Construction method of characteristic chromatogram of fructus psoraleae medicinal material and related application thereof

Abstract

The invention discloses a construction method of a characteristic chromatogram of a fructus psoraleae medicinal material and related application thereof, and relates to the technical field of medicine quality control. The construction method adopts specific chromatographic conditions to realize construction of the characteristic chromatogram, chemical components of the fructus psoraleae medicinal material can be accurately analyzed at the same time, quality evaluation of the fructus psoraleae medicinal material is realized, the detection process is quicker and easy to implement, and the method has the advantages of high detection sensitivity, good stability, accurate and reliable determination result and the like, and is suitable for popularization and application. The method is of great significance to quality control and clinical curative effect guarantee of the fructus psoraleae.

Description

Technical Field

[0001] The present invention relates to the technical field of drug quality control, and in particular to a method for constructing a characteristic spectrum of a psoralea corylifolia medicinal material and related applications thereof. Background Art

[0002] Psoralea corylifolia L. is the dried mature fruit of the leguminous plant Psoralea corylifolia L. When the fruit is ripe in autumn, the fruit sequence is harvested, dried in the sun, the fruit is rubbed out, and impurities are removed. It has the effects of warming the kidney and supporting yang, absorbing qi and relieving asthma, warming the spleen and stopping diarrhea; it can be used externally to eliminate wind and remove spots. It is used for kidney yang deficiency, impotence and spermatorrhea, enuresis and frequent urination, cold pain in the waist and knees, asthma caused by kidney deficiency, and diarrhea at dawn; it can be used externally to treat vitiligo and alopecia areata.

[0003] At present, researchers mainly control the content of psoralen and isopsoralen, which cannot achieve a comprehensive quality evaluation of Psoralea corylifolia. The construction of the existing public Psoralea corylifolia characteristic map has some shortcomings, such as long detection time, complex process, low efficiency, and inability to effectively achieve quality evaluation of Psoralea corylifolia.

[0004] In view of this, the present invention is proposed. Summary of the invention

[0005] The object of the present invention is to provide a method for constructing a characteristic spectrum of a Psoralea corylifolia medicinal material and related applications thereof.

[0006] The present invention is achieved in that:

[0007] In a first aspect, an embodiment of the present invention provides a method for constructing a characteristic spectrum of a Psoralea corylifolia medicinal material, comprising: subjecting a test solution and a reference solution to high performance liquid chromatography, respectively, to obtain a characteristic spectrum; the chromatographic conditions of the high performance liquid chromatography include: mobile phase A includes acetonitrile, and mobile phase B includes aqueous phosphoric acid; the elution conditions of the chromatography are as follows:

[0008] From 0 to 2 min, the volume percentage of mobile phase A changed from 8% to 12% to 17% to 21%, and the volume percentage of mobile phase B changed from 88% to 92% to 79% to 83%;

[0009] From 2 to 7 min, the volume percentage of mobile phase A changed from 17% to 21% to 34% to 38%, and the volume percentage of mobile phase B changed from 79% to 83% to 62% to 66%;

[0010] From 7 to 9 min, the volume percentage of mobile phase A changed from 34% to 38% to 73% to 77%, and the volume percentage of mobile phase B changed from 62% to 66% to 33% to 37%;

[0011] From 9 to 11 min, the volume percentage of mobile phase A changed from 73% to 77% to 91% to 95%, and the volume percentage of mobile phase B changed from 33% to 37% to 5% to 9%;

[0012] 11-12 min, the volume percentage of mobile phase A is maintained at 91%-95%, and the volume percentage of mobile phase B is maintained at 5%-9%;

[0013] 12-12.1 min, the volume percentage of mobile phase A changes from 91%-95% to 8%-12%, and the volume percentage of mobile phase B changes from 5%-9% to 88%-92%;

[0014] From 12.1 to 15 min, the volume percentage of mobile phase A is maintained at 8% to 12%, and the volume percentage of mobile phase B is maintained at 88% to 92%.

[0015] In a second aspect, an embodiment of the present invention provides a method for evaluating the quality of Psoralea corylifolia medicinal materials, which comprises: obtaining a chromatogram of a sample to be tested for high performance liquid chromatography detection; and performing quality evaluation on the sample to be tested based on the chromatogram and a characteristic spectrum constructed by the construction method described in the aforementioned embodiment.

[0016] In a third aspect, an embodiment of the present invention provides an application of the construction method described in the aforementioned embodiment in screening Psoralea corylifolia medicinal materials or preparing Psoralea corylifolia-related drugs.

