CN115856181A - Characteristic spectrum of linseed decoction pieces and preparation thereof as well as construction method and application of characteristic spectrum - Google Patents

Abstract

The invention belongs to the field of traditional Chinese medicine identification, and particularly provides a characteristic spectrum of linseed decoction pieces and a preparation thereof, and a construction method and application thereof, wherein the construction method comprises the following steps: (1) Mixing the linseed decoction piece preparation with the alcohol solution, performing ultrasonic treatment, and filtering to obtain a first test sample solution; (2) Mixing linseed decoction pieces with water, decocting, filtering, drying filtrate, mixing with an alcohol solution, performing ultrasonic treatment, and filtering to obtain a second test sample solution; (3) Respectively carrying out UPLC detection on the first sample solution and the second sample solution, screening peaks with consistent retention time, good peak shape and high resolution as characteristic peaks according to detection results, selecting S peaks with determined components and good peak shape from the characteristic peaks, and calculating the relative retention time of other characteristic peaks relative to the S peaks to obtain a characteristic spectrum. The constructed characteristic spectrum provided by the invention has good repeatability, high precision and good stability, and can be used for effectively identifying the quality of the linseed medicine.

Description

Characteristic spectrum of linseed decoction pieces and preparation thereof as well as construction method and application of characteristic spectrum

Technical Field

The invention belongs to the field of traditional Chinese medicine identification, and particularly relates to a characteristic spectrum of a linseed decoction piece and a preparation thereof, a construction method and application thereof, and particularly relates to a characteristic spectrum of a linseed decoction piece and a preparation thereof with high precision, a construction method and application thereof.

Background

The semen Lini is dry mature seed of Linum usitatissimum L.of Linaceae, has sweet taste and neutral nature, and has effects of moistening dryness, relaxing bowels, nourishing blood, and dispelling pathogenic wind, and can be used for treating constipation due to intestinal dryness, dry skin, pruritus, and alopecia. Researches show that the chemical components in the linseed mainly contain lignans, polyphenols, flavonoids, unsaturated fatty acids, saccharides, cyclic peptides and amino acid compounds, and the overall quality control of the effective components of the linseed is required to ensure the clinical curative effect of the linseed. The linseed standard decoction freeze-dried powder is a traditional Chinese medicine product prepared by extracting, concentrating, separating and drying linseed decoction pieces by a modern pharmaceutical technology, and meanwhile, the quality of the product is controlled by using a modern detection technology, wherein a linseed characteristic map method established by a high-efficiency liquid phase is used, so that the multi-component qualitative analysis can be performed on the linseed decoction pieces and the linseed preparation, the quality of the linseed is controlled, the consistency of the material base is ensured, and the tryptophan component in the linseed can be quantitatively determined, so that the stability and the effectiveness of the whole process are realized.

At present, in the qualitative and quantitative research on the linseed in the first part of the Chinese pharmacopoeia 2020, only physical and chemical identification and gas phase detection items on fatty oil exist, and a method for controlling the quality of the non-oil component content and the characteristic spectrum by using a high performance liquid chromatography is not available. CN114076806A discloses a linseed oil fingerprint detection method and application thereof, wherein the detection method adopts a fingerprint to carry out detection, and comprises the following steps: constructing the phytosterol control fingerprint spectra of different varieties of linseed oil by adopting a gas chromatography-mass spectrometry combined method; the detection conditions of the gas chromatograph comprise: a chromatographic column: 5% phenyl-95% methylpolysiloxane column; the split ratio is as follows: 38-43; temperature rising procedure: the initial temperature is 160-200 deg.C, the temperature is maintained for 0.5-2min, the temperature is raised to 280-320 deg.C at 1-8 deg.C/min, and the temperature is maintained for 20-30min. The method can be used for distinguishing and identifying the linseed oil of different varieties, can realize the quick and effective identification of the linseed oil, and has certain significance for quality control and adulteration identification of the linseed oil. However, the method adopts a gas chromatography-mass spectrometry detection method, so that the detection cost is high, the method is difficult to popularize industrially, and the fingerprint contains only the phytosterol component and is completely unrelated to the contents of other components.

