Quality detection method of nasal drops for treating allergic rhinitis

Abstract

The invention provides a quality detection method of nose drops for treating allergic rhinitis, and belongs to the field of medicine detection. The method specifically comprises the following steps: S1, preparing a test solution; s2, preparing a mixed reference substance solution; s3, respectively sucking the reference substance solution and the test solution, and carrying out detection analysis; and S4, generating a control fingerprint spectrum. According to the method, different types of components of all the medicinal materials can be presented on one map through one-time detection, the content of multiple components can be measured at the same time, and comprehensive and accurate control over the quality of the artemisia selengensis and scutellaria baicalensis nose drops is achieved. According to the quality evaluation method provided by the invention, chromatographic conditions are subjected to optimization screening and methodology verification, and the method has the advantages of being stable, good in repeatability and stable and reliable in measurement result.

Description

Technical Field

[0001] The invention belongs to the field of drug detection, and in particular relates to a quality detection method for nasal drops for treating allergic rhinitis. Background Art

[0002] The Chinese medicine compound preparation for treating allergic rhinitis is composed of six herbs, including Xinjiang Artemisia, Ephedra, Xanthium sibiricum, and Scutellaria baicalensis. It has the effects of clearing away heat and detoxifying, ventilating the lungs and opening the orifices, and is used for acute rhinitis, chronic rhinitis, and allergic rhinitis. Chinese patent 201710827980.6, entitled "Nasal preparation for treating rhinitis, its preparation method and application", discloses a nasal preparation for treating rhinitis. Compared with the prior art, the dosage of the nasal preparation for treating rhinitis of the invention is only 20% to 30% of the dosage of the prior art, and the nasal preparation for treating rhinitis is more effective in anti-inflammatory and anti-allergic aspects than the prior art; at the same time, the preparation process of the invention is simple, the efficacy is definite, and there are no toxic side effects. The drug directly acts on the surface of the nasal mucosa, has high bioavailability, fast onset, convenience, and good patient compliance. Different dosage forms can be prepared according to the needs of patients, and can be better used to treat acute, chronic and allergic rhinitis. Therefore, it is particularly important to explore the quality control method of the preparation.

[0003] At present, the implementation standards of this preparation only include routine inspection, thin layer identification method and determination of artemisinic acid and baicalin content, which cannot fully reflect the quality of the product. There are few literatures on the quality research of this preparation, and there has been no research report on the overall control of its quality. Traditional Chinese medicine compound contains multiple traditional Chinese medicines with complex chemical composition. With the improvement of the quality control requirements of traditional Chinese medicine, the content determination of a single or a few components cannot comprehensively and scientifically evaluate the quality of traditional Chinese medicine. The qualitative and quantitative analysis of multi-index components has become the key to the quality control of traditional Chinese medicine.

[0004] Traditional Chinese medicine fingerprint is a comprehensive and quantifiable quality analysis method that can characterize the overall chemical composition of traditional Chinese medicine and more comprehensively reflect the difference information of the chemical composition of traditional Chinese medicine. It is mainly used to identify the authenticity and quality of traditional Chinese medicines and Chinese patent medicines, and to evaluate the uniformity and stability of the quality of raw materials, semi-finished products and finished products of traditional Chinese medicine preparations. Traditional Chinese medicine and its preparations are multi-component complex systems. Therefore, the evaluation of their quality should adopt a detection method that is suitable for them and can provide rich identification information. The establishment of traditional Chinese medicine fingerprint will be able to more comprehensively reflect the types and quantities of chemical components contained in traditional Chinese medicine and its preparations, and then provide an overall description and evaluation of the quality of the medicine. By establishing a fingerprint combined with a multi-component content determination method and conducting a systematic analysis, it is not only possible to effectively evaluate the consistency of the quality of traditional Chinese medicine, but also to interpret the main factors that form the quality difference, and to evaluate the quality of traditional Chinese medicine more objectively and comprehensively.