[0017] The present invention has the following beneficial effects:

[0018] The embodiment of the present invention provides a method for constructing an HPLC characteristic spectrum of a Psoralea corylifolia medicinal material. The characteristic spectrum constructed based on the construction method can simultaneously and accurately analyze the chemical components of the Psoralea corylifolia medicinal material, realize the quality evaluation of the Psoralea corylifolia medicinal material, and the inspection process is faster and easier to implement. It has the advantages of high detection sensitivity, good stability, accurate and reliable measurement results, etc., which is of great significance to the quality control of Psoralea corylifolia and the guarantee of clinical efficacy. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0020] Figure 1 is the ultraviolet absorption spectrum of psoralen;

[0021] Figure 2 is the ultraviolet absorption spectrum of isopsoralen;

[0022] Figure 3 is the ultraviolet absorption spectrum of psoralen;

[0023] Figure 4 is the ultraviolet absorption spectrum of isopsoralen;

[0024] Figure 5 is the ultraviolet absorption spectrum of bakuchiol;

[0025] Figure 6 This is a comparison chart of different wavelengths of Psoralea corylifolia medicinal materials;

[0026] Figure 7 The test results of the chromatographic column at different temperatures;

[0027] Figure 8 The characteristic spectra of three batches of Psoralea corylifolia;

[0028] Fig. 9 This is the characteristic spectrum of Psoralea corylifolia reference medicinal material;

[0029] Fig.10 Characteristic spectra of 10 batches of medicinal materials;

[0030] Fig.11 The test results of 10 batches of finished medicinal materials;

[0031] Fig.12 See the figure for a comparison between the elution time of medicinal materials and the elution time of finished products. DETAILED DESCRIPTION

[0032] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0033] In response to the existing problems, the embodiments of the present application provide a new method for constructing a characteristic spectrum of Psoralea corylifolia medicinal materials. The constructed characteristic spectrum provides a more scientific basis for authenticity identification or quality evaluation of Psoralea corylifolia medicinal materials.

[0034] On the one hand, an embodiment of the present invention provides a method for constructing a characteristic spectrum of a Psoralea corylifolia medicinal material, comprising:

[0035] The test solution and the reference solution are respectively subjected to high performance liquid chromatography to obtain characteristic spectra;

[0036] The chromatographic conditions of the high performance liquid chromatography method include: mobile phase A includes acetonitrile, and mobile phase B includes aqueous phosphoric acid solution.

[0037] The elution conditions of the chromatography were as follows:

[0038] From 0 to 2 min, the volume percentage of mobile phase A changes from 8% to 12% to 17% to 21%, and the volume percentage of mobile phase B changes from 88% to 92% to 79% to 83%; from 2 to 7 min, the volume percentage of mobile phase A changes from 17% to 21% to 34% to 38%, and the volume percentage of mobile phase B changes from 79% to 83% to 62% to 66%; from 7 to 9 min, the volume percentage of mobile phase A changes from 34% to 38% to 73% to 77%, and the volume percentage of mobile phase B changes from 62% to 66% to 33% to 37%; from 9 to 11 min, the volume percentage of mobile phase A changes from 1.5% to 2.5% to 3.5% to 4.5% to 5.5% to 6.5% to 7.5% to 8.5% to 9.5% to 10.5% to 11.5% to 12.5% ​​to 13.5% to 14.5% to 15.5% to 16.5% to 17.5% to 18.5% to 19.5% to 20.5% to 21.5% to 22.5% to 23.5% to 24.5% to 25.5% to 26.5% to 27.5% to 28.5% to 29.5% to 30.5% to 31.5% to 32.5% to 33.5% to 34.5% to From 14:00 to 15:00, the volume percentage of mobile phase A changed from 73% to 77% to 91% to 95%, and the volume percentage of mobile phase B changed from 33% to 37% to 5% to 9%; from 11 to 12 minutes, the volume percentage of mobile phase A was maintained at 91% to 95%, and the volume percentage of mobile phase B was maintained at 5% to 9%; from 12 to 12.1 minutes, the volume percentage of mobile phase A changed from 91% to 95% to 8% to 12%, and the volume percentage of mobile phase B changed from 5% to 9% to 88% to 92%; from 12.1 to 15 minutes, the volume percentage of mobile phase A was maintained at 8% to 12%, and the volume percentage of mobile phase B was maintained at 88% to 92%.