At present, no method for simultaneously controlling the quality of the linseed decoction pieces and the linseed decoction piece preparations exists. Therefore, the problem to be solved is how to provide a characteristic spectrum of the linseed decoction pieces and the linseed decoction piece preparation and carry out effective quality control on the linseed decoction pieces and the linseed decoction piece preparation.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to provide a characteristic spectrum of a linseed decoction piece and a preparation thereof, a construction method and application thereof, and particularly provides a characteristic spectrum of a linseed decoction piece and a preparation thereof with high precision, a construction method and application thereof. The constructed characteristic spectrum provided by the invention has good repeatability, high precision and good stability, and can be used for effectively identifying the quality of the linseed medicine.

In order to achieve the purpose, the invention adopts the following technical scheme:

in a first aspect, the invention provides a construction method of a characteristic spectrum of linseed decoction pieces and a preparation thereof, and the construction method comprises the following steps:

(1) Mixing the linseed decoction piece preparation with the alcohol solution, performing ultrasonic treatment, and filtering to obtain a first test sample solution;

(2) Mixing and decocting linseed decoction pieces and water, filtering, drying filtrate, mixing the filtrate with an alcohol solution, carrying out ultrasonic treatment, and filtering to obtain a second test sample solution;

(3) Respectively carrying out UPLC detection on the first test solution and the second test solution, screening peaks which have consistent retention time, good peak shape and high resolution as characteristic peaks according to detection results, selecting the peaks with determined components and good peak shape as S peaks from the characteristic peaks, and calculating the relative retention time of other characteristic peaks relative to the S peaks to obtain the characteristic maps of the linseed decoction pieces and the preparations thereof.

The sequence of the step (1) and the step (2) is not distinguished.

According to the method, the characteristic spectrogram is constructed by the linseed decoction pieces and the linseed decoction piece preparation together, so that the obtained characteristic spectrogram can be used for identifying the linseed decoction pieces and the linseed decoction piece preparation; and the characteristic spectrum obtained by performing UPLC detection on the first test sample solution and the second test sample solution by adopting a specific method has good repeatability, high precision and good durability.

The linseed decoction piece preparation comprises linseed standard decoction freeze-dried powder, linseed formula granules and the like.

Preferably, the material-liquid ratio of the linseed decoction piece preparation in the step (1) to the alcohol solution is 1 (90-110) g/mL.

Preferably, the time of the ultrasound in the step (1) is 25-35min.

Preferably, the material-liquid ratio of the linseed decoction pieces to the water in the step (2) is 1 (90-110) g/mL.

Preferably, the decocting temperature in the step (2) is 95-100 ℃, and the time is 25-35min.

Preferably, the material-liquid ratio of the linseed decoction pieces to the alcohol solution in the step (2) is 1 (18-22) g/mL.

Preferably, the time of the ultrasound in the step (2) is 25-35min.

Preferably, the alcohol solution is a methanol aqueous solution, and the volume fraction of the methanol aqueous solution is 65-75%.

Wherein, the ratio of the material to the alcoholic solution of the linseed decoction piece preparation may be 1, 95g/mL, 1, 100g/mL, 1, 105g/mL or 1.

By controlling the parameters, the invention can fully extract the effective components in the linseed decoction piece preparation and the linseed decoction piece, and improve the accuracy of subsequent detection and characteristic spectrum.

Preferably, the filler of the chromatography column for UPLC detection in step (3) is octadecylsilane chemically bonded silica.

Preferably, the mobile phase for UPLC detection in step (3) includes mobile phase a and mobile phase B, where the mobile phase a is acetonitrile, the mobile phase B is an aqueous phosphoric acid solution, and the mass fraction of the aqueous phosphoric acid solution is 0.08-0.12%.

Preferably, the flow rate of the UPLC detection in the step (3) is 0.28-0.32mL/min, and the column temperature is 32-38 ℃.

Preferably, the wavelength of the UPLC detection in step (3) is 210-220nm, preferably 217nm.

The mass fraction of the phosphoric acid aqueous solution may be 0.08%, 0.09%, 0.1%, 0.11%, or 0.12%, the flow rate may be 0.28mL/min, 0.29mL/min, 0.3mL/min, 0.31mL/min, or 0.32mL/min, the column temperature may be 32 ℃, 33 ℃, 34 ℃, 35 ℃, 36 ℃, 37 ℃, or 38 ℃, the wavelength may be 210nm, 211nm, 212nm, 213nm, 214nm, 215nm, 216nm, 217nm, 218nm, 219nm, or 220nm, but the above-mentioned values are not limited thereto, and other values not listed in the above-mentioned range of values are also applicable.

