CN-122017088-A - UPLC-based construction method of characteristic spectrum of xipayi Gum fixing liquid and method for measuring content of characteristic components of UPLC-based construction method

CN122017088ACN 122017088 ACN122017088 ACN 122017088ACN-122017088-A

Abstract

The invention provides a UPLC-based construction method of a characteristic spectrum of a xipanyi gum-securing liquid and a method for measuring the content of characteristic components thereof. The construction method comprises the steps of taking gallic acid, methyl gallate, benzoic acid, 1,2,3, 6-tetragalloyl glucose and 1,2,3,4, 6-pentagalloyl glucose as reference substances, respectively adding water for dissolution to obtain reference substance solutions, mixing xipanyi gum fixing liquid with water, filtering to obtain test substance solutions, placing the test substance solutions and various reference substance solutions in an ultra-high performance liquid chromatograph, measuring to obtain test substance patterns and reference substance patterns, and establishing characteristic patterns by adopting a traditional Chinese medicine chromatographic fingerprint similarity evaluation system. The characteristic spectrum has good reproducibility and batch consistency and can be used as a standard for overall quality evaluation. The construction method can also be used for measuring the content of the characteristic components, and realizes the integrated detection of the construction of the characteristic spectrum and the measurement of the content of multiple indexes.

Inventors

CHEN JINCHENG
WANG CHANGHONG
MA XUAN
ZHAO LU
Qin Huaxia
CHENG XUEMEI
LI MANLIN
HUANG HAILONG

Assignees

新奇康药业股份有限公司

Dates

Publication Date: 20260512
Application Date: 20260408

Claims (10)