[0039] The chromatographic elution program provided in the embodiment of the present invention can quickly and effectively realize the construction of the HPLC characteristic spectrum of Psoralea corylifolia medicinal materials. The construction method is obtained by the inventor of the present application after creative labor and a large number of experiments. Compared with other elution programs, it has the advantages of shorter time and easier operation.

[0040] In some embodiments, from 0 to 2 min, the volume percentage of mobile phase A changes from 9% to 11% to 18% to 20%, and the volume percentage of mobile phase B changes from 89% to 91% to 80% to 82%; from 2 to 7 min, the volume percentage of mobile phase A changes from 18% to 20% to 35% to 37%, and the volume percentage of mobile phase B changes from 80% to 82% to 63% to 65%; from 7 to 9 min, the volume percentage of mobile phase A changes from 35% to 37% to 74% to 76%, and the volume percentage of mobile phase B changes from 63% to 65% to 34% to 36%; from 9 to 11 min, the volume percentage of mobile phase A changes from 18% to 20% to 35% to 37%, and the volume percentage of mobile phase B changes from 80% to 82% to 63% to 65%; The percentage changes from 74% to 76% to 92% to 94%, and the volume percentage of mobile phase B changes from 34% to 36% to 6% to 8%; from 11 to 12 minutes, the volume percentage of mobile phase A is maintained at 92% to 94%, and the volume percentage of mobile phase B is maintained at 6% to 8%; from 12 to 12.1 minutes, the volume percentage of mobile phase A changes from 92% to 94% to 9% to 11%, and the volume percentage of mobile phase B changes from 6% to 8% to 89% to 91%; from 12.1 to 15 minutes, the volume percentage of mobile phase A is maintained at 9% to 11%, and the volume percentage of mobile phase B is maintained at 89% to 91%.

[0041] In some embodiments, the chromatographic conditions used in the liquid chromatography include: using octadecylsilane bonded silica gel as a filler chromatographic column, a detection wavelength of 240 to 260 nm, a mobile phase flow rate of 0.6 to 0.7 mL / min, and a column temperature of 25 to 35°C.

[0042] In some embodiments, the filler particle size of the chromatographic column is 1 to 20 μm, specifically any one of 1, 1.5, 1.8, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20 μm or a range between any two thereof. The tube diameter of the chromatographic column can be 2.1 to 4.6 mm, specifically any one of 2, 2.1, 3, 3.6, 4, 4.6 mm or a range between any two thereof. The length of the chromatographic column can be 80 to 250 mm, specifically any one of 80, 100, 120, 140, 160, 180, 200, 220, 240, 250 mm or a range between any two thereof.

[0043] In some embodiments, the detection wavelength can be any one of 240, 242, 244, 245, 246, 248, 250, 252, 254, 255, 256, 248, and 260 nm, or a range between any two of them.

[0044] In some embodiments, the column temperature is any one of 25, 26, 28, 30, 32, 34, 35°C or a range between any two thereof.

[0045] In some embodiments, the mobile phase flow rate is any one of 0.6, 0.61, 0.62, 0.63, 0.64, 0.65, 0.66 and 0.67 mL / min or a range between any two thereof.

[0046] In some embodiments, the mobile phase B is a phosphoric acid aqueous solution with a phosphoric acid mass percentage of 0.01% to 0.1%. The phosphoric acid mass percentage in the mobile phase B can be any one of 0.01%, 0.02%, 0.04%, 0.05%, 0.06%, 0.08%, 0.1%, or a range between any two of them.

[0047] In some embodiments, the test sample includes: any one or more of Psoralea corylifolia medicinal materials and Psoralea corylifolia medicinal material extracts.

[0048] In some embodiments, the method for preparing the psoralea corylifolia extract comprises:

[0049] Mixing the powder of Psoralea corylifolia medicinal material with water and then electrically heating and refluxing;

[0050] The product after electric heating reflux is filtered, and the supernatant is taken as the extract.

[0051] In some embodiments, the mixing ratio of the psoralea corylifolia powder to water is 1-5 g:30-70 mL. The 1-5 g in the mixing ratio can be any one of 1, 2, 3, 4, and 5 g, or a range between any two of them; the 30-70 mL in the mixing ratio can be any one of 30, 40, 50, 60, and 70 mL, or a range between any two of them.

[0052] In some embodiments, the electric heating reflux time is 30 to 80 minutes, starting from the time when the solution begins to boil. The time can be any one of 30, 40, 50, 60, 70, 80 minutes or a range between any two of them.

[0053] In some embodiments, the method for preparing the test solution includes: mixing the Psoralea corylifolia medicinal material extract and methanol, ultrasonically treating the mixture, filtering the mixture, and taking the filtrate as the test solution.

