CN114563496B - Quantitative fingerprint analysis method for components in ginger, ginger and pinellia tuber percolate - Google Patents
Quantitative fingerprint analysis method for components in ginger, ginger and pinellia tuber percolate Download PDFInfo
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Abstract
The invention discloses a quantitative fingerprint spectrum analysis method for components in ginger and pinellia ternate percolate. The method comprises the following steps: (1) preparing a mixed solution of a test sample solution and a reference substance; (2) Analyzing the reference substance mixed solution and the sample solution by using a high performance liquid chromatograph to obtain a quantitative fingerprint of components in the ginger and pinellia tuber percolate. Wherein the chromatographic conditions are as follows: the instrument adopts a high performance liquid chromatography system, and a Waters Atlantis T3 chromatographic column (100 mm multiplied by 2.1mm,3.0 mu m) is adopted as a chromatographic column; a DAD detector is adopted; the flow rate is 0.30mL/min, the column temperature is 25 ℃, and the sample injection volume is 3-5 muL; pure water and acetonitrile are used as mobile phases; (3) Chromatographic peaks existing in chromatograms of ginger and pinellia ternate percolates of different batches are selected as common peaks, the component content of the percolates of different batches is calculated, the similarity is calculated, and scientific basis and technical support are provided for quality control of Jiang Jiang pinellia ternate percolates. The method makes up the deficiency of the detection method of the chemical components of the production intermediate of the bupleurum tenue capsule.
Description
Technical Field
The invention relates to a detection method of traditional Chinese medicines, and particularly relates to a quantitative fingerprint analysis method of components in ginger, ginger and pinellia tuber percolate.
Background
The Xiaochaihu capsule is a compound preparation prepared by seven medicinal materials of bupleurum, scutellaria, liquorice, codonopsis pilosula, chinese date, ginger pinellia and the like through decoction, percolation, concentration and other processes, and can be used for treating exogenous diseases, pathogenic factors attacking shaoyang syndrome, and symptoms of alternating cold and heat, fullness in chest and hypochondrium, inappetence, dysphoria with joy and vomiting, bitter taste in mouth, dry throat and the like. The midbody of the bupleurum tenue capsule comprises percolate of two medicinal materials such as ginger, ginger pinellia and the like, mixed decoction of five medicinal materials such as bupleurum, scutellaria, liquorice, codonopsis pilosula, chinese date and the like, and various concentrated solutions.
The internationally recognized concept of "quality-derived design" for pharmaceuticals suggests that the center of gravity for drug quality control should be shifted from the final product to the starting materials, intermediates, and pharmaceutical processes. Therefore, the quality control of the production intermediate of the bupleurum tenue capsule should be taken into consideration. However, the current research on the quality control of the bupleurum tenue capsules is only limited to finished products and medicinal materials, and the research on the quality control method of the intermediate is not reported.
In recent years, the magnitude transfer research from medicinal materials to decoction pieces to intermediates to preparations is widely used for the development of classical famous Chinese medicines and formula granules. By observing the quantity change of index components from medicinal materials to decoction pieces to intermediates to preparations, the quality standard of the intermediates can be determined, and illegal addition can be prevented from the perspective of supervision. In order to determine the value transfer rule of the index component, it is necessary to establish an analysis method of the index component in the intermediate. The analysis method can also be used for pharmaceutical process control and the like. The chemical composition of the intermediate is sometimes simpler than that of the formulation, in which case it is contemplated to use relatively easily achievable analytical conditions.
The percolates of ginger and ginger pinellia are important intermediates of the bupleurum tenue capsule. The intermediate is prepared by mixing rhizoma Zingiberis recens and rhizoma Pinelliae Preparata, soaking in 70% ethanol for 24 hr, and slowly percolating according to 2020 version of Chinese pharmacopoeia (one part). At present, no literature reports about the mass analysis method of the intermediate.