[0005] The traditional method of determining the content of multi-index components requires the use of a large number of reference substances, which are expensive and difficult to obtain. The quantitative analysis multi-components by single marker (QAMS) method utilizes the intrinsic functional relationship between the effective components of traditional Chinese medicines. By only measuring the content of common components that are cheap and easily available, it can simultaneously measure the content of multiple components. It has the characteristics of low cost and simple operation, and is suitable for the quality control of traditional Chinese medicine compound preparations. In order to comprehensively and effectively control the quality of the preparation, the present invention uses HPLC for the first time to establish the fingerprint of the traditional Chinese medicine compound preparation for the treatment of allergic rhinitis, and simultaneously uses the one-measurement-multiple-evaluation method to determine the content of 16 components in the preparation, so as to achieve comprehensive and accurate quality control. Summary of the invention

[0006] In view of this, the purpose of the present invention is to provide a quality detection method for nasal drops for treating allergic rhinitis, which can realize presenting different types of components of all medicinal materials in the preparation on a spectrum under the same chromatographic conditions, and simultaneously determining the contents of 16 components, so as to realize comprehensive and accurate control of the quality of Artemisia scutellariae nasal drops. The detection method is stable and reliable, has strong specificity and high accuracy, and can effectively control the quality of the product.

[0007] In order to achieve the above object, the present invention provides the following technical solutions:

[0008] The present invention provides a method for detecting the quality of nasal drops for treating allergic rhinitis, comprising the following steps:

[0009] S1. Prepare the test solution: accurately measure 2.0 mL of nasal drops, place in a 10 mL volumetric flask, add 50% methanol to dilute to the mark, shake well, filter, and take the filtrate;

[0010] S2. Prepare mixed reference solution: accurately weigh appropriate amounts of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, hyperoside, luteolin, isochlorogenic acid, 1,5-dicaffeoylquinic acid, isochlorogenic acid C, rosmarinic acid, baicalin, montanol, isochlorogenic acid, artemisinic acid, ammonium glycyrrhizinate, vitexin and imperatorin, and add 50% methanol to prepare a mixed reference solution;

[0011] The concentration of the reference substance in the mixed reference substance solution is 0.04 to 7.684 mg / mL;

[0012] S3. Pipette the mixed reference solution and the test solution separately, inject into the liquid chromatograph to detect, analyze and record the chromatogram for quality evaluation;

[0013] The liquid chromatography conditions were as follows:

[0014] Column: Agilent TCC18 Column, specifications: 250 × 4.6 mm, 5 μm;

[0015] Mobile phase: Mobile phase A is 0.1% phosphoric acid solution, mobile phase B is acetonitrile; mobile phase A and mobile phase B are gradient eluted: 0-10min, 97%-88% A; 10-30min, 88%-78% A; 55-95min, 70%-54% A; 95-115min, 54%-20% A; 115-120min, 20% A;

[0016] Volume flow rate 1.0mL / min;

[0017] Column temperature: 30°C;

[0018] Detection wavelength: 245nm;

[0019] Injection volume: 5 μL.

[0020] S4. Generate a reference fingerprint: export the chromatogram file of the nasal drops obtained in S3 from the instrument and import it into the similarity evaluation system of the chromatographic fingerprint of traditional Chinese medicine; select the chromatographic peaks existing in the chromatograms of different batches of nasal drops as common peaks; generate a reference fingerprint of the nasal drops using the average value calculation method.

[0021] Preferably, the nasal drops are the nasal preparations for treating rhinitis prepared in patent 201710827980.6.

[0022] Preferably, the specific concentrations of the components in the S2 mixed reference solution are: neochlorogenic acid 0.523 mg / mL, chlorogenic acid 0.756 mg / mL, cryptochlorogenic acid 1.046 mg / mL, hyperoside 0.489 mg / mL, olibanum 1.126 mg / mL, isochlorogenic acid B 1.246 mg / mL, 1,5-dicaffeoylquinic acid 0.876 mg / mL, isochlorogenic acid C 0.964 mg / mL, rosmarinic acid 0.222 mg / mL, baicalin 7.684 mg / mL, montana glycoside 0.0474 mg / mL, isochlorogenic acid 0.237 mg / mL, artemisinic acid 1.001 mg / mL, ammonium glycyrrhizinate 2.056 mg / mL, vitex luteolin 1.174 mg / mL, and imperatorin 0.627 mg / mL.

[0023] Preferably, 52 common peaks are identified in the chromatogram of S3, specifically: the fingerprint of S3 includes 52 common peaks, specifically: Peak 8 corresponding to neochlorogenic acid, Peak 10 corresponding to chlorogenic acid, Peak 11 corresponding to cryptochlorogenic acid, Peak 22 corresponding to hyperoside, Peak 23 corresponding to oleanolic acid, Peak 26 corresponding to isochlorogenic acid B, Peak 28 corresponding to 1,5-dicaffeoylquinic acid, Peak 29 corresponding to isochlorogenic acid C, Peak 30 corresponding to rosmarinic acid, Peak 32 corresponding to baicalin, Peak 35 corresponding to montanol, Peak 37 corresponding to isochlorogenic acid, Peak 39 corresponding to artemisinic acid, Peak 44 corresponding to ammonium glycyrrhizinate, Peak 46 corresponding to vitexin, and Peak 49 corresponding to imperatorin.

