Method for detecting 14 components in fructus psoraleae

Abstract

The invention provides a method for detecting 14 components in fructus psoraleae, and belongs to the technical field of analysis and detection. The method comprises the following steps: performing alcohol extraction on fructus psoraleae to obtain a to-be-detected solution; performing high performance liquid chromatography detection on the liquid to be detected to obtain a chromatogram; and comparing the chromatogram with a standard substance spectrogram to obtain 14 components in the fructus psoraleae, the fructus psoraleae is prepared from the following 14 components: psoralen glycoside, isopsoralen glycoside, psoralen, isopsoralen, neopsoralen isoflavone, psoralen A, imperatorin, psoralen, psoralen chalcone, psoralen, psoralen B, psoralen flavanone methyl ether, psoralen isoflavone A and bakuchiol. The 14 components in the fructus psoraleae can be simultaneously distinguished and measured by controlling the composition and proportion of the mobile phase, the reference wavelength and the type of the chromatographic column, the detection method is rapid and efficient, and theoretical support is provided for distinguishing the type of processed products according to the content of different components subsequently.

Description

Technical Field

[0001] The present invention relates to the technical field of analysis and detection, and in particular to a method for detecting 14 components in psoralea corylifolia. Background Art

[0002] Psoralea corylifolia, also known as Psoralea corylifolia, Psoralea corylifolia or Psoralea corylifolia, belongs to the Rosales order and is an annual upright herb of the Leguminosae family. Its fruit is used as medicine, which has the functions of nourishing the kidney and strengthening yang, nourishing the spleen and strengthening the stomach, and can treat skin diseases such as psoriasis. It has the effects of enhancing kidney function, improving soreness of the waist and knees and frequent urination at night, and is often used to relieve symptoms such as fear of cold and fatigue caused by insufficient kidney yang.

[0003] It is difficult to determine multiple components in Psoralea corylifolia using related methods. Summary of the Invention

[0004] In view of this, the object of the present invention is to provide a method for detecting 14 components in Psoralea corylifolia. The detection method of the present invention can simultaneously achieve qualitative detection of the 14 components in Psoralea corylifolia.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The present invention provides a method for detecting 14 components in Psoralea corylifolia, comprising the following steps:

[0007] Extracting Psoralea corylifolia with alcohol to obtain a test solution;

[0008] Performing high performance liquid chromatography on the test liquid to obtain a chromatogram;

[0009] The chromatogram was compared with the standard spectrum to obtain 14 components in the psoralea corylifolia, which included psoralen, isopsoralen, psoralen, isopsoralen, neopsoralen isoflavones, psoralen A, imperatorin, psoralenine, psoralea corylifolia chalcone, psoralen, psoralen B, psoralea corylifolia methyl ether, psoralea corylifolia isoflavone A (Corylifol A) and bakuchiol;

[0010] The chromatographic conditions of the high performance liquid chromatography (HPLC) detection include:

[0011] Column: Poroshell 120EC-C 18 ;

[0012] Mobile phase: a mixture of methanol-acetonitrile solution and formic acid aqueous solution, wherein the volume fraction of methanol in the methanol-acetonitrile solution is 10% and the volume fraction of formic acid in the formic acid aqueous solution is 0.1%;

[0013] Gradient elution: 0-10 min, 10 vol% to 30 vol% methanol acetonitrile solution; 10-20 min, 30 vol% to 45 vol% methanol acetonitrile solution; 20-45 min, 45 vol% to 70 vol% methanol acetonitrile solution; 45-55 min, 70 vol% to 90 vol% methanol acetonitrile solution; 55-60 min, 90 vol% methanol acetonitrile solution; 60-70 min, 90 vol% to 10 vol% methanol acetonitrile solution;

[0014] Detection wavelength is 246 nm, reference wavelength is 360 nm for 0-20 min, and reference wavelength is 0 nm for 20-70 min;

[0015] Volume flow rate: 1 mL / min;

[0016] Column temperature: 30℃;

[0017] Injection volume: 5 μL.

[0018] Preferably, the alcohol extraction of Psoralea fruit further comprises sieving, and the mesh size of the sieve is 355±13 μm.

[0019] Preferably, the extraction agent of the alcohol extraction comprises methanol.

