CN111679011B - Fingerprint spectrum construction and detection method and device for radix pseudostellariae oligosaccharide - Google Patents
Fingerprint spectrum construction and detection method and device for radix pseudostellariae oligosaccharide Download PDFInfo
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Abstract
The invention discloses a method and a device for constructing and detecting a fingerprint of radix pseudostellariae oligosaccharide, wherein the method comprises the following steps: extracting radix Pseudostellariae oligosaccharide with ethanol as extraction solvent by ultrasonic extraction method to obtain test solution; and performing HPLC analysis on the test solution according to HPLC chromatographic conditions, performing chromatographic peak identification on the obtained HPLC chromatogram according to a control chromatogram, constructing a fingerprint by taking the common chromatographic peak as a common peak, and performing similarity calculation on the to-be-detected HPLC chromatogram of the radix pseudostellariae oligosaccharide according to the fingerprint. The oligosaccharide component quality of the radix pseudostellariae containing medicinal material is effectively detected, and the quality monitoring of the medicinal material is facilitated.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicine analysis, in particular to a method and a device for constructing and detecting a fingerprint spectrum of radix pseudostellariae oligosaccharide.
Background
The statements in this section merely provide background information related to the present disclosure and may not necessarily constitute prior art.
Radix Pseudostellariae is dry root tuber of Pseudostellaria heterophylla (Miq.) Pax ex Pax et Hoffm, belonging to family Caryophyllaceae, and has sweet and slightly bitter taste and mild nature. Spleen and lung meridian entered; has effects of invigorating qi, invigorating spleen, promoting fluid production, and moistening lung, and can be used for treating spleen deficiency, fatigue, anorexia, asthenia after illness, deficiency of qi and yin, spontaneous perspiration, thirst, lung dryness, and tussiculation; modern researches show that radix pseudostellariae mainly contains cyclic peptide, triterpenoid saponin, sugar and other chemical components. In recent years, more and more researches show that the oligosaccharide contained in the traditional Chinese medicine is also an important active ingredient for the traditional Chinese medicine to exert the drug effect, for example, the oligosaccharide in the morinda officinalis has the activities of resisting depression, enhancing memory, resisting aging, improving reproductive function and the like; polygala tenuifolia oligosaccharide has antidepressant and neuroprotective activities; the rehmanniae radix oligosaccharide has effects of regulating sugar metabolism, enhancing mouse immunity, and promoting proliferation and differentiation of mouse bone marrow hematopoietic cells.
The radix pseudostellariae contains oligosaccharides such as maltose, raffinose, alpha-sophorose, ethanol-alpha-D-galactose and the like, and the plant source, shape, identification, inspection, extract and the like of the radix pseudostellariae are included in the first part of the Chinese pharmacopoeia of 2015 edition, but the inventor thinks that the quality of the components of the oligosaccharide of the radix pseudostellariae cannot be measured at present, the whole quality of medicinal materials is difficult to effectively control, the quality control of the medicinal materials is not facilitated, and the safety and the effectiveness of the medicine application cannot be ensured.
Disclosure of Invention
In order to solve the problems, the invention provides a method and a device for constructing and detecting a fingerprint of radix pseudostellariae oligosaccharide.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the invention provides a method for constructing a fingerprint spectrum of radix pseudostellariae oligosaccharide, which comprises the following steps:
extracting radix Pseudostellariae oligosaccharide with ethanol as extraction solvent by ultrasonic extraction method to obtain test solution;
and carrying out HPLC analysis on the sample solution according to HPLC chromatographic conditions, carrying out chromatographic peak identification on the obtained HPLC chromatogram according to a reference chromatogram, and constructing a fingerprint by taking the common chromatographic peak as a common peak.
In a second aspect, the invention provides a fingerprint spectrum detection method of radix pseudostellariae oligosaccharide, which comprises the following steps:
extracting the radix pseudostellariae oligosaccharide to be detected by using ethanol as an extraction solvent and adopting an ultrasonic extraction method to obtain a solution to be detected;
performing HPLC analysis on the solution to be detected according to HPLC chromatographic conditions to obtain a chromatogram of the HPLC to be detected;
and calculating and comparing the similarity of the HPLC chromatogram to be detected and the fingerprint spectrum according to the similarity.