The invention aims to develop a quantitative fingerprint detection method for embodying the components in the ginger and pinellia ternate percolate, and lays a good foundation for further improving the quality consistency of bupleurum tenue capsule products by enhancing the quality control of the ginger and pinellia ternate percolate intermediate.
Disclosure of Invention
The invention mainly aims to provide a quantitative fingerprint analysis method for components in ginger and ginger pinellia ternate percolate, and the method can make up the deficiency of a detection method for chemical components of a bupleurum tenue capsule production intermediate, so that the quality of the ginger and ginger pinellia ternate percolate can be detected and evaluated more comprehensively, and the quality control level of the bupleurum tenue capsule production intermediate can be improved.
In order to achieve the purpose, the invention adopts the following technical scheme:
one of the objects of the present invention: the ginger and pinellia ternate percolate is prepared by soaking ginger and ginger with 70% ethanol as solvent for 24 hr, slowly percolating, and collecting the percolate. The method for establishing the fingerprint comprises the following steps:
(1) Preparing a reference substance mixed solution and a test solution:
preparation of control mixed solution: precisely weighing uridine, adenine, 5-hydroxymethylfurfural and guanosine reference substances, placing the uridine, adenine, 5-hydroxymethylfurfural and guanosine reference substances into a volumetric flask, fixing the volume to a scale by using pure water, and shaking up to prepare a mixed reference substance solution containing 12.5 mu g/mL of uridine, 23.9 mu g/mL of adenine, 51.5 mu g/mL of 5-hydroxymethylfurfural and 21.5 mu g/mL of guanosine.
Preparation of a test solution: weighing 1.5mL of ginger and pinellia tuber percolate, centrifuging and concentrating at 35 ℃ until the percolate is dried, adding 1.0mL of pure water for redissolving, centrifuging at 12000rpm for 10min, and taking supernatant fluid to obtain the ginger and pinellia tuber percolate.
(2) High performance liquid chromatography assay
Injecting the reference substance mixed solution and the test substance solution in the step (1) into a liquid chromatography for analysis and determination, wherein the apparatus is as follows: an Agilent 1100 high performance liquid chromatography system (Agilent technologies, usa), a degasser (G1322A), a quaternary pump (G1311A), an autosampler (G1313A), a column oven (G1316A), a diode array detector (DAD G1315B) and an Agilent chromatography workstation (ChemStation version b.04.03); the chromatographic conditions are as follows: waters Atlantis T3 chromatography column (100 mm. Times.2.1mm, 3.0 μm); the mobile phase A is pure water, and the mobile phase B is acetonitrile; gradient elution is adopted, and the elution conditions are as follows: 0-7 min, the volume fraction of the mobile phase B is 0%; 7-15 min, the volume fraction of the mobile phase B is changed to 0-10%; 15-16 min, the volume fraction of the mobile phase B is changed to 10-12%; the volume fraction of the mobile phase B is changed to 12-25 percent within 16-20 min; the volume fraction of the mobile phase B is changed to 25-95% in 20-21 min; 21-26 min, wherein the volume fraction of the mobile phase B is 95%; flow rate: 0.30mL/min; column temperature: 25 ℃; detection wavelength: 260 nm-262 nm, and the injection volume is 3 mu L-5 mu L.
(3) Establishing quantitative fingerprint and evaluating quality
Taking 10 different batches of ginger, ginger and pinellia tuber percolates to prepare 10 different batches of test solution according to the step (1), carrying out high performance liquid chromatography according to the step (2), and recording a chromatogram; and determining a common peak according to the chromatograms of the 10 different batches by using the chromatographic peaks common to the different batches, and generating the comparison fingerprint. Meanwhile, the quality of the ginger and ginger pinellia tuber percolate is evaluated through the established fingerprint similarity and the measured content of the 4 components.
Preferably, the detection wavelength in the step (2) is 260nm, and under the detection wavelength, chromatographic peak separation is better, the baseline is more stable, the peak area is larger, and the response is better.