[0024] Preferably, the characteristic peak in the fingerprint is based on the baicalin peak as the reference peak, and the relative retention times of the 16 chromatographic peaks are 0.322, 0.419, 0.431, 0.712, 0.727, 0.741, 0.777, 0.814, 0.879, 1.000, 1.113, 1.279, 1.410, 1.777, 1.845, and 2.178, respectively.

[0025] Preferably, the quality evaluation in S3 is performed by importing the fingerprint into a traditional Chinese medicine chromatographic fingerprint similarity evaluation software to perform a similarity evaluation with a control fingerprint.

[0026] The present invention also provides application of the above-mentioned nasal drops quality detection method in quality control of artemisia scutellariae nasal drops.

[0027] Preferably, the quality detection method includes the application of fingerprint spectrum combined with multi-component content determination method in the quality control of Chinese medicinal materials or solid pharmaceutical preparations containing six herbs: Artemisia argyi, Ephedra sinica, Xanthium sibiricum, Scutellaria baicalensis, Licorice and Angelica dahurica.

[0028] Contains at least the following beneficial technical effects:

[0029] The present invention provides a quality evaluation method based on high performance liquid chromatography, presenting different types of components of all medicinal materials in the preparation on a spectrum, and simultaneously determining the contents of 16 components, thereby achieving comprehensive and accurate quality control of Artemisia Scutellariae Nasal Drops. The quality evaluation method provided by the present invention has the advantages of stable method, good repeatability, and stable and reliable measurement results after the chromatographic conditions have been optimized, screened, and verified by methodology.

[0030] The present invention establishes a fingerprint spectrum of Artemisia scutellariae nasal drops, which can more comprehensively reflect the types and quantities of chemical components contained in the preparation, and then comprehensively describe and evaluate the quality of the preparation;

[0031] The present invention establishes a method for determining the content of multiple components of Artemisia scutellariae nasal drops, and combines the fingerprint spectrum for systematic analysis, which can not only effectively evaluate the consistency of the quality of traditional Chinese medicine, but also explain the main factors that form the quality difference, and can evaluate the quality of traditional Chinese medicine more objectively and comprehensively.

[0032] The invention establishes a one-test-multiple-evaluation method for determining 16 components in artemisia nasal drops, takes baicalin as an internal reference (S), utilizes the intrinsic functional relationship between effective components of traditional Chinese medicine, and determines the content of baicalin, which is cheap and easily available, so as to simultaneously determine the contents of 16 components in the artemisia nasal drops. The method has the characteristics of low cost and simple operation, and can achieve comprehensive and accurate control of the quality of the artemisia nasal drops. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 It is the HPLC chromatogram of the mixed reference substance;

[0034] Figure 2 This is the HPLC chromatogram of the Artemisia Scutellariae nasal drops sample;

[0035] Figure 3 The fingerprints of Artemisia scutellariae nasal drops (S1-S5) and the reference spectrum (R);

[0036] Figure 4 These are the chromatograms of single-flavor drugs, corresponding flavor-deficient drugs, and Artemisia scutellariae nasal drops. DETAILED DESCRIPTION

[0037] The technical solution of the present invention is further described below through the accompanying drawings and embodiments.

[0038] Unless otherwise defined, technical or scientific terms used in the present invention shall have the common meanings understood by one having ordinary skills in the field to which the present invention belongs.

[0039] It is obvious to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the present invention can be implemented in other specific forms without departing from the main idea or essential features of the present invention. Therefore, from all points of view, the embodiments should be regarded as exemplary and non-restrictive, and the scope of the present invention is defined by the appended claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims are included in the present invention, and any reference numerals in the claims should not be regarded as limiting the claims involved.

[0040] In addition, it should be understood that although this specification is described according to the implementation modes, not every implementation mode includes only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation modes that can be understood by those skilled in the art. These other implementation modes are also covered within the protection scope of the present invention.

[0041] It should also be understood that the specific embodiments described above are only used to explain the present invention, and the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field, within the technical scope disclosed by the present invention, can make equivalent replacements or changes based on the technical solutions and inventive concepts of the present invention, which should be covered by the protection scope of the present invention / invention.