In a third aspect, the invention provides a fingerprint spectrum detection device for radix pseudostellariae oligosaccharide, which comprises:
the solution preparation unit is used for respectively extracting the radix pseudostellariae oligosaccharide and the radix pseudostellariae oligosaccharide to be detected by using ethanol as an extraction solvent and adopting an ultrasonic extraction method to obtain a test solution and a solution to be detected;
the sample solution analysis unit is used for carrying out HPLC analysis on the sample solution according to HPLC chromatographic conditions, carrying out chromatographic peak identification on the obtained HPLC chromatogram according to a comparison chromatogram, and constructing a fingerprint by taking the common chromatographic peak as a common peak;
the to-be-detected solution analysis unit is used for carrying out HPLC analysis on the to-be-detected solution according to HPLC chromatographic conditions to obtain a to-be-detected HPLC chromatogram;
and the detection unit is used for calculating and comparing the similarity between the HPLC chromatogram to be detected and the fingerprint spectrum through the similarity.
Compared with the prior art, the invention has the beneficial effects that:
the method is used for establishing the oligosaccharide fingerprint of the radix pseudostellariae based on HPLC analysis, effectively representing the quality of oligosaccharide components of the radix pseudostellariae, detecting the quality of the medicinal material containing the radix pseudostellariae, effectively determining the quality controllability of the medicinal material, being beneficial to quality monitoring of the medicinal material, ensuring the safety and effectiveness of medication, having good repeatability and stability of a detection result and solving the problem of the deficiency of the existing quality control technology.
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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 exemplary embodiments of the invention and together with the description serve to explain the invention and not to limit the invention.
Fig. 1 is a flowchart of a method for constructing a fingerprint of oligosaccharide from radix pseudostellariae provided in embodiment 1 of the present invention;
FIG. 2 is an HPLC chromatogram of water extraction of radix Pseudostellariae provided in example 1 of the present invention;
fig. 3 is an HPLC chromatogram of the radix pseudostellariae medicinal material extracted with 50% ethanol provided in embodiment 1 of the present invention;
fig. 4 is an HPLC chromatogram of the radix pseudostellariae medicinal material extracted with 70% ethanol provided in embodiment 1 of the present invention;
fig. 5 is an HPLC chromatogram of ethanol extraction of radix pseudostellariae medicinal material provided in embodiment 1 of the present invention;
FIG. 6 is an HPLC chromatogram obtained by elution method 1 provided in example 1 of the present invention;
FIG. 7 is an HPLC chromatogram obtained by elution method 2 provided in example 1 of the present invention;
fig. 8 is a reference fingerprint of 12 batches of radix pseudostellariae medicinal materials provided in embodiment 1 of the present invention;
fig. 9 is a fingerprint of 12 batches of radix pseudostellariae medicinal materials provided in embodiment 1 of the present invention.
The specific implementation mode is as follows:
the invention is further described with reference to the following figures and examples.
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. As used herein, the singular forms "a", "an", and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise, and it should be understood that the terms "comprises" and "comprising", and any variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.
The embodiments and features of the embodiments of the invention may be combined with each other without conflict.
Example 1
As shown in fig. 1, this embodiment provides a method for constructing a fingerprint of a pseudostellaria root oligosaccharide, including:
s1: extracting radix Pseudostellariae oligosaccharide with ethanol as extraction solvent by ultrasonic extraction method to obtain sample solution;
s2: and carrying out HPLC analysis on the test solution according to HPLC chromatographic conditions, carrying out chromatographic peak identification on the obtained HPLC chromatogram according to a reference chromatogram, and constructing a fingerprint by taking the common chromatographic peak as a common peak.
In this embodiment, comparing the extraction efficiencies of the four solvents, i.e., water, 50% ethanol, 70% ethanol, and 100% ethanol, on the pseudostellaria root oligosaccharide specifically includes:
pulverizing radix Pseudostellariae into fine powder, and sieving with 60-80 mesh sieve to obtain radix Pseudostellariae powder. Taking 1g of medicinal powder, adding 8-40ml of extraction solvent which is water, 10-70% ethanol or 10-70% methanol, and extracting by ultrasonic extraction or heating reflux extraction for 1-3 times for 20-60min. Mixing extractive solutions, concentrating under reduced pressure to 0.8-1ml, adding 4-5 times volume of anhydrous ethanol, placing in refrigerator at 4 deg.C overnight to precipitate polysaccharide, and filtering supernatant with 0.24 microporous membrane to obtain test solution;
as shown in fig. 2 to 5, the extraction yield was the highest when water and 50% ethanol were used as the extraction solvent, but the amount of impurities such as polysaccharides and proteins extracted was large when water was used as the extraction solvent, and therefore, in this example, 50% ethanol is preferably used as the extraction solvent for the oligosaccharide of radix pseudostellariae.