Preferably, the injection volume in step (2) is 3 μ L, and the peak area is larger and the response is better at this injection volume.
Preferably, the quantitative detection indexes in the step (3) are as follows: uridine, adenine, 5-hydroxymethylfurfural, guanosine. Under the detection conditions of the present invention, the above four components can be sufficiently separated.
Preferably, the number of common peaks in step (3) is 14.
The second object of the present invention is: provides a method for measuring the content of chemical components in ginger and ginger pinellia percolate.
The chromatographic conditions were as follows: the instrument comprises the following steps: agilent 1100 hplc; a chromatographic column: waters Atlantis T3 column (100 mm. Times.2.1mm, 3.0. Mu.m); the mobile phase A is pure water, and the mobile phase B is acetonitrile; gradient elution is adopted, and the elution conditions are as follows: 0-7 min, wherein the volume fraction of the mobile phase B is 0%; 7-15 min, the volume fraction of the mobile phase B is changed to 0-10%; 15-16 min, the volume fraction of the mobile phase B is changed to 10-12%; 16-20 min, the volume fraction of the mobile phase B is changed to 12-25%; 20-21 min, the volume fraction of the mobile phase B is changed to 25-95%; 21-26 min, wherein the volume fraction of the mobile phase B is 95%; flow rate: 0.30mL/min; column temperature: 25 ℃; detection wavelength: 260 nm-262 nm and the injection volume is 3-5 mu L.
Precisely weighing appropriate amounts of uridine, adenine, 5-hydroxymethylfurfural and guanosine reference substances, respectively adding pure water to dilute the reference substances into solutions with different concentrations, injecting the solutions into a high performance liquid chromatography to analyze, drawing a standard curve by taking the concentration of each compound as a horizontal coordinate and a peak area as a vertical coordinate, calculating a regression equation, and calculating the content of index components in samples of different batches.
The third object of the present invention is: provides an application of a quantitative fingerprint analysis method of components in ginger and pinellia tuber percolate in quality control of the ginger and pinellia tuber percolate.
Introducing chromatograms of test solution of 10 batches of ginger and pinellia ternate percolates into a traditional Chinese medicine chromatogram fingerprint similarity evaluation system (2012 edition of the national pharmacopoeia committee), generating comparison fingerprints, introducing the chromatograms of different batches of ginger and pinellia ternate percolates into similarity evaluation software, and calculating the similarity.
The invention has the beneficial effects that:
the invention selects uridine, adenine, 5-hydroxymethyl furfural and guanosine as reference substances to carry out quantitative detection, and can realize the quality control of the intermediate of the ginger and pinellia ternate percolate. The invention establishes the quantitative fingerprint spectrum of part of strong polar components in the ginger and ginger processed pinellia percolate for the first time, and the quantitative fingerprint spectrum can make up the deficiency of the existing detection method of the chemical components of the production intermediate of the bupleurum tenue capsule, so that the quality of the ginger processed pinellia and ginger processed percolate can be detected and evaluated more comprehensively, and the quality control level of the production intermediate can be improved. Among them, uridine controls energy balance and adjusts body temperature, and is one of the effective components of Xiao chai Hu capsule with the efficacy of relieving exterior syndrome and bringing down fever. Guanosine can enhance the release of macrophages to regulate the immunologic function, adenine can promote the proliferation of white blood cells, and the guanosine is a medicinal component for treating exogenous diseases by using the bupleurum tenue capsules. Rhizoma Pinelliae Preparata contains more uridine, adenine and guanosine. Therefore, it is necessary to detect three components of uridine, adenine and guanosine in the percolate of ginger and pinellia ternate, and the method can be further used for the quantity value transmission analysis of the bupleurum tenue capsules.