[0042] Example 1

[0043] 1 Materials

[0044] 1.1 Instruments

[0045] Agilent 1260 high performance liquid chromatograph (Agilent, USA, YF-17, YF-18); U3000 high performance liquid chromatograph (Thermo Fisher, YF-8); CPA225D electronic balance (Sartorius, d = 0.01 mg); AS20500BDT ultrasonic cleaner (frequency 60 kHz, power 500 W, Tianjin Aotesense Instrument Co., Ltd.); UPT-10T UPT experimental ultrapure water machine (Chengdu Ultrapure Technology Co., Ltd.); chromatographic column Agilent XDB C 18 (250×4.6mm, 5μm); Chromatographic column YMC-C 18 (250×4.6mm, 5μm); chromatographic column Agilent SB-C 18 (250×4.6mm, 5μm).

[0046] 1.2 Samples and Reagents

[0047] Chlorogenic acid (batch number 110753-202018, purity 96.1%), hyperoside (batch number 111521-201406, purity 100%), luteolin (batch number 111720-201307, purity 100%), rosmarinic acid (batch number 111871-202007, purity 98.1%), baicalin (batch number 110715-202122, purity 94.2%) %), montanol (111528-202112, purity 98.0%), ammonium glycyrrhizinate (batch number 110731-202122, purity 94.4%), vitex lutein (batch number 111554-201705, purity 98.3%), imperatorin (batch number 110826-201918, purity 99.0%), ferulic acid (batch number 110773-201313, purity 99.6%), liquiritin (batch number 111610-201908, purity 95.0%), isoimperatorin (batch number 110827-202113, purity 99.2%), rutin (batch number 100080-202012, purity 92.2%), apigenin (batch number 111901-202004, purity 99.4%), kaempferol (batch number 110861-202013 , purity 93.2%), ephedrine hydrochloride (batch number 171241-201809, purity 100%), pseudoephedrine hydrochloride (batch number 171237-201510, purity 99.8%), acetylcholine (batch number 111879-201102, purity 97.2%), and ursolic acid (110742-201823, purity 99.9%) were all purchased from China Food and Drug Inspection Institutes. Neochlorogenic acid (batch number DSTDX001504), isochlorogenic acid (batch number DSTDX001504), isochlorogenic acid B (batch number DSTDY003703), isochlorogenic acid C (batch number DSTDPS0068-0020), 1,5-dicaffeoylquinic acid (batch number DSTDE000602), cryptochlorogenic acid (batch number DST221220-035), isochlorogenic acid A (batch number DSTDE000602), with a purity greater than 98%, were purchased from Chengdu Dester Biotechnology Co., Ltd. Artemisia striata acid (batch number DZ20240302, purity 99.71%) was purchased from Xinjiang Institute of Physical and Chemical Technology, Chinese Academy of Sciences.

[0048] Artemisia scutellariae nasal drops, S1-S5, were prepared by Xinjiang Uygur Autonomous Region Institute of Pharmacy according to the method for preparing nasal preparations for treating rhinitis described in Patent 201710827980.6.

[0049] Acetonitrile, chromatography grade, Fisher, USA; water was Yibao purified water, and the other reagents were of analytical grade.

[0050] 2 Methods and Results

[0051] 2.1 Preparation of reference solution

[0052] Accurately weigh appropriate amounts of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, hyperoside, luteolin, isochlorogenic acid B, 1,5-dicaffeoylquinic acid, isochlorogenic acid C, rosmarinic acid, baicalin, montanol, isochlorogenic acid, artemisinic acid, ammonium glycyrrhizinate, vitexin and imperatorin reference substances, add 50% methanol to dissolve and make up to volume, and prepare mass concentrations of 0.523, 0.756, 1.046, 0.489, 1.126, and 1.666, respectively.

[0053] Reference substance stock solutions of 1.246, 0.876, 0.964, 0.222, 7.684, 0.0474, 0.237, 1.001, 2.056, 1.174, and 0.627 mg / mL.

[0054] 2.2 Preparation of test solution

[0055] Accurately measure 2.0 mL of Artemisia scutellariae nasal drops, place in a 10 mL volumetric flask, add 50% methanol to dilute to the scale, shake well, filter, and take the filtrate to obtain the product.