In step S1, the preparation of the sample solution includes: precisely weighing 1.0g of radix Pseudostellariae powder, placing in a triangular flask with a plug, adding 20ml of 50% ethanol solution, ultrasonically extracting for 2 times (30 min each time), filtering the extractive solutions, mixing, concentrating under reduced pressure to 1ml, adding 4ml of anhydrous ethanol, mixing uniformly, placing in a refrigerator at 4 deg.C, and standing overnight; the supernatant is filtered through a 0.22 μm filter membrane to obtain a test solution.
Preparation of control solutions: precisely weighing 10mg of sucrose reference substance and 10mg of raffinose reference substance, placing into a 5ml volumetric flask, adding a proper amount of ultrapure water for dissolving, diluting to scale, and shaking uniformly to obtain a mixed reference substance solution.
In the step S2, HPLC chromatographic conditions comprise:
(1) A chromatographic column: hydrophilic columns such as amino columns, XAmide, XIon, and the like;
(2) Column temperature: 15 to 35 ℃;
(3) Flow rate: 0.8-1.0ml/min;
(4) Sample introduction amount: 0.1-1.5mg;
(5) A detector: an evaporative light scattering detector;
(6) The mobile phase and elution mode are any one of the following four:
(1) 0-30min,75-55% acetonitrile, 30-32min,55-10% acetonitrile, 32-35min,10% acetonitrile;
(2) 0-30min,80-50% acetonitrile, 30-32min,50-10% acetonitrile, 32-35min,10% acetonitrile;
(3) 0-40min,75-50% acetonitrile, 40-42min,50-10% acetonitrile, 42-45min,10% acetonitrile;
(4) 0-30min,90-50% acetonitrile, 30-32min,50-10% acetonitrile, 32-35min,10% acetonitrile.
Performing HPLC analysis on the sample solution, performing gradient elution according to the above chromatographic conditions, and performing HPLC chromatogram with sample amount of 5 μ l, as shown in FIG. 6-FIG. 7;
constructing a standard fingerprint spectrum according to the chromatogram peak identification of a comparison chromatogram obtained by a comparison product solution; the mass spectrometry of chemical components in the HPLC chromatogram of the radix pseudostellariae comprises the following steps:
analyzing 8 main chromatographic peaks in an HPLC chromatogram of the radix pseudostellariae extract by adopting HPLC-DAD-Q-TOF/MS, identifying a common peak by using a reference substance, identifying and determining 2 chromatographic peaks as cane sugar and raffinose, wherein the analysis result of a high-resolution mass spectrum is shown in a table 1;
TABLE 1 HPLC-DAD-Q-TOF/MS measurement results of main chemical components in radix Pseudostellariae
In this embodiment, the checking of the constructed standard fingerprint specifically includes:
(1) And (3) checking the precision:
preparation of a test solution: precisely weighing 1.0g of radix Pseudostellariae powder, placing in a triangular flask with plug, adding 20ml of 50% ethanol solution, ultrasonically extracting for 2 times, each time for 30min, filtering the extractive solutions, mixing, concentrating under reduced pressure to 1ml, adding 4ml of anhydrous ethanol, mixing, placing in a refrigerator at 4 deg.C, and standing overnight; filtering the supernatant with 0.22 μm filter membrane to obtain test solution;
performing gradient elution according to chromatographic conditions, wherein the sample introduction amount is 5 mu l, obtaining the fingerprint of the radix pseudostellariae oligosaccharide, continuously introducing samples for 6 times, recording the retention time and the peak area of a common peak, taking the No. 2 peak as a reference peak, calculating the relative retention time, the relative peak area and the RSD of each common peak, wherein the calculation results are shown in tables 2 and 3, and the results show that the RSD value of the relative retention time is 0.05-0.11%, and the RSD value of the relative peak area is 0.85-2.59%; the result shows that the precision is good, and the requirement of fingerprint spectrum detection is met.