[0020] Preferably, the ratio of Psoralea fruit to extraction agent is 0.5-1 g: 25 mL.

[0021] Preferably, the time of the alcohol extraction is 30-60 min.

[0022] Preferably, the alcohol extraction further comprises passing through a microporous filter membrane, and the pore size of the microporous filter membrane is 0.45 μm.

[0023] Preferably, the alcohol extraction is performed under ultrasonic condition.

[0024] Preferably, the packing particle size of the chromatographic column is 4 μm, the inner diameter is 4.6 mm, and the length is 150 mm.

[0025] Preferably, the detection method further comprises:

[0026] According to the chromatogram and a predetermined standard curve, the content of 14 components in Psoralea fruit is obtained.

[0027] The invention provides a detection method for 14 components in psoralea corylifolia. The method comprises the following steps: performing alcohol extraction on psoralea corylifolia to obtain a test solution; performing high performance liquid chromatography detection on the test solution to obtain a chromatogram; and comparing the chromatogram with a spectrum of a standard product to obtain the 14 components in psoralea corylifolia. The 14 components in psoralea corylifolia include psoralen, isopsoralen, psoralen, isopsoralen, neopsoralen isoflavones, psoralen A, imperatorin, psoralenine, psoralea corylifolia chalcone, psoralen, psoralen B, psoralea corylifolia dihydroflavonoid methyl ether, psoralea corylifolia isoflavone A and bakuchiol.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] By controlling the composition and proportion of the mobile phase, the reference wavelength, and the type of chromatographic column, the present invention can simultaneously distinguish and determine 14 components in Psoralea corylifolia. The detection method is fast and efficient, providing theoretical support for subsequent differentiation of the types of processed products by the content of different components. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 is the HPLC spectrum of the mixed reference solution;

[0031] Figure 2 is the HPLC spectrum of the test solution;

[0032] Figures 1-2 Among them, 1 is psoralen, 2 is isopsoralen, 3 is psoralen, 4 is isopsoralen, 5 is psoralen isoflavone, 6 is psoralen A, 7 is imperatorin, 8 is psoralenine, 9 is psoralen chalcone, 10 is psoralen, 11 is psoralen B, 12 is psoralen dihydroflavonoid methyl ether, 13 is Corylifol A, and 14 is bakuchiol;

[0033] Figure 3 HPLC spectrum of the mixed reference solution in Comparative Example 1;

[0034] Figure 4 HPLC spectrum of the mixed reference solution in Comparative Example 2;

[0035] Figure 5 The HPLC spectrum of the test solution in Comparative Example 3 is shown below:

[0036] Figure 6 HPLC spectrum of the mixed reference solution in Comparative Example 4;

[0037] Figure 7 HPLC spectrum of the mixed reference solution in Comparative Example 5;

[0038] Figure 8 This is the HPLC spectrum of the mixed reference solution in Comparative Example 6;

[0039] Figure 9 This is the HPLC spectrum of the mixed reference solution in Comparative Example 7. DETAILED DESCRIPTION

[0040] The present invention provides a method for detecting 14 components in Psoralea corylifolia, comprising the following steps:

[0041] Extracting Psoralea corylifolia with alcohol to obtain a test solution;

[0042] Performing high performance liquid chromatography on the test liquid to obtain a chromatogram;

[0043] The chromatogram was compared with the standard spectrum to obtain 14 components in the psoralea corylifolia, which included psoralen, isopsoralen, psoralen, isopsoralen, neopsoralen isoflavones, psoralen A, imperatorin, psoralenine, psoralen chalcone, psoralenidine, psoralen B, psoralen dihydroflavonoid methyl ether, psoralen isoflavone A and bakuchiol;

[0044] The chromatographic conditions of the high performance liquid chromatography detection include:

[0045] Column: Poroshell 120EC-C 18 ;

[0046] Mobile phase: a mixture of methanol-acetonitrile solution and formic acid aqueous solution, wherein the volume fraction of methanol in the methanol-acetonitrile solution is 10% and the volume fraction of formic acid in the formic acid aqueous solution is 0.1%;