[0056] 2.3 Chromatographic conditions

[0057] Chromatographic column Agilent TC C 18 (250×4.6mm, 5μm), mobile phase was 0.1% phosphoric acid solution (A)-acetonitrile (B), gradient elution: 0-10min, 97%-88% A; 10-30min, 88%-78% A; 55-95min, 70%-54% A; 95-115min, 54%-20% A; 115-120min, 20% A; volume flow rate 1.0mg / mL; column temperature 30℃; detection wavelength 245nm; injection volume 5μL, see Figure 1 and Figure 2 , the corresponding relationships among the chromatographic peaks in the figure are: 1. Neochlorogenic acid; 2. Chlorogenic acid; 3. Cryptochlorogenic acid; 4. Hyperoside; 5. Oleuroside; 6. Isochlorogenic acid B; 7. 1,5-dicaffeoylquinic acid; 8. Isochlorogenic acid C; 9. Rosmarinic acid; 10. Baicalin; 11. Menghuaside; 12. Isochlorogenic acid; 13. Artemisinic acid; 14. Ammonium glycyrrhizinate; 15. Vitexin; 16. Imperatorin.

[0058] 2.4 Fingerprint Methodology Review

[0059] 2.4.1 Precision test

[0060] Take the test solution (S1) and inject it continuously for 6 times according to the chromatographic method under "2.3", taking Peak 32 (baicalin) as the reference peak. The results show that the relative retention time RSD of 52 common peaks is less than 0.17%, and the relative peak area RSD is less than 2.9% (n=6), indicating that the instrument has good precision.

[0061] 2.4.2 Stability test

[0062] Take the test solution (S1) and inject it according to the chromatographic method under "2.3". Determine at 0, 2, 4, 8, 12, and 24 hours respectively, and use Peak 32 (baicalin) as the reference peak. The results show that the relative retention time RSD of 52 common peaks is less than 0.18%, and the relative peak area RSD is less than 3.0% (n=6), indicating that the test solution is stable within 24 hours.

[0063] 2.4.3 Repeatability test

[0064] Take the same batch of samples (S1), prepare 6 test solutions according to the method under "2.2", and inject them according to the chromatographic conditions under "2.3". Take Peak 32 (baicalin) as the reference peak. The results show that the relative retention time RSD of 52 common peaks is less than 0.23%, and the relative peak area RSD is less than 2.9% (n=6), indicating that this method has good repeatability.

[0065] 2.5 Analysis and evaluation of fingerprints

[0066] 2.5.1 Fingerprint establishment and similarity evaluation

[0067] Five batches of Artemisia scutellariae nasal drops were taken, and the test solution was prepared according to the method under "2.2". The sample was injected according to the chromatographic method under "2.3". The chromatogram information was imported into the Chinese medicine fingerprint similarity software (2012 version) for chromatographic peak matching. The median method was used, and the time window was set to 0.2 min. After multi-point calibration and automatic peak matching, a reference spectrum (R) was generated. Peak 32 (baicalin peak) was used as the reference peak (peak S). A total of 52 common peaks were identified, see Figure 3 The HPLC fingerprints of 5 batches of Artemisia scutellariae nasal drops were imported into the similarity evaluation software of Chinese medicine chromatographic fingerprints, and the similarity was evaluated with the control fingerprints. The results showed that the similarities of each batch of samples were 0.942, 0.964, 0.938, 0.942, and 0.938, all of which were greater than 0.9, with good similarity, indicating that the production process of Artemisia scutellariae nasal drops was stable.

[0068] 2.5.2 Chromatographic peak attribution analysis

[0069] Take S1 sample, Artemisia striata medicinal material, Ephedra medicinal material, Angelica dahurica medicinal material, Xanthium sibiricum medicinal material, Licorice extract, Scutellaria baicalensis extract, negative sample lacking Artemisia striata, negative sample lacking Ephedra, negative sample lacking Angelica dahurica, negative sample lacking Xanthium sibiricum, negative sample lacking Licorice extract and negative sample lacking Scutellaria baicalensis extract, prepare the test solution according to the method under "2.2", and measure according to the chromatographic conditions of "2.3". By comparing the chromatograms of samples, single medicinal materials and negative samples, the ownership of 52 common peaks was determined. By comparing the retention time and spectral information of the chromatographic peaks with those of the reference, a total of 28 peaks were identified, see Table 1, Figure 4 .