TABLE 2 relative retention time of Tanshen fingerprint precision experiment
TABLE 3 relative peak area of Tanshenira fingerprint precision experiment
(2) And (3) repeatability experiment:
preparation of a test solution: precisely weighing 6 parts of the same batch of medicinal material samples, each part of the samples is 1.0g, placing the samples in a triangular flask with a plug, adding 20ml of 50% ethanol solution, carrying out ultrasonic extraction for 2 times, each time for 30min, filtering the extracting solutions, mixing, concentrating under reduced pressure to 1ml, adding 4ml of absolute ethanol, uniformly mixing, placing the mixture in a refrigerator at 4 ℃ and standing overnight; filtering the supernatant with 0.22 μm filter membrane to obtain test solution.
And respectively and sequentially carrying out sample introduction for 6 times, carrying out gradient elution according to chromatographic conditions, wherein the sample introduction amount is 5 mu l, taking the No. 2 peak as a reference peak, calculating the relative retention time, the relative peak area and the RSD of each common peak, wherein the calculation results are shown in tables 4 and 5, the results show that the RSD value of the relative retention time is 0.09-0.13%, the RSD value of the relative peak area is 1.07-3.74%, and experiments show that the repeatability is good.
TABLE 4 relative retention time of radix Pseudostellariae fingerprint method repeatability experiment
TABLE 5 relative peak area of radix Pseudostellariae fingerprint method repeatability experiment
(3) Stability test:
preparation of a test solution: precisely weighing 1.0g of radix Pseudostellariae powder, placing in a triangular flask with plug, adding 20ml of 50% ethanol solution, ultrasonically extracting for 2 times, each time for 30min, filtering the extractive solutions, mixing, concentrating under reduced pressure to 1ml, adding 4ml of anhydrous ethanol, mixing, placing in a refrigerator at 4 deg.C, and standing overnight; filtering the supernatant with 0.22 μm filter membrane to obtain test solution.
According to chromatographic conditions, samples are respectively injected at 0 hour, 2 hours, 4 hours, 8 hours, 16 hours and 24 hours, the sample injection amount is 5 mu l, the No. 2 peak is taken as a reference peak, the relative retention time RSD value and the relative peak area RSD value of each common peak are calculated, the calculation results are shown in tables 6 and 7, the results show that the RSD value of the relative retention time is 0.13% -0.54%, the RSD value of the relative peak area is 1.04% -3.99%, and the results show that the chromatographic peak stability of the radix pseudostellariae sample solution in 24 hours is good.
TABLE 6 relative retention time of radix Pseudostellariae fingerprint method stability test
TABLE 7 relative peak area of radix Pseudostellariae fingerprint method stability experiment
(4) And (3) similarity analysis:
preparation of a test solution: accurately weighing 12 radix Pseudostellariae medicinal materials from different sources, weighing 1.0g, placing in a triangular flask with a plug, adding 20ml 50% ethanol solution, ultrasonically extracting for 2 times, each time for 30min, filtering the extractive solutions, mixing, concentrating under reduced pressure to 1ml, adding 4ml anhydrous ethanol, mixing, placing in a refrigerator at 4 deg.C, and standing overnight; filtering the supernatant with 0.22 μm filter membrane to obtain test solution. The sample information of 12 batches of radix pseudostellariae medicinal materials is shown in a table 8;
analyzing the characteristic fingerprints of 12 batches of radix pseudostellariae medicinal materials according to chromatographic conditions, wherein the sample volume is 3 mu l, the 12 batches of radix pseudostellariae characteristic fingerprints are respectively introduced into software of a traditional Chinese medicine fingerprint similarity evaluation system (2012.130723 edition), the map of a sample S1 is set as a reference map, as shown in figure 8, the time window width is set to be 0.1min, so that peak identification, peak matching and similarity calculation are carried out, the similarity of the 12 batches of radix pseudostellariae characteristic maps is between 0.883 and 0.989, and the result shows that the sum of 8 common peak areas accounts for more than 90 percent of all peak areas of the original chromatogram; the results show that the radix pseudostellariae in different batches has small difference and good similarity. The map can be used as an evaluation standard of radix pseudostellariae by taking oligosaccharide parts as indexes, and can provide reference for quality control of different radix pseudostellariae decoction pieces.
TABLE 8 calculation results of radix Pseudostellariae fingerprint similarity of 12 batches
Example 2
The embodiment provides a method for detecting a fingerprint of radix pseudostellariae oligosaccharide, which adopts the fingerprint constructed in embodiment 1, and is applicable to quality inspection of radix pseudostellariae-containing medicinal materials and quality control of components of the radix pseudostellariae oligosaccharide, and the method specifically comprises the following steps:
s1: extracting the radix pseudostellariae oligosaccharide to be detected by using ethanol as an extraction solvent and adopting an ultrasonic extraction method to obtain a solution to be detected;
s2: performing HPLC analysis on the solution to be detected according to HPLC chromatographic conditions to obtain an HPLC chromatogram to be detected;
s3: and calculating and comparing the similarity of the HPLC chromatogram to be detected and the fingerprint spectrum according to the similarity.