[0054] In some embodiments, the ultrasonic treatment conditions include: power of 250-350 W, frequency of 30-50 kHz. The power can be any one of 250, 260, 280, 300, 320, 340, 350 W or a range between any two of them. The frequency can be any one of 30, 35, 40, 45, 50 kHz or a range between any two of them.

[0055] In some embodiments, in the test solution, 5 to 10 mL of methanol is added for every 3 mL of psoralea corylifolia extract. The amount of methanol added for every 3 mL of extract can be any one of 5, 6, 7, 8, 9, and 10 mL or a range between any two of them.

[0056] In some embodiments, the control substances include: psoralen, isopsoralen, psoralen, isopsoralen and bakuchiol.

[0057] In some embodiments, the reference substance solution comprises a reference substance and methanol.

[0058] In some embodiments, in the reference solution, 10-50 μg of psoralen, 10-50 μg of isopsoralen, 10-50 μg of psoralen, 10-50 μg of isopsoralen and 10-50 μg of bakuchiol are added per 1 mL of methanol. The mass of each reference added per 1 mL of methanol can be independently any one of 10, 20, 30, 40, 50 μg or any two thereof.

[0059] In some embodiments, the characteristic spectrum includes 5 characteristic peaks, with the chromatographic peak of characteristic peak 3 psoralen as the reference peak S, the relative retention time of each characteristic peak is within ±10% of the specified value, and the specified values ​​of characteristic peak 1 psoralen, characteristic peak 2 isopsoralen, characteristic peak 3 psoralen, characteristic peak 4 isopsoralen and characteristic peak 5 bakuchiol are: 0.36, 0.40, 1, 1.04 and 1.57, respectively.

[0060] In some embodiments, within ±10% of the specified value can be: within ±10%, ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2% or ±1% of the specified value.

[0061] In some embodiments, the theoretical plate number of the characteristic spectrum obtained should be no less than 2000 calculated based on the psoralen peak.

[0062] In some embodiments, the theoretical plate number of the characteristic spectrum obtained should be no less than 5000 calculated based on the psoralen peak.

[0063] On the other hand, an embodiment of the present invention provides a method for evaluating the quality of Psoralea corylifolia medicinal materials, comprising:

[0064] Obtaining a chromatogram of the sample to be tested for high performance liquid chromatography detection;

[0065] The quality of the sample to be tested is evaluated based on the chromatogram and the characteristic spectrum constructed by the construction method described in any of the above embodiments.

[0066] In some embodiments, the judgment criteria of the quality evaluation include: if the chromatogram of the sample to be tested has the 5 characteristic peaks described in any of the aforementioned embodiments, the sample to be tested is judged to be authentic or of high quality, otherwise it is a fake or of low quality. Specifically, the chromatogram of the sample to be tested has the 5 characteristic peaks described in any of the aforementioned embodiments, which can be judged according to the relative retention time of the peaks. If there are 5 peaks in the chromatogram of the sample to be tested, the 5 peaks correspond to the corresponding reference peaks of the reference substance, and the peak corresponding to the psoralen reference peak is the reference peak S, and the relative retention time of peak 1, peak 2, peak 4, peak 5 and peak S is calculated. If the relative retention time is within the range of ±10% of the specified value, and the specified value is 0.36 (peak 1), 0.40 (peak 2), 1.04 (peak 4), 1.57 (peak 5), then it is judged that the sample to be tested has 5 characteristic peaks.

[0067] In some embodiments, when the chromatogram of the sample to be tested has the five characteristic peaks described in any of the above embodiments, the judgment criteria also include: calculating the relative peak area of ​​characteristic peak 1 and characteristic peak 3 (S peak) and the relative peak area of ​​characteristic peak 2 and characteristic peak 3 in the chromatogram of the sample to be tested, if the relative peak area of ​​characteristic peak 1 and S peak is ≥2.05 and the relative peak area of ​​characteristic peak 2 and S peak is ≥1.86, the sample to be tested is judged to be authentic or of high quality, otherwise it is a counterfeit or of low quality. The calculation of the relative peak area of ​​characteristic peak 1 and characteristic peak 2 can further enhance the identification of counterfeit and inferior medicinal materials.

[0068] In addition, an embodiment of the present invention also provides the application of the construction method described in any of the above embodiments in screening Psoralea corylifolia medicinal materials or preparing Psoralea corylifolia-related drugs.