Preferably, the elution procedure of UPLC detection in step (3) is as follows:

when the time is 0-8min, the volume fraction of the mobile phase A is changed from 2% to 5% at a constant speed, and the volume fraction of the mobile phase B is changed from 98% to 95% at a constant speed;

when the time is 8-15min, the volume fraction of the mobile phase A is changed from 5% to 12% at a constant speed, and the volume fraction of the mobile phase B is changed from 95% to 88% at a constant speed;

at 15-21min, the volume fraction of the mobile phase A is changed from 12% to 14% at constant speed, and the volume fraction of the mobile phase B is changed from 88% to 86% at constant speed;

at the time of 21-35min, the volume fraction of the mobile phase A is changed from 14% to 18% at a constant speed, and the volume fraction of the mobile phase B is changed from 86% to 82% at a constant speed.

The specific elution procedure can effectively distinguish components in the sample, has high peak shape separation degree and obvious peak appearance, can obtain a characteristic peak with better peak shape, improves the accuracy of detection and characteristic spectrum, can obviously shorten the elution time and improve the detection efficiency.

In a second aspect, the invention provides the characteristic maps of the linseed decoction pieces and the preparations thereof obtained by the construction method, wherein in the characteristic maps, the number of characteristic peaks is 7, the characteristic maps are sorted from small to large according to retention time, and the relative retention times of a peak 1 as an S peak, and peaks 2, 3, 4, 5, 6 and 7 are respectively 1.59 + -10%, 1.71 + -10%, 1.92 + -10%, 2.00 + -10%, 3.14 + -10% and 4.27 + -10%.

The component of the No. 1 peak is tryptophan.

In a third aspect, the invention provides the application of the characteristic spectrum of the linseed decoction piece and the linseed preparation in quality judgment of the linseed medicine.

In a fourth aspect, the invention also provides a quality judgment method of the linseed medicine, which is characterized in that the linseed medicines of different batches are detected by adopting the UPLC detection method, the detection result is subjected to median processing, and the standard characteristic spectrum is obtained by combining the characteristic spectrum; and detecting the sample to be detected by adopting the UPLC detection method, judging the characteristic peak according to the characteristic spectrum, then judging the similarity or judging the relative retention time, and judging whether the sample to be detected is a qualified product or an unqualified product.

In the similarity judgment, calculating the similarity between the characteristic peak of the sample to be detected and the characteristic peak in the standard characteristic spectrum; and if the similarity is not lower than 0.85, determining that the sample to be detected is a qualified product, and if the similarity is lower than 0.85, determining that the sample to be detected is an unqualified product.

In the relative retention time judgment, calculating the relative retention time of each characteristic peak of the sample to be detected, and comparing the relative retention time with the relative retention time of each characteristic peak in the characteristic map, wherein if the relative retention time of the characteristic peak of the sample to be detected is out of the relative retention time range of the characteristic peak in the characteristic map, the sample to be detected is an unqualified product; and if the relative retention time of the characteristic peaks of all the samples to be detected falls within the relative retention time range of the characteristic peaks in the characteristic spectrum, determining that the samples to be detected are qualified products.

The semen Lini medicine is semen Lini decoction piece or semen Lini decoction piece preparation.

The method can effectively distinguish whether the sample to be detected is a qualified product or not by detecting the sample to be detected by using the prepared characteristic spectrum, and provides a new method for judging the quality of the linseed medicine.

Compared with the prior art, the invention has the following beneficial effects:

the invention provides a construction method of a characteristic spectrum of a linseed decoction piece and a preparation thereof, wherein the characteristic spectrum is constructed by the linseed decoction piece and the linseed decoction piece together, so that the obtained characteristic spectrum can identify the linseed decoction piece and the linseed decoction piece preparation, and can control the overall quality of linseed; the characteristic spectrum obtained by performing UPLC detection on the first test solution and the second test solution by adopting a specific method has good repeatability, high precision and good durability; the components in the sample can be effectively distinguished through a specific elution program, the peak shape separation degree is high, the peak appearance is obvious, the characteristic peak with better peak shape can be obtained, the accuracy of detection and a characteristic map is improved, the elution time can be obviously shortened, the detection efficiency is improved, the method is simple, convenient and quick, the industrial application is convenient, and the detection cost is reduced.