[0070] Table 1 Chromatographic peaks attribution

[0071]

[0072]

[0073] 2.6 Determination of multi-component content

[0074] 2.6.1 Linear Relationship

[0075] Accurately pipette the reference substance stock solution under "2.1" respectively, accurately measure different volumes, prepare a series of mixed reference substance solutions of different concentrations, and measure, and draw a standard curve with the reference substance concentration (μg / mL) as the horizontal axis (X) and the peak area as the vertical axis (Y). As a result, the 16 components have a good linear relationship within their concentration range, and the results are shown in Table 2.

[0076] Table 2 Linear range of 16 components

[0077]

[0078] 2.6.2 Precision test

[0079] Take the mixed reference solution and inject it continuously for 6 times according to the chromatographic method under "2.3". The RSDs of the peak areas of 16 components were measured to be 0.65%, 0.93%, 1.83%, 0.68%, 1.16%, 0.98%, 1.70%, 0.38%, 0.55%, 0.56%, 0.72%, 0.55%, 0.48%, 0.46%, 1.43% and 0.66%, respectively, indicating that the instrument has good precision. The results are shown in Table 3.

[0080] Table 3 Precision test results

[0081]

[0082]

[0083] 2.6.3 Stability test

[0084] The same test solution (S1) was taken and measured at 0, 2, 4, 8, 12, and 24 h according to the chromatographic conditions under "2.3". The RSDs of the peak areas of the 16 components were 0.83%, 1.21%, 1.77%, 1.34%,

[0085] 1.57%, 2.42%, 1.07%, 1.09%, 1.26%, 1.20%, 1.27%, 1.06%, 0.94%, 0.89%, 1.62%, and 1.25%, indicating that the solution is stable after 24 hours. The results are shown in Table 4.

[0086] Table 4 Stability test results

[0087]

[0088]

[0089] 2.6.4 Repeatability test

[0090] From the same batch of test samples (S1), 6 test sample solutions were prepared according to the method under "2.2", and the chromatographic conditions under "2.3" were used for determination. The RSDs of the contents of the 16 components were 2.55%, 2.91%, 1.71%, 0.90%, 1.76%, 1.31%, 1.02%, 1.18%, 2.92%, 2.48%, 2.66%, 1.93%, 2.87%, 2.33%, 1.40%, and 0.76%, respectively, indicating that this method has good repeatability. The results are shown in Table 5.

[0091] Table 5 Repeatability test results

[0092]

[0093]

[0094] 2.6.5 Sample recovery test

[0095] Accurately measure 6 portions of the test sample (S1) with known content, each portion is 1.0 mL, and add 16 component reference solution in a 1:1 ratio according to the content of each component. Prepare the test solution according to the method under "2.2", and determine it according to the chromatographic method under "2.3". The average recovery rates of the 16 components are 101.4%, 99.0%, 101.2%, 98.8%, 100.6%, 101.6%, 100.2%, and 101.6%, respectively. The clinical data were as follows: 99.6%, 100.8%, 99.8%, 101.5%, 97.5%, 102.0%, 101.4%, 99.3%, 101.7%, with RSDs of 1.9%, 2.0%, 2.7%, 0.8%, 2.9%, 1.4%, 2.4%, 1.8%, 2.3%, 2.0%, 2.8%, 1.8%, 2.7%, 1.2%, 2.7%, 1.3%, respectively. The results are shown in Table 6.

[0096] Table 6 Sample recovery test results

[0097]

[0098]

[0099] Example 2

[0100] 2.7.1 Establishment of QAMS

[0101] 2.7.1 Determination of relative correction factor (f)

[0102] Take the mixed reference solution and inject 1, 2, 4, 5, 8, 10, and 12 μL of it according to the chromatographic conditions under "2.1". Take baicalin as the internal reference (S) and calculate the content of neochlorogenic acid (A), chlorogenic acid (B), cryptochlorogenic acid (C), hyperoside (D), oleoside (E), isochlorogenic acid B (F), 1,5-dicaffeoylquinic acid (G), isochlorogenic acid C (H), rosmarinic acid (I), baicalin (J), montanol (K), isochlorogenic acid (L), artemisinic acid (M), and ammonium glycyrrhizinate according to the formula.

[0103] The relative correction factors f of feroxanthin (N), vitexin (O) and imperatorin (P) S / A 、f S / B 、f S / C 、f S / D 、f S / E 、f S / F 、f S / G 、f S / H 、f S / I 、f S / J 、f S / K 、f S / L 、f S / M 、f S / N 、fS / O 、f S / P , the results are shown in Table 7.