[0069] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0070] High performance liquid chromatograph: Agilent 1290 Infinity II high performance liquid chromatography (equipped with quaternary pump, column oven, autosampler, DAD detector);

[0071] Mettler Toledo XA-205 electronic balance (Mettler-Toledo Instrument Co., Ltd.);

[0072] Mettler Toledo AL204S 1 / 100,000 electronic analytical balance (Mettler-Toledo Instrument Co., Ltd.);

[0073] Pure water machine (Sichuan Youpu Ultrapure Technology Co., Ltd.).

[0074] Example 1

[0075] A method for constructing a characteristic spectrum of Psoralea corylifolia medicinal material comprises the following steps.

[0076] (1) Preparation of test solution: Prepare the extract of Psoralea corylifolia. Take about 2 g of crude powder of Psoralea corylifolia (passed through No. 2 sieve), weigh accurately, place in a stoppered conical flask, add 50 ml of pure water accurately, stopper tightly, weigh, heat and reflux for 60 minutes, cool, weigh again, make up the lost weight with pure water, filter, take 3 ml of the supernatant, place in a 10 ml volumetric flask, add 7 ml of methanol, ultrasonically treat (power 300 W, frequency 40 kHz) for 30 minutes, cool, make up to volume, shake well, filter, and take the filtrate.

[0077] (2) Preparation of reference medicinal material solution (a medicinal material standard substance used for identification and comparison of Chinese medicinal materials or finished medicines): Prepare the solution in the same way as the test sample.

[0078] (3) Preparation of reference solution: Take appropriate amount of psoralen, isopsoralen, psoralen, isopsoralen and bakuchiol reference substances, weigh accurately, and add methanol to prepare a mixed solution containing 30 μg of each substance per 1 ml.

[0079] (4) Construction of characteristic spectrum of the test sample: Chromatographic conditions: octadecylsilane bonded silica gel as filler chromatographic column (1.8 μm, 2.1×100 mm); acetonitrile as mobile phase A and 0.05% by mass phosphoric acid aqueous solution as mobile phase B for gradient elution;

[0080] The elution program is shown in Table 1.

[0081] Table 1 Elution program

[0082] Time (minutes) Mobile phase A (acetonitrile) Mobile phase B (0.05% phosphoric acid) 0～2 10％～19％ 90％～81％ 2～7 19％～36％ 81％～64％ 7～9 36％～75％ 64％～35％ 9～11 75％～93％ 35％～7％ 11～12 93％ 7％ 12～12.1 93％～10％ 7％～90％ 12.1～15 10％ 90％

[0083] Note: “%” in the table refers to volume percentage.

[0084] The detection wavelength was 250 nm and the mobile phase flow rate was 0.65 mL min -1 , the column temperature is 30°C. Under the above chromatographic conditions, 1-2 μL of the Psoralea corylifolia medicinal material sample is accurately aspirated, and ultra-high performance liquid chromatography is performed under the above chromatographic conditions to obtain a characteristic spectrum of the Psoralea corylifolia medicinal material (characteristic spectrum of the sample), and the theoretical plate number calculated based on the psoralen peak should be no less than 5000.

[0085] (5) Construction of the characteristic spectrum of the reference substance: Chromatographic conditions: octadecylsilane bonded silica gel as the filler column (1.8 μm, 2.1×100 mm); acetonitrile as mobile phase A and 0.05% phosphoric acid as mobile phase B for gradient elution; detection wavelength of 250 nm, mobile phase flow rate of 0.65 mL min -1 , the column temperature is 30° C. Under the above chromatographic conditions, 1 to 2 μL of the reference solution is accurately drawn and analyzed by ultra-high performance liquid chromatography to obtain a characteristic spectrum of the reference.

[0086] The chromatogram of the test sample should show 5 characteristic peaks, of which 5 peaks should correspond to the corresponding reference peaks of the reference substance. The peak corresponding to the psoralen reference peak is the reference peak S. Calculate the relative retention time of peak 1, peak 2, peak 4, peak 5 and peak S. The relative retention time should be within the range of ±2% of the specified value, and the specified value is 0.36 (peak 1), 0.40 (peak 2), 1.04 (peak 4), 1.57 (peak 5);

[0087] Calculate the relative peak areas of Peak 1, Peak 2, and Peak S. The relative peak areas should be within the specified range: not less than 2.05 (Peak 1) and not less than 1.86 (Peak 2).

[0088] Example 2

[0089] Based on the construction method provided in Example 1, the test solution was scanned in the entire band using a diode array detector, and the chromatograms of the test solution at wavelengths of 240 nm, 245 nm, 250 nm, 255 nm, and 260 nm were extracted respectively.