In the step S1: the preparation of the solution to be tested comprises the following steps: precisely weighing 1.0g of radix Pseudostellariae powder to be tested, placing into a triangular flask with a plug, adding 20ml of 50% ethanol solution, ultrasonically extracting for 2 times (30 min each time), filtering the extractive solutions, mixing, concentrating under reduced pressure to 1ml, adding 4ml of anhydrous ethanol, mixing uniformly, placing into a refrigerator at 4 ℃, and standing overnight; filtering the supernatant with 0.22 μm filter membrane to obtain test solution.
In the step S2: the HPLC chromatographic conditions comprise:
(1) And (3) chromatographic column: hydrophilic chromatography columns such as amino column, XAmide, XIon, etc.;
(2) Column temperature: 15-35 ℃;
(3) Flow rate: 0.8-1.0ml/min;
(4) Sample introduction amount: 0.1-1.5mg;
(5) A detector: an evaporative light scattering detector;
(6) The mobile phase and elution mode are any one of the following four:
(1) 0-30min,75-55% acetonitrile, 30-32min,55-10% acetonitrile, 32-35min,10% acetonitrile;
(2) 0-30min,80-50% acetonitrile, 30-32min,50-10% acetonitrile, 32-35min,10% acetonitrile;
(3) 0-40min,75-50% acetonitrile, 40-42min,50-10% acetonitrile, 42-45min,10% acetonitrile;
(4) 0-30min,90-50% acetonitrile, 30-32min,50-10% acetonitrile, 32-35min,10% acetonitrile.
Carrying out gradient elution on the solution to be detected according to the chromatographic conditions, wherein the sample volume is 5 mu l, and obtaining an HPLC chromatogram of the radix pseudostellariae oligosaccharide to be detected; and (3) guiding the HPLC chromatogram of the radix pseudostellariae to be detected into software of a traditional Chinese medicine fingerprint similarity evaluation system (2012.130723 edition) for similarity evaluation, and performing peak identification and peak matching similarity calculation with a standard fingerprint.
Accurately weighing 12 radix pseudostellariae medicinal material powders from different sources for similarity analysis, wherein the similarity of 12 batches of radix pseudostellariae feature maps is between 0.883 and 0.989, and the sum of 8 common peak areas accounts for more than 90 percent of all peak areas of an original chromatogram, so that the radix pseudostellariae from different batches has small difference and good similarity; the map can be used as an evaluation standard of the radix pseudostellariae by taking oligosaccharide parts as indexes, and can provide reference for quality control of different radix pseudostellariae decoction pieces.
Example 3
The embodiment provides a radix pseudostellariae oligosaccharide's fingerprint detection device, includes:
the solution preparation unit is used for respectively extracting the radix pseudostellariae oligosaccharide and the radix pseudostellariae oligosaccharide to be detected by using ethanol as an extraction solvent and adopting an ultrasonic extraction method to obtain a test solution and a solution to be detected;
the sample solution analysis unit is used for carrying out HPLC analysis on the sample solution according to HPLC chromatographic conditions, carrying out chromatographic peak identification on the obtained HPLC chromatogram according to a comparison chromatogram, and constructing a fingerprint by taking the common chromatographic peak as a common peak;
the to-be-detected solution analysis unit is used for carrying out HPLC analysis on the to-be-detected solution according to HPLC chromatographic conditions to obtain a to-be-detected HPLC chromatogram;
and the detection unit is used for calculating and comparing the similarity between the HPLC chromatogram to be detected and the fingerprint spectrum through the similarity.
It should be noted here that the above units are the same as those of the example and application scenario realized by the steps corresponding to embodiment 1 or embodiment 2, but are not limited to the disclosure of embodiment 1 or embodiment 2.
The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and various modifications and changes will occur to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Although the embodiments of the present invention have been described with reference to the accompanying drawings, it is not intended to limit the scope of the present invention, and it should be understood by those skilled in the art that various modifications and variations can be made without inventive efforts by those skilled in the art based on the technical solution of the present invention.