The 2020 edition (Chinese pharmacopoeia (one) stipulates that processing method of ginger processed pinellia tuber is to boil pinellia tuber, ginger decoction and alum thoroughly, and the method is heating processing. However, other processing methods are available in the market, and the whole process is not heated. For example, the alum-pickled ginger juice mixing method is a method of mixing pinellia ternate, alum powder and ginger juice uniformly at normal temperature. Because the heating conditions are different in different processing methods, the amount of the 5-hydroxymethylfurfural generated by degrading the carbohydrate in the ginger processed pinellia prepared according to the pharmacopoeia method is large. The content of the 5-hydroxymethyl furfural in the ginger and the ginger processed pinellia percolate can be detected to simply judge whether the ginger processed pinellia is heated or not, and then the processing method of the ginger processed pinellia is deduced. And 5-hydroxymethyl furfural can play an anti-inflammatory role by inhibiting related proteins of multiple pathways, and has pharmacological activity. Therefore, the content of the 5-hydroxymethylfurfural in the percolate needs to be detected.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, are included to provide a further understanding of the invention, and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description, serve to explain the invention without undue limitation of the invention.
FIG. 1 shows HPLC finger prints of percolate of ginger and pinellia tuber of different batches.
FIG. 2 shows HPLC control spectra of the percolate of rhizoma Zingiberis recens and rhizoma Pinelliae.
FIG. 3 is a graph of the percolate of the mixed control and ginger and pinellia tuber.
Detailed Description
It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the invention. Furthermore, it will be further understood that the terms "comprises" and/or "comprising," when used in this specification, specify the presence of stated features, steps, operations, and/or combinations thereof.
1 laboratory instruments and materials
1.1 instruments
Agilent 1100 high performance liquid chromatography system (Agilent technologies, usa), preparation degasser (G1322A), quaternary pump (G1311A), autosampler (G1313A), column oven (G1316A), diode array detector (DAD G1315B) and Agilent chromatography workstation (ChemStation b.04.03 edition). Electronic balance (AB 204-N, mettler Toledo). A small high speed centrifuge (AB 204-N, mettler Toledo). The fingerprint similarity evaluation software is a Chinese medicine chromatogram fingerprint similarity evaluation system (2012 edition of the national pharmacopoeia committee).
1.2 reagents
Acetonitrile, methanol (Merck, germany), absolute ethanol (batch No.: 20200710, yonghua chemical Co., ltd.), and water as Mili-Q ultrapure water.
Uridine control (uridine, purity: 99%, sigma, lot number: SLCB 8621), adenine control (purity: 99%, sigma, lot number: WXBD 1877V), 5-hydroxymethylfurfural control (purity: 98%, shanghai Po pharmaceutical science and technology development Limited, lot number: 170910), guanosine control (guanosine, purity: 98%, lot number: BCCB 9660). The ginger of different batches are purchased from the vegetable market or supermarket. 3 batches of ginger processed pinellia were purchased from Jiangsu Kaiyuan Chinese medicine, baohe Tang (Bozhou) pharmacy, and Jiaxing Oriental decoction pieces, with lots of 210203, C0131210031 and 210501, respectively.
2 conditions of the experiment
2.1 chromatographic conditions
Waters Atlantis T3 chromatography column (100 mm. Times.2.1mm, 3 μm); an acetonitrile (B) -water (a) mobile phase system was used, with gradient elution. The elution conditions were: 0-7 min, the volume fraction of the mobile phase B is 0%; 7-15 min, the volume fraction of the mobile phase B is changed to 0-10%; 15-16 min, the volume fraction of the mobile phase B is changed to 10-12%; 16-20 min, the volume fraction of the mobile phase B is changed to 12-25%; 20-21 min, the volume fraction of the mobile phase B is changed to 25-95%; 21-26 min, and the volume fraction of the mobile phase B is 95%. After each needle was removed, the column was equilibrated by running with 100% pure water for 14 min. Flow rate: 0.30mL/min; column temperature: 25 ℃; detection wavelength: 260nm, injection volume 3 uL.