Drawings

FIG. 1 is a feature map of example 1;

FIG. 2 is a characteristic map of comparative example 1;

FIG. 3 is a characteristic map of comparative example 2;

FIG. 4 is a characteristic map of comparative example 3;

FIG. 5 is a liquid phase result chart of the linseed standard decoction lyophilized powder detection test.

FIG. 6 is a liquid phase result chart of the flax seed decoction piece detection test.

Detailed Description

The technical solution of the present invention is further explained by the following embodiments. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitations of the present invention.

The instruments and reagents used in the following examples are as follows:

the instrument comprises the following steps: waters UPLC H-Class ultra high performance liquid chromatograph; electronic analytical balances (XP 26, ME104E, mettlerlitosan; BSA124S, sidolis; JY20002, shanghai Shunhui constant-level scientific instruments, inc.); KQ-500DB ultrasonic cleaning machine (kunshan ultrasonic electronic device factory);

reagent: acetonitrile (Fisher Chemical), phosphoric acid (Fisher Scientific) were chromatographically pure, methanol was analytically pure, and water was Drech's purified water.

Example 1

The embodiment provides a characteristic spectrum of linseed decoction pieces and a preparation thereof, and the specific construction method comprises the following steps:

taking a proper amount of linseed standard decoction freeze-dried powder, grinding, accurately weighing 0.1g, placing in a conical flask with a plug, accurately adding 10mL of 70% methanol aqueous solution, sealing the plug, weighing, carrying out ultrasonic treatment (power 250W, frequency 40 kHz) for 30min, taking out, cooling, weighing again, complementing the weight loss by 70% methanol aqueous solution, shaking up, filtering, and taking the subsequent filtrate to obtain a first sample solution;

taking a proper amount of linseed decoction pieces, grinding, precisely weighing 0.5g, adding 50mL of water, decocting for 30 minutes, filtering, evaporating filtrate to dryness, precisely adding 10mL of 70% methanol aqueous solution, sealing, weighing, carrying out ultrasonic treatment (power of 250W and frequency of 40 kHz) for 30 minutes, taking out, cooling, weighing again, complementing the weight loss by using 70% methanol aqueous solution, shaking uniformly, filtering, and taking a subsequent filtrate to obtain a second sample solution;

respectively sucking 1 mu L of the first sample solution and 1 mu L of the second sample solution, and injecting the first sample solution and the second sample solution into a high liquid chromatograph for determination.

The chromatographic conditions are as follows: using octadecylsilane chemically bonded silica as filler (column length is 100mm, inner diameter is 2.1mm, particle size is 1.7 μm), acetonitrile as mobile phase A, 0.1% phosphoric acid solution as mobile phase B, and performing gradient elution according to the specification in the following table; the detection wavelength was 217nm. The column temperature is 35 ℃; the flow rate was 0.3mL per minute.

The results were mapped to a characteristic map, as shown in FIG. 1.

According to the results shown in figure 1, 7 peaks which have consistent retention time, high separation degree and good peak shape and have peak-appearing time covering the whole elution process are selected as characteristic peaks and are respectively marked as No. 1-No. 7 peaks, the No. 1 peak (identified as a tryptophan peak) is taken as an S peak, the relative retention time of other peaks is calculated, and the results are as follows: 1.59 (peak 2), 1.71 (peak 3), 1.92 (peak 4), 2.00 (peak 5), 3.14 (peak 6), 4.27 (peak 7), namely obtaining the characteristic map.

Comparative example 1

The comparative example provides a characteristic spectrum of linseed decoction pieces and a preparation thereof, and the construction method is the same as that of example 1 except that the chromatographic conditions are as follows.

The chromatographic conditions are as follows: using octadecylsilane chemically bonded silica as filler (column length 250mm, inner diameter 4.6mm, particle size 5 μm), acetonitrile as mobile phase A, 0.1% phosphoric acid solution as mobile phase B, and performing gradient elution according to the specification in the following table; the detection wavelength was 260nm. The column temperature is 35 ℃; the flow rate was 1mL per minute.

The results are shown in FIG. 2.

Comparative example 2

The comparative example provides a characteristic spectrum of linseed decoction pieces and a preparation thereof, and the construction method is the same as that of example 1 except that the chromatographic conditions are as follows.

The chromatographic conditions are as follows: using octadecylsilane chemically bonded silica as filler (column length is 100mm, inner diameter is 2.1mm, particle size is 1.8 μm), acetonitrile as mobile phase A, 0.1% phosphoric acid solution as mobile phase B, and performing gradient elution according to the specification in the following table; the detection wavelength was 220nm. The column temperature is 35 ℃; the flow rate was 0.4mL per minute.