[0047] Gradient elution: 0-10 min, 10 vol%-30 vol% methanol-acetonitrile solution; 10-20 min, 30 vol%-45 vol% methanol-acetonitrile solution; 20-45 min, 45 vol%-70 vol% methanol-acetonitrile solution; 45-55 min, 70 vol%-90 vol% methanol-acetonitrile solution; 55-60 min, 90 vol% methanol-acetonitrile solution; 60-70 min, 90 vol%-10 vol% methanol-acetonitrile solution;

[0048] The detection wavelength was 246 nm, the reference wavelength for 0-20 min was 360 nm, and the reference wavelength for 20-70 min was 0 nm;

[0049] Volume flow rate: 1mL / min;

[0050] Column temperature: 30°C;

[0051] Injection volume: 5 μL.

[0052] The invention performs alcohol extraction on the psoralea corylifolia to obtain a liquid to be tested.

[0053] The present invention has no particular limitation on the source of the psoralea corylifolia, and commercially available products known to those skilled in the art may be used.

[0054] In the present invention, the alcohol pretreatment of Psoralea corylifolia preferably further comprises sieving, and the pore size of the sieving mesh is preferably 355±13 μm.

[0055] In the present invention, the extractant for the alcohol extraction preferably includes methanol.

[0056] In the present invention, the dosage ratio of the psoralea corylifolia to the extractant is preferably 0.5-1 g:25 mL, specifically 0.5 g:25 mL or 1 g:25 mL.

[0057] In the present invention, the alcohol extraction time is preferably 30 to 60 minutes, specifically 30, 40, 50 or 60 minutes.

[0058] In the present invention, the alcohol extraction is preferably carried out under ultrasonic conditions, and the present invention has no special limitation on the specific parameters of the ultrasonic conditions.

[0059] In the present invention, the alcohol extraction preferably further includes passing through a microporous filter membrane, and the pore size of the microporous filter membrane is preferably 0.45 μm.

[0060] In a specific embodiment of the present invention, the psoralea corylifolia is crushed and sieved, accurately weighed and placed in an EP tube, and ultrasonically extracted with methanol. The tube is taken out and naturally cooled, and the lost mass is supplemented with methanol. The tube is shaken and filtered through a microporous filter membrane to obtain the test solution.

[0061] After obtaining the liquid to be tested, the present invention performs high performance liquid chromatography on the liquid to be tested to obtain a chromatogram;

[0062] The chromatogram was compared with the standard spectrum to obtain 14 components in the psoralea corylifolia, which included psoralen, isopsoralen, psoralen, isopsoralen, neopsoralen isoflavones, psoralen A, imperatorin, psoralenine, psoralen chalcone, psoralenidine, psoralen B, psoralen dihydroflavonoid methyl ether, psoralen isoflavone A and bakuchiol;

[0063] The chromatographic conditions of the high performance liquid chromatography detection include:

[0064] Column: Poroshell 120EC-C 18 ;

[0065] Mobile phase: a mixture of methanol-acetonitrile solution and formic acid aqueous solution, wherein the volume fraction of methanol in the methanol-acetonitrile solution is 10% and the volume fraction of formic acid in the formic acid aqueous solution is 0.1%;

[0066] Gradient elution: 0-10 min, 10 vol%-30 vol% methanol-acetonitrile solution; 10-20 min, 30 vol%-45 vol% methanol-acetonitrile solution; 20-45 min, 45 vol%-70 vol% methanol-acetonitrile solution; 45-55 min, 70 vol%-90 vol% methanol-acetonitrile solution; 55-60 min, 90 vol% methanol-acetonitrile solution; 60-70 min, 90 vol%-10 vol% methanol-acetonitrile solution;

[0067] The detection wavelength was 246 nm, the reference wavelength for 0-20 min was 360 nm, and the reference wavelength for 20-70 min was 0 nm;

[0068] Volume flow rate: 1mL / min;

[0069] Column temperature: 30°C;

[0070] Injection volume: 5 μL.

[0071] In the present invention, the filler particle size of the chromatographic column is preferably 4 μm, the inner diameter is preferably 4.6 mm, and the length is preferably 150 mm.

[0072] After obtaining the chromatogram, the present invention preferably further comprises: obtaining the contents of the 14 components in the psoralea corylifolia according to the chromatogram and a predetermined standard curve.