2.2 preparation of control mix solution
Preparation of control mixed solution: precisely weighing uridine, adenine, 5-hydroxymethylfurfural and guanosine reference substances, placing the uridine, adenine, 5-hydroxymethylfurfural and guanosine reference substances into a volumetric flask, fixing the volume to a scale by using a pure water solution, and shaking the volume uniformly to prepare a mixed reference substance solution containing 12.5 mu g/mL of uridine, 23.9 mu g/mL of adenine, 51.5 mu g/mL of 5-hydroxymethylfurfural and 21.5 mu g/mL of guanosine. And (5) absorbing the reference solution, injecting into a high performance liquid chromatograph, and recording the chromatogram.
2.3 preparation of test solutions
Soaking rhizoma Pinelliae and rhizoma Zingiberis recens with 70% ethanol as solvent for 24 hr, slowly percolating, and collecting percolate. Weighing 1.5mL of ginger and pinellia tuber percolate, centrifuging at 35 ℃ and concentrating to dryness, adding 1.0mL of pure water for redissolving, centrifuging at 12000rpm for 10min, and taking supernatant for sample injection analysis. And absorbing the test solution, injecting the test solution into a high performance liquid chromatograph, and recording the chromatogram.
3 investigation of fingerprint spectrum research methodology
The methodological verification of the fingerprint mainly comprises the investigation of injection precision, repeatability and sample stability.
3.1 sample introduction precision test
The same sample solution is continuously injected for 6 times. And (3) taking uridine as a No. 11 peak as a control peak, and respectively calculating the ratio of the retention time of each common peak to the reference peak to the peak area to obtain the relative retention time of each common peak shown in table 1 and the relative peak area shown in table 2. The relative retention time RSD of each common peak is 0.06% -0.17%, and the relative peak area RSD is 1.00% -4.13%, which indicates that the precision of the instrument is good.
TABLE 1 relative retention times from precision tests
TABLE 2 relative peak areas from precision tests
3.2 method repeatability test
Six samples of the test solution prepared in parallel are taken and respectively subjected to sample injection analysis. And (3) taking uridine as a No. 11 peak as a control peak, and respectively calculating the ratio of the retention time of each common peak to the reference peak to the peak area to obtain the relative retention time of each common peak shown in table 3 and the relative peak area shown in table 4. The relative retention time RSD of each common peak is 0.07-0.30%, and the relative peak area RSD is 0.13-3.69%, which indicates that the method has good repeatability.
TABLE 3 relative retention times from repeated experiments
TABLE 4 relative peak areas from repeatability tests
3.3 sample stability test
And (3) taking the same sample solution, carrying out sample injection analysis on 0,3,6,9, 12, 18 and 24h, taking uridine No. 11 peak as a reference peak, and respectively calculating the retention time of each common peak and the reference peak and the ratio of the peak area to obtain the relative retention time of each common peak shown in table 5 and the relative peak area shown in table 6. The relative retention time RSD of each common peak is 0.13-0.93%, and the relative peak area RSD is 0.91-4.01%, which indicates that the stability of the test solution is good within 24 h.
TABLE 5 relative retention times from stability testing
TABLE 6 relative peak areas from stability test
Establishment and application example of fingerprint of ginger pinellia tuber percolate
HPLC (high performance liquid chromatography) images of different batches of ginger, ginger and pinellia tuber percolate samples and sample samples of the same batch of percolate at different percolation times are integrated according to the same method, and then are respectively introduced into software of a traditional Chinese medicine chromatogram fingerprint similarity evaluation system (2012) in an AIA format. And (4) performing chromatographic peak matching, wherein S1 is taken as a reference spectrum, and the time window width is 0.1 by using a median method. And (5) performing multipoint correction and performing Mark peak matching. Generating percolation HPLC finger print and control finger print of rhizoma Zingiberis recens and rhizoma Pinelliae of different batches, as shown in figure 1 and figure 2 respectively.