The results are shown in FIG. 3.

Comparative example 3

The comparative example provides a characteristic map of linseed decoction pieces and a preparation thereof, and the construction method is the same as that of example 1 except that the elution procedure is as follows.

The results are shown in FIG. 4.

The results show that the chromatographic peak separation degree is high by controlling the gradient elution procedure, the characteristic peak can be effectively obtained, and the accuracy of detection and characteristic spectrum is improved; the peak position of the map of the comparative example 1 is greatly different from that of the example 1, the base line is uneven, and a large number of peaks are overlapped; in comparative example 2, a large amount of peak overlap occurred around 20 min; in comparative example 3, the base line was also uneven around 20 to 33min, resulting in failure to confirm the characteristic peak.

And (3) repeatability test:

6 parts of linseed standard decoction freeze-dried powder is taken and measured according to the method provided by the embodiment 1, the relative retention time of each characteristic peak is calculated, and the result is as follows:

the relative retention time RSD of each characteristic peak can be found to be in the range of 0.8-1.0%, which indicates that the repeatability of the characteristic spectrum is better.

And (3) testing intermediate precision:

taking 6 parts of linseed standard decoction freeze-dried powder, measuring according to the method provided in example 1 (the ultra-high performance liquid chromatograph is replaced by another Waters UPLC H-Class, TUV detector), calculating the relative retention time of each characteristic peak, and obtaining the following results:

in a characteristic spectrum obtained by a Waters UPLC H-Class TUV detector, the relative retention time RSD of each characteristic peak is in the range of 0.6-1.3%, and the relative retention time RSD range between different instruments is in the range of 0.9-1.9%, which indicates that the relative peak area of the characteristic spectrum between different instruments meets the analysis requirement.

And (3) stability testing:

the test solution was prepared according to the method of example 1, and the test was performed according to the method of example 1 at 0, 2, 4, 6, 8, 12, and 24h after the preparation, and the relative retention time of each characteristic peak was calculated as follows:

the results show that the characteristic spectrum obtained by the construction method provided by the invention has good stability, the chemical components in the solution are stable within 24 hours, and the relative retention time RSD of each characteristic peak is within the range of 1.5-1.9%.

And (3) testing accuracy:

taking a proper amount of tryptophan reference substance, precisely weighing, and adding 70% methanol water solution to prepare a solution containing 253 μ g per 1mL. A linseed standard decoction freeze-dried powder (KL 190730-741600-01, tryptophan content of 9.85 mg/g) with known content is taken to be divided into 9 parts in parallel, each part is 0.05g, precisely weighed, 1mL (0.252547 mg tryptophan) of a tryptophan reference solution and 9mL (0.505094 mg tryptophan) of 70% methanol aqueous solution, 8mL (0.505094 mg tryptophan) of 70% methanol aqueous solution, 3mL (0.757642 mg tryptophan) of the tryptophan reference solution and 7mL (70% methanol aqueous solution) of 1. The recovery was calculated according to the following formula and the results were as follows:

the measured recovery rate range is 95.6-101.8%, the average recovery rate is 99.1%, the RSD value is 2.2%, the recovery rate requirement of methodology verification is met, and the characteristic spectrum obtained by the construction method is high in accuracy.

Precisely weighing a proper amount of tryptophan reference substance, placing the tryptophan reference substance in a 50mL volumetric flask, and adding methanol to prepare a solution containing 0.5123mg per 1mL to obtain a tryptophan reference substance mother solution as a linear 1; precisely sucking 1mL of tryptophan reference product mother liquor of the linear 1, placing the tryptophan reference product mother liquor in a 2mL volumetric flask, and adding methanol to the scale to obtain a linear 2; precisely sucking 1mL of tryptophan reference product mother liquor of the linear 1, placing the tryptophan reference product mother liquor in a volumetric flask of 5mL, and adding methanol to the scale to obtain a linear 3; precisely sucking 1mL of tryptophan reference product mother liquor of linear 1, placing the tryptophan reference product mother liquor in a 10mL volumetric flask, and adding methanol to the scale to obtain linear 4; precisely sucking 1mL of tryptophan reference product mother liquor of the linear 1, placing the tryptophan reference product mother liquor in a 20mL volumetric flask, and adding methanol to the scale to obtain a linear 5; 1mL of the tryptophan control stock solution of line 1 was pipetted precisely and placed in a 50mL volumetric flask and methanol added to the mark as line 6. Precisely sucking 1 μ L of the secondary filtrate, injecting into high performance liquid chromatograph, and mixingThe method of example 1 measures the peak area of the tryptophan chromatographic peak, takes the peak area of the tryptophan chromatographic peak as the ordinate and the tryptophan concentration as the abscissa, draws a standard curve, obtains the tryptophan regression equation of y =3099742.83x-18347.44 ² =0.999, the linear range is 0.0102-0.5123mg/mL, and the parameters in the construction method of the present invention are examined using the linear equation.