[0073] The present invention has no particular limitation on the method for drawing the predetermined standard curve, and any method well known to those skilled in the art may be used.

[0074] The present invention can simultaneously distinguish and determine the contents of 14 components of Psoralea corylifolia, and preferably effectively distinguish different processed products of Psoralea corylifolia according to the contents of the 14 components.

[0075] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions of the present invention. Obviously, the embodiments described are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0076] Example 1

[0077] Preparation of the test solution: Psoralea corylifolia was crushed and sieved, 0.5 g was accurately weighed and placed in an EP tube, 25 mL of methanol was added and ultrasonic extraction was performed for 30 min, the tube was taken out and naturally cooled, and the lost mass was supplemented with methanol, shaken well, and filtered through a microporous filter membrane (0.45 μm) to obtain the test solution.

[0078] Preparation of mixed reference solution: Accurately weigh psoralen, isopsoralen, psoralen, isopsoralen, neopsoralen isoflavones, psoralen A, imperatorin, psoralen, psoralen chalcone, psoralen, psoralen B, psoralen dihydroflavonoid methyl ether, Corylifol A and bakuchiol, and add methanol to prepare a mixed reference solution.

[0079] The test solution and the mixed reference solution are respectively subjected to high performance liquid chromatography detection, and the chromatographic conditions of the high performance liquid chromatography detection include:

[0080] Column: Poroshell 120EC-C 18 (4 μm, 4.6 mm × 150 mm);

[0081] Mobile phase: a mixture of methanol-acetonitrile solution and formic acid aqueous solution, wherein the volume fraction of methanol in the methanol-acetonitrile solution is 10% and the volume fraction of formic acid in the formic acid aqueous solution is 0.1%;

[0082] Gradient elution: 0-10 min, 10 vol%-30 vol% methanol-acetonitrile solution; 10-20 min, 30 vol%-45 vol% methanol-acetonitrile solution; 20-45 min, 45 vol%-70 vol% methanol-acetonitrile solution; 45-55 min, 70 vol%-90 vol% methanol-acetonitrile solution; 55-60 min, 90 vol% methanol-acetonitrile solution; 60-70 min, 90 vol%-10 vol% methanol-acetonitrile solution;

[0083] The detection wavelength was 246 nm, the reference wavelength for 0-20 min was 360 nm, and the reference wavelength for 20-70 min was 0 nm;

[0084] Volume flow rate: 1mL / min;

[0085] Column temperature: 30°C;

[0086] Injection volume: 5 μL.

[0087] Figure 1 is the HPLC spectrum of the mixed reference solution, Figure 2 is the HPLC spectrum of the test solution, Figures 1-2 Among them, 1 is psoralen, 2 is isopsoralen, 3 is psoralen, 4 is isopsoralen, 5 is psoralen isoflavone, 6 is psoralen A, 7 is imperatorin, 8 is psoralen, 9 is psoralea corylifolia chalcone, 10 is psoralen, 11 is psoralea corylifolia B, 12 is psoralea corylifolia dihydroflavonoid methyl ether, 13 is Corylifol A, and 14 is bakuchiol. It can be seen that the present invention can simultaneously produce 14 ingredients in psoralea corylifolia.

[0088] Example 2 Quantitative Analysis

[0089] Linear relationship determination

[0090] Accurately draw the reference solution, dilute it step by step, and obtain mixed reference solution of different mass solubility. Accurately draw 5 μL of the mixed reference solution of 6 concentrations and inject it into the liquid chromatograph. Inject the sample according to the chromatographic conditions in Example 1. The injection volume (mg·mL -1 ) as the abscissa and the peak area as the ordinate to draw a standard curve, perform linear regression, and obtain the regression equation. Table 1 shows the regression equations and linear ranges of 14 compounds in Psoralea corylifolia.

