousandth), electric constant temperature water bath, etc.

[0071] 2. Reagents and reference substances:

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

[0073] Reference substances: Rehmannia glutinosa D, salidroside, phellodendronine hydrochloride, paeoniflorin, privet glycosides, berberine hydrochloride and paeonol, etc.

[0074] 3. Specific operation process and operation plan:

[0075] Determined by high performance liquid chromatography (General Rules 0512 of Part IV of the 2020 edition of the Chinese Pharmacopoeia).

[0076] Example 1

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

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

[0079] Preparation of reference solution: Take phellodendronine hydrochloride, berberine hydrochloride, schisandra methanol, paeoniflorin, paeonol and terunculoside respectively, add methanol to dissolve them, and obtain the corresponding reference solution.

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

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

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

[0083] Acetonitrile was used as mobile phase A, 0.05 mol / L ammonium acetate aqueous solution was used as mobile phase B, and gradient elution was performed using 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°C; flow rate: 0.8 mL / min; detector: DAD detector. -15 to 0 min is the time to equilibrate the column.

[0087] On the basis of the above chromatographic conditions, the components were first screened by multi-wavelength, and the peak positioning of the fingerprint of Jingling oral liquid was studied.

[0088] Accurately pipette 10 μL of the test solution, reference solution, and single-drug-deficient negative sample solution, respectively, inject them into the liquid chromatograph for qualitative analysis, and record the chromatogram.

[0089] The components are screened by multiple wavelengths, and the results are Figure 1-12 As shown, the chromatograms of the test sample, the reference sample and the negative sample lacking a single drug are attributed to the medicinal flavors of Phellodendron chinense, Polygala tenuifolia, Fructus Ligustri Lucidi, Paeonia suffruticosa moutan and Schisandra chinensis, and the chemical components identified are Phellodendron chinense hydrochloride, berberine hydrochloride, Schisandra chinensis methanol, paeoniflorin, paeonol and privetrin.

[0090] Based on the results of the above peak positioning study, this embodiment sets the detection wavelengths to 220nm, 320nm, 237nm, and 293nm, and starts full wavelength scanning at the same time as the final confirmed wavelength detection method. The wavelength switching method is shown in Table 2:

[0091] Table 2

[0092]

[0093]

[0094] Accurately pipette 10 μL of the test solution, reference solution, and single-drug-deficient negative sample solution, respectively, and inject them into the liquid chromatograph. Use the above-mentioned confirmed detection method for qualitative analysis and record the chromatogram.

[0095] according to Figure 13-14 The content shown shows that there are 22 common chromatographic peaks in the liquid fingerprint of Jingling oral liquid, among which Peak 12 is the chromatographic peak with the same retention time as the reference substance, terebinthrin, and is defined as the reference peak. Compared with the reference peak retention time (counted as 1), the relative retention times of the chromatographic peaks are: 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 (Huangbai alkaloid hydrochloride) 0.628±10%, Peak 10 (peoniflorin) 0.737±10%, Peak 11 0.762±10%. ±10%, Peak 12 (special privet 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 (schisandrin 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] As shown in Table 3, peaks 1 and 22 are derived from Schisandrae 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 amurense. The medicinal flavors attributed to the compounds are Polygala tenuifolia, Phellodendron amurense, Ligustrum lucidum, Paeonia suffruticosa, and Schisandrae Chinensis, and the chemical components identified are phellodendrine hydrochloride, berberine hydrochloride, paeonol, paeoniflorin, schisandrin A, and terebutrin.

[0101] Example 2

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

[0103] Table 4 Calculation results of fingerprint spectrum precision relative retention time

[0104]

[0105]

[0106] Table 5 Precision inspection 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] Among them, R: control fingerprint, 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 of the test sample presents chromatographic peaks with the same retention time as the reference chromatographic peaks, and presents 22 main chromatographic peaks. The relative retention time RSD is less than 2.0%. Compared with the generated control fingerprint, the similarity is greater than 0.90, indicating that the instrument has good precision.

[0110] Example 3

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

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

[0113]

[0114]

[0115] Table 7 Stability test similarity evaluation results

[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] Among them, R: control fingerprint, 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 of the test sample presents chromatographic peaks with the same retention time as the reference chromatographic peaks, and presents 22 main chromatographic peaks. The relative retention time RSD is less than 2.0%. Compared with the generated control fingerprint, the similarity is greater than 0.90, indicating that the test 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 inspection methods of the test solution in Example 1, and calculate the relative retention time and similarity. The specific measurement results are shown in Tables 8-9.