1. A construction method of a characteristic map of a xipanyi gingival strengthening solution, which is characterized by comprising the following steps: a) Preparing reference solution by using gallic acid, methyl gallate, benzoic acid, 1,2,3, 6-tetragalloyl glucose and 1,2,3,4, 6-pentagalloyl glucose as reference, and dissolving in water to obtain the reference solution; wherein the reference substance solution comprises gallic acid solution, methyl gallate solution, benzoic acid solution, 1,2,3, 6-tetragalloyl glucose solution and 1,2,3,4, 6-pentagalloyl glucose; b) Preparing a test solution, namely mixing the xipanyi gum fixing liquid with water and then filtering to obtain the test solution; c) And (3) placing the sample solution and the plurality of reference substance solutions in an ultra-high performance liquid chromatograph, determining to obtain a sample pattern and a reference substance pattern, and establishing the characteristic pattern by adopting a traditional Chinese medicine chromatographic fingerprint similarity evaluation system.
2. The method according to claim 1, wherein the concentration of the gallic acid solution is 0.8-1.0 mg/mL; Preferably, the concentration of the methyl gallate solution is 0.3-0.6 mg/mL; preferably, the concentration of the benzoic acid solution is 0.3-0.6 mg/mL; preferably, the concentration of the 1,2,3, 6-tetragalloyl glucose solution is 0.4-0.6 mg/mL; Preferably, the concentration of the 1,2,3,4, 6-pentagalloylglucose solution is 0.4-0.6 mg/mL.
3. The method of claim 1, wherein the chromatographic conditions of the ultra-high performance liquid chromatograph include: the chromatographic column is Waters ACQUITY UPLC HSS T, the specification is 2.1X100 mm, the grain diameter is 1.8 μm, the inorganic acid is used as mobile phase A, the acetonitrile is used as mobile phase B, the flow rate is 0.3-0.5 mL/min, the column temperature is 33-38 ℃, the sample injection amount is 1-2 mu L, and the gradient elution is carried out; preferably, the elution conditions of the gradient elution are: The volume fraction of the mobile phase A is reduced from 97% to 92% after 0-4 min, the volume fraction of the mobile phase B is increased from 3% to 8%, and the detection wavelength is 240 nm; 4-6.2 min, the volume fraction of the mobile phase A is reduced from 92% to 90%, the volume fraction of the mobile phase B is increased from 8% to 10%, and the detection wavelength is 240 nm; 6.2-14.6 min, the volume fraction of the mobile phase A is reduced from 90% to 85%, the volume fraction of the mobile phase B is increased from 10% to 15%, and the detection wavelength is 240 nm; 14.6-20.8 min, the volume fraction of the mobile phase A is reduced from 85% to 82%, the volume fraction of the mobile phase B is increased from 15% to 18%, and the detection wavelength is 275nm; and 20.8-30 min, the volume fraction of the mobile phase A is reduced from 82% to 70%, the volume fraction of the mobile phase B is increased from 18% to 30%, and the detection wavelength is 275 nm.
4. A method of construction according to claim 3, wherein the mineral acid comprises phosphoric acid; Preferably, the inorganic acid has a mass-volume concentration of 0.1% to 0.3%; preferably, the volume fraction of the cetirizine gum fixing solution in the test solution is 15% -25%.
5. A characteristic map of a xipanyi gingival strengthening solution constructed by the construction method of any one of claims 1-4; preferably, 7 characteristic peaks are included in the characteristic spectrum, wherein the peak 1 is the peak of gallic acid, the peak 4 is the peak of benzoic acid, the peak 5 is the peak of 1,2,3, 6-tetragalloylglucose, the peak 6 is the peak of 1,2,3,4, 6-pentagalloylglucose, the peak 7 is pentagalloylglucose, the peak 8 is the first unknown peak, and the peak 9 is the second unknown peak; Preferably, the peak No. 4 is taken as a locating peak, the relative retention time is 1.000, and the relative retention time of the other 6 characteristic peaks is as follows: the relative retention time of the No.1 peak is 0.123-0.151; the relative retention time of the No. 5 peak is 1.076-1.315; the relative retention time of the No. 6 peak is 1.361-1.663; the relative retention time of the peak 7 is 1.477-1.805; the relative retention time of the peak 8 is 1.642-2.006; the relative retention time of the No. 9 peak is 1.671-2.043; Further preferably, the relative peak area is 1.000 with respect to the peak No. 4, and the relative peak areas of the other 6 characteristic peaks are: The relative peak area of the No. 1 peak is 0.453-0.854; The relative peak area of the No. 5 peak is 0.436-0.778; The relative peak area of the No. 6 peak is 0.115-0.375; the relative peak area of the No. 7 peak is 0.444-0.854; the relative peak area of the No. 8 peak is 0.223-0.407; the relative peak area of the No. 9 peak is 0.205-0.432.
6. A method for detecting the quality of a xipanyi gingival retraction fluid, which is characterized by comprising the following steps: the construction method of any one of claims 1-4 is utilized to construct a characteristic spectrum of the to-be-detected xipanyi fixed gum solution, obtain the characteristic spectrum of the to-be-detected xipanyi fixed gum solution, and compare the characteristic spectrum of the to-be-detected xipanyi fixed gum solution with a comparison characteristic spectrum obtained under the same characteristic spectrum detection condition to realize quality detection of the to-be-detected xipanyi fixed gum solution.
7. The method according to claim 6, wherein the number of characteristic peaks in the control characteristic map is 7; Wherein, the peak 1 is the peak of the gallic acid, the peak 4 is the peak of the benzoic acid, the peak 5 is the peak of the 1,2,3, 6-tetragalloyl glucose, the peak 6 is the peak of the 1,2,3,4, 6-pentagalloyl glucose, the peak 7 is pentagalloyl glucose; Preferably, the peak No. 4 is taken as a locating peak, the relative retention time is 1.000, and the relative retention time of the other 6 characteristic peaks is as follows: the relative retention time of the No.1 peak is 0.123-0.151; the relative retention time of the No. 5 peak is 1.076-1.315; the relative retention time of the No. 6 peak is 1.361-1.663; the relative retention time of the peak 7 is 1.477-1.805; the relative retention time of the peak 8 is 1.642-2.006; the relative retention time of the No. 9 peak is 1.671-2.043; Further preferably, the relative peak area is 1.000 with respect to the peak No. 4, and the relative peak areas of the other 6 characteristic peaks are: The relative peak area of the No. 1 peak is 0.453-0.854; The relative peak area of the No. 5 peak is 0.436-0.778; The relative peak area of the No. 6 peak is 0.115-0.375; the relative peak area of the No. 7 peak is 0.444-0.854; the relative peak area of the No. 8 peak is 0.223-0.407; the relative peak area of the No. 9 peak is 0.205-0.432.
8. A method for measuring the content of characteristic components of a xipanyi gum fixing solution is characterized in that the characteristic components are selected from any one or more of gallic acid, benzoic acid, 1,2,3, 6-tetragalloylglucose or 1,2,3,4, 6-pentagalloylglucose; the assay method comprises: 1) Placing the reference substance solution and the sample solution into an ultra-high performance liquid chromatograph for chromatographic detection to obtain a reference substance map and a sample map; 2) Comparing the reference substance spectrum with the sample spectrum to obtain a common peak; 3) Calculating the peak area of a common peak between the reference substance spectrum and the sample spectrum; 4) Calculating the content of the characteristic components in the sample solution by adopting an external standard method; Wherein the preparation method of the sample solution is the preparation method of the sample solution in the construction method of any one of claims 1 to 4; the preparation method of the reference substance solution comprises the steps of dissolving the characteristic components by adopting water to obtain the reference substance solution.
9. The method according to claim 8, wherein the chromatographic condition of the ultra-high performance liquid chromatograph is the chromatographic condition in the construction method according to claim 3 or 4.
10. Use of the method for constructing a characteristic pattern of a xipanyi gingival retraction fluid according to any one of claims 1-4 or the characteristic pattern according to claim 5 or the quality detection method according to claim 6 or 7 or the assay method according to claim 8 or 9 for quality control of a xipanyi gingival fluid.