[0091] Table 1 Regression equations and linear ranges of 14 compounds in Psoralea corylifolia

[0092] Compound Regression equation r <![CDATA[线性范围(mg·mL -1 )]]> Psoralen y=563.18x+8.3862 0.9993 0.003814～0.1907 Isopsoralen y=499.65x+7.7653 0.9992 0.003372～0.1686 Psoralen y=167.75x+2.912 0.9993 0.002368～0.1184 Isopsoralen y=275.49x+4.2673 0.9993 0.003136～0.1568 New Psoralea corylifolia Isoflavones y=361.45x+7.4881 0.9992 0.003488～0.1744 Psoralen y=290.72x+6.025 0.9994 0.004684～0.2342 Imperatorin y=226.49x+3.1376 0.9994 0.001928～0.0964 Psoralea corylifolia y=591.56x+15.174 0.9991 0.003104～0.1552 Psoralea corylifolia y=139.77x+1.8653 0.9993 0.00304～0.152 Psoralea corylifolia chalcone y=263.53x+4.4311 0.9994 0.002868～0.1434 Psoralen y=164.79x+0.3431 0.9991 0.003488～0.1744 Psoralea corylifolia methyl ether y=137.86x-26.382 0.9996 0.002044～0.1022 Corylifol A y=95.642x+1.6842 0.9995 0.00198～0.099 Bakuchiol y=1807.4x+86.99 0.9991 0.030656～1.5328

[0093] Precision test

[0094] 5 μL of the test solution in Example 1 was taken and injected 6 times according to the chromatographic conditions in Example 1. The relative standard deviation (RSD) of the peak area of ​​each component was <2.00%, indicating that the instrument had good precision.

[0095] Stability test

[0096] 5 μL of the test solution in Example 1 was injected at 0, 2, 4, 8, 12, and 24 h after preparation under the chromatographic conditions in Example 1. The RSDs of the peak areas were all <2.00%, indicating that the test solution had good stability within 24 h.

[0097] Repeatability test

[0098] The test solution in Example 1 was injected according to the chromatographic conditions in Example 1, the peak areas were recorded and the mass fractions of the components were calculated. The RSDs of the peak areas were all <2.00%, indicating that the method had good repeatability.

[0099] Sample recovery test

[0100] Six portions of psoralea corylifolia powder of known content were precisely pipetted and added to 100% of the corresponding reference solution. Samples were injected and tested according to the chromatographic conditions described in Example 1. Peak areas were recorded, and the recoveries of the 14 components were calculated. The average recoveries for the 14 components ranged from 98.53% to 102.68%, with RSDs of 1.5% to 2.5%, demonstrating the high accuracy of this method.

[0101] Comparative Example 1

[0102] The mixed reference solution was prepared as in Example 1, and the mixed reference solution was subjected to HPLC detection. The chromatographic conditions of HPLC detection included:

[0103] Chromatographic column: Agilent C18 column (5 μm, 4.6 mm × 250 mm);

[0104] The mobile phase was a mixture of acetonitrile and 0.1 vol% formic acid in water;

[0105] Gradient elution: 0-5 min, 5 vol% acetonitrile; 5-10 min, 5 vol%-10 vol% acetonitrile; 10-20 min, 10 vol%-25 vol% acetonitrile; 20-30 min, 25 vol%-45 vol% acetonitrile; 30-45 min, 45 vol%-90 vol% acetonitrile; 45-55 min, 90 vol%-2 vol% acetonitrile; 55-60 min, 2 vol% acetonitrile;

[0106] Detection wavelength: 246 nm; reference wavelength: 360 nm;

[0107] Volume flow rate 1.0 mL / min;

[0108] Column temperature: 30°C; injection volume: 10 μL.

[0109] Figure 3 This is the HPLC spectrum of the mixed reference solution in Comparative Example 1. It can be seen that some components have inverted peaks and bakuchiol cannot be determined.

[0110] Comparative Example 2

[0111] The mixed reference solution was prepared as in Example 1, and the mixed reference solution was subjected to HPLC detection. The chromatographic conditions of HPLC detection included:

[0112] Chromatographic column: Agilent C18 column (5 μm, 4.6 mm × 250 mm);

[0113] The mobile phase was a mixture of acetonitrile and water;

[0114] Gradient elution: 0-5 min, 20 vol%-30 vol% acetonitrile; 5-20 min, 30 vol%-35 vol% acetonitrile; 20-40 min, 35 vol%-45 vol% acetonitrile; 40-50 min, 45 vol%-55 vol% acetonitrile; 50-70 min, 55 vol%-75 vol% acetonitrile; 70-80 min, 75 vol%-95 vol% acetonitrile;

[0115] Detection wavelength: 246 nm; reference wavelength: 360 nm;

[0116] Volume flow rate 1.0 mL / min;

[0117] Column temperature: 30°C; injection volume: 10 μL.