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

[0122]

[0123] Table 9 Repeatability test similarity evaluation results

[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] Among them, R: control fingerprint, 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 of the test sample presents chromatographic peaks with the same retention time as the reference chromatographic peaks, and presents 22 main chromatographic peaks. The relative retention time RSD is less than 2.0%. Compared with the generated control fingerprint, the similarity is greater than 0.90, indicating that the repeatability of this quality detection method is good.

[0127] Example 5

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

[0129] The results showed that the results were basically consistent under different flow rates and column temperatures, and the common peaks in the chromatograms were sharp, symmetrical, and had good separation, indicating that the method had good durability under different flow rates and column temperatures.

[0130] Example 6

[0131] Testing of multiple batches of samples: Jingling oral liquid is an exclusive product of Liaoning Dongfangren Pharmaceutical Co., Ltd. (National Medicine Standard No. Z10910056). In this experiment, 10 different batches of Jingling oral liquid samples were taken and inspected using the quality inspection method in Example 1. The inspection results showed that 22 common peaks could be detected in all 10 batches of samples, and 6 chemical components (phellodendron hydrochloride, berberine hydrochloride, schisandra methanol, paeoniflorin, paeonol, and privet glycosides) could be identified and attributed to 5 medicinal materials (phellodendron, polygala tenuifolia, privet fruit, moutan bark, and schisandra chinensis). The test results were all qualified, further indicating that this quality inspection method has a good identification effect and can be used for qualitative detection of Jingling oral liquid.

[0132] Comparative Example 1

[0133] Study the test results under different mobile phase conditions: The preliminary exploratory tests showed that when 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, the response values ​​of the chromatographic peaks were low, the separation was small, the chemical components identified were small, or the medicinal flavors were small. After continuous screening and optimization of chromatographic conditions, acetonitrile-ammonium acetate aqueous solution was finally selected as the mobile phase system, and it was found that this mobile phase system had excellent results when used for the detection of Jingling oral liquid and could meet the actual detection needs.

[0134] The embodiments of the present application are described in detail above in conjunction with the accompanying drawings, but the present application is not limited to the above embodiments, and various changes can be made within the knowledge scope of ordinary technicians in the relevant technical field without departing from the purpose of the present application.

Claims

1. A quality detection method for Jingling oral liquid, characterized in that: High performance liquid chromatography was used for detection, and the chromatographic conditions used were: ZORBAX Eclipse Plus C18 was used as the packing material of the chromatographic column; Acetonitrile was used as mobile phase A, and 0.04-0.06 mol / L ammonium acetate aqueous solution was used as mobile phase B for gradient elution; The conditions of the gradient elution are: 0-50min, mobile phase A 5%→28%, mobile phase B 95%→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%; 80-83min, mobile phase A 40%→70%, mobile phase B 60%→40%; 83-85min, mobile phase A 70%→20%, mobile phase B 40%→80%; 85-90min, mobile phase A 20%→5%, mobile phase B 80%→95%.

2. The quality inspection method according to claim 1, characterized in that: The following steps are involved: Weigh phellodendronine hydrochloride, berberine hydrochloride, schisandra methanol, paeoniflorin, paeonol, and ligustri lucidifolia glycosides respectively, add solvent to dissolve, and prepare each reference substance solution; Weigh Jingling oral liquid and dissolve it in solvent to prepare the test solution; On the basis of the 12 medicinal materials of Jingling oral liquid, a single medicinal material is removed, and the remaining medicinal materials are mixed in equal mass, and water is added for extraction to obtain a negative sample lacking a single medicinal material; the negative sample lacking a single medicinal material is taken, evaporated to dryness, and dissolved in a solvent to obtain a negative sample solution lacking a single medicinal material; The reference substance solution, the test substance solution and the single-drug-deficient negative sample solution are injected into a high performance liquid chromatograph for detection, chromatograms of the reference substance, the test substance and the single-drug-deficient negative sample are obtained, and the chromatograms are analyzed.

3. The quality inspection method according to claim 2, characterized in that: 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 drug, identifying the drug components based on the chromatograms of the reference sample and the test sample, or establishing a liquid fingerprint of Jingling oral liquid from the chromatogram of the test sample.

4. The quality inspection method according to claim 2, characterized in that: In the preparation of the reference substance solution, the test substance solution and the single-drug-deficient negative sample solution, the solvent is methanol or a methanol solution.

5. The quality detection method according to any one of claims 1 to 3, characterized in that: The quality detection method adopts multi-wavelength detection; the wavelengths of the multi-wavelength detection include 220nm, 237nm, 293nm and 320nm.

6. The quality inspection method according to any one of claims 1 to 3, characterized in that: The specification of the chromatographic column is 250×4.6 mm, and the filler particle size of the chromatographic column is 5 μm.

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

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

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

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

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