[0090] Results Figure 1 to Figure 6 .in, Figure 1 is the ultraviolet absorption spectrum of psoralen; Figure 2 is the ultraviolet absorption spectrum of isopsoralen; Figure 3 is the ultraviolet absorption spectrum of psoralen; Figure 4 is the ultraviolet absorption spectrum of isopsoralen; Figure 5 is the ultraviolet absorption spectrum of bakuchiol; Figure 6 This is a comparison chart of different wavelengths of Psoralea corylifolia medicinal materials.

[0091] The results show that when the detection wavelength is 250nm, the chromatographic peak information is larger, the chromatogram baseline is more stable, and the peak area is larger.

[0092] Example 3

[0093] Based on the construction method provided in Example 1, verification was performed at column temperatures of 25°C, 30°C, and 35°C, respectively. The results are shown in Table 1. Figure 7 .

[0094] Table 1 Column temperature test - characteristic peak relative retention time ratio

[0095]

[0096] The results showed that when the column temperatures were 25℃, 30℃ and 35℃, the RSD of the relative retention time of each characteristic peak was 0.00%-0.75%. When the column temperature was 30℃, the chromatogram peak shape was good and the separation was moderate.

[0097] Example 4

[0098] Based on the construction method provided in Example 1, verification was performed when the mobile phase flow rates were 0.6 ml / min, 0.65 ml / min, and 0.7 ml / min, respectively, as shown in Table 2.

[0099] Table 2 Flow rate test - characteristic peak relative retention time ratio

[0100]

[0101] The results showed that when the flow rates were 0.6mL / min, 0.65mL / min, and 0.7mL / min, respectively, the relative retention time RSD of each characteristic peak was 0.00%-4.28%. When the flow rate was 0.65mL / min, the chromatogram peak shape was better and the separation was moderate.

[0102] Example 5

[0103] Precision test.

[0104] Take the Psoralea corylifolia medicinal material test solution (batch number: medicinal material 1), and inject it 6 times continuously according to the method of Example 1, 1 μl each time, and calculate the retention time and peak area of ​​each peak. See Table 3 and Table 4 for details.

[0105] Table 3 Precision characteristic peak retention time test results

[0106]

[0107] Table 4 Precision test - characteristic peak area

[0108]

[0109]

[0110] The results showed that the RSD values ​​of the retention times of the peaks were 0.1706% to 0.36%, and the RSD values ​​of the peak areas were 0.17% to 0.77%, indicating that the instrument had good precision.

[0111] Example 6

[0112] Repeatability test.

[0113] 6 portions of Psoralea corylifolia (batch number: medicinal material 1) were accurately weighed and prepared and measured according to the method of Example 1. The results are shown in Tables 5 and 6.

[0114] Table 5 Repeatability test - characteristic peak relative retention time ratio

[0115]

[0116] Table 6 Repeatability test - characteristic peak relative peak area ratio The results showed that the RSD of the relative retention time of each characteristic peak was between 0.00% and 0.69%, and the RSD of the relative peak area of ​​each characteristic peak was between 0.00% and 2.17%. The method had good repeatability.

[0117] Example 7

[0118] Stability test.

[0119] Based on the experimental conditions of Example 1, the same test solution was taken and measured at 0h, 2h, 4h, 8h, 16h, and 24h. See Table 7 and Table 8 for details.

[0120] Table 7 Stability test - characteristic peak relative retention time ratio Table 8 Stability test - characteristic peak area ratio The results showed that the RSD of the corresponding characteristic peak retention time was between 0.00% and 0.28%, and the RSD of the characteristic peak area was between 0.00% and 3.77%, indicating that the method had good stability.

[0121] In summary, the RSD of the relative retention time of each characteristic peak meets the requirements in the above investigations, and this method is good.

[0122] Example 8

[0123] The characteristic spectra of three batches of samples were measured by the method of Example 1, and the relative retention time and relative peak area were calculated. Figure 8 , Fig. 9 , Tables 9 and 10. Figure 8 The characteristic spectra of three batches of medicinal materials and the control medicinal materials are shown in Figure 2. Fig. 9 It is the characteristic spectrum of the reference medicinal material.