[0118] Figure 4 This is the HPLC spectrum of the mixed reference solution in Comparative Example 2. It can be seen that some components have inverted peaks, and components such as psoralen and isopsoralen cannot be determined.

[0119] Comparative Example 3

[0120] The test solution was prepared as in Example 1, and the test solution was subjected to HPLC detection. The chromatographic conditions of HPLC detection included:

[0121] Column: Poroshell 120EC-C 18 (4 μm, 4.6 mm × 150 mm);

[0122] The mobile phase was a mixture of acetonitrile and 0.1 vol% formic acid in water;

[0123] Gradient elution: 0-10 min, 2 vol%-10 vol% acetonitrile; 10-15 min, 10 vol%-30 vol% acetonitrile; 15-30 min, 30 vol%-35 vol% acetonitrile; 30-40 min, 35 vol%-45 vol% acetonitrile; 40-50 min, 45 vol%-55 vol% acetonitrile; 50-70 min, 55 vol%-75 vol% acetonitrile; 70-100 min, 75 vol% acetonitrile;

[0124] Detection wavelength: 246 nm; reference wavelength: 360 nm;

[0125] Volume flow rate 1.0 mL / min;

[0126] Column temperature: 30°C; injection volume: 10 μL.

[0127] Figure 5 This is the HPLC spectrum of the test solution in Comparative Example 3. It can be seen that some components have inverted peaks, and components such as psoralen and isopsoralen cannot be determined.

[0128] Comparative Example 4

[0129] The mixed reference solution was prepared as in Example 1, and the mixed reference solution was subjected to HPLC detection. The chromatographic conditions of HPLC detection included:

[0130] Column: Poroshell 120EC-C 18 (4 μm, 4.6 mm × 150 mm);

[0131] Mobile phase: a mixture of methanol and 0.1 vol% formic acid aqueous solution;

[0132] Gradient elution: 0-10 min, 30 vol%-40 vol% methanol; 10-30 min, 40 vol%-90 vol% methanol; 30-50 min, 90 vol% methanol;

[0133] The detection wavelength is 246 nm; the reference wavelength is 360 nm;

[0134] Volume flow rate: 1mL / min;

[0135] Column temperature: 30°C;

[0136] Injection volume: 5 μL.

[0137] Figure 6 This is the HPLC spectrum of the mixed reference solution in Comparative Example 4. It can be seen that some components have inverted peaks, the peak shapes are generally poor, and no bakuchiol is observed.

[0138] Comparative Example 5

[0139] The mixed reference solution was prepared as in Example 1, and the mixed reference solution was subjected to HPLC detection. The chromatographic conditions of HPLC detection included:

[0140] Column: Poroshell 120EC-C 18 (4 μm, 4.6 mm × 150 mm);

[0141] Mobile phase: methanol-water mixture;

[0142] Gradient elution: 0-10 min, 30 vol%-40 vol% methanol; 10-30 min, 40 vol%-90 vol% methanol; 30-50 min, 90 vol% methanol;

[0143] The detection wavelength is 246 nm; the reference wavelength is 360 nm;

[0144] Volume flow rate: 1mL / min;

[0145] Column temperature: 30°C;

[0146] Injection volume: 5 μL.

[0147] Figure 7 This is the HPLC spectrum of the mixed reference solution in Comparative Example 5. It can be seen that some components have inverted peaks, the peak shapes are generally poor, and no bakuchiol is observed.