4.1 calibration of common peaks
The percolate sample is calibrated to 14 common peaks, and 7 components in the percolate sample are identified by comparison of a reference substance. Wherein, peaks No. 3, 5, 6, 8, 10, 11 and 14 are guanine, xanthine, guanosine, adenine, 5-hydroxymethylfurfural, uridine and adenosine respectively. The peak 11 (uridine) with moderate retention time, stable peak area, good separation from adjacent chromatographic peaks and stable baseline is selected as a reference peak.
4.2 evaluation of similarity of percolate of ginger and pinellia Tuber and application example
And (3) introducing the percolate samples of 10 batches respectively by using a traditional Chinese medicine chromatogram fingerprint similarity evaluation system (2012) and calculating the fingerprint similarity. The fingerprint similarity results are shown in table 7, the similarity is between 0.937 and 0.984, and the content difference of the compounds in different batches of ginger and ginger pinellia percolate is small.
TABLE 7 fingerprint similarity
If the enterprise sets the internal control standard of the similarity of the fingerprint map to be not less than 0.90. Preparing a new batch of ginger and pinellia ternate percolate, preparing a sample and detecting to obtain a fingerprint. The similarity of the fingerprint is calculated and found to be 0.888 which is lower than the internal control standard. The quality of the percolate of the batch is considered to be greatly different from that of the percolate of the normal batch, and the subsequent production is not recommended to be released.
5 content determination methodological verification
The chromatographic condition for measuring the content of the nucleoside component in the percolate is the same as the chromatographic method of the fingerprint, and the methodological verification of the content measurement comprises the investigation of the linear range, precision and stability of each content measurement component and the sample adding and recycling experiment. 4 components (uridine, adenine, 5-hydroxymethylfurfural and guanosine) with high map response of the intermediate are selected for content determination, and the map of the mixed reference substance and the ginger and ginger pinellia ternate percolate is shown in figure 3.
5.1 Linear and Range inspection
Mixed reference substance solutions with different concentrations are taken, and 3 mu L of the mixed reference substance solutions are injected respectively for analysis. And (4) taking the peak area measured by each component as a vertical coordinate and the concentration as a horizontal coordinate to prepare a standard curve so as to obtain a linear regression equation and an analysis range. The detection limit and the quantification limit are respectively determined according to the concentration of each component when the peak signal-to-noise ratio is about 3:1 and 10. The regression equation, linear range, detection limit and quantification limit results for uridine, adenine, 5-hydroxymethylfurfural and guanosine are shown in Table 8.
TABLE 8 standard curve and linear range of ginger processed pinellia
5.2 sample introduction precision, repeatability and stability test
And (3) sample injection precision experiment: and taking the same sample solution, carrying out continuous sample introduction for 6 times, and calculating to obtain the RSD value of each component peak area. And (3) repeatability experiment: taking six parts of test solution prepared in parallel, respectively carrying out sample injection analysis, and calculating to obtain the RSD value of each component. Sample stability test: and (3) taking the same sample solution, performing sample injection analysis on 0,3,6,9, 12, 15, 18, 21 and 24 hours, and calculating to obtain the RSD value of each component peak area. The test results of sample injection precision, repeatability and stability are shown in table 9, wherein the precision, the repeatability and the stability RSD are all less than 5%, the requirements of Chinese pharmacopoeia are met, the method is good in linearity, and the sample solution is stable within 24 hours.
TABLE 9 ginger processed pinellia ingredient precision, repeatability, sample stability
5.3 sample application recovery test
Respectively taking 9 parts of sample solution with known content, dividing the sample solution into 3 groups, setting the concentration levels of high, medium and low, controlling the ratio of the addition amount of a reference substance to the amount of the component to be detected in the test sample to be respectively 1.5, 1.0 and 1. Precisely adding 0.8mL of test solution and a proper amount of reference solution, diluting to a constant volume in a 5mL volumetric flask, centrifuging at 12000rpm for 10min, and taking the supernatant for sample injection analysis. The sample-adding recovery experiment result is shown in table 10, the average recovery rate of each component meets the requirement, and the RSD value is less than 5%, so that the optimized method is accurate and reliable, and can be used for measuring the content of 4 strong-polarity components in the percolate of ginger pinellia ternate and ginger.