The linseed standard decoction lyophilized powder was taken, the detection was performed according to the method in example 1, and the column temperature, flow rate, and chromatography column (respectively replacing the stationary phase with Waters absorbance)

HSS T3(2.1mm×100mm，1.8μm)、Waters CORTECS/>

T3 (2.1 mm. Times.100mm, 1.6 μm) and Waters ACQUITY->

BEH Shield RP18 (2.1 mm. Times.100mm, 1.7 μm)), the mass fraction of the phosphoric acid aqueous solution was varied, and the durability of the construction method was examined, and the results were as follows:

/>

it can be seen that the method provided by the present invention has good durability for different column temperatures, flow rates, chromatographic columns and phosphoric acid mass fractions, and RSD values of not more than 0.86%.

A linseed standard decoction freeze-dried powder detection test comprises the following steps:

taking 15 batches of linseed standard decoction freeze-dried powder, respectively detecting according to the method in the embodiment 1, fitting a detection spectrum to obtain a standard characteristic spectrum, judging a characteristic peak according to the characteristic spectrum obtained in the embodiment 1, and calculating the similarity, wherein the results are shown in the following table and a figure 5:

the similarity between the characteristic map of 15 batches of linseed standard decoction freeze-dried powder and the reference characteristic map is 0.982-0.999, which are all higher than 0.85, and the flaxseed standard decoction freeze-dried powder is qualified.

Detection test of linseed decoction pieces:

taking 15 batches of linseed decoction pieces, respectively detecting according to the method in the embodiment 1, fitting a detection spectrum to obtain a standard characteristic spectrum, judging a characteristic peak according to the characteristic spectrum obtained in the embodiment 1, and calculating the similarity, wherein the results are shown in the following table and figure 6:

the similarity between the characteristic spectrum of 15 batches of flaxseed decoction pieces and the reference characteristic spectrum is 0.933-0.996, is higher than 0.85, and is qualified product

The detection tests fully prove that the characteristic spectrum and the construction method thereof provided by the invention can effectively judge the quality of the linseed medicine and provide a new method for judging the quality of the linseed medicine.

The applicant states that the present invention is illustrated by the above examples to show the characteristics of the flaxseed drink and its preparation, and the construction method and application thereof, but the present invention is not limited to the above examples, i.e. it is not meant that the present invention must be implemented by the above examples. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.

The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.

It should be noted that the various technical features described in the above embodiments can be combined in any suitable manner without contradiction, and the invention is not described in any way for the possible combinations in order to avoid unnecessary repetition.

Claims (10)

1. A construction method of a characteristic spectrum of linseed decoction pieces and a preparation thereof is characterized by comprising the following steps:

(1) Mixing the linseed decoction piece preparation with the alcohol solution, carrying out ultrasonic treatment, and filtering to obtain a first test sample solution;

(2) Mixing and decocting linseed decoction pieces and water, filtering, drying filtrate, mixing the filtrate with an alcohol solution, carrying out ultrasonic treatment, and filtering to obtain a second test sample solution;

(3) Respectively carrying out UPLC detection on the first test solution and the second test solution, screening peaks which have consistent retention time, good peak shape and high resolution as characteristic peaks according to detection results, selecting the peaks with determined components and good peak shape as S peaks from the characteristic peaks, and calculating the relative retention time of other characteristic peaks relative to the S peaks to obtain the characteristic maps of the linseed decoction pieces and the preparations thereof;

the sequence of the step (1) and the step (2) is not distinguished.

2. The construction method according to claim 1, wherein the material-to-liquid ratio of the linseed decoction piece preparation to the alcohol solution in the step (1) is 1 (90-110) g/mL;

preferably, the time of the ultrasound in the step (1) is 25-35min.