[0148] Comparative Example 6

[0149] The mixed reference solution was prepared as in Example 1, and the mixed reference solution was subjected to HPLC detection. The chromatographic conditions of HPLC detection included:

[0150] Chromatographic column: Poroshell 120EC-C 18 (4 μm, 4.6 mm x 150 mm);

[0151] Mobile phase: mixture of methanol acetonitrile solution and formic acid aqueous solution, the concentration of methanol in the methanol acetonitrile solution is 10 vol%, the concentration of formic acid in the formic acid aqueous solution is 10 vol%;

[0152] Gradient elution: 0-10 min, 10 vol%-30 vol% of methanol acetonitrile solution; 10-20 min, 30 vol%-45 vol% of methanol acetonitrile solution; 20-45 min, 45 vol%-70 vol% of methanol acetonitrile solution; 45-55 min, 70 vol%-90 vol% of methanol acetonitrile solution;

[0153] Detection wavelength: 246 nm, reference wavelength: 360 nm;

[0154] Volume flow rate: 1 mL / min;

[0155] Column temperature: 30°C;

[0156] Injection volume: 5 μL.

[0157] Figure 8 The HPLC spectrum of the mixed reference solution in Comparative Example 6 shows that the peak shape is well separated, but there is a reverse peak.

[0158] Comparative Example 7

[0159] The mixed reference solution was prepared as in Example 1, and the mixed reference solution was detected by high performance liquid chromatography, the chromatographic conditions of the high performance liquid chromatography detection were the same as in Example 1, except that there was no reference wavelength.

[0160] Figure 9 The HPLC spectrum of the mixed reference solution in Comparative Example 7 shows that the reverse peak is improved, but the peak shape in the front part is poor, and there is a split peak.

[0161] The above only describes the preferred embodiments of the present application, and does not limit the present application in any form. It should be noted that for ordinary skilled persons in the art, several improvements and refinements can be made without departing from the principles of the present application, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. A method for detecting 14 components in Psoralea corylifolia, characterized in that: The following steps are involved: Extracting Psoralea corylifolia with alcohol to obtain a test solution; Performing high performance liquid chromatography on the test liquid to obtain a chromatogram; The chromatogram was compared with the standard spectrum to obtain 14 components in the psoralea corylifolia, which included psoralen, isopsoralen, psoralen, isopsoralen, neopsoralen isoflavones, psoralen A, imperatorin, psoralenine, psoralen chalcone, psoralenidine, psoralen B, psoralen dihydroflavonoid methyl ether, psoralen isoflavone A and bakuchiol; The chromatographic conditions of the high performance liquid chromatography detection include: Column: Poroshell 120EC-C 18 ; Mobile phase: a mixture of methanol-acetonitrile solution and formic acid aqueous solution, wherein the volume fraction of methanol in the methanol-acetonitrile solution is 10% and the volume fraction of formic acid in the formic acid aqueous solution is 0.1%; Gradient elution: 0-10 min, 10 vol%-30 vol% methanol-acetonitrile solution; 10-20 min, 30 vol%-45 vol% methanol-acetonitrile solution; 20-45 min, 45 vol%-70 vol% methanol-acetonitrile solution; 45-55 min, 70 vol%-90 vol% methanol-acetonitrile solution; 55-60 min, 90 vol% methanol-acetonitrile solution; 60-70 min, 90 vol%-10 vol% methanol-acetonitrile solution; The detection wavelength was 246 nm, the reference wavelength for 0-20 min was 360 nm, and the reference wavelength for 20-70 min was 0 nm; Volume flow rate: 1mL / min; Column temperature: 30°C; Injection volume: 5 μL.

2. The detection method according to claim 1, wherein The alcohol pretreatment of Psoralea corylifolia also includes sieving, and the pore size of the sieve is 355±13 μm.

3. The detection method according to claim 1, wherein The extractant for the alcohol extraction includes methanol.

4. The detection method according to claim 1 or 3, characterized in that The dosage ratio of the psoralea corylifolia to the extractant is 0.5-1g:25mL.

5. The detection method according to claim 1, wherein The alcohol extraction time is 30 to 60 minutes.

6. The detection method according to claim 1, characterized in that After the alcohol extraction, the product is filtered through a microporous membrane, and the pore size of the microporous membrane is 0.45 μm.

7. The detection method according to claim 1, characterized in that The alcohol extraction is carried out under ultrasonic conditions.

8. The detection method according to claim 1, wherein The chromatographic column has a filler particle size of 4 μm, an inner diameter of 4.6 mm, and a length of 150 mm.

9. The detection method according to claim 1, wherein Also includes: According to the chromatogram and the predetermined standard curve, the contents of the 14 components in the psoralea corylifolia are obtained.

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