[0124] Table 9 Relative retention time of three batches of Psoralea corylifolia

[0125]

[0126]

[0127] Table 10 Relative peak areas of three batches of Psoralea corylifolia medicinal materials According to the principle of stable relative retention time and the fact that each batch of samples can be detected and the peak area is relatively large, a total of 5 peaks with good repeatability were selected as characteristic peaks. The relative retention time RSD of the 5 characteristic peaks of 3 batches of Psoralea corylifolia medicinal materials was between 0.00% and 0.17%, and the relative retention time RSD of the 5 characteristic peaks during the methodological investigation was between 0.00% and 2.16%, indicating that the method has good repeatability.

[0128] Example 9

[0129] A method for evaluating the quality of psoralea corylifolia medicinal materials, comprising:

[0130] (1) Obtaining a chromatogram of the sample to be tested by high performance liquid chromatography, the sample detection process is the same as the detection of the test sample in Example 1;

[0131] (2) Performing quality evaluation on the sample to be evaluated based on the chromatogram and the characteristic spectrum constructed by the construction method provided in Example 1.

[0132] The quality evaluation criteria include:

[0133] Determine whether there are 5 characteristic peaks in the chromatogram of the sample to be tested: calculate the relative retention time of peak 1, peak 2, peak 4, peak 5 and peak S. If the relative retention time is within the range of ±2% of the specified value, the specified value is 0.36 (peak 1), 0.40 (peak 2), 1.04 (peak 4), 1.57 (peak 5), then it is determined that there are 5 characteristic peaks in the chromatogram, otherwise it does not;

[0134] If it does not have five characteristic peaks, the sample to be tested is judged to be authentic or of high quality;

[0135] If there are 5 characteristic peaks, and the relative peak area between peak 1 and peak S is ≥2.05, and the relative peak area between peak 2 and peak S is ≥1.86, then the sample to be tested is judged to be authentic or of high quality; otherwise, it is a fake or of low quality.

[0136] Example 10

[0137] The quality evaluation method provided in Example 9 was used to evaluate the quality of 10 batches of Psoralea corylifolia medicinal materials, and the relative retention time and relative peak area were calculated. The results are shown below: Fig.10 , Table 11 and Table 12, Fig.10 The characteristic spectra of 10 batches of medicinal materials. After the 10 batches of psoralea corylifolia medicinal materials were processed into formula granules, the quality of the psoralea corylifolia formula granules under the national drug standard of Chinese medicine formula granules was tested. The results are shown below. Fig.11 and Table 13. Comparison of elution time of medicinal materials and finished products is shown in Fig.12 .

[0138] Table 11 Relative retention time of ten batches of Psoralea corylifolia

[0139]

[0140] Table 12 Relative peak areas of ten batches of Psoralea corylifolia medicinal materials

[0141]

[0142]

[0143] Table 13 Quality evaluation results

[0144] Batch number of medicinal materials Example 9 Finished product batch number Finished product quality test results YD1 conform to DT1 Comply with national standards YD2 conform to DT2 Comply with national standards YD3 conform to DT3 Comply with national standards YD4 conform to DT4 Comply with national standards YD5 conform to DT5 Comply with national standards YD6 conform to DT6 Comply with national standards YD7 conform to DT7 Comply with national standards YD8 conform to DT8 Comply with national standards YD9 conform to DT9 Comply with national standards YD10 conform to DT10 Comply with national standards

[0145] From the results, it can be seen that the quality evaluation method provided by the invention is stable and reliable for identifying Psoralea corylifolia medicinal materials, and the testing of the preparations produced by it meets the national standards.

[0146] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for constructing a characteristic spectrum of Psoralea corylifolia medicinal material, It is characterized in that It includes: The test solution and the reference solution are respectively subjected to high performance liquid chromatography to obtain characteristic spectra; The chromatographic conditions of the high performance liquid chromatography method include: mobile phase A includes acetonitrile, and mobile phase B includes aqueous phosphoric acid solution; The elution conditions of the chromatography were as follows: From 0 to 2 min, the volume percentage of mobile phase A changed from 8% to 12% to 17% to 21%, and the volume percentage of mobile phase B changed from 88% to 92% to 79% to 83%; From 2 to 7 min, the volume percentage of mobile phase A changed from 17% to 21% to 34% to 38%, and the volume percentage of mobile phase B changed from 79% to 83% to 62% to 66%; From 7 to 9 min, the volume percentage of mobile phase A changed from 34% to 38% to 73% to 77%, and the volume percentage of mobile phase B changed from 62% to 66% to 33% to 37%; From 9 to 11 min, the volume percentage of mobile phase A changed from 73% to 77% to 91% to 95%, and the volume percentage of mobile phase B changed from 33% to 37% to 5% to 9%; 11-12 min, the volume percentage of mobile phase A is maintained at 91%-95%, and the volume percentage of mobile phase B is maintained at 5%-9%; 12-12.1 min, the volume percentage of mobile phase A changes from 91%-95% to 8%-12%, and the volume percentage of mobile phase B changes from 5%-9% to 88%-92%; From 12.1 to 15 min, the volume percentage of mobile phase A is maintained at 8% to 12%, and the volume percentage of mobile phase B is maintained at 88% to 92%.