TABLE 10 sample recovery test results
5.4 content determination results and application examples
The quantitative component content measurement results of 21 sample solutions in the percolation process of ginger processed pinellia and ginger are shown in table 11. When ginger, ginger and pinellia tuber are percolated, the concentration of strong polar components (uridine, adenine, 5-hydroxymethylfurfural and guanosine) is reduced along with the increase of the percolation time.
If the enterprise regards the adenine content in the percolate as being lower than 1.0 mug/mL as the percolation end point, when the percolation is carried out for 509 minutes, the detection finds that the adenine content is 0.89 mug/mL and is already lower than 1.0 mug/mL, the percolation process of the ginger and pinellia ternata can be considered to reach the end point, and the percolation can be stopped. Similarly, criteria may be set for uridine, 5-hydroxymethylfurfural and guanosine for percolation process quality control.
TABLE 11 detection results of quantitative component contents of 21 test sample solutions in ginger, ginger and pinellia ternate percolation curve
Claims (4)
1. A quantitative fingerprint analysis method for components in ginger, ginger and pinellia tuber percolate is characterized by comprising the following steps:
(1) Preparation of control mixed solution and test solution
Preparing a mixed reference substance solution: taking uridine, adenine, 5-hydroxymethylfurfural and guanosine as reference substances, and preparing a reference substance mixed solution;
preparing a test solution: weighing ginger, ginger and pinellia tuber percolate, centrifuging, concentrating to dry, adding a solvent for redissolving, shaking uniformly, and centrifuging and filtering to obtain a test solution;
(2) High performance liquid chromatography assay
Injecting the reference substance mixed solution and the test substance solution in the step (1) into a liquid chromatography for analysis and determination, wherein the chromatographic conditions are as follows: a chromatographic column: waters Atlantis T3 chromatographic column, column length 100mm × inner diameter 2.1mm, particle size 3.0 μm; the mobile phase A is pure water, and the mobile phase B is acetonitrile; gradient elution is adopted, and the elution conditions are as follows: 0-7 min, the volume fraction of the mobile phase B is 0%; 7-15 min, the volume fraction of the mobile phase B is changed to 0-10%; 15-16 min, the volume fraction of the mobile phase B is changed to 10-12%; the volume fraction of the mobile phase B is changed to 12-25 percent within 16-20 min; 20-21 min, the volume fraction of the mobile phase B is changed to 25-95%; 21-26 min, the volume fraction of the mobile phase B is 95%; flow rate: 0.30mL/min; column temperature: 25 ℃;
(3) Establishing quantitative fingerprint and evaluating quality
Preparing a plurality of different batches of ginger, ginger and pinellia tuber percolates according to the method in the step (1) to obtain different batches of test solution, performing high performance liquid chromatography analysis according to the step (2), and recording a chromatogram; determining common peaks and quantitative detection indexes according to the obtained chromatograms of different batches; and evaluating the quality of the ginger and ginger pinellia tuber percolate according to the established fingerprint spectrum similarity and the content of each component.
2. The quantitative fingerprint analysis method for the components in the ginger and pinellia ternate percolate according to claim 1, wherein a solvent adopted in the process of adding the solvent for redissolution in the preparation process of a test solution is pure water.
3. The quantitative fingerprint analysis method for the components in the ginger and pinellia tuber percolate according to claim 1, wherein the chromatographic conditions further comprise: detection wavelength: 260 nm-262 nm and the injection volume is 3 mu L-5 mu L.
4. The quantitative fingerprint analysis method for the components in the ginger and pinellia tuber percolate according to claim 1, wherein the fingerprint contains 14 peaks in total.
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