[0104] f S / i =f S / f i =(A s ×C i ) / (A i ×C s )

[0105] A s is the peak area of ​​the internal reference, C s is the internal reference concentration, A i is the peak area of ​​the component to be measured, C i is the concentration of the component to be tested.

[0106] Table 7 The f using baicalin as internal reference si Measurement results

[0107]

[0108]

[0109] 2.7.2 Effect of different volume flow rates on f S / i Impact

[0110] The 16 components f were investigated at volume flow rates of 0.9, 1.0, and 1.1 mL / min. S / i The results show that f S / i The RSDs of the two components are all less than 3%, indicating that the volume flow rate has a significant influence on the f S / i There was no significant effect. The results are shown in Table 8.

[0111] Table 8 Effect of different volume flow rates on f S / i Impact

[0112]

[0113] 2.7.3 Effect of different column temperatures on f S / i Impact

[0114] The effects of column temperature on the 9 components f at 28, 30, and 32 °C were investigated. S / i The results show that at different column temperatures, f S / i The RSDs of the two components were all less than 3%, indicating that the column temperature had a significant effect on the f S / i There was no significant effect. The results are shown in Table 9.

[0115] Table 9 Effect of different column temperatures on f S / i Impact

[0116]

[0117] 2.7.4 Effects of different instruments on f S / i Impact

[0118] The effects of three chromatographic systems, Thermo Scitific Ultimate 3000 and Agilent 1260 HPLC, on the performance of each component were investigated. S / i The results are shown in Table 10. The results show that f S / i Good adaptability under different instruments.

[0119] Table 10 Effects of different instruments on f S / i Impact

[0120]

[0121]

[0122] 2.7.5 Effect of different chromatographic columns on f S / i Impact

[0123] Investigated Agilent TC-C 18 (250×4.6mm, 5μm), YMC-Pack-ODS-A C 18 (250×4.6mm, 5μm), Cosmosil C 18 (250×4.6mm, 5μm) 3 kinds of chromatographic columns for 16 components f S / i The results show that f S / i The RSDs of the two columns were all less than 3%, indicating that different chromatographic columns had good effects on the f S / i The effect is small, and the results are shown in Table 11.

[0124] Table 11 Effect of different chromatographic columns on f S / i Impact

[0125]

[0126]

[0127] 2.7.6 Investigation of chromatographic peak location of the component to be measured

[0128] Take the mixed reference solution, use different instruments and different chromatographic columns, and measure according to the chromatographic conditions under "2.1". Calculate the relative retention time (t i / s ), locate the chromatographic peaks of each component, t i / s =t R(i) / t R(s) (i is the component to be tested, s is the internal reference), the results are as follows: i / s The RSDs of the i / sThe chromatographic peaks of each component were located and the results are shown in Table 12.

[0129] Table 12 Effect of different chromatographic columns and instruments on t i / s Impact

[0130]

[0131]

[0132] 2.8 Comparison of the results between QAMS and external standard method (ESM)

[0133] The ESM and QAMS methods were used to determine the contents of 16 components in different batches of Artemisia and Scutellaria baicalensis nasal drops. The results showed that there was no significant difference in the contents measured by the two methods (P>0.05), indicating that the QAMS method can be used for the determination of multi-component contents of Artemisia and Scutellaria baicalensis nasal drops. The results are shown in Table 13.

[0134] Table 13 QAMS and ESM measurement results

[0135]

[0136]

[0137]

[0138] In summary, the present invention provides a quality evaluation method based on high performance liquid chromatography, which can present different types of components of all medicinal materials on one spectrum by one detection under specific sample solution and chromatographic conditions, and can simultaneously perform content determination of multiple components, thereby achieving comprehensive and accurate quality control of Artemisia scutellariae nasal drops. The quality evaluation method provided by the present invention has the advantages of stable method, good repeatability, and stable and reliable measurement results after optimized screening and methodological verification of chromatographic conditions.

[0139] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present invention.