2. The construction method according to claim 1, It is characterized in that The chromatographic conditions adopted by the liquid chromatography include: using octadecylsilane bonded silica gel as a chromatographic column filler, a detection wavelength of 240 to 260 nm, a mobile phase flow rate of 0.6 to 0.7 mL / min, and a column temperature of 25 to 35° C.; Optionally, the detection wavelength is 245-255 nm; Optionally, the column temperature is 28-32°C; Optionally, the flow rate of the mobile phase is 0.63-0.67 mL / min.

3. The construction method according to claim 1, It is characterized in that The mobile phase B is a phosphoric acid aqueous solution with a phosphoric acid mass percentage of 0.01% to 0.1%.

4. The construction method according to claim 1, It is characterized in that The test samples include: any one or more of psoralea corylifolia medicinal materials and psoralea corylifolia medicinal material extracts; Optionally, the method for preparing the psoralea corylifolia medicinal material extract comprises: Mixing the powder of Psoralea corylifolia medicinal material with water and then electrically heating and refluxing; The product after electric heating and reflux is filtered, and the supernatant is taken as the extract; Optionally, the mixing ratio of the Psoralea corylifolia medicinal material powder and water is 1-5g:30-70mL.

5. The construction method according to claim 4, It is characterized in that The preparation method of the test solution comprises: mixing the extract of psoralea corylifolia and methanol, ultrasonically treating the mixture, filtering the mixture, and taking the filtrate as the test solution; Optionally, the conditions of the ultrasonic treatment include: power of 250-350W and frequency of 30-50kHz; Optionally, in the test solution, 5 to 10 mL of methanol is added for every 3 mL of Psoralea corylifolia medicinal material extract.

6. The construction method according to any one of claims 1 to 5, It is characterized in that The reference substances include: psoralen, isopsoralen, psoralen, isopsoralen and bakuchiol; Optionally, the reference substance solution includes a reference substance and methanol; Optionally, in the reference solution, 10 to 50 μg of psoralen, 10 to 50 μg of isopsoralen, 10 to 50 μg of psoralen, 10 to 50 μg of isopsoralen and 10 to 50 μg of bakuchiol are added per 1 mL of methanol.

7. The construction method according to claim 6, It is characterized in that The characteristic spectrum includes 5 characteristic peaks, with the chromatographic peak of characteristic peak 3 psoralen as the reference peak S, the relative retention time of each characteristic peak is within ±10% of the specified value, and the specified values ​​of characteristic peak 1, characteristic peak 2, characteristic peak 3, characteristic peak 4 and characteristic peak 5 are: 0.36, 0.40, 1, 1.04 and 1.57 respectively.

8. A method for evaluating the quality of Psoralea corylifolia medicinal materials, It is characterized in that It includes: Obtaining a chromatogram of the sample to be tested for high performance liquid chromatography detection; The quality of the sample to be tested is evaluated according to the chromatogram and the characteristic spectrum constructed by the construction method according to any one of claims 1 to 7.

9. The quality evaluation method according to claim 8, It is characterized in that The judgment criteria for the quality evaluation include: if the chromatogram of the sample to be tested has the five characteristic peaks described in claim 7, the sample to be tested is judged to be authentic or of high quality, otherwise it is a fake or of low quality; Optionally, when the chromatogram of the sample to be tested has the five characteristic peaks described in claim 7, the judgment criteria also include: calculating the relative peak areas of characteristic peak 1 and characteristic peak 3, and the relative peak areas of characteristic peak 2 and characteristic peak 3 in the chromatogram of the sample to be tested; if the relative peak areas of characteristic peak 1 and characteristic peak 3 are ≥2.05 and the relative peak areas of characteristic peak 2 and characteristic peak 3 are ≥1.86, the sample to be tested is judged to be authentic or of high quality, otherwise it is a counterfeit or of low quality.

10. Use of the construction method according to any one of claims 1 to 7 in screening Psoralea corylifolia medicinal materials or in the preparation of Psoralea corylifolia-related drugs.

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