3. The construction method according to claim 1 or 2, characterized in that the material-liquid ratio of the linseed decoction pieces to the water in the step (2) is 1 (90-110) g/mL;

preferably, the decocting temperature in the step (2) is 95-100 ℃, and the time is 25-35min;

preferably, the material-liquid ratio of the linseed decoction pieces to the alcohol solution in the step (2) is 1 (18-22) g/mL;

preferably, the time of the ultrasound in the step (2) is 25-35min.

4. The method of any one of claims 1-3, wherein the alcohol solution is an aqueous methanol solution having a volume fraction of 65-75%.

5. The method for constructing the UPLC detection column of any one of claims 1-4, wherein the filler of the UPLC detection column of step (3) is octadecylsilane chemically bonded silica.

6. The construction method according to any one of claims 1 to 5, wherein the mobile phase of the UPLC detection in step (3) comprises mobile phase A and mobile phase B, wherein the mobile phase A is acetonitrile, the mobile phase B is an aqueous phosphoric acid solution, and the mass fraction of the aqueous phosphoric acid solution is 0.08-0.12%;

preferably, the flow rate of the UPLC detection in the step (3) is 0.28-0.32mL/min, and the column temperature is 32-38 ℃;

preferably, the wavelength of the UPLC detection in step (3) is 210-220nm, preferably 217nm.

7. The construction method according to claim 6, wherein the elution procedure of the UPLC detection in step (3) is as follows:

when the time is 0-8min, the volume fraction of the mobile phase A is changed from 2% to 5% at a constant speed, and the volume fraction of the mobile phase B is changed from 98% to 95% at a constant speed;

when the time is 8-15min, the volume fraction of the mobile phase A is changed from 5% to 12% at a constant speed, and the volume fraction of the mobile phase B is changed from 95% to 88% at a constant speed;

at 15-21min, the volume fraction of the mobile phase A is changed from 12% to 14% at constant speed, and the volume fraction of the mobile phase B is changed from 88% to 86% at constant speed;

at the time of 21-35min, the volume fraction of the mobile phase A is changed from 14% to 18% at a constant speed, and the volume fraction of the mobile phase B is changed from 86% to 82% at a constant speed.

8. A characteristic map of linseed decoction pieces and a preparation thereof obtained by the construction method according to any one of claims 1 to 7, wherein in the characteristic map, the number of characteristic peaks is 7, the retention times are sorted from small to large, and the relative retention times of the No. 1 peak as the S peak, the No. 2 peak, the No. 3 peak, the No. 4 peak, the No. 5 peak, the No. 6 peak and the No. 7 peak are 1.59 +/-10%, 1.71 +/-10%, 1.92 +/-10%, 2.00 +/-10%, 3.14 +/-10% and 4.27 +/-10%, respectively;

the component of the No. 1 peak is tryptophan.

9. The use of the characteristic spectrum of the linseed decoction pieces and the preparation thereof according to claim 8 in quality judgment of linseed drugs.

10. A quality judgment method of linseed drugs is characterized in that linseed drugs of different batches are detected by the UPLC detection method of any one of claims 1-7, the detection result is subjected to median processing, and a standard characteristic spectrum is obtained by combining the characteristic spectrum of claim 8; detecting a sample to be detected by adopting the UPLC detection method of any one of claims 1-7, judging a characteristic peak according to the characteristic spectrum of claim 8, and then judging the similarity or the relative retention time to judge whether the sample to be detected is a qualified product or an unqualified product;

in the similarity judgment, calculating the similarity between the characteristic peak of the sample to be detected and the characteristic peak in the standard characteristic spectrum; when the similarity is not lower than 0.85, the sample to be detected is a qualified product, and when the similarity is lower than 0.85, the sample to be detected is an unqualified product;

in the relative retention time judgment, calculating the relative retention time of each characteristic peak of the sample to be detected, and comparing the relative retention time with the relative retention time of each characteristic peak in the characteristic map of claim 8, wherein if the relative retention time of the characteristic peak of the sample to be detected is out of the range of the relative retention time of the characteristic peak in the characteristic map of claim 8, the sample to be detected is an unqualified product; if the relative retention time of the characteristic peaks of all samples to be tested falls within the relative retention time range of the characteristic peaks in the characteristic spectrum of claim 8, determining that the samples to be tested are qualified products;

the semen Lini medicine is semen Lini decoction piece or semen Lini decoction piece preparation.

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