Claims (7)
1. A fingerprint construction method of radix pseudostellariae oligosaccharide is characterized by comprising the following steps:
extracting the radix pseudostellariae oligosaccharide by an ultrasonic extraction method by taking 10-70% ethanol as an extraction solvent to obtain a test solution;
performing HPLC analysis on the sample solution according to HPLC chromatographic conditions, performing chromatographic peak identification on the obtained HPLC chromatogram according to a reference chromatogram, and constructing a fingerprint by taking the common chromatographic peak as a common peak;
the HPLC chromatographic conditions include:
a chromatographic column: selecting an XIon hydrophilic chromatographic column;
column temperature: 15-35 ℃;
flow rate: 0.8-1.0ml/min;
sample injection amount: 0.1-1.5mg;
a detector: an evaporative light scattering detector;
gradient elution is adopted as an elution mode;
the gradient elution is:
0-40min,75-50% acetonitrile, 40-42min,50-10% acetonitrile, 42-45min and 10% acetonitrile.
2. The method for constructing fingerprint of radix pseudostellariae oligosaccharide as claimed in claim 1, wherein the preparation of the test solution comprises: taking 1.0g of radix Pseudostellariae medicinal powder, adding 8-40ml of 10% -70% ethanol extraction solvent, performing ultrasonic extraction or heating reflux extraction for 1-3 times, extracting for 20-60min, mixing extractive solutions, concentrating under reduced pressure to 0.8-1ml, and adding 4-5 times volume of anhydrous ethanol.
3. The method for constructing the fingerprint spectrum of the radix pseudostellariae oligosaccharide as claimed in claim 2, wherein the preparation of the test solution comprises the following steps: taking 1.0g radix Pseudostellariae powder, placing in triangular flask with plug, adding 20ml 50% ethanol solution, ultrasonic extracting for 2 times, each for 30min, filtering the extractive solutions, mixing, concentrating under reduced pressure to 1ml, and adding 4ml anhydrous ethanol.
4. The fingerprint construction method of pseudostellaria heterophylla oligosaccharide as claimed in claim 3, wherein the sample solution is obtained by adding absolute ethyl alcohol, mixing, standing at 4 deg.C, and filtering the supernatant with 0.22 μm filter membrane.
5. The fingerprint spectrum construction method of pseudostellaria heterophylla oligosaccharide as claimed in claim 1, wherein HPLC analysis is performed on the test solution by HPLC-DAD-Q-TOF/MS.
6. A fingerprint spectrum detection method of radix pseudostellariae oligosaccharide is characterized by comprising the following steps:
extracting the pseudostellaria root oligosaccharide to be detected by using 10-70% ethanol as an extraction solvent and adopting an ultrasonic extraction method to obtain a solution to be detected;
performing HPLC analysis on the solution to be detected according to HPLC chromatographic conditions to obtain an HPLC chromatogram to be detected; the HPLC chromatographic conditions comprise:
and (3) chromatographic column: selecting an XIon hydrophilic chromatographic column;
column temperature: 15-35 ℃;
flow rate: 0.8-1.0ml/min;
sample introduction amount: 0.1-1.5mg;
a detector: an evaporative light scattering detector;
gradient elution is adopted as an elution mode;
the gradient elution is:
0-40min,75-50% acetonitrile, 40-42min,50-10% acetonitrile, 42-45min,10% acetonitrile;
and calculating and comparing the similarity of the HPLC chromatogram to be detected and the fingerprint spectrum through the similarity.
7. The fingerprint detection method of radix pseudostellariae oligosaccharide as claimed in claim 6, wherein a traditional Chinese medicine fingerprint similarity evaluation system is adopted to perform similarity calculation on an HPLC chromatogram to be detected;
and the similarity calculation comprises the similarity calculation of peak identification and peak matching of the HPLC chromatogram to be detected and the fingerprint.
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| CN108254470A (en) * | 2016-12-28 | 2018-07-06 | 上海医药工业研究院 | In glutinous rehmannia while carbohydrate content measure and its fingerprint map construction method |
| CN110082466A (en) * | 2018-12-11 | 2019-08-02 | 浙江省食品药品检验研究院 | A kind of detection method for Rehmannia glutinosa Processing methods quality |
| CN110596271A (en) * | 2019-09-25 | 2019-12-20 | 广东一方制药有限公司 | UPLC characteristic spectrum construction method and identification method of radix pseudostellariae medicinal material and radix pseudostellariae standard decoction |
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