Description

UPLC-based construction method of characteristic spectrum of xipayi Gum fixing liquid and method for measuring content of characteristic components of UPLC-based construction method Technical Field The invention relates to the technical field of medicine detection, in particular to a UPLC-based construction method of a characteristic spectrum of a xipayi Gum fixing liquid and a characteristic component content determination method thereof. Background The xipayi Gum fixing liquid has the effects of strengthening teeth, fixing gum, clearing blood, relieving pain, relieving sore throat and deodorizing, and is clinically mainly used for treating oral diseases such as periodontitis, gingivitis, canker sore and the like. Currently, the quality standard of the Xepai Gum fixing liquid is mainly gallic acid content measurement and identification, a High Performance Liquid Chromatography (HPLC) method is established such as Wu Haodong and the contents of gallic acid (GALLIC ACID, GA) and ellagic acid (ELLAGIC ACID, EA) in the Xepai Gum fixing liquid slag are simultaneously measured, a large amount of active ingredient residues in the gallic slag are found, the method has a large utilization value, and a method for simultaneously measuring gallic acid, 1-galloylglucose (1-GALLIC ACID glucose, 1-GG), methyl gallate (METHYL GALLATE, MG), 1,2,3, 6-tetragalloylglucose (1, 2,3,6-tetragalacylglucose, teGG) and 1,2,3,4,6-pentagalloylglucose (1, 2,3,4,6-pentagalloylglucose, PGG) in the Xepai Gum fixing liquid by using HPLC is reported such as Huo Shixia, and the problem of synchronously measuring a plurality of gallic tannins in the gallic acid is solved. However, when preparing a sample solution, methanol is adopted to treat the xipanyi gum fixing liquid in the research, so that the polyphenol components in the xipanyi gum fixing liquid undergo methyl esterification reaction in the sample treatment process, the generation of methyl gallate is promoted, the quality and the original appearance of the preparation cannot be truly reflected, and Peng Yan is used for measuring the content of gallic acid in the xipanyi gum fixing liquid by using a differential spectrophotometry, so that the method is simple, quick and low in cost. However, the method only measures single components of gallic acid, is difficult to comprehensively characterize the quality of the gallic acid, and Qin Yi is strong in using an ultraviolet spectrophotometry to measure the content of total tannins in the Xipayi Gum liquid, but the method has no separation function and has poor specificity. The characteristic spectrum is an important technical means for evaluating the quality of the traditional Chinese medicine, and is combined with cluster analysis and orthorhombic partial least square method discriminant analysis, and the characteristic spectrum is focused on selecting representative key characteristic information in the spectrum, so that the characteristic spectrum is used as an important identification method for the quality control of the traditional Chinese medicine, and the integrity and the ambiguity characteristic of the traditional Chinese medicine can be fully reflected. The characteristic spectrum becomes an ideal traditional Chinese medicine quality evaluation means under the condition that the substance basis is not clear or the price of a reference substance is high. Ultra high performance liquid chromatography (UPLC) has better separation effect, peak capacity and sensitivity than High Performance Liquid Chromatography (HPLC). Therefore, based on the problems of single index, solvation effect of sample pretreatment and insufficient integrity in the above study on the quality control of the Sipay gum-securing liquid, the study characterizes the quality of the Sipay gum-securing liquid from the whole level by establishing an overall quality control strategy of an ultra-high performance liquid chromatography-characteristic spectrum and a multi-index component content measuring method. Disclosure of Invention The invention mainly aims to provide a UPLC-based construction method of a characteristic spectrum of a xipanyi gum-fixing liquid and a characteristic component content measurement method thereof, so as to solve the problem of single quality control index of the xipanyi gum-fixing liquid in the prior art. In order to achieve the above object, according to a first aspect of the present invention, there is provided a method for constructing a characteristic map of a xipanyi gingival retraction fluid, the method comprising: a) Preparing reference substance solutions by respectively dissolving gallic acid, methyl gallate, benzoic acid, 1,2,3, 6-tetragalloyl glucose and 1,2,3,4, 6-pentagalloyl glucose in water to obtain reference substance solutions, wherein the reference substance solutions comprise gallic acid solution, methyl gallate solution, benzoic acid solution, 1,2,3, 6-tetragalloyl glucose solution and 1,2,3,4, 6-pentagalloyl glucose;