Claims

1. A method for detecting the quality of nasal drops for treating allergic rhinitis, characterized in that: The following steps are involved: S1. Prepare the test solution: accurately measure 2.0 mL of nasal drops, place in a 10 mL volumetric flask, add 50% methanol to dilute to the mark, shake well, filter, and take the filtrate; S2. Prepare mixed reference solution: accurately weigh appropriate amounts of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, hyperoside, luteolin, isochlorogenic acid, 1,5-dicaffeoylquinic acid, isochlorogenic acid C, rosmarinic acid, baicalin, montanol, isochlorogenic acid, artemisinic acid, ammonium glycyrrhizinate, vitexin and imperatorin, and add 50% methanol to prepare a mixed reference solution; The concentration of the reference substance in the mixed reference substance solution is 0.04 to 7.68 mg / mL; S3. Pipette the mixed reference solution and the test solution separately, inject into the liquid chromatograph to detect, analyze and record the chromatogram for quality evaluation; The liquid chromatography conditions were as follows: Column: Agilent TCC 18 Column, specifications: 250 × 4.6 mm, 5 μm; Mobile phase: Mobile phase A is 0.1% phosphoric acid solution, mobile phase B is acetonitrile; mobile phase A and mobile phase B are gradient eluted: 0-10min, 97%-88%A; 10-30min, 88%-78%A; 55-95min, 70%-54%A; 95-115min, 54%-20%A; 115-120min, 20%A; Volume flow rate 1.0mL / min; Column temperature: 30°C; Detection wavelength: 245nm; Injection volume: 5 μL; S4. Generate a reference fingerprint: export the chromatogram file of the nasal drops obtained in S3 from the instrument and import it into the similarity evaluation system of the chromatographic fingerprint of traditional Chinese medicine; select the chromatographic peaks existing in the chromatograms of different batches of nasal drops as common peaks; generate a reference fingerprint of the nasal drops using the average value calculation method.

2. The quality inspection method according to claim 1, characterized in that: The nasal drops are nasal preparations for treating rhinitis prepared in patent 201710827980.

6.

3. The quality inspection method according to claim 2, characterized in that: The specific concentrations of the components in the S2 mixed reference solution are: neochlorogenic acid 0.523 mg / mL, chlorogenic acid 0.756 mg / mL, cryptochlorogenic acid 1.046 mg / mL, hyperoside 0.489 mg / mL, luteolin 1.126 mg / mL, isochlorogenic acid B 1.246 mg / mL, 1,5-dicaffeoylquinic acid 0.876 mg / mL, isochlorogenic acid C 0.964 mg / mL, rosmarinic acid 0.222 mg / mL, baicalin 7.684 mg / mL, montana glycoside 0.0474 mg / mL, isochlorogenic acid 0.237 mg / mL, artemisia selengensis acid 1.001 mg / mL, ammonium glycyrrhizinate 2.056 mg / mL, vitex luteolin 1.174 mg / mL, and imperatorin 0.627 mg / mL.

4. The quality inspection method according to claim 2, characterized in that: The fingerprint spectrum of S3 includes 52 common peaks, specifically: Peak 8 corresponding to neochlorogenic acid, Peak 10 corresponding to chlorogenic acid, Peak 11 corresponding to cryptochlorogenic acid, Peak 22 corresponding to hyperoside, Peak 23 corresponding to oleoside, Peak 26 corresponding to isochlorogenic acid B, Peak 28 corresponding to 1,5-dicaffeoylquinic acid, Peak 29 corresponding to isochlorogenic acid C, Peak 30 corresponding to rosmarinic acid, Peak 32 corresponding to baicalin, Peak 35 corresponding to montanol, Peak 37 corresponding to isochlorogenic acid, Peak 39 corresponding to artemisinic acid, Peak 44 corresponding to ammonium glycyrrhizinate, Peak 46 corresponding to vitexin, and Peak 49 corresponding to imperatorin.

5. The quality inspection method according to claim 2, characterized in that: The characteristic peaks in the fingerprint spectrum use baicalin peaks as reference peaks, and the relative retention times of the 16 chromatographic peaks are 0.322, 0.419, 0.431, 0.712, 0.727, 0.741, 0.777, 0.814, 0.879, 1.000, 1.113, 1.279, 1.410, 1.777, 1.845, and 2.178, respectively.

6. The quality inspection method according to claim 2, characterized in that: The quality evaluation in S3 is performed by importing the fingerprint into the traditional Chinese medicine chromatographic fingerprint similarity evaluation software and performing similarity evaluation with the control spectrum.

7. Use of the nasal drops quality detection method according to any one of claims 1 to 6 in the quality control of Artemisia scutellariae nasal drops.

8. The use according to claim 7, characterized in that: The quality detection method includes a fingerprint spectrum combined with a multi-component content determination method, and is applied to the quality control of Chinese medicinal raw materials or solid pharmaceutical preparations containing six herbs: Artemisia argyi, ephedra, Xanthium sibiricum, Scutellaria baicalensis, Licorice and Angelica dahurica.

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